@article {pmid40251382, year = {2025}, author = {Al-Amin, HM and Gyawali, N and Graham, M and Alam, MS and Lenhart, A and Xi, Z and Rašić, G and Beebe, NW and Hugo, LE and Devine, GJ}, title = {Fitness compatibility and dengue virus Inhibition in a Bangladeshi strain of Aedes aegypti infected with the Wolbachia strain wAlbB.}, journal = {Scientific reports}, volume = {15}, number = {1}, pages = {13425}, pmid = {40251382}, issn = {2045-2322}, mesh = {Animals ; *Aedes/virology/microbiology/genetics ; *Wolbachia/physiology ; *Dengue Virus/physiology ; Bangladesh ; *Dengue/transmission/virology/prevention & control ; *Mosquito Vectors/virology/microbiology ; Genetic Fitness ; Female ; Mosquito Control ; }, abstract = {Dengue cases in Bangladesh have surged in recent years. The existing insecticide-based control program is challenged by issues of insufficient household coverage and high levels of insecticide resistance in the primary dengue virus (DENV) vector, Aedes aegypti. A more sustainable, effective alternative could be the implementation of a Wolbachia-mediated disease management strategy. Hence, we created and characterised a Wolbachia-infected Ae. aegypti strain with a Dhaka wild-type genetic background, and compared its reproductive compatibility, maternal inheritance, fitness, and virus-blocking ability to the parental strains (Dhaka wild-type and wAlbB2-F4). The new Ae. aegypti strain wAlbB2-Dhaka demonstrated complete cytoplasmic incompatibility with the wild-type and complete maternal transmission, retaining levels of pyrethroid resistance of the Dhaka wild-type. No significant fitness costs were detected during laboratory comparison. Compared to the wild-type, wAlbB2-Dhaka mosquitoes demonstrated a significantly reduced genome copies of DENV in the bodies (44.4%, p = 0.0034); a two-fold reduction in dissemination to legs and wings (47.6%, p < 0.0001); and > 13-fold reduction of DENV in saliva expectorates (proxy of transmission potential) (92.7%, p < 0.0001) 14 days after ingesting dengue-infected blood. Our work indicates that the wAlbB2-Dhaka strain could be used for Ae. aegypti suppression or replacement strategies for dengue management in Bangladesh.}, } @article {pmid40249197, year = {2025}, author = {Hubert, J and Glowska-Patyniak, E and Dowd, SE and Klimov, PB}, title = {Cardinium disrupts Wolbachia-host dynamics in the domestic mite Tyrophagus putrescentiae: evidence from manipulative experiments.}, journal = {mSystems}, volume = {}, number = {}, pages = {e0176924}, doi = {10.1128/msystems.01769-24}, pmid = {40249197}, issn = {2379-5077}, abstract = {UNLABELLED: We investigated the tripartite interactions between two intracellular bacterial symbionts, Cardinium and Wolbachia in Tyrophagus putrescentiae. Cultures of Tyrophagus putrescentiae are typically single-infected by one intracellular symbiont. However, co-infection can be experimentally induced by mixing single-infected cultures, resulting in 10% of mite individuals being double-infected (Cardinium + Wolbachia) and a corresponding reduction in host fitness. Here, we assembled the genomes of Cardinium and Wolbachia and analyzed their gene expression in parental single-infected and mixed mite cultures using population-level samples (ranging from 7,500 to 10,000 mites). Wolbachia interacts more extensively with its mite host than Cardinium in single-infected cultures. However, in mixed cultures, (i) Wolbachia exhibited reduced regulation of the host compared with Cardinium; (ii) the gene expression profile of Cardinium shifted, increasing its interactions with the host, whereas the gene expression profile of Wolbachia remained unchanged; and (iii) Wolbachia genes exhibited a loss of interactions with mite gene expression, as indicated by reduced correlations (for example with host MAPK, endocytosis, and calcium signaling pathways). The experiments show that at the mite population level, symbiont infection disrupts gene expression interaction between the two symbionts and their host in different ways. Wolbachia was more influenced by Cardinium gene expression than vice versa. Cardinium can inhibit the growth of Wolbachia by disrupting its interaction with the host, leading to a loss of Wolbachia's influence on mite immune and regulatory pathways. The reasons for responses are due to co-infection or the reduced frequency of Wolbachia single-infected individuals due to the analyses of population-level samples.

IMPORTANCE: We found that Cardinium disrupts the interaction between Wolbachia and mite host. In Wolbachia single-infected cultures, strong correlations exist between symbiont and host gene expressions. Interestingly, although Cardinium can also interact with the host, this interaction appears weaker compared with Wolbachia in single-infected cultures. These results suggest that both symbionts affect mite host gene expression, particularly in immune and regulatory pathways. In mixed samples, Cardinium appears to outcompete Wolbachia by disrupting its host interaction. It indicates competition between these two intracellular symbionts in mite populations. Wolbachia belongs to a mite-specific supergroup Q, distinct from the more commonly studied Wolbachia supergroups. As these mite-specific bacteria exhibit pathogen-blocking effects, our findings may have relevance for other systems, such as ticks and tick-borne diseases. The study sheds light on intracellular symbiont interaction within a novel mite-symbiont model.}, } @article {pmid40243374, year = {2025}, author = {Kustra, MC and Carrier, TJ}, title = {Microbes as manipulators of egg size and developmental evolution.}, journal = {mBio}, volume = {}, number = {}, pages = {e0365524}, doi = {10.1128/mbio.03655-24}, pmid = {40243374}, issn = {2150-7511}, abstract = {UNLABELLED: Marine invertebrates mainly reproduce by energy-poor eggs that develop into feeding larvae or energy-rich eggs that develop into non-feeding larvae. Evolutionary transitions between these developmental modes have been studied in detail, yet the evolutionary factor(s) responsible for these switches remains elusive. Here, we use theoretical models to support the premise that microbes with the capacity to manipulate host reproduction may be one possible factor. Our model predicts that microbial manipulators could create a sperm-limited environment that selects for larger eggs by shifting the host's sex ratio toward female dominance and, as a result, drive an evolutionary transition in the developmental mode for marine invertebrates. The loss of a microbial manipulator could then recover the ancestral egg size and developmental mode. We also suggest more than a dozen genera of marine invertebrates from throughout the world's oceans that fit the framework of a microbe-induced evolutionary transition between these predominant developmental modes. We anticipate that microbial manipulators have a yet-to-be-appreciated influence on the developmental evolution of marine invertebrates. We find it paramount to understand whether evolutionary transitions in developmental mode occur with and without microbial manipulators as well as whether the underlying mechanisms of these manipulations are convergent with terrestrial systems.

IMPORTANCE: Microbes that manipulate animal reproduction are widespread on land, and their evolutionary influence is widely acknowledged. Relatives of these manipulators are increasingly found in the ocean, but uniquely with taxa that recently underwent a transition in developmental evolution from feeding to non-feeding larvae. Here, we present theoretical models supporting that microbial manipulators could create a sperm-limited environment that selects for larger eggs by shifting the host's sex ratio toward female dominance and, as a result, drive an evolutionary transition in the developmental mode for free-spawning marine invertebrates. This theoretical model provides a complementary viewpoint to the theory regarding the evolutionary process that marine invertebrates undergo to transition between developmental modes as well as a fruitful opportunity to compare with terrestrial systems.}, } @article {pmid40240454, year = {2025}, author = {Leroy, E and Gao, S and Gonzalez, M and Ellies-Oury, MP and Tuda, M}, title = {Wolbachia infection facilitates adaptive increase in male egg size in response to environmental changes.}, journal = {Scientific reports}, volume = {15}, number = {1}, pages = {13213}, pmid = {40240454}, issn = {2045-2322}, support = {19K06840//JSPS/ ; }, mesh = {Animals ; *Wolbachia/physiology ; Male ; Female ; *Ovum/microbiology/physiology ; *Coleoptera/microbiology/physiology ; Climate Change ; Symbiosis ; *Adaptation, Physiological ; Environment ; Temperature ; Longevity ; }, abstract = {Under challenging conditions such as maladapted biotic and abiotic conditions, females can plastically adjust their egg size (gamete or zygote size) to counteract fitness declines early in life. Recent evidence suggests that endosymbionts may enhance this egg-size plasticity. Possible endosymbionts' modification of impact of multiple stressors is not well explored. Therefore, this study aims to test (1) whether Wolbachia infection influences the plasticity of parental investment in egg size under suboptimal environmental conditions and (2) whether the plasticity depends on the sex of eggs. We used three lines of the azuki bean beetle (Callosobruchus chinensis): a line coinfected with the wBruCon and wBruOri Wolbachia strains, a cured line infected solely with the wBruCon, and an uninfected (cured) line. These lines were subjected to either a control environment or a simulated climate change environment (elevated temperature and carbon dioxide levels, eT&CO2) to examine Wolbachia infection effects on parental investment in their offspring (egg size) and its subsequent impact on offspring fitness, including survival, development, and adult lifespan under starvation. After two days of eT&CO2 exposure, coinfected parents increased male egg size only. Larger eggs developed faster in both sexes and exhibited higher survival. However, offspring adult lifespan was not influenced by egg size but by environment, sex, Wolbachia infection, and development time: eT&CO2 reduced male lifespan but not female lifespan, the singly-infected line females lived longer than coinfected and uninfected line females, and shorter development time linked to longer lifespan. The negative correlation between development time and lifespan was higher under eT&CO2 but not sex-specific. This study is the first to demonstrate sex-specific egg size plasticity associated with Wolbachia infection in species with sex determination systems other than haplodiploid.}, } @article {pmid40237471, year = {2025}, author = {Njogu, AK and Logozzo, F and Conner, WR and Shropshire, JD}, title = {Counting rare Wolbachia endosymbionts using digital droplet PCR.}, journal = {Microbiology spectrum}, volume = {}, number = {}, pages = {e0326624}, doi = {10.1128/spectrum.03266-24}, pmid = {40237471}, issn = {2165-0497}, abstract = {Wolbachia is the most widespread animal-associated intracellular microbe, living within the cells of over half of insect species. Since they can suppress pathogen replication and spread rapidly through insect populations, Wolbachia is at the vanguard of public health initiatives to control mosquito-borne diseases. Wolbachia's abilities to block pathogens and spread quickly are closely linked to their abundance in host tissues. The most common method for counting Wolbachia is quantitative polymerase chain reaction (qPCR), yet qPCR can be insufficient to count rare Wolbachia, necessitating tissue pooling and consequently compromising individual-level resolution of Wolbachia dynamics. Digital droplet PCR (ddPCR) offers superior sensitivity, enabling the detection of rare targets and eliminating the need for sample pooling. Here, we report three ddPCR assays to measure total Wolbachia abundance, Wolbachia abundance adjusted for DNA extraction efficiency, and Wolbachia density relative to host genome copies. Using Drosophila melanogaster with wMel Wolbachia as a model, we show these ddPCR assays can reliably detect as few as 7 to 12 Wolbachia gene copies in a 20 µL reaction. The designed oligos are homologous to sequences from at least 106 Wolbachia strains across supergroup A and 53 host species from the Drosophila, Scaptomyza, and Zaprionus genera, suggesting broad utility. These highly sensitive ddPCR assays are expected to significantly advance Wolbachia-host interactions research by enabling the collection of molecular data from individual insect tissues. Their ability to detect rare Wolbachia will be especially valuable in applied and natural field settings where pooling samples could obscure important variation.IMPORTANCEWolbachia bacteria live inside the cells of many animals, especially insects. In many insect species, almost every individual carries Wolbachia. How common Wolbachia becomes within a population often depends on how much of it is present in the insect's body. Therefore, accurately measuring Wolbachia levels is crucial for understanding how these bacteria interact with their hosts and spread. However, traditional molecular assays can lack the sensitivity needed for accurate, individual-level quantification of rare Wolbachia. Here, we present three highly sensitive digital droplet PCR assays for Wolbachia detection, offering superior sensitivity compared to existing methods. These assays will be useful for studies that measure Wolbachia abundance and related phenotypes in individual insects, providing enhanced resolution and improving efforts to characterize the mechanisms that govern phenotypic variation.}, } @article {pmid40229230, year = {2025}, author = {Sewade, W and Polat, C and Kasap, OE}, title = {Molecular evidence of Wolbachia and Orthoflavivirus infection in field-collected mosquitoes in three provinces of Türkiye.}, journal = {Tropical medicine & international health : TM & IH}, volume = {}, number = {}, pages = {}, doi = {10.1111/tmi.14099}, pmid = {40229230}, issn = {1365-3156}, abstract = {BACKGROUND: Mosquitoes transmit various pathogens causing diseases like Zika, Dengue, West Nile and Chikungunya. They also harbour insect-specific viruses (ISVs) and Wolbachia, which can block arbovirus transmission. This study investigated the prevalence of Orthoflavivirus and Wolbachia in mosquito populations from three provinces in Türkiye.

METHODS: Mosquitoes were collected using CDC Miniature Light traps in 2022-2023. Morphologically identified specimens were pooled (1-10 individuals) and screened for Orthoflavivirus and Wolbachia via PCR and confirmed by Sanger sequencing. Infection prevalence was estimated using the maximum likelihood method. Mosquito taxa richness across provinces was estimated using the abundance-based, non-parametric Chao1 index.

RESULTS: Among 8766 mosquitoes (11 taxa) collected, Culex perexiguus, Ochlerotatus caspius and Anopheles claviger were most abundant. Anopheles flavivirus (AnFV) detected in one Oc. caspius pool, while Wolbachia sequences belonging to supergroup B were detected in An. claviger, Cx. pipiens s.l., Cx. perexiguus and Oc. caspius, with an overall infection prevalence of 0.0119 (95% CI: 0.008-0.0161). The richest mosquito fauna was detected in Ankara, followed by Adana, and Çankırı.

CONCLUSION: This study provides new insights into mosquito richness and the prevalence of Orthoflavivirus and Wolbachia in Türkiye, contributing to vector surveillance and the potential use of Wolbachia in mosquito control strategies.}, } @article {pmid40227227, year = {2025}, author = {Arai, H and Katsuma, S and Matsuda-Imai, N and Lin, SR and Inoue, MN and Kageyama, D}, title = {Prophage-encoded Hm-oscar gene recapitulates Wolbachia-induced male-killing in the tea tortrix moth Homona magnanima.}, journal = {eLife}, volume = {13}, number = {}, pages = {}, pmid = {40227227}, issn = {2050-084X}, support = {19J13123//Japan Society for the Promotion of Science/ ; 21J00895//Japan Society for the Promotion of Science/ ; 22K14902//Japan Society for the Promotion of Science/ ; 23H02229//Japan Society for the Promotion of Science/ ; 24H02293//Japan Society for the Promotion of Science/ ; 22H00366//Japan Society for the Promotion of Science/ ; 24H02289//Japan Society for the Promotion of Science/ ; 21KK0105//Japan Society for the Promotion of Science/ ; }, mesh = {Animals ; *Wolbachia/genetics ; Male ; *Moths/microbiology/physiology/genetics ; *Prophages/genetics ; Female ; Symbiosis ; }, abstract = {Wolbachia are maternally transmitted bacterial symbionts that are ubiquitous among arthropods. They can hijack host reproduction in various ways, including male-killing (MK), where the sons of infected mothers are killed during development. The recent discovery of MK-associated Wolbachia genes, i.e., oscar in Ostrinia moths and wmk in Drosophila flies, stimulates our interest in the diversity and commonality of MK mechanisms, which remain largely unclear. We recently discovered that a Wolbachia symbiont of the moth Homona magnanima carries an MK-associated prophage region encoding homologs of oscar (Hm-oscar) and wmk (wmk-1-4). Here, we investigated the effects of these genes in the native host. Upon transient overexpression, Hm-oscar, but not wmk, induced male lethality in H. magnanima, in contrast to our observations in Drosophila, where the wmk homologs, but not Hm-oscar, killed the males. Hm-oscar disrupted sex determination in male embryos by inducing a female-type doublesex splicing and impaired dosage compensation, recapitulating the Wolbachia phenotype. Cell-based transfection assays confirmed that Hm-oscar suppressed the function of masculinizer, the primary male sex determinant involved in lepidopteran dosage compensation. Our study highlights the conserved roles of oscar homologs in Wolbachia-induced lepidopteran MK and argues that Wolbachia have evolved multiple MK mechanisms in insects.}, } @article {pmid40218346, year = {2025}, author = {Esteves-Guimarães, J and Montoya-Alonso, JA and Matos, JI and Ramalheira, E and Carretón, E and Rodríguez-Escolar, I and Balmori-de la Puente, A and Collado-Cuadrado, M and Morchón, R and Fontes-Sousa, AP}, title = {Raising Awareness of Canine, Feline and Human Dirofilariosis in Aveiro, Portugal: A One Health Perspective.}, journal = {Animals : an open access journal from MDPI}, volume = {15}, number = {7}, pages = {}, doi = {10.3390/ani15070952}, pmid = {40218346}, issn = {2076-2615}, support = {UIDB/04308/2020 and UIDP/04308/2020//Foundation for Science and Technology/ ; LANZADERA_2024//General Foundation of the University of Salamanca/ ; }, abstract = {Climatic and various socio-geographical variables have significantly influenced the global spread of Dirofilaria immitis. The coastal district of Aveiro, Portugal, marked by its unique hydrographic structure, Ria de Aveiro, and a concerning rise in heartworm disease, was the focus of our study. We aimed to update the prevalence of D. immitis in dogs and the seroprevalence in cats and humans, correlating these data with epidemiological information. A total of 430 dogs were sampled for D. immitis antigens, and 426 cats and 398 humans for D. immitis and Wolbachia sp. antibodies. In addition, we developed and validated an infection risk map for D. immitis with the geolocation of positive samples. Our results indicate a canine prevalence of 4.7%, peaking at 16.7% in Vagos. Feline and human seroprevalences were 8.9% (26.7% in Espinho) and 3.0% (Vagos presented the most alarming results), respectively. Positive samples were found in both high- and low-risk areas highlighting the need for chemoprophylaxis in all municipalities. Risk factors identified included lack of vaccination and internal deworming in cats, while dogs faced risks from inadequate vaccination and outdoor exposure. Our study identifies Aveiro as an endemic area, with a need for control measures to address this public health threat.}, } @article {pmid40209710, year = {2025}, author = {Arai, H and Wijonarko, A and Katsuma, S and Naka, H and Kageyama, D and Hornett, EA and Hurst, GDD}, title = {Evolution of Wolbachia male-killing mechanism within a host species.}, journal = {Current biology : CB}, volume = {}, number = {}, pages = {}, doi = {10.1016/j.cub.2025.03.027}, pmid = {40209710}, issn = {1879-0445}, abstract = {Male-killing bacterial symbionts, prevalent in arthropods, skew population sex ratios by selectively killing male progeny, profoundly impacting ecology and the evolution of their hosts. Male killing is a convergently evolved trait, with microbes evolving diverse male-killing mechanisms across host species with widely divergent sex determination pathways. A common evolutionary response to male-killing presence is the spread of suppressor mutations that restore male survival. In this study, we demonstrate the evolution of a novel male-killing mechanism that is insensitive to an existing male-killing suppressor. Hypolimnas bolina butterflies from Yogyakarta, Indonesia, showed extreme female-biased population sex ratio associated with high prevalence of a male-killing Wolbachia. This strain, wBol1Y, shared a very recent common ancestor with the previously characterized "suppressed" male-killing strain in the species, wBol1, but it retained its male-killing ability in the presence of the male-killing suppressor. The genome of wBol1Y differed from the suppressed wBol1 in carrying an additional prophage that included strong candidate genes for male killing. In vitro and in vivo data demonstrated that wBol1Y feminized splicing and expression of lepidopteran sex determination pathway genes and that the gene Hb-oscar-present on wBol1Y's unique prophage insert-was sufficient to disrupt the male sex determination pathway. Our study demonstrates that the diversity of male-killing mechanisms is a product both of interaction with varying insect sex determination systems and the evolution of male killing within a host species. Our data indicate that the male killer and host may be involved in escalating arms races, where spreading male-killing suppression drives the evolution of additional systems that reestablish male killing by the symbiont.}, } @article {pmid40207924, year = {2025}, author = {Hendricks, A and Philips, TK and Engl, T and Plarre, R( and Martinson, VG}, title = {The bacterial microbiome in spider beetles and deathwatch beetles.}, journal = {Microbiology spectrum}, volume = {}, number = {}, pages = {e0198124}, doi = {10.1128/spectrum.01981-24}, pmid = {40207924}, issn = {2165-0497}, abstract = {UNLABELLED: The beetle family Ptinidae contains a number of economically important pests, such as the cigarette beetle Lasioderma serricorne, the drugstore beetle Stegobium paniceum, and the diverse spider beetles. Many of these species are stored product pests, which target a diverse range of food sources, from dried tobacco to books made with organic materials. Despite the threat that the 2,200 species of Ptinidae beetles pose, fewer than 50 have been surveyed for microbial symbionts, and only a handful have been screened using contemporary genomic methods. In this study, we screen 116 individual specimens that cover most subfamilies of Ptinidae, with outgroup beetles from closely related families Dermestidae, Endecatomidae, and Bostrichidae. We used 16S ribosomal RNA gene amplicon data to characterize the bacterial microbiomes of these specimens. The majority of these species had never been screened for microbes. We found that, unlike in their sister family, Bostrichidae, that has two mutualistic bacteria seen in most species, there are no consistent bacterial members of ptinid microbiomes. For specimens which had Wolbachia infections, we did additional screening using multilocus sequence typing and showed that our populations have different strains of Wolbachia than noted in previous publications.

IMPORTANCE: Ptinid beetles are both household pests of pantry goods and economic pests of dried goods warehouses and cultural archives, such as libraries and museums. Currently, the most common pest control measures for ptinid beetles are phosphine and/or heat treatments. Many ptinid beetles have been observed to have increasing resistance to phosphine, and heat treatments are not appropriate for many of the goods commonly infested by ptinids. Pest control techniques focused on symbiotic bacteria have been shown to significantly decrease populations and often have the beneficial side effect of being more specific than other pest control techniques. This survey provides foundational information about the bacteria associated with diverse ptinid species, which may be used for future control efforts.}, } @article {pmid40202691, year = {2025}, author = {Pimentel, AC and Cesar, CS and Martins, AHB and Martins, M and Cogni, R}, title = {Wolbachia Offers Protection Against Two Common Natural Viruses of Drosophila.}, journal = {Microbial ecology}, volume = {88}, number = {1}, pages = {24}, pmid = {40202691}, issn = {1432-184X}, mesh = {Animals ; *Wolbachia/physiology ; *Drosophila melanogaster/virology/microbiology ; *Drosophila/virology/microbiology ; Symbiosis ; Female ; }, abstract = {Wolbachia pipientis is a maternally transmitted endosymbiont infecting more than half of terrestrial arthropod species. Wolbachia can express parasitic phenotypes such as manipulation of host reproduction and mutualist phenotypes such as protection against RNA virus infections. Because Wolbachia can invade populations by reproductive manipulation and block virus infection, it is used to modify natural insect populations. However, the ecological importance of virus protection is not yet clear, especially due to scarce information on Wolbachia protection against viruses that are common in nature. We used systemic infection to investigate whether Wolbachia protects its host by suppressing the titer of DMELDAV and DMelNora virus, two viruses that commonly infect Drosophila melanogaster flies in natural populations. Antiviral protection was tested in three systems to assess the impact of Wolbachia strains across species: (1) a panel of Wolbachia strains transfected into Drosophila simulans, (2) two Wolbachia strains introgressed into the natural host D. melanogaster, and (3) two native Wolbachia strains in their natural hosts Drosophila baimaii and Drosophila tropicalis. We showed that certain Wolbachia strains provide protection against DMelNora virus and DMELDAV, and this protection is correlated with Wolbachia density, which is consistent with what has been observed in protection against other RNA viruses. Additionally, we found that Wolbachia does not protect its original host, D. melanogaster, from DMELDAV infection. While native Wolbachia can reduce DMELDAV titers in D. baimaii, this effect was not detected in D. tropicalis. Although the Wolbachia protection-induced phenotype seems to depend on the virus, the specific Wolbachia strain, and the host species, our findings suggest that antiviral protection may be one of the mutualistic effects that helps explain why Wolbachia is so widespread in arthropod populations.}, } @article {pmid40196488, year = {2025}, author = {Lindsey, AR and Lue, CH and Davis, JS and Borjon, LJ and Mauthner, SE and Fricke, LC and Eads, L and Murphy, M and Drown, MK and Faulk, C and Buffington, ML and Tracey, WD}, title = {Genomics and reproductive biology of Leptopilina malgretoutensis Buffington, Lue, Davis & Tracey sp. nov. (Hymenoptera: Figitidae): An asexual parasitoid of Caribbean Drosophila.}, journal = {bioRxiv : the preprint server for biology}, volume = {}, number = {}, pages = {}, doi = {10.1101/2025.03.28.645512}, pmid = {40196488}, issn = {2692-8205}, abstract = {Drosophila and parasitic wasps in the genus Leptopilina have long been a model for understanding host-parasite interactions. Indeed, parasitic wasps are important drivers of ecological and evolutionary processes broadly, but we are generally lacking information about the diversity, natural history, and evolution of these relationships. We collected insects from the Caribbean Island of Saint Lucia, home to the eastern Caribbean ' dunni ' subgroup of Drosophila : a clade long appreciated for its recent patterns of speciation and adaptation. Here we present an integrative approach that incorporates natural history, taxonomy, physiology, and genomics to describe Leptopilina malgretoutensis Buffington, Lue, Davis & Tracey sp. nov. (Hymenoptera: Figitidae), a virulent parasitoid of dunni group flies, especially Drosophila antillea. Leptopilina malgretoutensis is nested within an early-branching clade of Leptopilina , offering insights into the evolution of this important genus of Drosophila parasitoids. We present a high-quality assembly for this wasp's 1Gbp genome, and for its bacterial endosymbiont: Wolbachia strain " w Lmal". Furthermore, we show that w Lmal induces parthenogenesis in the wasp, and that these wasps are reliant upon their Wolbachia infections to produce female offspring. Finally, comparison to historical museum specimens indicate that Leptopilina malgretoutensis had been collected approximately 40 years prior from the nearby island of Guadeloupe and were also asexually reproducing. This work represents one of only a handful of studies in which field biology, taxonomy, systematics, genomics, and experimental biology are integrated into a species description: showcasing the possibilities for biodiversity research in the genomic era.}, } @article {pmid40178596, year = {2025}, author = {Arnce, LR and Bubnell, JE and Aquadro, CF}, title = {Comparative Analysis of Drosophila Bam and Bgcn Sequences and Predicted Protein Structural Evolution.}, journal = {Journal of molecular evolution}, volume = {}, number = {}, pages = {}, pmid = {40178596}, issn = {1432-1432}, support = {R01-GM095793//National Institute of Health (US)/ ; }, abstract = {The protein encoded by the Drosophila melanogaster gene bag of marbles (bam) plays an essential role in early gametogenesis by complexing with the gene product of benign gonial cell neoplasm (bgcn) to promote germline stem cell daughter differentiation in males and females. Here, we compared the AlphaFold2 and AlphaFold Multimer predicted structures of Bam protein and the Bam:Bgcn protein complex between D. melanogaster, D. simulans, and D. yakuba, where bam is necessary in gametogenesis to that in D. teissieri, where it is not. Despite significant sequence divergence, we find very little evidence of significant structural differences in high confidence regions of the structures across the four species. This suggests that Bam structure is unlikely to be a direct cause of its functional differences between species and that Bam may simply not be integrated in an essential manner for GSC differentiation in D. teissieri. Patterns of positive selection and significant amino acid diversification across species is consistent with the Selection, Pleiotropy, and Compensation (SPC) model, where detected selection at bam is consistent with adaptive change in one major trait followed by positively selected compensatory changes for pleiotropic effects (in this case perhaps preserving structure). In the case of bam, we suggest that the major trait could be genetic interaction with the endosymbiotic bacteria Wolbachia pipientis. Following up on detected signals of positive selection and comparative structural analysis could provide insight into the distribution of a primary adaptive change versus compensatory changes following a primary change.}, } @article {pmid40176883, year = {2025}, author = {Safaei, S and Derakhshan-Sefidi, M and Karimi, A}, title = {Wolbachia: A bacterial weapon against dengue fever- a narrative review of risk factors for dengue fever outbreaks.}, journal = {New microbes and new infections}, volume = {65}, number = {}, pages = {101578}, doi = {10.1016/j.nmni.2025.101578}, pmid = {40176883}, issn = {2052-2975}, abstract = {Arboviruses constitute the largest known group of viruses and are responsible for various infections that impose significant socioeconomic burdens worldwide, particularly due to their link with insect-borne diseases. The increasing incidence of dengue fever in non-endemic regions underscores the urgent need for innovative strategies to combat this public health threat. Wolbachia, a bacterium, presents a promising biological control method against mosquito vectors, offering a novel approach to managing dengue fever. We systematically investigated biomedical databases (PubMed, Web of Science, Google Scholar, Science Direct, and Embase) using "AND" as a Boolean operator with keywords such as "dengue fever," "dengue virus," "risk factors," "Wolbachia," and "outbreak." We prioritized articles that offered significant insights into the risk factors contributing to the outbreak of dengue fever and provided an overview of Wolbachia's characteristics and functions in disease management, considering studies published until December 25, 2024. Field experiments have shown that introducing Wolbachia-infected mosquitoes can effectively reduce mosquito populations and lower dengue transmission rates, signifying its potential as a practical approach for controlling this disease.}, } @article {pmid40171161, year = {2025}, author = {Dupuis, B and Pocquet, N and Failloux, AB}, title = {Understanding the role of trehalose in interactions between Wolbachia and Aedes aegypti.}, journal = {Frontiers in cellular and infection microbiology}, volume = {15}, number = {}, pages = {1547873}, pmid = {40171161}, issn = {2235-2988}, mesh = {*Wolbachia/metabolism/physiology ; *Aedes/microbiology ; Animals ; *Trehalose/metabolism ; Mosquito Vectors/microbiology/virology ; Symbiosis ; Host-Pathogen Interactions ; }, abstract = {Mosquito-borne diseases such as chikungunya, dengue, and Zika represent a major burden on global public health. To fight against these arboviruses, vector control strategies are a priority. One existing strategy is based on the use of an endosymbiotic bacterium, Wolbachia, which reduces the transmission of arboviruses by the mosquito Aedes aegypti via a pathogen blocking effect. Wolbachia in Ae. aegypti disrupts several pathways of the host's metabolism. Trehalose is a carbohydrate circulating mainly in insect hemolymph and plays a role in numerous mechanisms as energy source or stress recovery molecule and in chitin synthesis. This study explores the importance of trehalose in the interactions between Wolbachia and Ae. aegypti, and attempts to understand the pathogen blocking effect.}, } @article {pmid40162749, year = {2025}, author = {Brunner, A and Gauliard, C and Tutagata, J and Bordenstein, SR and Bordenstein, SR and Trouche, B and Reveillaud, J}, title = {Wolbachiaand its pWCP plasmid show differential dynamics during the development of Culex mosquitoes.}, journal = {Microbiology spectrum}, volume = {}, number = {}, pages = {e0004625}, doi = {10.1128/spectrum.00046-25}, pmid = {40162749}, issn = {2165-0497}, abstract = {Mosquitoes are major vectors of pathogens such as arboviruses and parasites, causing significant health impacts each year. Wolbachia, an intracellular bacterium widely distributed among arthropods, represents a promising vector control solution. This bacterium can reduce the transmission of dengue, Zika, and chikungunya arboviruses and manipulate the reproduction of its host through its prophage WO. Although research on the Wolbachia mobilome primarily focuses on WO and the phenotypes it induces, the function of Wolbachia plasmid pWCP, recently discovered and reported to be strikingly conserved worldwide, remains unknown. In this study, we analyzed the presence and abundance of pWCP as well as Wolbachia in two different species of Culex mosquitoes, one of the most widespread genera in the world and a vector of numerous diseases. We compared the relative densities of the bacterium and its mobile genetic element in Culex pipiens molestus and Culex quinquefasciatus, a facultatively autogenous and an anautogenous species, respectively, throughout their development from the larval stage L1 to the adult individual specimen using quantitative Polymerase Chain Reaction (PCR). Our results suggest that 2-5 copies of pWCP occur in Wolbachia cells on average, and the plasmid co-replicates with Wolbachia cells. Moreover, Wolbachia and pWCP exhibit differential levels of abundance at specific development stages throughout the mosquito's life cycle in each species. These findings indicate important, and likely beneficial, roles for the plasmid in the bacterium's biology in different mosquito species as well as complex interaction dynamics between Wolbachia and its host during its life cycle.IMPORTANCEMosquitoes of the Culex genus are critical vectors for numerous diseases, causing significant public health concerns. The intracellular bacterium Wolbachia has emerged as a promising vector control solution due to its ability to interfere with pathogen transmission and manipulate mosquito reproduction. However, unlike the extensively studied WO phage, the biological significance and function of Wolbachia's pWCP plasmid, a recently discovered and strikingly conserved mobile genetic element in Culex species, remain unknown. This study investigates the developmental dynamics of pWCP and Wolbachia in two Culex mosquito species, Culex pipiens molestus and Culex quinquefasciatus across their life cycle. In general, the abundance levels of Wolbachia and the plasmid were found to vary across life stages and differ between the two species. However, a relatively small number of pWCP copies were observed per Wolbachia cell, together with a co-replication of the plasmid with the bacterium for most developmental stages. Altogether, these findings suggest a likely beneficial and non-parasitic role for pWCP in Wolbachia's biology, which may contribute to the intricate interactions between the bacterium and its mosquito hosts.}, } @article {pmid40161648, year = {2025}, author = {Nelson, E and Pereira, TN and Ribeiro, EMC and Mafra E Silva, BD and Camillo, C and Rodrigues da Costa, T and Teixeira, MM and Ko, AI and Cummings, DAT and Moreira, LA}, title = {Comparison of oviposition and adult trapping to monitor w Mel introgression for Wolbachia- based vector control.}, journal = {bioRxiv : the preprint server for biology}, volume = {}, number = {}, pages = {}, doi = {10.1101/2025.03.10.642347}, pmid = {40161648}, issn = {2692-8205}, abstract = {UNLABELLED: Wolbachia introgression into Aedes aegypti mosquito populations has been shown to be effective in preventing dengue and is being evaluated for WHO prequalification. Monitoring the long-term introgression of Wolbachia (w Mel)-positive Aedes aegypti mosquitoes, however, requires labor-intensive and costly BG-Sentinel traps (BG-traps). More affordable alternatives, such as using oviposition traps (ovitraps), have not been fully evaluated. Ae. aegypti eggs and adults were collected from 124 ovitraps and 237 BG-traps, respectively, across 12 clusters in Belo Horizonte, Brazil from March to May 2023 as part of the EVITA Dengue trial. We used a qPCR assay to detect w Mel in a sample of L3-L4 stage larvae (up to 29) that were reared from eggs in ovitraps and adults from BG-traps (up to 10 per BG-trap). We used mixed effects models to compare estimates of cluster-level w Mel introgression from ovitrap and BG-trap data over time. Among 3,675 larvae reared from ovitraps, w Mel prevalence was 0.50 (95% CI: 0.48-0.51). Among 1,244 adult Ae. aegypti tested from BG-traps, w Mel prevalence was 0.45 (95% CI: 0.42-0.48). Cluster-level w Mel introgression in larvae and adults was highly correlated (Spearman's r = 0.70, p = 6.71e-06). Multivariate analysis found that ovitrap estimates of introgression were associated with BG-trap estimates in the same month when models incorporated the previous month's ovitrap w Mel-positive count, the proportion of w Mel in ovitraps in the current and previous month, and Ae. aegypti abundance. Leveraging this model, predicted w Mel introgression from ovitrap data were highly correlated with observed introgression from BG-trap data (r s,counts =0.98, p=1.53e-14; r s,prevalences =0.82, p=0.11e-05) and provided greater precision than crude ovitrap-based estimates. These findings indicate that ovitrap-based monitoring represents a low cost, more efficient approach to evaluating introgression as the Wolbachia -based interventions are scaled up and implemented broadly in high burden regions for dengue and other arboviral diseases.

AUTHOR SUMMARY: Dengue fever is a major global health burden, and one promising way to control it is by releasing Aedes aegypti mosquitoes infected with a bacteria called Wolbachia . This bacteria reduces the mosquitoes' ability to spread dengue. However, monitoring the success of Wolbachia in mosquito populations over time requires expensive and labor-intensive traps. Our study explored a more affordable alternative: using oviposition traps (ovitraps), which collect mosquito eggs instead of adults. In Belo Horizonte, Brazil, we compared data from eggs collected in ovitraps with data from standard mosquito traps (BG-traps) that catch adult mosquitoes. By analyzing the mosquitoes for Wolbachia , we found that the egg-based method provided reliable estimates of Wolbachia levels in the mosquito population. These results suggest that ovitraps could be a cost-effective and efficient way to monitor Wolbachia 's spread. This approach could help improve dengue prevention efforts, making it easier for public health programs to track and expand this control strategy in areas where dengue is a major concern.}, } @article {pmid40155909, year = {2025}, author = {Lim, JT and Mailepessov, D and Chong, CS and Dickens, B and Lai, YL and Ng, Y and Lu, D and Lee, C and Tan, LY and Chain, G and Ho, SH and Chang, CC and Ma, P and Bansal, S and Lee, V and Sim, S and Tan, CH and Ng, LC}, title = {Adjacent spillover efficacy of Wolbachia for control of dengue: emulation of a cluster randomised target trial.}, journal = {BMC medicine}, volume = {23}, number = {1}, pages = {184}, pmid = {40155909}, issn = {1741-7015}, mesh = {*Wolbachia/physiology ; *Dengue/prevention & control/transmission ; Humans ; Animals ; *Aedes/microbiology/virology ; Male ; Female ; Adult ; Young Adult ; Adolescent ; Middle Aged ; Child ; Child, Preschool ; Aged ; Mosquito Control/methods ; Infant ; }, abstract = {BACKGROUND: Matings between male Aedes aegypti mosquitoes infected with wAlbB strain of Wolbachia and wild-type females yield non-viable eggs, thereby suppressing Ae. aegypti abundance in the field. We evaluated the spillover efficacy of releasing wAlbB-infected Ae. aegypti male mosquitoes to suppress dengue in sites adjacent to release sites (spillover sites).

METHODS: The protocol of a two-arm cluster-randomised test-negative controlled trial (cRCT) was specified and emulated using a nationally representative dengue test-negative/positive database of 454,437 individuals reporting for febrile illness to primary or secondary care in public healthcare institutions. Spillover intervention sites were defined by geolocating locations which were adjacent to, i.e. shared geographical borders with, actual Wolbachia intervention sites. We built a cohort of individuals who resided in spillover sites versus a comparator control group who resided in sites which did not receive Wolbachia interventions. We emulated a constrained randomisation protocol used in cRCTs to balance dengue risk between spillover and control arms in the pre-intervention period. We matched individuals reporting for testing in intervention and control groups by calendar time and a high-dimensional battery of sociodemographic, environmental and anthropogenic variables. Intention-to-treat analysis was conducted to estimate the protective efficacy against dengue given spillover Wolbachia exposure.

RESULTS: The final cohort consisted of 2354 matched individuals residing in Wolbachia spillover and control sites for at least 3 months in the study period. Compared to the controls, individuals residing in spillover sites for 3 or more months were associated with a 45% (OR: 0.55, 95% CI: 0.42‒0.74) reduction in risk of contracting dengue. Higher durations of spillover Wolbachia exposure also modestly increased protective efficacies. Compared to the control arm, the proportion of virologically confirmed dengue cases was lower in the spillover arm overall and across each subgroup. Protective efficacies were found across all years, age and sex subgroups.

CONCLUSIONS: Our results demonstrated the potential of Wolbachia-mediated sterility for reducing the risk of contracting dengue even in sites which were not directly treated by the intervention.}, } @article {pmid40151487, year = {2025}, author = {Shishkina, OD and Gruntenko, NE}, title = {Symbiosis of intracellular bacteria Wolbachia with insects: a hundred years of study summarized.}, journal = {Vavilovskii zhurnal genetiki i selektsii}, volume = {29}, number = {1}, pages = {79-91}, doi = {10.18699/vjgb-25-10}, pmid = {40151487}, issn = {2500-0462}, abstract = {Wolbachia pipientis is an α-proteobacterium, which is a widespread intracellular symbiont in a number of Arthropoda and some Nematoda species. With insects, W. pipientis forms a symbiont-host system characterized by very close interactions between its components. The mutual effects of Wolbachia on the host and the host on Wolbachia are important biotic factors for both components of this symbiotic system. Wolbachia is able to affect both host reproduction and somatic organ function. Due to its prevalence among insects and a wide variety of both negative (cytoplasmic incompatibility and androcide are among the most well-known examples) and positive (increasing resistance to biotic and abiotic factors, providing vitamins and metabolites) effects on the host organism, Wolbachia is of great interest for both entomologists and microbiologists. The diversity of host phenotypes induced by Wolbachia provides a broad choice of evolutionary strategies (such as reproductive parasitism or mutually beneficial symbiont-host relationships) that it utilizes. The influence of Wolbachia is to be considered in the design of any experiment conducted on insects. The application of sequencing technologies has led to new approaches being created to study the existing relationships within the Wolbachia-insect system, but interpretation of the data obtained is challenging. Nevertheless, the prospects for the use of the whole-genome analysis data to study Wolbachia-host coevolution are beyond doubt. Ongoing projects to introduce Wolbachia strains, which provide antiviral host defense, into insect populations to control the spread of RNA-viruses are actively pursued, which could result in saving many human lives. The aim of this brief review is to summarize the data collected by scientists over the past hundred years of Wolbachia studies and the current understanding of its genetic diversity and mechanisms of interaction with the host, including those based on transcriptome analysis.}, } @article {pmid40146382, year = {2025}, author = {Nain, D and Rana, A and Raychoudhury, R and Sen, R}, title = {Parasite-Induced Replacement of Host Microbiota: Impact of Xenos gadagkari Parasitization on the Microbiota of Polistes wattii.}, journal = {Microbial ecology}, volume = {88}, number = {1}, pages = {20}, pmid = {40146382}, issn = {1432-184X}, support = {DST/INSPIRE/03/2021/000175, IF200146//Department of Science and Technology, Ministry of Science and Technology, India/ ; 1061830779//Council of Scientific and Industrial Research, India/ ; CRG-2021/007010//Science and Engineering Research Board/ ; CRG-2021/007010//Science and Engineering Research Board/ ; }, mesh = {Animals ; *Wasps/microbiology/physiology ; *Microbiota ; Female ; Male ; *RNA, Ribosomal, 16S/genetics ; Host-Parasite Interactions ; Seasons ; Bacteria/genetics/classification/isolation & purification ; }, abstract = {The study of microbiota of social insects under different ecological conditions can provide important insights into the role of microbes in their biology and behavior. Polistes is one of the most widely distributed and extensively studied genera of social wasps, yet a comprehensive study on the microbiota of any species of Polistes or any primitively eusocial wasp is missing. Polistes wattii is an Asian wasp, which hibernates in winter and exhibits a biannual nest founding strategy. It is often parasitized by the strepsipteran endoparasite/parasitoid Xenos gadagkari, which changes the morpho-physiology and behavior of their hosts. In this study, we employ 16S rRNA amplicon sequencing, using the Oxford Nanopore platform, to study the microbial community of P. wattii and investigate the effects of seasonality, sex, and Xenos parasitism. We show that the microbiota differs in females from solitary foundress spring nests and multiple foundress summer nests. The microbiota also differs in males and females. Finally, we show that X. gadagkari parasitism replaces and homogenizes the microbiota of P. wattii. Unlike the unparasitized wasps, the microbiota of X. gadagkari parasitoids and parasitized wasps are dominated by Wolbachia and Providencia. Although the normal microbiota of P. wattii resembles that of highly eusocial vespid wasps, we show that the microbiota of parasitized P. wattii becomes more like the microbiota of strepsipterans. Therefore, it appears that X. gadagkari and other such strepsipteran parasitoids may have a bigger impact on the biology of their hosts than previously thought.}, } @article {pmid40144380, year = {2025}, author = {Gokhman, VE and Ryabinin, AS and Bykov, RA and Ilinsky, YY}, title = {The lowest chromosome number in the family Pteromalidae (Hymenoptera: Chalcidoidea): the karyotype and other genetic features of Pachycrepoideus vindemmiae (Rondani, 1875).}, journal = {Vavilovskii zhurnal genetiki i selektsii}, volume = {29}, number = {1}, pages = {108-112}, doi = {10.18699/vjgb-25-12}, pmid = {40144380}, issn = {2500-0462}, abstract = {Various genetic features of the hitman strain of the widespread parasitoid of Drosophilidae (Diptera), Pachycrepoideus vindemmiae (Rondani, 1875) (Pteromalidae, Pachyneurinae) were studied. This strain was established and is maintained at the Institute of Cytology and Genetics of the Siberian Branch of the Russian Academy of Sciences (Novosibirsk, Russia). An analysis of air-dried chromosome preparations from prepupae of this parasitoid showed that it has n = 4 and 2n = 8 in males and females, respectively, which is the lowest known chromosome number in the family Pteromalidae. All chromosomes in the karyotype of this species are metacentric. The first and second chromosomes are of similar size, the remaining ones are substantially shorter. The same results were obtained for an additional strain of this species kept at the Moscow State University (Moscow, Russia). A comparison of the DNA sequence of the barcoding region of the mitochondrial cytochrome c oxidase (COI) gene of the hitman strain of P. vindemmiae with those available from the GenBank and BoLD databases demonstrated that this strain clustered together with conspecifics originating from China, Turkey and Italy. Despite certain endosymbionts being previously reported for the genus Pachycrepoideus Ashmead, 1904 as well as for P. vindemmiae itself, the hitman strain turned out to be free of endosymbiotic bacteria in the genera Arsenophonus Gherna et al., 1991, Cardinium Zchori-Fein et al., 2004, Rickettsia da Rocha-Lima, 1916, Spiroplasma Saglio et al., 1973 and Wolbachia Hertig, 1936. The above-mentioned results improve our knowledge of various genetic features of parasitoids of the family Pteromalidae and those of P. vindemmiae in particular.}, } @article {pmid40142534, year = {2025}, author = {Wang, S and Wang, X and Basit, A and Wei, Q and Zhao, K and Zhao, Y}, title = {Interactions Between Endosymbionts Wolbachia and Rickettsia in the Spider Mite Tetranychus turkestani: Cooperation or Antagonism?.}, journal = {Microorganisms}, volume = {13}, number = {3}, pages = {}, doi = {10.3390/microorganisms13030642}, pmid = {40142534}, issn = {2076-2607}, support = {No. 32260676,No. 31860508//National Natural Science Foundation of China/ ; No. 2022CB002 - 06//the Program for Young Leading Scientists in Science and Technology of XPCC/ ; No. 2024DA018//the Natural Science Foundation of XPCC/ ; }, abstract = {Maternally inherited endosymbionts are widespread in arthropods, with multiple symbionts commonly co-existing within a single host, potentially competing for or sharing limited host resources and space. Wolbachia and Rickettsia, two maternally-inherited symbionts in arthropods, can co-infect hosts, yet research on their combined impacts on host reproduction and interaction remains scarce. Tetranychus turkestani (Acari: Tetranychidae) is an important agricultural pest mite, characterized by rapid reproduction, a short life cycle, and being difficult to control. Wolbachia and Rickettsia are two major endosymbiotic bacteria present in T. turkestani. This study used diverse parthenogenetic backcross and antibiotic screening to explore the reproductive effects of these two symbionts on T. turkestani. The results show that single Rickettsia infection induced male killing in the amphigenesis of T. turkestani, leading to arrhenotokous embryo death and fewer offspring. Single Wolbachia infection induced strong cytoplasmic incompatibility (CI). During dual infection, CI intensity decreased because Rickettsia's male-killing effect antagonized the Wolbachia-induced CI. Dual-infected mites had increased oviposition, lower mortality, a higher female-to-male ratio, and more offspring, thus enhancing T. turkestani's fitness. These findings will be helpful for understanding the nature of host-endosymbiont interactions and the potential for evolutionary conflicts, offering insights into their co-evolutionary relationship.}, } @article {pmid40142501, year = {2025}, author = {Wang, X and Wang, S and Basit, A and Wei, Q and Zhao, K and Liu, F and Zhao, Y}, title = {Metabolomics Provides New Insights into the Mechanisms of Wolbachia-Induced Plant Defense in Cotton Mites.}, journal = {Microorganisms}, volume = {13}, number = {3}, pages = {}, doi = {10.3390/microorganisms13030608}, pmid = {40142501}, issn = {2076-2607}, support = {32260676//National Natural Science Foundation of China/ ; 31860508//National Natural Science Foundation of China/ ; 2022CB002-06//China Corps strong youth science and technology leading talent project/ ; 2024DA018//Natural Science Foundation of the China Corps/ ; }, abstract = {Endosymbiotic bacteria play a significant role in the co-evolution of insects and plants. However, whether they induce or inhibit host plant defense responses remains unclear. In this study, non-targeted metabolomic sequencing was performed on cotton leaves fed with Wolbachia-infected and uninfected spider mites using parthenogenetic backcrossing and antibiotic treatment methods. A total of 55 differential metabolites were identified, which involved lipids, phenylpropanoids, and polyketides. KEGG pathway enrichment analysis revealed seven significantly enriched metabolic pathways. Among them, flavonoid and flavonol biosynthesis, glycerophospholipid metabolism, and ether lipid metabolism showed extremely significant differences. In Wolbachia-infected cotton leaves, the flavonoid biosynthesis pathway was significantly up-regulated, including quercetin and myricetin, suggesting that the plant produces more secondary metabolites to enhance its defense capability. Glycerophosphocholine (GPC) and sn-glycerol-3-phosphoethanolamine (PE) were significantly down-regulated, suggesting that Wolbachia may impair the integrity and function of plant cell membranes. The downregulation of lysine and the upregulation of L-malic acid indicated that Wolbachia infection may shorten the lifespan of spider mites. At various developmental stages of the spider mites, Wolbachia infection increased the expression of detoxification metabolism-related genes, including gene families such as cytochrome P450, glutathione S-transferase, carboxylesterase, and ABC transporters, thereby enhancing the detoxification capability of the host spider mites. This study provides a theoretical basis for further elucidating the mechanisms by which endosymbiotic bacteria induce plant defense responses and expands the theoretical framework of insect-plant co-evolution.}, } @article {pmid40137770, year = {2025}, author = {Moretti, R and Lim, JT and Ferreira, AGA and Ponti, L and Giovanetti, M and Yi, CJ and Tewari, P and Cholvi, M and Crawford, J and Gutierrez, AP and Dobson, SL and Ross, PA}, title = {Exploiting Wolbachia as a Tool for Mosquito-Borne Disease Control: Pursuing Efficacy, Safety, and Sustainability.}, journal = {Pathogens (Basel, Switzerland)}, volume = {14}, number = {3}, pages = {}, doi = {10.3390/pathogens14030285}, pmid = {40137770}, issn = {2076-0817}, mesh = {*Wolbachia/physiology/genetics ; Animals ; *Mosquito Vectors/microbiology/virology ; *Vector Borne Diseases/prevention & control/transmission ; Humans ; *Mosquito Control/methods ; Female ; Male ; Culicidae/microbiology/virology ; Mosquito-Borne Diseases ; }, abstract = {Despite the application of control measures, mosquito-borne diseases continue to pose a serious threat to human health. In this context, exploiting Wolbachia, a common symbiotic bacterium in insects, may offer effective solutions to suppress vectors or reduce their competence in transmitting several arboviruses. Many Wolbachia strains can induce conditional egg sterility, known as cytoplasmic incompatibility (CI), when infected males mate with females that do not harbor the same Wolbachia infection. Infected males can be mass-reared and then released to compete with wild males, reducing the likelihood of wild females encountering a fertile mate. Furthermore, certain Wolbachia strains can reduce the competence of mosquitoes to transmit several RNA viruses. Through CI, Wolbachia-infected individuals can spread within the population, leading to an increased frequency of mosquitoes with a reduced ability to transmit pathogens. Using artificial methods, Wolbachia can be horizontally transferred between species, allowing the establishment of various laboratory lines of mosquito vector species that, without any additional treatment, can produce sterilizing males or females with reduced vector competence, which can be used subsequently to replace wild populations. This manuscript reviews the current knowledge in this field, describing the different approaches and evaluating their efficacy, safety, and sustainability. Successes, challenges, and future perspectives are discussed in the context of the current spread of several arboviral diseases, the rise of insecticide resistance in mosquito populations, and the impact of climate change. In this context, we explore the necessity of coordinating efforts among all stakeholders to maximize disease control. We discuss how the involvement of diverse expertise-ranging from new biotechnologies to mechanistic modeling of eco-epidemiological interactions between hosts, vectors, Wolbachia, and pathogens-becomes increasingly crucial. This coordination is especially important in light of the added complexity introduced by Wolbachia and the ongoing challenges posed by global change.}, } @article {pmid40134009, year = {2025}, author = {Zhang, J and Lv, T and Tan, S and Yu, L and Chi, Y and Chen, J and Fan, X and Lu, X}, title = {Screening helper T lymphocyte epitopes based on IFN-γ/IL-10 ratio for developing a novel multi-epitope vaccine candidate using Wolbachia surface protein as an adjuvant against visceral leishmaniasis.}, journal = {Parasites & vectors}, volume = {18}, number = {1}, pages = {116}, pmid = {40134009}, issn = {1756-3305}, support = {82404961//National Natural Science Foundation of China/ ; 226-2024-00001//Fundamental Research Funds for the Central Universities/ ; }, mesh = {*Epitopes, T-Lymphocyte/immunology ; *Interferon-gamma/metabolism/immunology ; *Leishmaniasis Vaccines/immunology ; *Adjuvants, Immunologic ; *Leishmaniasis, Visceral/prevention & control/immunology ; Animals ; *Interleukin-10/metabolism/immunology ; *Wolbachia/immunology ; Membrane Proteins/immunology ; Humans ; Bacterial Proteins/immunology ; Th1 Cells/immunology ; Molecular Docking Simulation ; T-Lymphocytes, Helper-Inducer/immunology ; Toll-Like Receptor 4/immunology ; Toll-Like Receptor 2/immunology ; Leishmania donovani/immunology ; }, abstract = {BACKGROUND: Visceral leishmaniasis (VL) is the most lethal form of leishmaniasis. In terms of anti-leishmanial vaccines, favorable immune responses are Th1 responses that primarily produce interferon gamma (IFN-γ) and activate macrophages for leishmanicidal effects. The selection of IFN-γ-inducing epitopes in silico may reduce the steps of pre-clinical evaluation and increase the certainty of the better-designed vaccine. Wolbachia surface protein (WSP) derived from Wolbachia bacteria that have been reported to reside in sandflies can trigger TLR2 and TLR4 activation to favor Th1 immune responses, which may serve as a potential adjuvant candidate for the Leishmania vaccine. Therefore, in this study, helper T lymphocyte epitopes that may induce favorable immune responses were identified, and WSP was served as an adjuvant to design a novel multi-epitope vaccine candidate.

METHODS: Leishmania hemoglobin receptor (HbR), kinetoplastid membrane protein-11 (KMP-11), glycoprotein of 63 kDa (Gp63), thiol-specific antioxidant antigen (TSA), and sterol 24-c-methyltransferase (SMT) were analyzed by immunoinformatics to screen helper T lymphocyte and cytotoxic T lymphocyte epitopes. The antigenicity, toxicity, allergenicity, and IFN-γ-inducing epitope potential of T epitopes were predicted. The immune simulation was performed to calculate IFN-γ/interleukin (IL)-10 ratios to predict the immune responses induced by the helper T lymphocyte epitopes. Molecular docking and molecular dynamics simulations were carried out to analyze the interactions and stability of the docked complexes. The immune simulation of a multi-epitope vaccine candidate was carried out to predict its immunogenicity.

RESULTS: Some helper T lymphocyte epitopes that were predicted with the potential of inducing Th1 responses and cytotoxic T lymphocyte epitopes were selected to develop a novel multi-epitope vaccine candidate with WSP as an adjuvant. It was found in molecular docking and interaction analysis that TLR2 and TLR4 can recognize WSP, supporting the potential of adjuvant for the Leishmania vaccine. The results from immune simulation demonstrated that the multi-epitope vaccine candidate induced obvious cytokine (IFN-γ, IL-12, and IL-2) secretion and Th1 as well as memory T cell production, similar to that of Leish-111f.

CONCLUSIONS: Our vaccine candidate may interact with TLR2 and TLR4 and exhibit good immunogenicity, favoring Leishmania clearance. Our strategy provides a novel multi-epitope vaccine candidate and references for other vaccine developments.}, } @article {pmid40128692, year = {2025}, author = {Leitner, M and Murigneux, V and Etebari, K and Asgari, S}, title = {Wolbachia elevates host methyltransferase expression and alters the m[6]A methylation landscape in Aedes aegypti mosquito cells.}, journal = {BMC microbiology}, volume = {25}, number = {1}, pages = {164}, pmid = {40128692}, issn = {1471-2180}, support = {DP210101791//Australian Research Council/ ; }, mesh = {*Wolbachia/genetics ; Animals ; *Aedes/microbiology/genetics ; *Methyltransferases/genetics/metabolism ; *Adenosine/analogs & derivatives/metabolism ; Methylation ; Insect Proteins/genetics/metabolism ; }, abstract = {Wolbachia pipientis is an intracellular endosymbiotic bacterium that blocks the replication of several arboviruses in transinfected Aedes aegypti mosquitoes, yet its antiviral mechanism remains unknown. For the first time, we employed Nanopore direct RNA sequencing technology to investigate the impact of wAlbB strain of Wolbachia on the host's N[6]-methyladenosine (m[6]A) machinery and post-transcriptional modification landscape. Our study revealed that Wolbachia infection elevates the expression of genes involved in the mosquito's m[6]A methyltransferase complex. However, knocking down these m[6]A-related genes did not affect Wolbachia density. Nanopore sequencing identified 1,392 differentially modified m[6]A DRACH motifs on mosquito transcripts, with 776 showing increased and 616 showing decreased m[6]A levels due to Wolbachia. These m[6]A sites were predominantly enriched in coding sequences and 3'-untranslated regions. Gene Ontology analysis revealed that genes with reduced m[6]A levels were over-represented in functional GO terms associated with purine nucleotide binding functions critical in the post-transcriptional modification process of m[6]A. Differential gene expression analysis of the Nanopore data uncovered that a total of 643 protein-coding genes were significantly differentially expressed, 427 were downregulated, and 216 were upregulated. Several classical and non-classical immune-related genes were amongst the downregulated DEGs. Notably, it revealed a critical host factor, transmembrane protein 41B (TMEM41B), which is required for flavivirus infection, was upregulated and methylated in the presence of Wolbachia. Indeed, there is a strong correlation between gene expression being upregulated in genes with both increased and decreased levels of m[6]A modification, respectively. Our findings underscore Wolbachia's ability to modulate many intracellular aspects of its mosquito host by influencing post-transcriptional m[6]A modifications and gene expression, and it unveils a potential link behind its antiviral properties.}, } @article {pmid40126663, year = {2025}, author = {Karkuzhali, N and Edward, YSJT and Chitra, N and Senthilkumar, M and Ramalingam, J}, title = {Unveiling the diversity of gut microbes in green lacewings (Chrysopidae: Neuroptera) and their role as protagonist in nutrition.}, journal = {Archives of microbiology}, volume = {207}, number = {5}, pages = {100}, pmid = {40126663}, issn = {1432-072X}, mesh = {Animals ; *Gastrointestinal Microbiome/physiology ; Bacteria/metabolism/genetics/classification ; Insecta/microbiology ; Larva/microbiology ; }, abstract = {Green lacewings (Chrysopidae; Neuroptera) plays a crucial role as predators against insect pests in diverse cropping systems. Larval chrysopids are predatory on mealybugs, aphids, scales, whiteflies, mites and eggs of many arthropods. Adults are palynoglycophagous and feed on nectar, pollen, and honeydew secreted by aphids. Many insects cannot synthesize necessary vitamins and amino acids on their own and depend on gut microbes. Microbes associated with chrysopid gut help them with balanced nutrition and ecological fitness to withstand extreme stresses, especially adult gut microbiota, which constitutes an indispensable part of nutrients in addition to reproduction. Except for yeast, microbes such as bacteria in the chrysopid larval and adult gut have not been extensively studied. This review aims to seek a comprehensive overview of the gut microbes present in the chrysopids and their role in improving the fitness of chrysopids through adequate nutrition. This will pave the way for further research on understanding the microbe-mediated metabolic activities, their role in toxin production, and the development of probiotic feed from the novel gut microbiota for improving the productivity of laboratory-reared chrysopids used in augmentative biological control of major pests in agricultural ecosystems.}, } @article {pmid40126013, year = {2025}, author = {Hubert, J and Glowska-Patyniak, E and Dowd, SE and Klimov, PB}, title = {A novel Erwiniaceae gut symbiont modulates gene expression of the intracellular bacterium Cardinium in the stored product mite Tyrophagus putrescentiae.}, journal = {mSphere}, volume = {}, number = {}, pages = {e0087924}, doi = {10.1128/msphere.00879-24}, pmid = {40126013}, issn = {2379-5042}, abstract = {We examined host and bacterial gene expression profiles in the stored product mite Tyrophagus putrescentiae co-infected with Wolbachia (wTPut) and Cardinium (cTPut) while varying the presence of the Erwiniaceae symbiont (SLS). SLS, a novel symbiont in the family Erwiniaceae, with a genome size of 1.7 Mb, is found in 16% of mite species in infected cultures. In addition, SLS was detected in mite feces but not in their eggs. Although Wolbachia expression remained unchanged, the presence or absence of SLS significantly affected Cardinium expression. It indicated that the effect of Wolbachia on SLS was neutral. In SLS-positive samples, Cardinium exhibited 29 upregulated and 48 downregulated genes compared to SLS-negative samples. Furthermore, Cardinium gene expression strongly correlated with mite KEGG gene expression in SLS-positive samples. Positive Spearman's correlations between Cardinium gene expression and mite KEGG immune and regulatory pathways were doubled in SLS-positive compared to SLS-negative samples. The diversity of expressed genes in the mite host decreased in the presence of SLS. Cardinium had more interacting genes to mite host in SLS-positive samples than without SLS. Transposases are the most affected Cardinium genes, showing upregulation in the presence of SLS. Correlation analyses revealed interactions between Cardinium and SLS via mite immune and regulatory pathways, including lysosome, ubiquitin-mediated proteolysis, PIK3_Akt, and cGMP-PKG. The results showed that Cardinium indirectly affects the gut symbionts of mites.IMPORTANCEThis study introduces a new model to analyze interactions between intracellular bacterial symbionts, gut bacterial symbionts, and their mite hosts. Using gene expression correlations, we investigated how the intracellular Cardinium responds to the novel Erwiniaceae gut symbiont in the mold mite Tyrophagus putrescentiae. The data showed that both mite and Cardinium gene expression are different in the samples with and without Erwiniaceae symbionts. In the presence of Erwiniaceae symbionts, Cardinium increased the interaction with the mite host in terms of changes in gene expression. The mite immune and regulatory pathway gene expression is differently correlated to Cardinium genes in relation to Erwiniaceae symbionts. As a well-known producer of allergens, T. putrescentiae physiology and thus its allergen production are influenced by both symbionts, potentially affecting the release of allergens into human environments.}, } @article {pmid40110431, year = {2025}, author = {Bednarikova, S and Danek, O and Dundarova, H and Nemcova, M and Piacek, V and Zukalova, K and Zukal, J and Pikula, J}, title = {Filariasis of parti-colored bats: phylogenetic analysis, infection prevalence, and possible vector mite identification.}, journal = {Frontiers in veterinary science}, volume = {12}, number = {}, pages = {1546353}, pmid = {40110431}, issn = {2297-1769}, abstract = {INTRODUCTION: The epidemiology of filarial infections is a neglected area of bat research, with little information on filarial species diversity, life cycles, host ranges, infection prevalence and intensity, parasite pathogenicity, or competent vectors. Furthermore, molecular data for filarial worms are largely lacking.

METHODS: Here, we examined 27 cadavers of parti-colored bat (Vespertilio murinus) from Czech rescue centers for filarial infection using gross necropsy. We also used nested polymerase chain reactions targeting partial mitochondrial cytochrome c oxidase subunit I (COI) partial gene to detect and genotype filarial parasites within organs, and ectoparasites of V. murinus from Russian and Slovak summer bat colonies. Samples with mixed filarial infections were cloned to extract separate sequences. The COI gene sequences were then subjected to phylogenetic analysis and a phylogenetic tree constructed. Adult filarial worms were also screened for the bacterial symbiont Wolbachia, using a standard PCR targeting the partial 16S rRNA gene.

RESULTS: Two filarial nematode species were identified in single and mixed V. murinus infections, Litomosa sp. and a species of Onchocercidae. Adult Litomosa sp. and a species of Onchocercidae. Adult Litomosa sp. nematodes were only recorded during necropsy of the abdominal, thoracic, and gravid uterine cavities of four bats. Molecular screening of organs for filarial DNA revealed prevalences of 81.5, 51.9 and 48.1% in Litomosa sp., Onchocercid sp. and co-infected bats, respectively. Adult Litomosa sp. worms proved negative for Wolbachia. The macronyssid mite Steatonyssus spinosus, collected in western Siberia (Russia), tested positive for Onchocercid sp. and mixed microfilarial infection.

DISCUSSION: Our results revealed high prevalence, extensive geographic distribution and a potential vector of filarial infection in V. murinus. Our data represent an important contribution to the field of bat parasitology and indicate the need for a taxonomic revision of bat-infecting filarial nematodes based on both morphological and molecular methods.}, } @article {pmid40108258, year = {2025}, author = {Szymkowiak, P and Konecka, E and Rutkowski, T and Pecyna, A and Szwajkowski, P}, title = {Alien spiders in a palm house with the first report of parthenogenetic Triaeris stenaspis (Araneae: Oonopidae) infected by Wolbachia from new supergroup X.}, journal = {Scientific reports}, volume = {15}, number = {1}, pages = {9512}, pmid = {40108258}, issn = {2045-2322}, mesh = {Animals ; *Spiders/microbiology/physiology ; *Phylogeny ; *Wolbachia/genetics/physiology/classification ; RNA, Ribosomal, 16S/genetics ; Symbiosis ; Poland ; Parthenogenesis ; }, abstract = {Palm houses in Europe serve as urban biodiversity hot spots for alien spiders. As a result of several years of research in the Poznań Palm House, we documented the occurrence of 14 spider species, 9 of which were alien to Europe: Coleosoma floridanum, Hasarius adansoni, Howaia mogera, Ostearius melanopygius, Parasteatoda tabulata, Parasteatoda tepidariorum, Scytodes fusca, Spermophora kerinci and Triaeris stenaspis. The most abundant species was C. floridanum (39.9%). Three spider species were recorded for the first time in Poland: C. floridanum, S. fusca and S. kerinci. We studied the occurrence of endosymbiotic Wolbachia and Cardinium in parthenogenetic T. stenaspis and recorded for the first time the occurrence of Wolbachia in this spider. The endosymbiont was characterized based on the sequences of six bacterial housekeeping genes: 16S rRNA, coxA, fbpA, ftsZ, gatB and hcpA. Our phylogenetic reconstruction of Wolbachia supergroups revealed that the bacteria recovered from the spider formed distinct lineages in relation to all known supergroups. We assigned it to a novel supergroup X with unique sequences within the 16S rRNA and ftsZ genes. We discussed faunistic results in terms of long-term survival rates and the risk of invasion of alien species of spiders.}, } @article {pmid40101296, year = {2025}, author = {Ohm, JR and Lynd, A and McGowan, A and Cupid, A and Bellot, V and Le, JQ and Kakani, E and Livni, J and Crawford, JE and White, BJ}, title = {Mark-Release-Recapture of Packed and Shipped Aedes aegypti with Wolbachia: Implications for Conducting Remote Incompatible Insect Technique Programs.}, journal = {The American journal of tropical medicine and hygiene}, volume = {}, number = {}, pages = {}, doi = {10.4269/ajtmh.24-0262}, pmid = {40101296}, issn = {1476-1645}, abstract = {Male mosquitoes containing the endosymbiont Wolbachia (Wb+) can be used as a tool to suppress wild mosquito populations through a technique termed incompatible insect technique (IIT). IIT programs reduce wild mosquitoes via incompatible matings between released males and wild females to reduce the number of viable offspring produced in the next generation. Successful programs rely on regular release of incompatible males to outcompete wild males for female mates. Past IIT programs have relied on local production of Wb+ males to support regular releases of incompatible males. Here, we evaluated the survival and dispersal of packed and shipped Wb+ Aedes aegypti males in mark-release-recapture studies at a release site in the British Virgin Islands (BVI), separated by over 3,600 miles from the centralized production facility. Released mosquitoes were recaptured using BG-Sentinel 2 traps collected daily for up to 7 days after release. Wb+ male mosquitoes packed and shipped from a centralized production facility performed similarly to males that were locally reared in the BVI in survival, dispersal, and recapture rates. Our results support the conclusion that packing and shipping live Wb+ male mosquitoes does not impact their ability to survive and disperse in release sites and suggests that IIT mosquito control programs can feasibly be conducted nearly anywhere in the world without the need for local mosquito production facilities.}, } @article {pmid40084540, year = {2025}, author = {Detcharoen, M and Nilsai, A and Thaochan, N and Nuansuwon, C}, title = {Low Wolbachia incidence in Bactrocera and Zeugodacus species from Thailand and genome analysis of Wolbachia associated with Zeugodacus apicalis.}, journal = {Journal of economic entomology}, volume = {}, number = {}, pages = {}, doi = {10.1093/jee/toaf054}, pmid = {40084540}, issn = {1938-291X}, support = {SCI6402027S//Prince of Songkla University/ ; }, abstract = {Wolbachia are bacterial endosymbionts found widely in arthropods and filarial nematodes. Infecting about half of all arthropod species, Wolbachia manipulate their hosts in various ways, including cytoplasmic incompatibility. Here, we investigated Wolbachia diversity in Bactrocera and Zeugodacus, two prevalent tephritid fruit fly genera, using molecular methods. Wolbachia was only detected in Zeugodacus apicalis (de Meijere) (Diptera: Tephritidae) and not in the other 7 studied species. This newly discovered strain, named wZap, belongs to supergroup B with a 1.3 Mb genome containing 1,248 genes. Phylogenetic analysis of its cytoplasmic incompatibility factor genes cifA and cifB revealed their placement within the Type I clade. Given the presence of cif genes in the wZap genome, further research into their roles in fruit flies could be crucial for developing pest control strategies that exploit CI mechanisms.}, } @article {pmid40078832, year = {2023}, author = {Al Noman, A and Das, D and Nesa, Z and Tariquzzaman, M and Sharzana, F and Rakibul Hasan, M and Riaz, BK and Sharower, G and Rahman, MM}, title = {Importance of Wolbachia-mediated biocontrol to reduce dengue in Bangladesh and other dengue-endemic developing countries.}, journal = {Biosafety and health}, volume = {5}, number = {2}, pages = {69-77}, doi = {10.1016/j.bsheal.2023.03.003}, pmid = {40078832}, issn = {2590-0536}, abstract = {Mosquito-borne diseases, particularly dengue and chikungunya have become global threats, infecting millions of people worldwide, including developing countries of Southeast Asia and Latin America. Bangladesh, like many other developing countries, is experiencing frequent dengue outbreaks. This article, therefore, critically discussed the current status of dengue disease, vector control approaches, and the need for Wolbachia-mediated intervention in Bangladesh and other dengue-endemic developing countries. In this narrative review study, relevant literature was searched from major databases and search engines such as PubMed, BanglaJol, World Health Organization (WHO)/European Centre for Disease Prevention and Control (ECDC) and Google Scholar. Considering the selection criteria, our search strategies finally involved 55 related literature for further investigation. Findings showed that current vector control strategies could not render protection for an extended period, and the disease burden of arboviruses is increasing. The impoverished outbreak preparedness, urbanization, climate change, and less efficacy of existing control methods have made people susceptible to vector-borne diseases. Hence, Wolbachia, a naturally occurring endosymbiont of many mosquito species that can potentially limit virus transmission through several host genetic alterations, would be a potential alternative for dengue prevention. We also critically discussed the challenges and prospects of Wolbachia-based dengue control in developing countries. The evidence supporting the efficacy and safety of this intervention and its mechanism have also been elucidated. Empirical evidence suggests that this introgression method could be an eco-friendly and long-lasting dengue control method. This review would help the policymakers and health experts devise a scheme of Wolbachia-based dengue control that can control mosquito-borne diseases, particularly dengue in Bangladesh and other developing countries.}, } @article {pmid40069226, year = {2025}, author = {Carcauzon, V and Cordonin, C and Sebbane, F and Soarimalala, V and Goodman, SM and Tortosa, P}, title = {Malagasy flea microbiota results from a combination of vertically transmitted and environmentally acquired microbes.}, journal = {Scientific reports}, volume = {15}, number = {1}, pages = {8461}, pmid = {40069226}, issn = {2045-2322}, mesh = {Animals ; *Siphonaptera/microbiology ; Madagascar ; *Microbiota/genetics ; Insect Vectors/microbiology ; Plague/transmission/microbiology/epidemiology ; Host Specificity ; RNA, Ribosomal, 16S/genetics ; Seasons ; Bacteria/genetics/classification/isolation & purification ; }, abstract = {Fleas (Insecta, Siphonaptera) are the known vectors of serious bacterial pathogens, such as Yersinia pestis and Rickettsia typhi. The microbiota of fleas has been poorly investigated although it has a known influence on vector competence. Here, we report and analyse the microbiota of 577 flea specimens from Madagascar, a hotspot of plague transmission. Importantly, endemic Malagasy fleas show low host specificity, allowing addressing the importance of vertebrate host species in microbiota composition, as well as that of abiotic variables. We describe through Illumina sequencing of 2 hypervariable regions of 16 S rDNA the bacterial composition of 577 flea specimens of Madagascar. We address the importance of biotic (mammalian host and flea species) and abiotic (season and sampling site) variables on bacterial community composition. Bacterial composition appears driven by flea species and season, but interestingly not by the flea's vertebrate hosts. These results suggest that the flea microbiota is at least in part acquired before they become adult, possibly in the immature off-host stages. Taken together, our results suggest that the microbiota of sampled fleas are composed of bacterial taxa with vertical transmission, such as Wolbachia which are prevalent in the present dataset, together with several bacterial taxa for which the occurrence is driven by environment factors, especially season and habitat. Given the importance of the microbiota in vector competence, we discuss the epidemiological consequences of environmentally-driven acquisition of microbiota in fleas on plague transmission in Madagascar.}, } @article {pmid40067786, year = {2025}, author = {Darío Velez, I and Uribe, A and Barajas, J and Uribe, S and Ángel, S and David Suaza-Vasco, J and Sebastian Duran Ahumada, J and Camila Mejia Torres, M and Patricia Arbeláez, M and Santacruz-Sanmartin, E and Duque, L and Martínez, L and Posada, T and Cristina Patiño, A and Milena Gonzalez, S and Lucía Velez, A and Ramírez, J and Salazar, M and Gómez, S and Osorio, JE and Iturbe-Ormaetxe, I and Dong, Y and Muzzi, FC and Rances, E and Johnson, PH and Smithyman, R and Col, B and Green, BR and Frossard, T and Brown-Kenyon, J and Joubert, DA and Grisales, N and Ritchie, SA and Denton, JA and Gilles, JRL and Anders, KL and Kutcher, SC and Ryan, PA and O'Neill, SL}, title = {Correction: Large-scale releases and establishment of wMel Wolbachia in Aedes aegypti mosquitoes throughout the Cities of Bello, Medellín and Itagüí, Colombia.}, journal = {PLoS neglected tropical diseases}, volume = {19}, number = {3}, pages = {e0012936}, doi = {10.1371/journal.pntd.0012936}, pmid = {40067786}, issn = {1935-2735}, abstract = {[This corrects the article DOI: 10.1371/journal.pntd.0011642.].}, } @article {pmid40065479, year = {2025}, author = {Pomari, E and Voronin, D and Alvarez-Martinez, MJ and Arsuaga, M and Bottieau, E and Luzón-García, MP and Nickel, B and Rubio, JM and Salas-Coronas, J and Salvador, F and Soriano-Pérez, MJ and Sulleiro, E and Goterris, L and Van Esbroeck, M and van Hellemond, JJ and Wammes, LJ and Zammarchi, L and Lavezzari, D and Degani, M and Tais, S and Held, J and Gobbi, F and Tamarozzi, F}, title = {Wolbachia bacteria in Mansonella perstans isolates from patients infected in different geographical areas: a pilot study from the ESCMID Study Group for Clinical Parasitology.}, journal = {Parasites & vectors}, volume = {18}, number = {1}, pages = {97}, pmid = {40065479}, issn = {1756-3305}, support = {Individual reserach grant to EP//European Society of Clinical Microbiology and Infectious Diseases/ ; Fondi "Ricerca Corrente" - L2 to IRCCS Sacro Cuore Don Calabria Hospital, Negrar//Ministero della Salute/ ; to DV//Division of Intramural Research (DIR) of NIAID/NIH USA/ ; }, mesh = {*Wolbachia/genetics/isolation & purification/drug effects/classification ; *Mansonella/genetics/isolation & purification/drug effects ; Humans ; Pilot Projects ; *Mansonelliasis/epidemiology/parasitology ; Animals ; *Phylogeny ; Male ; Female ; Africa, Western/epidemiology ; Microfilariae/drug effects/isolation & purification ; Adult ; Polymerase Chain Reaction ; }, abstract = {BACKGROUND: Mansonella perstans is a vector-borne filarial parasite widely endemic in sub-Saharan Africa, with sporadic cases in Latin America. Infection is often overlooked; treatment is not standardized, and effectiveness of common regimes is difficult to ascertain. Anti-Wolbachia macrofilaricidal treatment with doxycycline has been applied, but there are scant and contrasting reports about the presence of Wolbachia in M. perstans isolates from different geographical locations. Taking advantage of a network of European centres expert in traveller and migrant health, we aimed to expand the knowledge concerning the distribution of Wolbachia in M. perstans to contribute to the design of optimal treatment approaches.

METHODS: We analysed 19 samples of concentrated microfilariae or whole blood from M. perstans-infected patients who reported having resided or travelled in one or more of 10 West African countries. Wolbachia was detected by PCR targeting 16S and ftsZ genes and phylogenetic analysis of M. perstans was performed based on COX1 gene sequencing.

RESULTS: Wolbachia was identified in 14/19 (74%) samples. With the possible inaccuracy deriving from potential origin of infection being identified retrospectively from routine clinical visit's documents, this study identified Wolbachia in M. perstans from Burkina Faso, Equatorial Guinea, Republic of Guinea and Senegal for the first time to our knowledge. Furthermore, Wolbachia might also be present in M. perstans from Democratic Republic of the Congo, Mali, Niger and Nigeria.

CONCLUSIONS: The retrieval of Wolbachia-positive and Wolbachia-negative M. perstans samples can either be explained by technical limitations or reflect the real existence of Wolbachia-positive and Wolbachia-negative M. perstans populations. However, this latter hypothesis was not supported by our phylogenetic analysis. Our results suggest that doxycycline could be used for the treatment of M. perstans infection upfront or, if possible, after ascertaining the presence of Wolbachia by PCR performed on concentrated microfilariae using two targets to avoid false-negative results.}, } @article {pmid40044916, year = {2025}, author = {Zheng, B and Yang, H and Elaydi, S and Yu, J}, title = {wStri spread dynamics in Nilaparvata lugens via discrete mathematical models.}, journal = {Journal of mathematical biology}, volume = {90}, number = {4}, pages = {38}, pmid = {40044916}, issn = {1432-1416}, support = {12371484//National Natural Science Foundation of China/ ; 12331017//National Natural Science Foundation of China/ ; }, mesh = {Animals ; *Hemiptera/microbiology/physiology/growth & development ; *Wolbachia/physiology/pathogenicity ; *Models, Biological ; *Mathematical Concepts ; *Pest Control, Biological/methods/statistics & numerical data ; Population Dynamics/statistics & numerical data ; Computer Simulation ; Female ; }, abstract = {Wolbachia, an intracellular bacterium, is well-known for inducing cytoplasmic incompatibility, which has become a promising and environmentally sustainable strategy for controlling pest populations. The strain wStri, specifically identified in Nilaparvata lugens (brown planthopper), has shown potential for such biocontrol applications. In this study, we develop a comprehensive discrete mathematical model to analyze the dynamics of wStri spread in a mixed population of wStri-infected, wLug-infected, and uninfected Nilaparvata lugens under both constant and periodically varying environmental conditions. Under a constant environment, the model identifies the critical threshold necessary for the successful establishment of wStri within the population. Our analysis reveals that the model exhibits a strong Allee effect, where a population must exceed a certain critical density-the Allee threshold-for the wStri strain to persist and spread. Below this threshold, the wStri strain is likely to be eliminated, failing in pest control efforts. When the environment varies periodically, the model transforms into a non-autonomous periodic discrete model, introducing additional complexity. In this scenario, we derive sufficient conditions that ensure the composition of finitely many Allee maps continues to function as an Allee map. Furthermore, we prove that a unique periodic orbit exists within such a periodic environment. This orbit is characterized as unstable and acts as a threshold, determining whether wStri will establish itself in the population or die out over time. The findings from this model provide critical insights into the conditions under which wStri can be effectively used to control Nilaparvata lugens, particularly in environments that are not constant but fluctuate periodically. These insights have significant implications for the practical deployment of Wolbachia-based biocontrol methods in pest management strategies.}, } @article {pmid40032901, year = {2025}, author = {Brar, G and Ngor, L and McFrederick, QS and Torson, AS and Rajamohan, A and Rinehart, J and Singh, P and Bowsher, JH}, title = {High abundance of lactobacilli in the gut microbiome of honey bees during winter.}, journal = {Scientific reports}, volume = {15}, number = {1}, pages = {7409}, pmid = {40032901}, issn = {2045-2322}, mesh = {Animals ; Bees/microbiology ; *Gastrointestinal Microbiome ; *Lactobacillus/genetics/isolation & purification ; *Seasons ; *RNA, Ribosomal, 16S/genetics ; }, abstract = {Honey bee gut microbiota play specific roles in promoting host growth and physiology by regulating the immune system, behavior, metabolism, and neurological processes. While the gut microbiota of honey bee queens, workers, and larvae has been extensively studied, less is known about the composition of gut microbiota in the winter worker bees. This study investigates the dynamics of the gut microbiota in overwintering adult worker bees, focusing on two commercial bee strains: Bolton™ bees and Mann Lake™ bees. These Apis mellifera strains were investigated under different storage conditions (indoor storage at 6 °C and outdoor storage in natural conditions) during the winter months (October, November, and December). Utilizing 16S rRNA gene amplicon sequencing, we characterized the microbial composition of the whole gut. We observed the Lactobacilli dominated in all the overwintering honey bee guts with a significantly higher abundance of unclassified Lactobacillus species in November, while Lactobacillus apis showed significantly higher abundance in October. Bolton bees exhibited significantly higher abundance levels of Bartonella (denoted as uncultured) and Bifidobacterium, along with an unexpected presence of Wolbachia. In contrast, Mann Lake bees demonstrated an increased abundance of Commensalibacter. Our results suggest that Shannon diversity is influenced by the month rather than by the bee strain or storage conditions. We also found significant differences in Bray Curtis diversity index by month. Overall, taxonomical abundance was not affected by whether the hives were stored outside or in constant temperature indoor storage. However, various bacterial species showed differences in abundance across different months, with slight variations observed between bee strains. Given the potential benefits of the honey bee gut microbiome for health and nutrition, our data suggests that the genus Lactobacillus may play a significant role in bee health during winter and overwintering storage.}, } @article {pmid40023240, year = {2025}, author = {Diesbourg, EE and Kidd, KA and Perrotta, BG}, title = {Effects of municipal wastewater effluents on the invertebrate microbiomes of an aquatic-riparian food web.}, journal = {Environmental pollution (Barking, Essex : 1987)}, volume = {}, number = {}, pages = {125948}, doi = {10.1016/j.envpol.2025.125948}, pmid = {40023240}, issn = {1873-6424}, abstract = {Municipal wastewater effluents (MWWEs) contain antimicrobials and other contaminants that can alter the microbiomes of exposed aquatic animals, potentially negatively impacting host health. Contaminants and nutrients from MWWEs may be transferred across the aquatic - riparian boundary by aquatic insects, potentially altering the microbiomes of both prey and consumers. We evaluated host microbiome compositions of several taxa of freshwater larval and adult insects and riparian spiders at sites upstream and downstream of three wastewater treatment plants. Host microbiome compositions were analyzed by sequencing the 16S rRNA gene and MWWE exposure was assessed using stable carbon (δ[13]C) and nitrogen (δ[15]N) isotopes and effluent-associated bacteria. Most downstream insects and riparian spiders were enriched in δ[13]C and δ[15]N, indicating exposure to MWWEs and transfer of MWWE-derived nutrients to riparian consumers. Within sites, insect microbiomes varied after metamorphosis with a greater proportion of endosymbionts and effluent-associated bacteria and decreased alpha diversity in adults, and the microbiomes of Tetragnathidae spiders were dominated by endosymbionts (mainly Rickettsia and Wolbachia) compared to all other taxa. Downstream, Larval caddisfly (Hydropsychidae) microbiomes had a significantly lower proportion of endosymbionts (Rickettsia) and higher diversity, and Araneidae spiders also had higher diversity. However, there were no significant downstream changes in endosymbiont proportions or alpha diversity of larval and adult chironomids, larval and adult mayflies, larval stoneflies, or Tetragnathidae spiders. Most downstream invertebrates (except larval Chironomidae, adult Diptera, and Tetragnathidae spiders) had altered beta diversity (community compositions); however, host taxonomy explained more of the variation in microbiome composition than site or the interaction between them did. Overall, MWWE bacteria and nutrients were incorporated into most insect larvae and retained throughout metamorphosis, however there were taxa-dependent alterations in downstream insect microbiomes and minimal microbiome alterations to their riparian spider predators.}, } @article {pmid40024849, year = {2025}, author = {Calle-Tobón, A and Rojo-Ospina, R and Zuluaga, S and Giraldo-Muñoz, JF and Cadavid, JM}, title = {Corrigendum to "Evaluation of Wolbachia infection in Aedes aegypti suggests low prevalence and highly heterogeneous distribution in Medellín, Colombia" [Acta Tropica, Volume 260, December 2024, 107423].}, journal = {Acta tropica}, volume = {}, number = {}, pages = {107572}, doi = {10.1016/j.actatropica.2025.107572}, pmid = {40024849}, issn = {1873-6254}, } @article {pmid40018116, year = {2024}, author = {Kumar, G and Baharia, R and Singh, K and Gupta, SK and Joy, S and Sharma, A and Rahi, M}, title = {Addressing challenges in vector control: a review of current strategies and the imperative for novel tools in India's combat against vector-borne diseases.}, journal = {BMJ public health}, volume = {2}, number = {1}, pages = {e000342}, pmid = {40018116}, issn = {2753-4294}, abstract = {Vector-borne diseases (VBDs) exert a substantial burden across the world, especially in tropical countries. Malaria, chikungunya, dengue, visceral leishmaniasis, lymphatic filariasis and Japanese encephalitis are among the public health concerns for India. One of the major pillars for the containment of VBDs is vector control and different tools have been employed for several decades. These range from chemical insecticides used in indoor residual sprays, space sprays, fogging, treated bednets and larvicides to biological control methods such as larvivorus fishes and environmental control and modification measures such as source reduction. However, these methods are increasingly becoming less effective due to several reasons such as insecticide resistance, outdoor biting, behavioural changes in vectors for biting and resting, climate change, movement of population, vector incursion to newer areas and others. It is essential to develop and test new tools for vector control to surmount these challenges. Though focusing on India's public health concerns, the new tools enumerated here can be tested by any country with similar epidemiological and environmental conditions. The promising new vector control tools are insecticide-treated nets with synergist and/or pyrrole chlorfenapyr, alternatives/additions to synthetic pyrethroids like neonicotinoids, clothianidin for indoor residual spray, newer formulations such as Bacillus sphaericus for use in larvicides, attractive toxic sugar baits, especially to curtail outdoor transmission, endectocides like ivermectin for use in animals/humans, insecticidal paints, spatial repellents, insecticide-treated wearables and others. Genetic modification technologies (Sterile Insect Technique/Incompatible Insect Technique/Wolbachia transfection) are also upcoming strategies. Among the six VBDs, India is committed to the elimination of three (malaria, visceral leishmaniasis and lymphatic filariasis) and it will require additional and/or novel tools to overcome the roadblocks in our current journey to the goal of control/elimination of these VBDs.}, } @article {pmid40012073, year = {2025}, author = {Chu, Y and Sakamoto, K and Evans, CC and Dzimianski, MT and Fricks, C and Mansour, A and DiCosty, U and McCall, S and McCall, JW and Nelson, CT and Moorhead, AR}, title = {Real-time PCR and immunohistochemistry detection of Wolbachia in adult Dirofilaria immitis from dogs treated with doxycycline and ivermectin.}, journal = {Parasites & vectors}, volume = {18}, number = {1}, pages = {78}, pmid = {40012073}, issn = {1756-3305}, abstract = {BACKGROUND: Wolbachia is present in all life stages of Dirofilaria immitis. Wolbachia surface protein (WSP) can be highly immunogenic and induce acute inflammatory reactions in the host upon worm death. To avoid the abrupt release of Wolbachia and its antigens from deceased parasites, the American Heartworm Society (AHS) has recommended using doxycycline (DOXY) and having a 1-month wait period between the DOXY treatment and the adulticidal process for Wolbachia elimination. Studies have shown that the 28 day, 10 mg/kg twice daily (BID) administration of DOXY can effectively clear Wolbachia in the bloodstream of the host. The 1-month wait period is hypothesized to allow for further reduction of Wolbachia. However, the levels of Wolbachia in adult parasites after the DOXY treatment remain unknown.

METHODS: Forty-five purposely bred dogs were intravenously transplanted with 20 Dirofilaria immitis adults, consisting of 12 females and 8 males. The dogs were divided into nine groups of five dogs each. Two groups each received 5, 7.5, or 10 mg/kg DOXY BID orally for 28 days, and ivermectin (IVM) monthly (6 µg/kg.) Three groups remained untreated as controls. Study animals were necropsied on day 0, day 30, and day 60, following the start of treatment. Adult worms were collected at necropsy and preserved for analysis. Quantitative polymerase chain reaction (qPCR) and immunohistochemistry for WSP were performed on worms collected at each time point. The data were analyzed using a linear mixed model (LMM). Multiple comparisons were adjusted using Tukey's test.

RESULTS: The qPCR results showed that all treatment doses significantly reduced Wolbachia levels compared with the control groups at 30 and 60 days. The intradose comparison indicated a significant decrease on day 60 compared with day 30. No significant differences were found between different doses on the two examination dates. Immunohistochemistry indicated the markedly reduced presence of Wolbachia in treatment groups.

CONCLUSIONS: All DOXY dosages can be considered effective in reducing Wolbachia on both tested dates (30 and 60 days). On the basis of the further reduction of Wolbachia levels in adult D. immitis, the 1-month rest period in the AHS heartworm treatment guidelines is beneficial. Wolbachia can still be detected on day 60 in all dosage groups.}, } @article {pmid40009197, year = {2025}, author = {Kelleher, LA and Ramalho, MO}, title = {Impact of Species and Developmental Stage on the Bacterial Communities of Aphaenogaster Ants.}, journal = {Current microbiology}, volume = {82}, number = {4}, pages = {157}, pmid = {40009197}, issn = {1432-0991}, support = {7513312112//West Chester University/ ; }, mesh = {*Ants/microbiology ; Animals ; *Bacteria/classification/genetics/isolation & purification ; *RNA, Ribosomal, 16S/genetics ; *Microbiota ; Symbiosis ; DNA, Bacterial/genetics ; Pennsylvania ; Phylogeny ; Ecosystem ; Biodiversity ; }, abstract = {Ants are distributed across the globe and there are currently over 14,000 described species. Due to the high diversity between species, ants are considered vital keystone species to many ecosystems. They provide basic ecosystem services such as: seed dispersal, soil bioturbation, decomposition, and pest control. Within these ecosystems ants form complex symbiotic relationships with plants, fungi, and bacteria. Studying the interaction between ants and their bacteria is important because of the crucial role that microbes play in the overall health of ants. Aphaenogaster Mayr, 1853, which is a globally distributed ant genus, remains understudied in terms of their bacterial community. This study aims to determine the taxonomic composition and abundance of the Aphaenogaster bacterial community and to determine if development stage and species impact the bacterial community composition. For this study, ants from several colonies were collected from the Gordon Natural Area in West Chester, Pennsylvania, USA. DNA was then extracted from the ants in all stages of development and the 16S rRNA gene was amplified and sequencing following the NGS amplicon approach. The findings from this study reveal that species and development stage have a significant impact upon the bacterial community composition and abundance of Aphaenogaster ants, and Wolbachia is highly associated with these ants.}, } @article {pmid40005669, year = {2025}, author = {Duan, YX and Zhuang, YH and Wu, YX and Huang, TW and Song, ZR and Du, YZ and Zhu, YX}, title = {Wolbachia Infection Alters the Microbiota of the Invasive Leaf-Miner Liriomyza huidobrensis (Diptera: Agromyzidae).}, journal = {Microorganisms}, volume = {13}, number = {2}, pages = {}, doi = {10.3390/microorganisms13020302}, pmid = {40005669}, issn = {2076-2607}, support = {BK20231330//Natural Science Foundation of Jiangsu Province/ ; XCX20240707//the 2024 College Student Innovation and Entrepreneurship Training Program of Yangzhou University/ ; }, abstract = {Microbe-microbe interactions within a host drive shifts in the host's microbiota composition, profoundly influencing host physiology, ecology, and evolution. Among these microbes, the maternally inherited endosymbiont Wolbachia is widespread in the invasive pest Liriomyza huidorbrensis (Diptera: Agromyzidae). However, its influence on the host microbiota remains largely unexplored. In the study presented herein, we investigated the bacterial communities of Wolbachia wLhui-infected (wLhui+) and -uninfected lines (wLhui-) of L. huidorbrensis using 16S rRNA gene high-throughput sequencing. For both leaf-miner lines, Bacteroidota was the dominant phylum (relative abundance: 59.18%), followed by Pseudomonadota (36.63%), Actinomycetota (2.42%), and Bacillota (0.93%). We found no significant differences in alpha-diversity indices between the wLhui+ and wLhui- lines (p > 0.05). However, principal coordinates analysis revealed significant differences in microbiota composition between the wLhui+ and wLhui- lines (PERMANOVA: p < 0.001), explaining 76.70% of the variance in microbiota composition. Correlation network analysis identified robust negative and positive associations between Wolbachia and several genera, suggesting that Wolbachia shapes microbial composition through competitive or cooperative interactions with specific taxa. Overall, our study suggests that Wolbachia plays a key role in shaping the leaf-miner microbiome, potentially affecting host fitness.}, } @article {pmid40003849, year = {2025}, author = {Gwiazdowska, A and Rutkowski, R and Sielezniew, M}, title = {Conservation Genetics of the Endangered Danube Clouded Yellow Butterfly Colias myrmidone (Esper, 1780) in the Last Central European Stronghold: Diversity, Wolbachia Infection and Balkan Connections.}, journal = {Insects}, volume = {16}, number = {2}, pages = {}, doi = {10.3390/insects16020220}, pmid = {40003849}, issn = {2075-4450}, support = {EZ.271.3.7.2021//General Directorate of the Polish State Forests/ ; }, abstract = {The Danube Clouded Yellow (Colias myrmidone) has experienced one of the most dramatic declines among European butterflies. To estimate genetic diversity in the last population in Poland that has survived in the Knyszyn Forest (KF), we analyzed mitochondrial (COI) and nuclear (EF-1α) polymorphisms in individuals sampled in 2014 and 2022. The results were compared with genetic data obtained in 2014 from a recently extirpated nearby population (Czerwony Bór, CB). Because mtDNA polymorphisms in insects can be modulated by endosymbionts, the samples were screened for Wolbachia. The polymorphism of EF-1α indicated that diversity was gradually decreasing. The KF experienced rapid demographic processes, manifested by a significant change in allele frequency. The small differentiation in nuclear markers between the KF and CB in 2014 suggests that the regional population used to be genetically uniform. Four COI haplotypes that were identified in this study probably belong to two different haplogroups. Wolbachia was detected only in individuals with one specific haplotype, and the prevalence was female-biased, suggesting the induction of two reproductive manipulations. The most common COI haplotype found in Poland was the same as that reported from other parts of Europe, not only for C. myrmidone but also C. caucasica. These results allow us to question the distinctiveness of each taxa.}, } @article {pmid40003778, year = {2025}, author = {M'madi, SA and Zan Diarra, A and Bérenger, JM and Hasnaoui, B and Parola, P}, title = {Identification of Bed Bugs from Comoros, Using Morphological, Matrix-Assisted Laser Desorption Ionisation Time-of-Flight Mass Spectrometry, and Molecular Biology Tools, and the Detection of Associated Bacteria.}, journal = {Insects}, volume = {16}, number = {2}, pages = {}, doi = {10.3390/insects16020148}, pmid = {40003778}, issn = {2075-4450}, support = {ANR-10- IAHU-03//Institut Hospitalo-Universitaire (IHU) Méditerranée Infection, the French National Research Agency under the "Investissements d'avenir" programme/ ; }, abstract = {After virtually disappearing from domestic dwellings in the Western world at the end of the Second World War, bed bugs have re-emerged in recent years. Few studies, however, have been carried out on these insects in tropical islands. In this study, we focussed on describing bed bug specimens collected from dwellings in a high-altitude village in Grande Comore, an island in the Comoros, in the Indian Ocean. We also aimed to detect the bacteria associated with them. Using MALDI-TOF MS coupled with molecular biology, we were able to confirm that the C. hemipterus species (the tropical bug) was the bug infesting these homes. Interestingly, the results also show that MALDI-TOF MS can differentiate between the developmental stages of bed bugs (immature and adult). Screening for bacteria was carried out using qPCR, regular PCR, and sequencing, with only Wolbachia DNA being found. Widespread surveys throughout the country are needed to ascertain the level of bed bug infestation, with a view to implementing appropriate control measures.}, } @article {pmid40003744, year = {2025}, author = {Price, G and Simard, A and McGraw, BA}, title = {Evaluation of Bacterial Communities of Listronotus maculicollis Kirby Reared on Primary and Secondary Host Plants.}, journal = {Insects}, volume = {16}, number = {2}, pages = {}, doi = {10.3390/insects16020114}, pmid = {40003744}, issn = {2075-4450}, abstract = {The annual bluegrass weevil (Listronotus maculicollis Kirby) is a devastating insect pest of annual bluegrass (Poa annua L.) and, to a lesser extent, creeping bentgrass (Agrostis stolonifera L.) on golf courses. Listronotus maculicollis-reared A. stolonifera, a comparatively tolerant host, incurs fitness costs, including longer developmental periods and reduced larval survivorship. This study sought to characterize microbiota diversity in L. maculicollis adults and larvae reared on P. annua and A. stolonifera cultivars (Penncross & A4) to explore whether intrinsic factors, such as microbial community composition, vary across host plants and developmental stages, potentially influencing host suitability. Alpha diversity analyses showed adults feeding on A4 exhibited higher bacterial species richness than their offspring reared on the same cultivar. Beta diversity analysis revealed significant dissimilarities between L. maculicollis adults and offspring regardless of host. Pseudomonas sp. was consistently abundant in larvae across all turfgrasses, indicating a potential association with larval development. Elevated levels of Wolbachia sp., known for insect reproductive manipulation, were observed in adults, but appear to be unrelated to host plant effects. The most prevalent bacterium detected was Candidatus Nardonella, a conserved endosymbiont essential for cuticular hardening in weevils. Given the role of cuticular integrity in insecticide resistance, further investigations into insect-microbe-plant interactions could guide the development of targeted pest management strategies, reducing resistance and improving control measures for L. maculicollis.}, } @article {pmid39998220, year = {2025}, author = {Kwak, Y and Argandona, JA and Miao, S and Son, TJ and Hansen, AK}, title = {A dual insect symbiont and plant pathogen improves insect host fitness under arginine limitation.}, journal = {mBio}, volume = {}, number = {}, pages = {e0358824}, doi = {10.1128/mbio.03588-24}, pmid = {39998220}, issn = {2150-7511}, abstract = {Some facultative bacterial symbionts are known to benefit insects, but nutritional advantages are rare among these non-obligate symbionts. Here, we demonstrate that the facultative symbiont Candidatus Liberibacter psyllaurous enhances the fitness of its psyllid insect host, Bactericera cockerelli, by providing nutritional benefits. L. psyllaurous, an unculturable pathogen of solanaceous crops, also establishes a close relationship with its insect vector, B. cockerelli, increasing in titer during insect development, vertically transmitting through eggs, and colonizing various tissues, including the bacteriome, which houses the obligate nutritional symbiont, Carsonella. Carsonella supplies essential amino acids to its insect host but has gaps in some of its essential amino acid pathways that the psyllid complements with its own genes, many of which have been acquired through horizontal gene transfer (HGT) from bacteria. Our findings reveal that L. psyllaurous increases psyllid fitness on plants by reducing developmental time and increasing adult weight. In addition, through metagenomic sequencing, we reveal that L. psyllaurous maintains complete pathways for synthesizing the essential amino acids arginine, lysine, and threonine, unlike the psyllid's other resident microbiota, Carsonella, and two co-occurring Wolbachia strains. RNA sequencing reveals the downregulation of a HGT collaborative psyllid gene (ASL), which indicates a reduced demand for arginine supplied by Carsonella when the psyllid is infected with L. psyllaurous. Notably, artificial diet assays show that L. psyllaurous enhances psyllid fitness on an arginine-deplete diet. These results corroborate the role of L. psyllaurous as a beneficial insect symbiont, contributing to the nutrition of its insect host.IMPORTANCEUnlike obligate symbionts that are permanently associated with their hosts, facultative symbionts rarely show direct nutritional contributions, especially under nutrient-limited conditions. This study demonstrates, for the first time, that Candidatus Liberibacter psyllaurous, a facultative symbiont and a plant pathogen, enhances the fitness of its Bactericera cockerelli host by supplying an essential nutrient arginine that is lacking in the plant sap diet. Our findings reveal how facultative symbionts can play a vital role in helping their insect hosts adapt to nutrient-limited environments. This work provides new insights into the dynamic interactions between insect hosts, their symbiotic microbes, and their shared ecological niches, broadening our understanding of symbiosis and its role in shaping adaptation and survival.}, } @article {pmid39976626, year = {2025}, author = {Sharpe, SR and Madhav, M and Klein, MJ and Blasdell, KR and Paradkar, PN and Lynch, SE and Eagles, D and López-Denman, AJ and Ahmed, KA}, title = {Characterisation of the virome of Culicoides brevitarsis Kieffer (Diptera: Ceratopogonidae), a vector of bluetongue virus in Australia.}, journal = {The Journal of general virology}, volume = {106}, number = {2}, pages = {}, doi = {10.1099/jgv.0.002076}, pmid = {39976626}, issn = {1465-2099}, mesh = {Animals ; *Ceratopogonidae/virology ; *Virome ; *Bluetongue virus/genetics/classification/isolation & purification ; *Insect Vectors/virology ; *Phylogeny ; High-Throughput Nucleotide Sequencing ; Genome, Viral ; Australia ; New South Wales ; RNA, Viral/genetics ; }, abstract = {Culicoides spp., a common biting midge genus, are haematophagous insects that can transmit pathogens to humans and other animals. Some species transmit arboviruses, including bluetongue virus, epizootic haemorrhagic disease virus, African horse sickness virus and Schmallenberg virus to vertebrates, which can be detrimental to livestock and wild animals. Culicoides spp. can also have a diversity of insect-specific viruses (ISVs) that can only be transmitted between insects and others related to known arboviruses. For Culicoides brevitarsis and other Culicoides spp. in Australia, the virome is largely unexplored. We used high-throughput sequencing to characterise the virome of C. brevitarsis collected from Casino, New South Wales, Australia. For virus detection, the total RNA was extracted from pools of C. brevitarsis followed by rRNA depletion and Illumina short-read-based RNA sequencing. The reads were quality-checked, filtered and assembled into contigs, compared with the non-redundant protein and conserved domain databases for viral detection and genome organisation, respectively. The phylogenetic analysis was used to further characterise the viruses. We detected new virus diversity including ten viruses belonging to eight different families with complete or near-complete coding regions. Seven of these were novel virus species belonging to the families: Chuviridae, Orthomyxoviridae, Peribunyaviridae, Qinviridae, Rhabdoviridae and Solemoviridae. In addition, the novel Peribunyaviridae virus should also be considered part of a new genus. Whilst most of the detected viruses grouped into families with viruses that can infect insects, animals or both, the novel species of Solemoviridae was closely related to an economically important plant pathogen, the sugarcane yellow leaf virus. Our quantitative PCR-based screening confirmed the absence of any Wolbachia endosymbiont within the collected samples. Furthermore, we detected fragments of three more virus families known to infect fungi and plants. The detection of potential arboviruses and ISVs in Culicoides spp. is important in understanding virus epidemiology.}, } @article {pmid39975058, year = {2025}, author = {Johnson, RM and Breban, MI and Nolan, BL and Sodeinde, A and Ott, IM and Ross, PA and Gu, X and Grubaugh, ND and Alex Perkins, T and Brackney, DE and Vogels, CBF}, title = {Implications of successive blood feeding on Wolbachia -mediated dengue virus inhibition in Aedes aegypti mosquitoes.}, journal = {bioRxiv : the preprint server for biology}, volume = {}, number = {}, pages = {}, doi = {10.1101/2025.02.06.636928}, pmid = {39975058}, issn = {2692-8205}, abstract = {Dengue virus (DENV) is a mosquito-borne virus that poses a continued and increasing threat to public health. A promising strategy to mitigate the burden of DENV is introgression of the virus-inhibiting Wolbachia pipientis bacterium into Aedes aegypti populations in the field. While previous studies on Wolbachia -mediated virus inhibition have typically assessed viral replication following a single bloodmeal, the main vector of DENV, Ae. aegypti , feeds frequently, often biting multiple hosts per gonotrophic cycle and promptly attempting to refeed following egg laying. Previously, we demonstrated that successive blood feeding reduces the extrinsic incubation period (EIP) and shortens the time it takes for a mosquito to be able to transmit viruses to a new host. With this in mind, we investigated the impact of successive blood meals on DENV serotype 2 (DENV-2) in Ae. aegypti in the presence or absence of Wolbachia (w AlbB and w MelM). We found that both WT and Wolbachia transinfected had increased DENV-2 dissemination 7 days post-infection as well as higher body titers of DENV-2 in the double-fed groups. Using these empirical data in a binomial regression model, we estimated that successive feeding increased the probability of WT and Wolbachia transinfected mosquitoes surviving the EIP. When we estimated the odds of surviving the EIP for mosquitoes with Wolbachia relative to WT mosquitoes, successive feeding increased the chances of WT mosquitoes surviving the EIP more than in mosquitoes with Wolbachia , indicating a strong inhibitory effect of Wolbachia even in the context of natural frequent blood feeding behavior. Our work shows that mosquito feeding behavior should be considered when assessing the inhibitory effects of Wolbachia on DENV.}, } @article {pmid39971081, year = {2025}, author = {Ma, Z and Gao, J and Wang, G and Zhao, M and Xing, D and Zhao, T and Zhang, H}, title = {Effects of Wolbachia on mitochondrial DNA variation in Aedes albopictus (Diptera: Culicidae).}, journal = {Acta tropica}, volume = {}, number = {}, pages = {107561}, doi = {10.1016/j.actatropica.2025.107561}, pmid = {39971081}, issn = {1873-6254}, abstract = {Wolbachia species are symbiotic bacteria that are commonly found in arthropods and nematodes and live inside their cells. In nature, endosymbiont-host interactions and dynamics are complex, often depending on environmental conditions and evolutionary history. Both Wolbachia and mitochondrial DNA are maternally inherited in cells, and after a long period of coexistence, the presence of Wolbachia may have an impact on mitochondrial sequence diversity, thereby confounding mtDNA-based host phylogeny. The universal and typing primers for the wsp gene were used for PCR amplification, the number of positive samples was counted, and the infection pattern was analysed. The mitochondrial DNA diversity of four groups (Wolbachia-infected and uninfected samples, as well as between singly and double infected samples.) was analysed. PACo and ParaFitGlobal tests were used to explore evolutionary associations. The overall prevalence of Wolbachia in the 22 natural populations was 94.2%, with Type A, Type B and A × B mixed infections detected in Aedes albopictus and coinfection between wAlbA and wAlbB prevalent. The mitochondrial DNA haplotype associated with Wolbachia (Hap1) became the dominant haplotype and was the most abundant and widely distributed in the population. The linkage map showed the predominant haplotype, Hap1, was more closely associated with wAlbA than with wAlbB. Neutral evolution deviated significantly from zero. The diversity of mtDNA COI genes associated with Wolbachia infection was reduced. Wolbachia infection may lead to the selective sweep of mitochondrial DNA in Ae. albopictus.}, } @article {pmid39966714, year = {2025}, author = {Jain, A and Li, T and Huston, DC and Kaur, J and Trollip, C and Wainer, J and Hodda, M and Linsell, K and Riley, IT and Toktay, H and Olowu, EA and Edwards, J and Rodoni, B and Sawbridge, T}, title = {Insights from draft genomes of Heterodera species isolated from field soil samples.}, journal = {BMC genomics}, volume = {26}, number = {1}, pages = {158}, pmid = {39966714}, issn = {1471-2164}, abstract = {BACKGROUND: The nematode phylum includes many species key to soil food webs with trophic behaviours extending from feeding on microbes to macrofauna and plant roots. Among these, the plant parasitic cyst nematodes retain their eggs in protective cysts prolonging their survival under harsh conditions. These nematodes, including those from the genus Heterodera, cause significant economic losses in agricultural systems. Understanding of nematode diversity and ecology has expanded through application of genomic research, however, for Heterodera species there are very few available whole genome sequences. Sequencing and assembling Heterodera genomes is challenging due to various technical limitations imposed by the biology of Heterodera. Overcoming these limitations is essential for comprehensive insights into Heterodera parasitic interactions with plants, population studies, and for Australian biosecurity implications.

RESULTS: We hereby present draft genomes of six species of which Heterodera australis, H. humuli, H. mani and H. trifolii are presently recorded in Australia and two species, H. avenae and H. filipjevi, currently absent from Australia. The draft genomes were sequenced from genomic DNA isolated from 50 cysts each using an Illumina NovaSeq short read sequencing platform. The data revealed disparity in sequencing yield between species. What was previously identified as H. avenae in Australia using morphological traits is now confirmed as H. australis and may have consequences for wheat breeding programs in Australia that are breeding for resistance to H. avenae. A multigene phylogeny placed the sequenced species into taxonomic phylogenetic perspective. Genomic comparisons within the Avenae species group revealed orthologous gene clusters within the species, emphasising the shared and unique features of the group. The data also revealed the presence of a Wolbachia species, a putative bacterial endosymbiont from Heterodera humuli short read sequencing data.

CONCLUSION: Genomic research holds immense significance for agriculture, for understanding pest species diversity and the development of effective management strategies. This study provides insight into Heterodera, cyst nematode genomics and the associated symbionts and this work will serve as a baseline for further genomic analyses in this economically important nematode group.}, } @article {pmid39958813, year = {2025}, author = {Abe, J and Takahashi, JI and Tsuchida, K}, title = {High Microsatellite but No Mitochondrial DNA Variation in an Invasive Japanese Mainland Population of the Parasitoid Wasp Melittobia sosui.}, journal = {Ecology and evolution}, volume = {15}, number = {2}, pages = {e71026}, pmid = {39958813}, issn = {2045-7758}, abstract = {Invasive populations are predicted to have reduced genetic diversity due to bottleneck events. The parasitoid wasp Melittobia sosui was previously identified only in the subtropical area of the southern Japanese islands and Taiwan but was recently found in the temperate area of the Japanese mainland. The distribution of this species may have recently expanded northward due to factors such as climatic events and global warming. The population genetics of both the native and invasive regions were investigated using mitochondrial and nuclear microsatellite DNA. As expected, mitochondrial variation was observed in the native region but not in the invasive region, which had only one haplotype. However, the two regions exhibited similar levels of microsatellite variation, and an average of 43% and 38% of alleles were uniquely found in the native and invasive populations, respectively. The difference in genetic variation between mitochondrial and microsatellite DNA in the invasive populations may be explained by the faster mutation rate of microsatellites, as well as the population structure of Melittobia, in which the subdivision into small inbreeding lineages may facilitate the accumulation of mutations. The high proportion of private alleles suggests that the mainland population diverged from the native populations at least 100 years ago, ruling out the possibility that the mainland population was established recently. The present study suggests that M. sosui might have already existed on the mainland but at a low frequency or that the mainland population was derived from a ghost population that diverged from the native populations more than 100 years ago.}, } @article {pmid39957355, year = {2025}, author = {Prabhu, D and Dharshini, MKD and Rajamanikandan, S and Padmavathi, AR and Velusamy, P and Gopinath, SCB}, title = {Potential Anti-Filarial Molecules Against ATP Binding Site of MurE Enzyme: A Molecular Docking and Dynamics Approach to Combat Lymphatic Filariasis.}, journal = {Biotechnology and applied biochemistry}, volume = {}, number = {}, pages = {}, doi = {10.1002/bab.2727}, pmid = {39957355}, issn = {1470-8744}, support = {//KAHE-Seed money research Grant/ ; //Department of Science and Technology, Ministry of Science and Technology, India/ ; }, abstract = {Lymphatic filariasis (LF) is a mosquito-borne disease caused by parasitic nematodes Brugia malayi, Brugia timori, and Wuchereria bancrofti. The drugs available are effective in several cases, and the absence of vaccination is the crucial factor hindering the elimination of LF. The UDP-N-acetylmuramoyl-L-alanyl-D-glutamate-2,6-diaminopimelate ligase (MurE) plays an important role in the peptidoglycan biosynthesis of Wolbachia endosymbiont B. malayi, which are reported to be a vital drug target for bacterial and endosymbiotic hosts. Thus, we selected the ATP binding cavity of MurE as the potential site to screen inhibitors. The MurE structure was modeled using AlphaFold due to the absence of an experimental structure. Structure-based screening identified five potent phytochemicals targeting the ATP binding site with higher Glide scores and affinity. The top five phytochemicals CID 311, CID 445713, CID 441626, CID 39077, and CID 10814 showed a docking score of -16.812, -16.117, -15.668, -15.324, and -13.442 kcal/mol, respectively. Further, the molecular dynamics simulations depicted the binding stability of the phytochemical inhibitors bound to the MurE complex. Moreover, ADME assessment and Density Functional Theory analyses of the predicted compounds have shown acceptable pharmacokinetic properties and high reactivity with the drug target of MurE.}, } @article {pmid39937795, year = {2025}, author = {Kittayapong, P and Ninphanomchai, S and Thayanukul, P and Yongyai, J and Limohpasmanee, W}, title = {Comparison on the quality of sterile Aedes aegypti mosquitoes produced by either radiation-based sterile insect technique or Wolbachia-induced incompatible insect technique.}, journal = {PloS one}, volume = {20}, number = {2}, pages = {e0314683}, doi = {10.1371/journal.pone.0314683}, pmid = {39937795}, issn = {1932-6203}, mesh = {Animals ; *Aedes/microbiology/radiation effects ; *Wolbachia/physiology/radiation effects ; Male ; Female ; *Mosquito Control/methods ; Mosquito Vectors/microbiology/radiation effects ; Longevity/radiation effects ; Sexual Behavior, Animal/radiation effects ; Infertility, Male/microbiology/etiology ; Pest Control, Biological/methods ; }, abstract = {Novel and alternative vector control approaches using a sterile male-based release to suppress Aedes aegypti mosquito vectors have recently been tested in the field in many countries. These approaches included the sterile insect technique (SIT), incompatible insect technique (IIT), and a combination of both techniques. In this study, we conducted a series of experiments to compare the quality between radiation-based and Wolbachia-induced sterile males in terms of flight ability, sterility, mating competitiveness, survival rate, and longevity. Aedes aegypti mosquitoes irradiated at 50 Gy (SIT) and those trans-infected with wAlbB Wolbachia (IIT) were used for quality comparison. Our results showed that irradiated and Wolbachia trans-infected males were not significantly different in flight ability (p > 0.05) and both could induce sterility in wild-type females. In addition, although irradiation at 50 Gy or Wolbachia trans-infection reduced male mating competitiveness, combined irradiation and Wolbachia wAlbB trans-infection increased male competitiveness at the one-to-one ratio. Increasing the number of sterile males released could compensate for reduced competitiveness but it does not make them more competitive. Irradiation did not affect the survival and longevity of irradiated males, but it showed significant negative impacts on females (p < 0.05); while the opposite was observed in the case of Wolbachia infection, i.e., with significant increase in the survival rate of Wolbachia trans-infected males (p < 0.05), but both survival and longevity were reduced in Wolbachia trans-infected females with no significant impacts (p > 0.05). In conclusion, neither irradiation nor Wolbachia trans-infection significantly affected the quality of sterile males except their mating competitiveness; but this could compensate by increasing the number of sterile males released. Sterility could be induced by either 50 Gy irradiation or wAlbB trans-infection. Mating competitiveness results showed that a higher number of sterile males produced by irradiation need to be released in comparison to those produced by Wolbachia trans-infection. Our results should be useful for planning SIT, IIT, or a combination for Ae. aegypti vector control.}, } @article {pmid39933580, year = {2025}, author = {Kaur, R and Bordenstein, SR}, title = {Cytoplasmic incompatibility factor proteins from Wolbachia prophage are costly to sperm development in Drosophila melanogaster.}, journal = {Proceedings. Biological sciences}, volume = {292}, number = {2040}, pages = {20243016}, doi = {10.1098/rspb.2024.3016}, pmid = {39933580}, issn = {1471-2954}, support = {/NH/NIH HHS/United States ; //Penn State/ ; }, mesh = {Animals ; *Wolbachia/physiology ; *Drosophila melanogaster/physiology/microbiology ; Male ; *Spermatozoa/physiology ; *Prophages/physiology/genetics ; Symbiosis ; Female ; Cytoplasm/metabolism ; }, abstract = {The symbiosis between arthropods and Wolbachia bacteria is globally widespread, largely due to selfish-drive systems that favour the fitness of symbiont-transmitting females. The most common drive, cytoplasmic incompatibility (CI), is central to arboviral control efforts. In Drosophila melanogaster carrying wMel Wolbachia deployed in mosquito control, two prophage genes in Wolbachia, cifA and cifB, cause CI that results in a paternal-effect lethality of embryos in crosses between Wolbachia-bearing males and aposymbiotic females. While the CI mechanism by which Cif proteins alter sperm development has recently been elucidated in D. melanogaster and Aedes aegypti mosquitoes, the Cifs' extended impact on male reproductive fitness such as sperm morphology and quantity remains unclear. Here, using cytochemical, microscopic and transgenic assays in D. melanogaster, we demonstrate that both CifA and CifB cause a significant portion of defects in elongating spermatids, culminating in malformed mature sperm nuclei. Males expressing Cifs have reduced spermatid bundles and sperm counts, and transgenic expression of Cifs can occasionally result in no mature sperm formation. We reflect on Cifs' varied functional impacts on the Host Modification model of CI as well as host evolution, behaviour and vector control strategies.}, } @article {pmid39934832, year = {2025}, author = {Perlmutter, JI and Atadurdyyeva, A and Schedl, ME and Unckless, RL}, title = {Wolbachia enhances the survival of Drosophila infected with fungal pathogens.}, journal = {BMC biology}, volume = {23}, number = {1}, pages = {42}, pmid = {39934832}, issn = {1741-7007}, abstract = {BACKGROUND: Wolbachia bacteria of arthropods are at the forefront of basic and translational research on multipartite host-symbiont-pathogen interactions. These vertically transmitted microbes are the most widespread endosymbionts on the planet due to factors including host reproductive manipulation and fitness benefits. Importantly, some strains of Wolbachia can inhibit viral pathogenesis within and between arthropod hosts. Mosquitoes carrying the wMel Wolbachia strain of Drosophila melanogaster have a greatly reduced capacity to spread viruses like dengue and Zika to humans. While significant research efforts have focused on viruses, relatively little attention has been given to Wolbachia-fungal interactions despite the ubiquity of fungal entomopathogens in nature.

RESULTS: Here, we demonstrate that Wolbachia increase the longevity of their Drosophila melanogaster hosts when challenged with a spectrum of yeast and filamentous fungal pathogens. We find that this pattern can vary based on host genotype, sex, and fungal species. Further, Wolbachia correlates with higher fertility and reduced pathogen titers during initial fungal infection, indicating a significant fitness benefit. Finally, RNA sequencing results show altered expression of many immune and stress response genes in the context of Wolbachia and fungal infection, suggesting host immunity may be involved in the mechanism.

CONCLUSIONS: This study demonstrates Wolbachia's protective role in diverse fungal pathogen interactions and determines that the phenotype is broad, but with several variables that influence both the presence and strength of the phenotype. It also is a critical step forward to understanding how symbionts can protect their hosts from a variety of pathogens.}, } @article {pmid39932664, year = {2025}, author = {da Fonseca Meireles, S and Ramalho, MO and Montenegro, H and do Nascimento Neto, JF and da Silva, JS and Cruz, DLV and Roque, RA and Rafael, MS}, title = {Do the microbiota of larval breeding site and the blood meal influence the composition and diversity of bacterial communities in the midgut of Mansonia humeralis (Diptera: Culicidae) from the western Amazon?.}, journal = {Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology]}, volume = {}, number = {}, pages = {}, pmid = {39932664}, issn = {1678-4405}, abstract = {Mansonia humeralis is a neotropical mosquito common in floodplain ecosystems and a potential host of Mayaro virus in the Amazon region. Studies show that bacterial communities associated with the midgut of mosquito vectors can mediate insecticide detoxification and modulate vector competence. Understanding the characteristics of the intestinal microbiota, and the factors that shape them, is essential for effective mosquito control. This study investigated the influence of bacterial communities in water from the larval environment on the composition and bacterial diversity in Ma. humeralis larval intestines and characterized the midgut microbiota of adult females with and without apparent blood. Samples were obtained from the Jirau Hydroelectric Plant, Madeira River, AM, Brazil, and their bacteria characterized using the 16S rRNA molecular marker. A total of 402 bacterial Amplicon Sequencing Variants (ASVs) were identified to genus level. The sample groups (water, larvae, blood + and blood-) had distinct bacterial communities, especially the larvae, probably due to the intense feeding activity which occurs at this stage. The phylum Proteobacteria was dominant in water, blood + and blood- (52.5; 99 and 98%), respectively, while Firmicutes dominated in larvae (45%). The most abundant taxa in water were: ASV Comamonadaceae (11.7%) and hgcI_clade (9.6%), in larvae: ASV Synergistaceae (11.1%) and ASV Rhodocyclaceae (7.8%), in blood-: Serratia (46.9%) and Asaia (12.6%) and, in blood+: ASV Enterobacteriaceae (54.5%) and Serratia (21.1%). Some 19% of the taxonomic groups from water were also recorded in larvae, while 46.7% of blood- microbiota were present in blood+. Wolbachia, Acinetobacter and Enterobacter bacteria were recorded at low frequency in all samples. This result provides a powerful tool for understanding the ecology of the intestinal microbiota of Ma. humeralis and the relationships with the ambient microbiota derived from larval filter, and adult blood, feeding. The results will also be useful for future studies which focusing the development of potential tools for the biological control of this vector.}, } @article {pmid39925617, year = {2025}, author = {Halford, G and Maes, D and Yung, CJ and Whiteford, S and Bourn, NAD and Bulman, CR and Goffart, P and Hodgson, JA and Saccheri, IJ}, title = {Genomic Monitoring of a Reintroduced Butterfly Uncovers Contrasting Founder Lineage Survival.}, journal = {Evolutionary applications}, volume = {18}, number = {2}, pages = {e70074}, pmid = {39925617}, issn = {1752-4571}, abstract = {Genetic factors can have a major influence on both short- and long-term success of reintroductions. Genomic monitoring can give a range of insights into the early life of a reintroduced population and ultimately can help to avoid wasting limited conservation resources. In this study, we characterise the genetic diversity of a reintroduced Carterocephalus palaemon (Chequered Skipper butterfly) population in England with respect to the spatial genetic structure and diversity of the source populations in south Belgium. We aim to evaluate the success of the reintroduction, including the effectiveness of the donor sampling strategy, and assess genetic vulnerabilities that may affect the population's future. We also use an isolation-by-distance approach to make quantitative inferences about dispersal, and we explore covariance between host mitochondrial and Wolbachia genomes. We find that, four generations following the initial release, the reintroduced population, founded by 66 wild-caught adults, has an effective size of c. 33, yet has retained similar levels of genomic heterozygosity to those in the source subpopulations in Belgium and shows low levels of inbreeding. However, the restricted number of founders and variance in reproductive success among the surviving families have resulted in a higher level of kinship, likely to result in somewhat higher rates of inbreeding in the future. Furthermore, there is a distinct split between two source landscapes in Belgium, and all genomic evidence suggests that the reintroduced population is descended from only one of these landscapes (called Fagne). We discuss potential causes behind these results, including whether Wolbachia strains are causing genetic incompatibility between clades. We conclude that a conservative strategy for any further translocations would prefer Fagne sites as sources because of the strong evidence of their ability to survive. However, our results warrant further investigation into the reasons for the divergence found in Belgium.}, } @article {pmid39914231, year = {2025}, author = {Páez-Triana, L and Martinez, D and Patiño, LH and Muñoz, M and Sandoval-Ramírez, CM and Pinilla León, JC and Ramirez, JD}, title = {Exploring endosymbionts and pathogens in Rhipicephalus sanguineus and Ctenocephalides felis felis with Oxford Nanopore Technology.}, journal = {Research in veterinary science}, volume = {185}, number = {}, pages = {105562}, doi = {10.1016/j.rvsc.2025.105562}, pmid = {39914231}, issn = {1532-2661}, abstract = {Fleas and ticks play a crucial role in public health as vectors of multiple diseases affecting humans and animals. Several rickettsial pathogens and endosymbionts are transmitted by fleas and ticks. Therefore, understanding this group of microorganisms is essential for fully grasping the spectrum of pathogens transmitted by vectors and the interactions between endosymbiotic microorganisms and their hosts. This study evaluated the presence and diversity of Rickettsiales species in fleas and ticks collected from the Santander department in Colombia. For the methodology a 16S gene amplification approach through Oxford Nanopore sequencing technologies in Rhipicephalus sanguineus and Ctenocephalides felis felis was used. Our findings revealed the presence of multiple pathogenic and endosymbiotic microorganisms, particularly from the Rickettsia and Wolbachia groups. We observed a clear association between Rickettsia species and ticks, while Wolbachia was predominantly found in fleas. Additionally, other important microorganisms were identified, including Anaplasma phagocytophilum, Rickettsia conorii, and different strains of Wolbachia that serve as endosymbionts in various arthropods. These results underscore the importance of fleas and ticks in the transmission of both pathogenic and endosymbiotic microorganisms. The distinct patterns of association between specific pathogens and vectors provide insight into their transmission dynamics. Identifying pathogens such as Anaplasma phagocytophilum and Rickettsia conorii further highlights the need for continued research into vector-borne diseases in Colombia. Understanding the interactions between endosymbionts and pathogenic microorganisms in these vectors could lead to the development of more effective strategies for controlling diseases transmitted by fleas and ticks.}, } @article {pmid39909190, year = {2025}, author = {Carbonara, M and Perles, L and Venco, L and Gabrielli, S and Barrs, VR and Miró, G and Papadopoulos, E and Lima, C and Bouhsira, E and Baneth, G and Pantchev, N and Iatta, R and Mendoza-Roldan, JA and Decaro, N and Schunack, B and Benelli, G and Otranto, D}, title = {Dirofilaria spp. infection in cats from the Mediterranean basin: diagnosis and epidemiology.}, journal = {International journal for parasitology}, volume = {}, number = {}, pages = {}, doi = {10.1016/j.ijpara.2025.01.011}, pmid = {39909190}, issn = {1879-0135}, abstract = {Dirofilaria immitis and Dirofilaria repens, causing heartworm disease and subcutaneous dirofilariosis, respectively, are zoonotic mosquito-borne filarioids infecting a plethora of hosts including cats. Only fragmented data are available on the diagnosis and epidemiology of feline dirofilariosis. We assessed the occurrence of both nematode infections, their risk factors and clinicopathological abnormalities in cats, from six countries of the Mediterranean Basin. In addition, Wolbachia spp. endosymbionts were assessed in Dirofilaria spp.-positive animals. Blood and sera samples were obtained from cats with outdoor access from Spain (n=354), Portugal (n=287), Italy (n=125), Greece (n=116), Israel (n=101) and France (n=100). Cat sera were tested by both direct antigenic (SNAP test, commercial ELISA kit) and indirect antibodies (in-house ELISA) serological tools, and blood samples by real time and conventional PCR targeting Dirofilaria spp. DNA, followed by sequencing. A statistical analysis was run to assess the link between Dirofilaria spp. infection and independent variables, as well as among feline immunodeficiency virus (FIV) and/or feline leukaemia virus (FeLV) co-infections, and clinicopathological abnormalities. Overall, 3.8% (i.e., 41/1,083) cats scored positive for Dirofilaria spp. infection with prevalences ranging from 2% in Israel to 7.8% in Greece. Of the 41 positive cats, 16 were infected by D. immitis (by SNAP test and/or PCR) and two by D. repens (by PCR); the remaining animals were antibody-positive for Dirofilaria spp. using the in-house ELISA. Wolbachia DNA was detected in one D. immitis-infected cat. Nematode positivity was significantly associated with age, breed, hyporexia, dandruff, and dyspnoea. This study provides data on the prevalence of Dirofilaria spp. infection in cats from the Mediterranean Basin, as well as new insights on its diagnosis, revealing the importance of performing strategic chemoprophylactic treatments for cats living in areas where the infection is also endemic in dogs.}, } @article {pmid39896488, year = {2025}, author = {Chappell, L and Peguero, R and Conner, WR and Fowler, S and Cooper, B and Pfarr, K and Hoerauf, A and Lustigman, S and Sakanari, J and Sullivan, W}, title = {Fexinidazole and Corallopyronin A target Wolbachia -infected sheath cells present in filarial nematodes.}, journal = {bioRxiv : the preprint server for biology}, volume = {}, number = {}, pages = {}, doi = {10.1101/2025.01.23.634442}, pmid = {39896488}, issn = {2692-8205}, abstract = {The discovery of the endosymbiotic bacteria Wolbachia as an obligate symbiont of filarial nematodes has led to antibiotic-based treatments for filarial diseases. While lab and clinical studies have yielded promising results, recent animal studies reveal that Wolbachia levels may rebound following treatment with suboptimal doses of the antibiotic rifampicin. Previous work showed that a likely source of the bacterial rebound in females were dense clusters of Wolbachia in ovarian tissue. The number, size, and density of these Wolbachia clusters were not diminished despite antibiotic treatment. Here we define the cellular characteristics of the Wolbachia clusters in Brugia pahangi (wBp) and identify drugs that also target them. We have evidence that the Wolbachia clusters originate from newly formed sheath cells adjacent to the ovarian Distal Tip Cells. The dramatically enlarged volume of an infected sheath cell is strikingly similar to endosymbiont-induced bacteriocytes found in many insect species. Ultrastructural analysis reveals that the clustered Wolbachia present within the sheath cells exhibit a distinct morphology and form direct connections with the oocyte membrane and possibly the cytoplasm. This includes membrane-based channels providing a connection between Wolbachia -infected sheath cells and oocytes. We also determined that the Wolbachia within the sheath cells are either quiescent or replicating at a very low rate. Screens of known antibiotics and other drugs revealed that two drugs, Fexinidazole and Corallopyronin A, significantly reduced the number of clustered Wolbachia located within the sheath cells.}, } @article {pmid39892716, year = {2025}, author = {Dudzic, JP and McPherson, AE and Taylor, KE and Eben, A and Abram, PK and Perlman, SJ}, title = {Candidate DNA and RNA viruses of Drosophila suzukii from Canada and Germany, and their interactions with Wolbachia.}, journal = {Journal of invertebrate pathology}, volume = {}, number = {}, pages = {108274}, doi = {10.1016/j.jip.2025.108274}, pmid = {39892716}, issn = {1096-0805}, abstract = {Some species of insects harbour strains of the endosymbiotic bacteria Wolbachia that do not cause obvious reproductive manipulations, and so it is unclear why they persist in host populations. There is some evidence that some of these endosymbionts may provide their hosts with protection against viruses, which would help to explain their persistence, but few studies have explored associations between Wolbachia and naturally occurring, common viruses in natural populations. Here, we asked whether individuals of the invasive vinegar fly Drosophila suzukii infected with the wSuz strain of Wolbachia were less likely to be infected by naturally occurring viruses in its invaded range, in western North America and in Europe. First, using next-generation sequencing, we conducted a virome survey of adult and larval D. suzukii in British Columbia, Canada, finding eight candidate RNA viruses and two candidate DNA viruses, all but one have not been reported previously. Only the previously described Teise virus, an RNA virus, was abundant in our virome survey. We then screened individual flies from British Columbia and Germany for Teise virus and Wolbachia. Wolbachia-infected D. suzukii from the field were not less likely to be infected by Teise virus. Overall, our results do not provide conclusive evidence that wSuz provides strong protection for D. suzukii against viruses that are common in natural populations. However, the other viruses that we discovered in this study, particularly the novel candidate Drosophila nudivirus, deserve further characterization in terms of their pathogenicity to D. suzukii and the frequency and dynamics of infection in wild populations.}, } @article {pmid39888481, year = {2025}, author = {Yao, RK and Gomgnimbou, MK and Coulibaly, IZ and Essoh, CY and Traoré, I and Amara, MF and Ako, BA and Diabate, A and Bilgo, E}, title = {Molecular detection of Wolbachia sp. and Cytoplasmic incompatibility factors (CifA/B) in wild caught mosquitoes in Côte d'Ivoire.}, journal = {Molecular biology reports}, volume = {52}, number = {1}, pages = {181}, pmid = {39888481}, issn = {1573-4978}, support = {ref/letter acceptation CEA/ITECH-MTV du 04/02/2021 à YAO R. Karlhis//CEA/ITECH-MTV/ ; ref/letter acceptation CEA/ITECH-MTV du 04/02/2021 à AMARA Miriam Félicité//CEA/ITECH-MTV/ ; 218771/Z/19/Z/WT_/Wellcome Trust/United Kingdom ; }, mesh = {*Wolbachia/genetics/isolation & purification ; Animals ; Cote d'Ivoire ; *Mosquito Vectors/microbiology/genetics ; *RNA, Ribosomal, 16S/genetics ; Culicidae/microbiology ; Aedes/microbiology/genetics ; Culex/microbiology ; Symbiosis ; Cytoplasm/microbiology ; Female ; }, abstract = {BACKGROUND: Wolbachia is an endosymbiont bacterium known to stimulate host immunity against arboviruses and protozoa. Côte d'Ivoire is in a malaria-endemic region, and has experienced several dengue epidemics in recent decades as well. In order to help reduce the transmission of pathogens by mosquito vectors, we studied the prevalence of Wolbachia and the distribution of Cytoplasmic incompatibility factors (Cif) genes in different mosquito species caught in the wild in Cote d'Ivoire.

METHODS AND RESULTS: Mosquitoes of the genera Anopheles, Aedes, Culex, Eretmapodites and Mansonia were captured in five cities. Mosquitoes were collected at larval stage in breeding sites and adults were captured using BG sentinel traps. The mosquitoes were identified morphologically and Wolbachia and Cif were screened using qPCR targeting the 16s rRNA gene and the CifA, B genes. A total of 518 mosquito samples belonging to 15 species and 4 genera were examined. 60% of the species were infected with Wolbachia. The three medically important mosquito species Aedes aegypti, Anopheles gambiae s.l. and Culex quinquefasciatus had a prevalence of 12.84%, 13.46% and 72.64% respectively. The Wolbachia strains infecting the different mosquito species of the genus Culex encoded 98.46% for the CifA gene and 77.69% for the CifB gene.

CONCLUSION: The presence of Wolbachia and CifA, B genes in mosquitoes of different species in Côte d'Ivoire offer a promising opportunity to reduce the competence of mosquito vectors. Characterization of Wolbachia strains and cytoplasmic incompatibility factors will provide a better understanding of these endosymbionts, enabling the development of vector control strategies.}, } @article {pmid39887637, year = {2025}, author = {Cagatay, NS and Akhoundi, M and Izri, A and Brun, S and Hurst, GDD}, title = {Prevalence of Heritable Symbionts in Parisian Bedbugs (Hemiptera: Cimicidae).}, journal = {Environmental microbiology reports}, volume = {17}, number = {1}, pages = {e70054}, doi = {10.1111/1758-2229.70054}, pmid = {39887637}, issn = {1758-2229}, support = {//TÜBİTAK [The Scientific and Technological Research Council of Türkiye]/ ; }, mesh = {Animals ; *Symbiosis ; *Bedbugs/microbiology/genetics ; *Wolbachia/genetics/isolation & purification/classification ; Paris ; DNA, Mitochondrial/genetics ; Haplotypes ; Prevalence ; }, abstract = {Like many insects, the biology of bedbugs is impacted by a range of partner heritable microbes. Three maternally inherited symbionts are recognised: Wolbachia (an obligate partner), Symbiopectobacterium purcellii strain SyClec, and Candidatus Tisiphia sp. (facultative symbionts typically present in some but not all individuals). Past work had examined the presence of these heritable microbes from established laboratory lines, but not from broader field samples. We therefore deployed targeted endpoint PCR assays to determine the symbiont infection status for 50 bedbugs collected from 10 districts of Paris during the 2023 outbreak. All three symbionts were found to be broadly present across Cimex lectularius samples, with the Symbiopectobacterium-Candidatus Tisiphia-Wolbachia triple infection most commonly observed. A minority of individuals lacked either one or both facultative symbionts. Five mtDNA haplotypes were observed across the COI barcode region, and triple infections were found in all mtDNA haplotypes, indicating that symbiont infection is not a recent invasion event. We conclude that the Parisian bedbug outbreak was one in which the host's secondary symbionts were present at high-frequency coinfections, and facultative symbionts are an important but uncharacterised component of bedbug populations.}, } @article {pmid39881623, year = {2025}, author = {Kryukova, NA and Polenogova, OV and Rotskaya, UN and Zolotareva, KA and Chertkova, EA}, title = {Wolbachia does not give an advantage to the ectoparasitoid Habrobracon hebetor (Say, 1836) when it develops on an infected host.}, journal = {Bulletin of entomological research}, volume = {}, number = {}, pages = {1-10}, doi = {10.1017/S0007485324000890}, pmid = {39881623}, issn = {1475-2670}, abstract = {The effect of Wolbachia on the viability and antimicrobial activity of the ectoparasitoid Habrobracon hebetor was evaluated in laboratory experiments. Two lines of the parasitoid, Wolbachia-infected (W+) and Wolbachia-free (W-), were used. Parasitoid larvae were fed with a host orally infected with a sublethal dose of Bacillus thuringiensis (Bt) and on the host uninfected with Bt. Parasitoid survival was assessed at developmental stages from second-instar larvae to adults. At all developmental stages, there were no statistically significant differences in survival between lines W+ and W-, regardless of host Bt infection. In both W+ and W- lines, the expression of lysozyme-like proteins, antimicrobial peptides (AMPs), and Hsp70 genes was analysed in fourth-instar larvae fed with an infected and uninfected host. In addition, lysozyme-like activity and antibacterial activity were evaluated. The expression of AMPs was significantly higher in W- larvae and did not get induced during the feeding on the Bt-infected host. mRNA expression of lysozyme-like proteins and lysozyme activity were significantly higher in W+ larvae than in W- larvae and did not get induced when the larvae were fed with the infected host. In whole-body homogenates of H. hebetor larvae fed with the uninfected host, antibacterial activity against gram-positive bacteria (Bacillus cereus and Bacillus subtilis) was significantly higher in the W+ line and did not get induced during the feeding with the Bt-infected host. Therefore, there is no obvious immunostimulatory effect of Wolbachia in H. hebetor larvae when they feed on a host infected with an entomopathogenic bacterium.}, } @article {pmid39865193, year = {2025}, author = {Chavarria, X and Choi, JH and Oh, S and Kim, M and Kang, D and Lee, IY and Jang, YS and Yi, MH and Yong, TS and Kim, JY}, title = {Metabarcoding for the Monitoring of the Microbiome and Parasitome of Medically Important Mosquito Species in Two Urban and Semi-urban Areas of South Korea.}, journal = {Current microbiology}, volume = {82}, number = {3}, pages = {102}, pmid = {39865193}, issn = {1432-0991}, support = {NRF-2020R1I1A2074562//National Research Foundation of Korea (KR)/ ; 6-2023-0065//Yonsei University College of Medicine/ ; HI23C1527//Ministry of Health and Welfare/ ; }, mesh = {Animals ; Republic of Korea ; *Microbiota ; *DNA Barcoding, Taxonomic ; *Bacteria/classification/genetics/isolation & purification ; *Aedes/microbiology ; Culex/microbiology ; Mosquito Vectors/microbiology ; RNA, Ribosomal, 16S/genetics ; RNA, Ribosomal, 18S/genetics ; }, abstract = {Interactions between microbial communities and the host can modulate mosquito biology, including vector competence. Therefore, future vector biocontrol measures will utilize these interactions and require extensive monitoring of the mosquito microbiome. Metabarcoding strategies will be useful for conducting vector monitoring on a large scale. We used 16S and 18S rRNA gene metabarcoding through iSeq100 sequencing to characterize the microbiome and eukaryome of Aedes albopictus (Skuse 1894) and Culex pipiens (Linnaeus 1758), two globally important vectors present in South Korea. Mosquitoes were collected from an urban and a semi-urban location in South Korea. Bacterial alpha and beta diversities varied by population. Pseudomonadota dominated the microbiomes of both species. The microbiome composition varied by population and was dominated by different taxa. At the genus level, Wolbachia sp. was the most enriched genus in Cx. pipiens, followed by Aeromonas sp. In Ae. Albopictus, the most abundant group was Enterococcus sp. The gregarine parasite Ascogregarina taiwanensis was highly prevalent in Ae. Albopictus and its absence was marked by the presence of seven bacterial taxa. To our knowledge, this is the first characterization of the microbiome of Ae. albopictus and Cx. pipiens in these regions of South Korea and contributes to the current information on the microbiome of mosquito species, which can be used in further studies to assess pathogen-microbiome and microbiome-microbiome interactions.}, } @article {pmid39859599, year = {2024}, author = {Fallon, AM}, title = {Resistance of Wolbachia to Trimethoprim: Insights into Genes Encoding Dihydrofolate Reductase, Thymidylate Synthase and Serine Hydroxymethyltransferase in the Rickettsiales.}, journal = {Insects}, volume = {16}, number = {1}, pages = {}, doi = {10.3390/insects16010018}, pmid = {39859599}, issn = {2075-4450}, abstract = {Bacterial and eukaryotic dihydrofolate reductase (DHFR) enzymes are essential for DNA synthesis and are differentially sensitive to the competitive inhibitors trimethoprim and methotrexate. Unexpectedly, trimethoprim did not reduce Wolbachia abundance, and the wStri DHFR homolog contained amino acid substitutions associated with trimethoprim resistance in E. coli. A phylogenetic tree showed good association of DHFR protein sequences with supergroup A and B assignments. In contrast, DHFR is not encoded by wFol (supergroup E) and wBm (supergroup D) or by genomes of the closely related genera Anaplasma, Ehrlichia, Neorickettsia, and possibly Orientia. In E. coli and humans, DHFR participates in a coupled reactions with the conventional thymidylate synthase (TS) encoded by thyA to produce the dTMP required for DNA synthesis. In contrast, Wolbachia and other Rickettsiales express the unconventional FAD-TS enzyme encoded by thyX, even when folA is present. The exclusive use of FAD-TS suggests that Wolbachia DHFR provides a supplementary rather than an essential function for de novo synthesis of dTMP, possibly reflecting the relative availability of, and competing demands for, FAD and NAD coenzymes in the diverse intracellular environments of its hosts. Whether encoded by thyA or thyX, TS produces dTMP by transferring a methyl group from methylene tetrahydrofolate to dUMP. In the Rickettsiales, serine hydroxymethyltransferase (SMHT), encoded by a conserved glyA gene, regenerates methylene tetrahydrofolate. Unlike thyA, thyX lacks a human counterpart and thus provides a potential target for the treatment of infections caused by pathogenic members of the Rickettsiales.}, } @article {pmid39845033, year = {2024}, author = {Luo, QC and Li, YY and Ren, YS and Yang, XH and Zhu, DH}, title = {Phage WO diversity and evolutionary forces associated with Wolbachia-infected crickets.}, journal = {Frontiers in microbiology}, volume = {15}, number = {}, pages = {1499315}, pmid = {39845033}, issn = {1664-302X}, abstract = {INTRODUCTION: Phage WO represents the sole bacteriophage identified to infect Wolbachia, exerting a range of impacts on the ecological dynamics and evolutionary trajectories of its host. Given the extensive prevalence of Wolbachia across various species, phage WO is likely among the most prolific phage lineages within arthropod populations. To examine the diversity and evolutionary dynamics of phage WO, we conducted a screening for the presence of phage WO in Wolbachia-infected cricket species from China.

METHODS: The presence of phage WO was detected using a PCR-based methodology. To elucidate the evolutionary forces driving phage WO diversity, analyses of intragenic recombination were conducted employing established recombination techniques, and horizontal transmission was investigated through comparative phylogenetic analysis of the phages and their hosts.

RESULTS AND DISCUSSION: Out of 19 cricket species infected with Wolbachia, 18 species were found to harbor phage WO. Notably, 13 of these 18 cricket species hosted multiple phage types, with the number of types ranging from two to 10, while the remaining five species harbored a single phage type. Twelve horizontal transmission events of phage WO were identified, wherein common phage WO types were shared among different Wolbachia strains. Notably, each phage WO horizontal transfer event was associated with distinct Wolbachia supergroups, specifically supergroups A, B, and F. Previous studies have found that four Wolbachia strains infect two to five species of crickets. However, among these cricket species, in addition to the shared phage WO types, all harbored species-specific phage WO types. This suggests that Wolbachia in crickets may acquire phage WO types through horizontal viral transfer between eukaryotes, independent of Wolbachia involvement. Furthermore, nine putative recombination events were identified across seven cricket species harboring multiple phage types. These findings suggest that horizontal transmission and intragenic recombination have played a significant role in the evolution of the phage WO genome, effectively enhancing the diversity of phage WO associated with crickets.}, } @article {pmid39840979, year = {2025}, author = {Lejarre, Q and Scussel, S and Esnault, J and Gaudillat, B and Duployer, M and Mavingui, P and Tortosa, P and Cattel, J}, title = {Development of the Incompatible Insect Technique targeting Aedes albopictus: introgression of a wild nuclear background restores the performance of males artificially infected with Wolbachia.}, journal = {Applied and environmental microbiology}, volume = {}, number = {}, pages = {e0235024}, doi = {10.1128/aem.02350-24}, pmid = {39840979}, issn = {1098-5336}, abstract = {The bacterium Wolbachia pipientis is increasingly studied for its potential use in controlling insect vectors or pests due to its ability to induce Cytoplasmic Incompatibility (CI). CI can be exploited by establishing an opportunistic Wolbachia infection in a targeted insect species through trans-infection and then releasing the infected males into the environment as sterilizing agents. Several host life history traits (LHT) have been reported to be negatively affected by artificial Wolbachia infection. Wolbachia is often considered the causative agent of these detrimental effects, and the importance of the host's genetic origins in the outcome of trans-infection is generally overlooked. In this study, we investigated the impact of host genetic background using an Aedes albopictus line recently trans-infected with wPip from the Culex pipiens mosquito, which exhibited some fitness costs. We measured several LHTs including fecundity, egg hatch rate, and male mating competitiveness in the incompatible line after four rounds of introgression aiming at restoring genetic diversity in the nuclear genome. Our results show that introgression with a wild genetic background restored most fitness traits and conferred mating competitiveness comparable to that of wild males. Finally, we show that introgression leads to faster and stronger population suppression under laboratory conditions. Overall, our data support that the host genome plays a decisive role in determining the fitness of Wolbachia-infected incompatible males.IMPORTANCEThe bacterium Wolbachia pipientis is increasingly used to control insect vectors and pests through the Incompatible Insect Technique (IIT) inducing a form of conditional sterility when a Wolbachia-infected male mates with an uninfected or differently infected female. Wolbachia artificial trans-infection has been repeatedly reported to affect mosquitoes LHTs, which may in turn compromise the efficiency of IIT. Using a tiger mosquito (Aedes albopictus) line recently trans-infected with a Wolbachia strain from Culex pipiens and displaying reduced fitness, we show that restoring genetic diversity through introgression significantly mitigated the fitness costs associated with Wolbachia trans-infection. This was further demonstrated through experimental population suppression, showing that introgression is required to achieve mosquito population suppression under laboratory conditions. These findings are significant for the implementation of IIT programs, as an increase in female fecundity and male performance improves mass rearing productivity as well as the sterilizing capacity of released males.}, } @article {pmid39836703, year = {2025}, author = {Sánchez-González, L and Crawford, JE and Adams, LE and Brown, G and Ryff, KR and Delorey, M and Ruiz-Valcarcel, J and Nazario, N and Borrero, N and Miranda, J and Mitchell, SN and Howell, PI and Ohm, JR and Behling, C and Wasson, B and Eldershaw, C and White, BJ and Rivera-Amill, V and Barrera, R and Paz-Bailey, G}, title = {Incompatible Aedes aegypti male releases as an intervention to reduce mosquito population-A field trial in Puerto Rico.}, journal = {PLoS neglected tropical diseases}, volume = {19}, number = {1}, pages = {e0012839}, doi = {10.1371/journal.pntd.0012839}, pmid = {39836703}, issn = {1935-2735}, abstract = {Mosquito-transmitted viruses such as dengue are a global and growing public health challenge. Without widely available vaccines, mosquito control is the primary tool for fighting the spread of these viruses. New mosquito control technologies are needed to complement existing methods, given current challenges with scalability, acceptability, and effectiveness. A field trial was conducted in collaboration with the Communities Organized to Prevent Arboviruses project in Ponce, Puerto Rico, to measure entomological and epidemiological effects of reducing Aedes aegypti populations using Wolbachia incompatible insect technique. We packed and shipped Wolbachia-males from California and released them into 19 treatment clusters from September 2020 to December 2020. Preliminary evaluation revealed sub-optimal Wolbachia-male densities and impact on the wild-type population. In 2021, we shifted to a phased release strategy starting in four clusters, reducing the mosquito population by 49% (CI 29-63%). We describe the investigation into male quality and other factors that may have limited the impact of Wolbachia-male releases. Laboratory assays showed a small but significant impact of packing and shipping on male fitness. However, mark-release-recapture assessments suggest that male daily survival rates in the field may have been significantly impacted. We compared induced-sterility levels and suppression of the wild population and found patterns consistent with mosquito population compensation in response to our intervention. Analysis of epidemiological impact was not possible due to very low viral transmission rates during the intervention period. Our entomological impact data provide evidence that Wolbachia incompatible-male releases reduced Ae. aegypti populations, although efficacy will be maximized when releases are part of an integrated control program. With improvement of shipping vessels and shipped male fitness, packing and shipping male mosquitoes could provide a key solution for expanding access to this technology. Our project underscores the challenges involved in large and complex field effectiveness assessments of novel vector control methods.}, } @article {pmid39829853, year = {2025}, author = {Namias, A and Martinez, J and Boussou, I and Terretaz, K and Conner, W and Justy, F and Makoundou, P and Perriat-Sanguinet, M and Labbé, P and Sicard, M and Landmann, F and Weill, M}, title = {Recombination, truncation and horizontal transfer shape the diversity of cytoplasmic incompatibility patterns.}, journal = {bioRxiv : the preprint server for biology}, volume = {}, number = {}, pages = {}, doi = {10.1101/2025.01.06.631454}, pmid = {39829853}, issn = {2692-8205}, abstract = {Wolbachia are endosymbiotic bacteria inducing various reproductive manipulations of which cytoplasmic incompatibility (CI) is the most common. CI leads to reduced embryo viability in crosses between males carrying Wolbachia and uninfected females or those carrying an incompatible symbiont strain. In the mosquito Culex pipiens , the Wolbachia w Pip causes highly complex crossing patterns. This complexity is linked to the amplification and diversification of the CI causal genes, cidA and cidB , with polymorphism located in the CidA-CidB interaction regions. We previously showed correlations between the identity of gene variants and CI patterns. However, these correlations were limited to specific crosses, and it is still unknown whether c id gene polymorphism in males' and females' Wolbachia can explain and predict the wide range of crossing types observed in C. pipiens . Taking advantage of a new method enabling full-gene acquisition, we sequenced complete cid repertoires from 45 w Pip strains collected worldwide. We demonstrated that the extensive diversity of cid genes arises from recombination and horizontal transfers. We uncovered further cidB polymorphism outside the interface regions and strongly correlated with CI patterns. Most importantly, we showed that in every w Pip genome, all but one cidB variant are truncated. Truncated cidB s located in palindromes are partially or completely deprived of their deubiquitinase domain, crucial for CI. The identity of the sole full-length cidB variant seems to dictate CI patterns, irrespective of the truncated cidBs present. Truncated CidBs exhibit reduced toxicity and stability in Drosophila cells, which potentially hinders their loading into sperm, essential for CI induction.}, } @article {pmid39824916, year = {2025}, author = {Wong, WJ and Tan, CH and Verkaik, MG and Ng, LC and Hoffmann, AA and Chong, CS}, title = {Suppression of Aedes aegypti may not affect sympatric Aedes albopictus populations: findings from two years of entomological surveillance in Singapore.}, journal = {Scientific reports}, volume = {15}, number = {1}, pages = {2253}, pmid = {39824916}, issn = {2045-2322}, mesh = {Animals ; *Aedes/physiology/virology ; Singapore ; *Mosquito Vectors/virology ; *Mosquito Control/methods ; Wolbachia/physiology ; Dengue/transmission/prevention & control ; Dengue Virus/physiology ; }, abstract = {Globally, multiple trials have successfully demonstrated the effectiveness of novel tools, such as the sterile and incompatible insect techniques, in suppressing Aedes aegypti populations. However, there is concern that Aedes albopictus, another arbovirus-competent vector, may occupy the niches vacated by Ae. aegypti in areas where these species occur in sympatry. Here we investigate these concerns within the context of a Wolbachia-based Ae. aegypti suppression programme in highly urban Singapore, where general mosquito management is sustained through environmental management. Using nationally representative and longitudinal Aedes surveillance data, we show (1) no consistent association of increase in Ae. albopictus abundance with Ae. aegypti population suppression within Wolbachia-Aedes release sites, and (2) no significant change in the vertical spatial distribution of Ae. albopictus in high-rise residential apartment blocks even after two years of Ae. aegypti suppression. Finally, we report that dengue viruses were less prevalent in field Ae. albopictus than in Ae. aegypti, which is consistent with previous findings that Ae. albopictus is a lesser vector than Ae. aegypti in Singapore. Together, these results indicate that suppression of Ae. aegypti in the community may not be directly associated with any increase of Ae. albopictus population or capacity. Nonetheless, the risk of increase in Ae. albopictus population is anticipated if the reduction in disease transmission leads to less aggressive source reduction efforts and environmental management for mosquito control.}, } @article {pmid39819374, year = {2025}, author = {Asselin, A and Johnson, K}, title = {The infectivity of virus particles from Wolbachia-infected Drosophila.}, journal = {BMC microbiology}, volume = {25}, number = {1}, pages = {25}, pmid = {39819374}, issn = {1471-2180}, mesh = {Animals ; *Wolbachia/physiology ; *Drosophila melanogaster/microbiology/virology ; *Dicistroviridae/physiology/pathogenicity ; *Virus Replication ; Nodaviridae/physiology/pathogenicity ; Virion ; Symbiosis ; Female ; Insect Viruses/physiology/pathogenicity ; }, abstract = {Viruses transmitted by arthropods pose a huge risk to human health. Wolbachia is an endosymbiotic bacterium that infects various arthropods and can block the viral replication cycle of several medically important viruses. As such, it has been successfully implemented in vector control strategies against mosquito-borne diseases, including Dengue virus. Whilst the mechanisms behind Wolbachia-mediated viral blocking are not fully characterised, it was recently shown that viruses grown in the presence of Wolbachia in some Dipteran cell cultures are less infectious than those grown in the absence of Wolbachia. Here, we investigate the breadth of this mechanism by determining if Wolbachia reduces infectivity in a different system at a different scale. To do this, we looked at Wolbachia's impact on insect viruses from two diverse virus families within the whole organism Drosophila melanogaster. Drosophila C virus (DCV; Family Dicistroviridae) and Flock House virus (FHV; Famliy Nodaviridae) were grown in adult D. melanogaster flies with and without Wolbachia strain wMelPop. Measures of the physical characteristics, infectivity, pathogenicity, and replicative properties of progeny virus particles did not identify any impact of Wolbachia on either DCV or FHV. Therefore, there was no evidence that changes in infectivity contribute to Wolbachia-mediated viral blocking in this system. Overall, this is consistent with growing evidence that the mechanisms behind Wolbachia viral blocking are dependent on the unique tripartite interactions occurring between the host, the Wolbachia strain, and the infecting virus.}, } @article {pmid39809813, year = {2025}, author = {Mewis, V and Wendt, M and Schmitt, T}, title = {Phylogeographic analyses reveal recent dispersal and multiple Wolbachia infections of the bright-eyed ringlet Erebia oeme within the European mountain systems.}, journal = {Scientific reports}, volume = {15}, number = {1}, pages = {1956}, pmid = {39809813}, issn = {2045-2322}, mesh = {*Wolbachia/genetics/classification/isolation & purification ; Animals ; *Phylogeography ; *Phylogeny ; Europe ; DNA, Mitochondrial/genetics ; Genetic Variation ; }, abstract = {The genus Erebia comprises numerous species in Europe. Due to preference of cold environments, most species have disjunct distributions in the European mountain systems. However, their biogeographical patterns may differ significantly. The Bright-eyed ringlet Erebia oeme is widespread in high-altitude grasslands of the European high mountains, hence showing a disjunct distribution pattern. Over its distribution, E. oeme shows high morphological variability indicating pronounced intraspecific differentiation. We analysed two mitochondrial (COI, Cytb) and eight nuclear markers as well as the Wolbachia surface protein-coding gene (WSP). A total of four lineages were identified: two Balkan lineages, one Slovenian lineage as well as one lineage containing all other individuals (Alps, Massif Central, Pyrenees). COI data only indicate a fifth lineage in the southern Carpathians. The region of origin of E. oeme is most likely the western Balkans. From here, E. oeme spread to the eastern Balkan area and further to the southern Carpathians as well as to Slovenia, from where it rapidly crossed the Alps to Massif Central and Pyrenees. Wolbachia was found to be highly prevalent with multiple strains, partly within one population. However, recent Wolbachia impact on the mitochondrial DNA and consequently influence on the intraspecific genetic structure was not detected.}, } @article {pmid39794995, year = {2024}, author = {Glowska-Patyniak, E and Kaszewska-Gilas, K and Laniecka, I and Olechnowicz, J and Ostrowska, K and Dmuchowska, W and Schmidt, BK and Hubert, J and Trzebny, A}, title = {First Detection of Wolbachia in Namibian Bird Ectoparasites (Acariformes: Syringophilidae) with a Description of New Quill Mite Species.}, journal = {Animals : an open access journal from MDPI}, volume = {15}, number = {1}, pages = {}, doi = {10.3390/ani15010052}, pmid = {39794995}, issn = {2076-2615}, support = {UMO-2021/03/Y/NZ8/00060//National Science Centre of Poland/ ; }, abstract = {Wolbachia is a common intracellular bacterial genus that infects numerous arthropods and filarial nematodes. In arthropods, it typically acts as a reproductive parasite, leading to various phenotypic effects such as cytoplasmic incompatibility, parthenogenesis, feminization, or male-killing. Quill mites (Acariformes: Syringophilidae) are a group of bird parasites that have recently attracted increasing interest due to the detection of unique phylogenetic lineages of endosymbiotic bacteria and potentially pathogenic taxa. Our study used an unbiased 16S rRNA gene amplicon sequencing approach to examine several populations of Namibian quill mites for the presence of bacteria that could affect their biology. We detected Wolbachia in two mite populations collected from two species of larks. However, we did not find any other endosymbiotic bacteria or any that could be of epidemiological importance. Since the mite taxa we tested were previously unknown to science, we conducted comprehensive morphological and molecular systematic analyses on them. Our research revealed two new quill mite species of the genus Syringophilopsis Kethley, 1970 which parasitize three sub-Saharan alaudids, i.e., Syringophilopsis erythrochlamys sp. n. from the dune lark Calendulauda erythrochlamys (Strickland, HE) and S. christinae sp. n. from the Karoo long-billed lark Certhilauda subcoronata (Smith) and spike-heeled lark Chersomanes albofasciata (de Lafresnaye, NFAA). In addition, we provided the African reed warbler Acrocephalus baeticatus (Vieillot, LJP) as a new host for S. acrocephali Skoracki, 1999. Our study expands the knowledge on parasite diversity and provides new insights into Wolbachia infection among quill mites in Africa.}, } @article {pmid39773184, year = {2025}, author = {Ribeiro, P and Butenko, A and Linke, D and Ghanavi, HR and Meier, JI and Wahlberg, N and Matos-Maraví, P}, title = {Pervasive horizontal transmission of Wolbachia in natural populations of closely related and widespread tropical skipper butterflies.}, journal = {BMC microbiology}, volume = {25}, number = {1}, pages = {5}, pmid = {39773184}, issn = {1471-2180}, support = {n.014/2022/P//Grantová Agentura Jihoceska Univerzita/ ; n.014/2022/P//Grantová Agentura Jihoceska Univerzita/ ; GACR 22-35084J//Grantová Agentura České Republiky/ ; GACR 22-35084J//Grantová Agentura České Republiky/ ; GACR 22-35084J//Grantová Agentura České Republiky/ ; 220540/Z/20/A/WT_/Wellcome Trust/United Kingdom ; }, mesh = {*Wolbachia/genetics/classification/isolation & purification ; Animals ; *Butterflies/microbiology ; *Symbiosis ; *Phylogeny ; *Whole Genome Sequencing ; Genome, Bacterial/genetics ; }, abstract = {BACKGROUND: The endosymbiotic relationship between Wolbachia bacteria and insects has been of interest for many years due to their diverse types of host reproductive phenotypic manipulation and potential role in the host's evolutionary history and population dynamics. Even though infection rates are high in Lepidoptera and specifically in butterflies, and reproductive manipulation is present in these taxa, less attention has been given to understanding how Wolbachia is acquired and maintained in their natural populations, across and within species having continental geographical distributions.

RESULTS: We used whole genome sequencing data to investigate the phylogenetics, demographic history, and infection rate dynamics of Wolbachia in four species of the Spicauda genus of skipper butterflies (Lepidoptera: Hesperiidae), a taxon that presents sympatric and often syntopic distribution, with drastic variability in species abundance in the Neotropical region. We show that infection is maintained by high turnover rates driven mainly by pervasive horizontal transmissions, while also presenting novel cases of double infection by distantly related supergroups of Wolbachia in S. simplicius.

CONCLUSIONS: Our results suggest that Wolbachia population dynamics is host species-specific, with genetic cohesiveness across wide geographical distributions. We demonstrate that low coverage whole genome sequencing data can be used for an exhaustive assessment of Wolbachia infection in natural populations of butterflies, as well as its dynamics in closely related host species. This ultimately leads to a better understanding of the endosymbiotic population dynamics of Wolbachia and its effects on the host's biology and evolution.}, } @article {pmid39772178, year = {2024}, author = {Branda, F and Cella, E and Scarpa, F and Slavov, SN and Bevivino, A and Moretti, R and Degafu, AL and Pecchia, L and Rizzo, A and Defilippo, F and Moreno, A and Ceccarelli, G and Alcantara, LCJ and Ferreira, A and Ciccozzi, M and Giovanetti, M}, title = {Wolbachia-Based Approaches to Controlling Mosquito-Borne Viral Threats: Innovations, AI Integration, and Future Directions in the Context of Climate Change.}, journal = {Viruses}, volume = {16}, number = {12}, pages = {}, doi = {10.3390/v16121868}, pmid = {39772178}, issn = {1999-4915}, mesh = {*Wolbachia/physiology ; *Climate Change ; Animals ; *Mosquito Control/methods ; *Mosquito Vectors/microbiology/virology ; *Vector Borne Diseases/prevention & control/transmission ; Humans ; Zika Virus Infection/prevention & control/transmission ; Culicidae/virology/microbiology ; Dengue/prevention & control/transmission ; Aedes/microbiology/virology ; Ecosystem ; }, abstract = {Wolbachia-based mosquito control strategies have gained significant attention as a sustainable approach to reduce the transmission of vector-borne diseases such as dengue, Zika, and chikungunya. These endosymbiotic bacteria can limit the ability of mosquitoes to transmit pathogens, offering a promising alternative to traditional chemical-based interventions. With the growing impact of climate change on mosquito population dynamics and disease transmission, Wolbachia interventions represent an adaptable and resilient strategy for mitigating the public health burden of vector-borne diseases. Changes in temperature, humidity, and rainfall patterns can alter mosquito breeding habitats and extend the geographical range of disease vectors, increasing the urgency for effective control measures. This review highlights innovations in Wolbachia-based mosquito control and explores future directions in the context of climate change. It emphasizes the integration of Wolbachia with other biological approaches and the need for multidisciplinary efforts to address climate-amplified disease risks. As ecosystems shift, Wolbachia interventions could be crucial in reducing mosquito-borne diseases, especially in vulnerable regions. AI integration in Wolbachia research presents opportunities to enhance mosquito control strategies by modeling ecological data, predicting mosquito dynamics, and optimizing intervention outcomes. Key areas include refining release strategies, real-time monitoring, and scaling interventions. Future opportunities lie in advancing AI-driven approaches for integrating Wolbachia with other vector control measures, promoting adaptive, data-driven responses to climate-amplified disease transmission.}, } @article {pmid39769589, year = {2024}, author = {Martín-Park, A and Contreras-Perera, Y and Che-Mendoza, A and Pérez-Carrillo, S and Pavía-Ruz, N and Villegas-Chim, J and Trujillo-Peña, E and Bibiano-Marín, W and Medina-Barreiro, A and González-Olvera, G and Navarrete-Carballo, J and Puerta-Guardo, H and Ayora-Talavera, G and Delfín-González, H and Palacio-Vargas, J and Correa-Morales, F and Bezerra, HSDS and Coelho, G and Vazquez-Prokopec, G and Xi, Z and Manrique-Saide, P and Gómez-Dantes, H}, title = {Recommendations for Implementing Innovative Technologies to Control Aedes aegypti: Population Suppression Using a Combination of the Incompatible and Sterile Insect Techniques (IIT-SIT), Based on the Mexican Experience/Initiative.}, journal = {Insects}, volume = {15}, number = {12}, pages = {}, doi = {10.3390/insects15120987}, pmid = {39769589}, issn = {2075-4450}, support = {YUC201703-01-556//Fondo Mixto Consejo Nacional de Ciencia y Tecnología (CONACYT) (México)-Gobierno del Es-tado de Yucatán/ ; AID-OAA-F-16-00082//U.S. Agency for International Development/ ; APS-7200AA20APS00013//U.S. Agency for International Development/ ; }, abstract = {The future of Aedes aegypti control emphasizes the transition from traditional insecticides toward more sustainable and multisectoral integrated strategies, like using Wolbachia-carrying mosquitoes for population suppression or replacement. We reviewed the integration of the successful Mexican initiative, "Mosquitos Buenos", with the key challenges outlined in the PAHO guidelines for incorporating innovative approaches into vector control programs. These challenges include establishing essential infrastructure, training personnel, managing field operations, and fostering community support. Our experience provides critical evidence to support the strategic National Plan for implementing and integrating IIT-SIT technologies to control Ae. aegypti and dengue. Furthermore, this experience serves as a foundation for other countries in the region interested in adopting these technologies. It underscores the importance of strategic planning, multisectoral collaboration, continuous evaluation, and scaling up innovative tools to ensure their long-term effectiveness and sustainability in urban areas where Aedes vectors and the diseases they transmit are endemic.}, } @article {pmid39763920, year = {2024}, author = {Arnce, LR and Bubnell, JE and Aquadro, CF}, title = {Comparative Analysis of Drosophila Bam and Bgcn Sequences and Predicted Protein Structural Evolution.}, journal = {bioRxiv : the preprint server for biology}, volume = {}, number = {}, pages = {}, doi = {10.1101/2024.12.17.628990}, pmid = {39763920}, issn = {2692-8205}, abstract = {The protein encoded by the Drosophila melanogaster gene bag of marbles (bam) plays an essential role in early gametogenesis by complexing with the gene product of benign gonial cell neoplasm (bgcn) to promote germline stem cell daughter differentiation in males and females. Here, we compared the AlphaFold2 and AlphaFold Multimer predicted structures of Bam protein and the Bam:Bgcn protein complex between D. melanogaster, D. simulans, and D. yakuba , where bam is necessary in gametogenesis to that in D. teissieri , where it is not. Despite significant sequence divergence, we find very little evidence of significant structural differences in high confidence regions of the structures across the four species. This suggests that Bam structure is unlikely to be a direct cause of its functional differences between species and that Bam may simply not be integrated in an essential manner for GSC differentiation in D. teissieri . Patterns of positive selection and significant amino acid diversification across species is consistent with the Selection, Pleiotropy, and Compensation (SPC) model, where detected selection at bam is consistent with adaptive change in one major trait followed by positively selected compensatory changes for pleiotropic effects (in this case perhaps preserving structure). In the case of bam , we suggest that the major trait could be genetic interaction with the endosymbiotic bacteria Wolbachia pipientis . Following up on detected signals of positive selection and comparative structural analysis could provide insight into the distribution of a primary adaptive change versus compensatory changes following a primary change.}, } @article {pmid39755093, year = {2025}, author = {Gao, T and Zhang, Y and Sun, W and Li, Q and Huang, X and Zhi, D and Zi, H and Ji, R and Long, Y and Gong, C and Yang, Y}, title = {The symbiont Wolbachia increases resistance to bifenthrin in Ectropis grisescens by regulating the host detoxification function.}, journal = {Ecotoxicology and environmental safety}, volume = {289}, number = {}, pages = {117666}, doi = {10.1016/j.ecoenv.2025.117666}, pmid = {39755093}, issn = {1090-2414}, abstract = {The global issue of insecticide resistance among pests is a major concern. Ectropis grisescens Warren (Lepidoptera: Geometridae), is a highly destructive leaf-eating pest distributed in tea plantations throughout China and Japan, and has exhibited resistance to various insecticides. Recent studies suggest that insect symbionts play a role in influencing insecticide resistance, however, their specific involvement in E. grisescens remains unclear. Here, we initially selected appropriate antibiotic mixtures at a concentration of 300 μg ml[-1]. The bioassay results showed that the insecticide susceptibility of the E. grisescens population treated with antibiotic mixtures significantly increased exposed to bifenthrin. Comparative analysis revealed that the LC50 value, survival rate, P450 enzyme activity, and relative content of Wolbachia in the E. grisescens population treated with 300 μg ml[-1] tetracycline were notably lower than those treated with other antibiotics (ampicillin, gentamicin, and streptomycin). Moreover, the population treated with 2.5 mg ml[-1] tetracycline exhibited even greater reductions in these parameters than the 300 μg ml[-1] tetracycline-treated group. Additionally, 16S rRNA sequencing results showed a significant decrease in xenobiotics metabolism by cytochrome P450 in the E. grisescens population treated with 2.5 mg ml[-1] tetracycline. Transcriptome analysis showed a significant down-regulation of two cytochrome P450 genes in E. grisescens population without Wolbachia. These results suggest that Wolbachia may contribute to the resistance of E. grisescens to bifenthrin by regulating cytochrome P450 genes, providing a foundation for further study on the mechanism of symbiont-mediated host detoxification metabolism in insect pests.}, } @article {pmid39747462, year = {2025}, author = {Wu, K and Vu, ED and Ghosh, S and Mishra, R and Bonning, BC}, title = {Continuous cell lines derived from the Asian citrus psyllid, Diaphorina citri, harbor viruses and Wolbachia.}, journal = {Scientific reports}, volume = {15}, number = {1}, pages = {124}, pmid = {39747462}, issn = {2045-2322}, support = {2022-70029-38506//National Institute of Food and Agriculture/ ; }, abstract = {The Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae), is a major pest of global citriculture. In the Americas and in Asia, D. citri vectors the phloem-limited bacterium, Candidatus Liberibacter asiaticus (CLas), which causes the fatal citrus disease huanglongbing, or citrus greening. Cell lines derived from D. citri can provide insight into both the basic biology of this pest and D. citri-associated pathogens including CLas. We previously identified CLG#2 as the optimal medium for long-term growth of D. citri primary cell cultures. Here we report on the establishment and characterization of three continuous D. citri cell lines, Dici1, Dici3, and Dici5, that have been passaged for > 40 times. Based on morphological and transcriptomic data, the Dici1 and Dici3 cell lines include undifferentiated and neurogenic progenitor cells. Dici1 and Dici5 are infected with Wolbachia. Both Dici1 and Dici5 are infected with D. citri reovirus, and Dici5 is also infected with D. citri-associated C virus. Dici3 is free of both Wolbachia and virus infection. These cell lines provide an ideal platform for the study of inter-microbial relationships as well as microbe interaction with host insect cells.}, } @article {pmid39740726, year = {2024}, author = {Bhattacharyya, J and Roelke, DL}, title = {Wolbachia-Based Mosquito Control: Environmental Perspectives on Population Suppression and Replacement Strategies.}, journal = {Acta tropica}, volume = {}, number = {}, pages = {107517}, doi = {10.1016/j.actatropica.2024.107517}, pmid = {39740726}, issn = {1873-6254}, abstract = {Mosquito-borne diseases pose a significant threat to global health, and traditional mosquito control methods often fall short of effectiveness. A promising alternative is the biological control strategy of transinfecting mosquitoes with Wolbachia, a bacterium capable of outcompeting harmful pathogens and reducing the ability of mosquitoes to transmit diseases. However, Wolbachia infections are sensitive to abiotic environmental factors such as temperature and humidity, which can affect their densities in mosquitoes and, consequently, their ability to block pathogens. This review evaluates the effectiveness of different Wolbachia strains transinfected into mosquitoes in reducing mosquito-borne diseases. It explores how Wolbachia contributes to mosquito population control and pathogen interference, highlighting the importance of mathematical models in understanding Wolbachia transmission dynamics. Additionally, the review addresses the potential impact on arboviral transmission and the challenges posed by environmental fluctuations in mosquito control programs.}, } @article {pmid39739581, year = {2024}, author = {Kagemann, CH and Bubnell, JE and Colocho, GM and Arana, D and Aquadro, CF}, title = {Wolbachia pipientis Modulates Germline Stem Cells and Gene Expression Associated with Ubiquitination and Histone Lysine Trimethylation to Rescue Fertility Defects in Drosophila.}, journal = {Genetics}, volume = {}, number = {}, pages = {}, doi = {10.1093/genetics/iyae220}, pmid = {39739581}, issn = {1943-2631}, abstract = {Wolbachia pipientis are maternally transmitted endosymbiotic bacteria commonly found in arthropods and nematodes. These bacteria manipulate reproduction of the host to increase their transmission using mechanisms, such as cytoplasmic incompatibility, that favor infected female offspring. The underlying mechanisms of reproductive manipulation by W. pipientis remain unresolved. Interestingly, W. pipientis infection partially rescues female fertility in flies containing hypomorphic mutations of bag of marbles (bam) in D. melanogaster, which plays a key role in germline stem cell (GSC) daughter differentiation. Using RNA-seq, we find W. pipientis infection in bam hypomorphic females results in differential expression of many of bam's genetic and physical interactors and enrichment of ubiquitination and histone lysine methylation genes. We find that W. pipientis also rescues the fertility and GSC functions of a subset of these genes when knocked down with RNAi in a wildtype bam genotype. Our results show that W. pipientis interacts with ubiquitination and histone lysine methylation genes which could be integral to the mechanism by which W. pipientis modulates GSC gene function.}, } @article {pmid39738989, year = {2024}, author = {Bassini-Silva, R and Calchi, AC and Castro-Santiago, AC and Marocco, JC and Dorigoni, L and de Quadros, RM and André, MR and Barros-Battesti, DM and Dowling, APG and Labruna, MB and Jacinavicius, FC}, title = {Molecular evidence of Wolbachia in bat-associated mite Periglischrus Iheringi Oudemans, 1902 (Mesostigmata: Spinturnicidae) from Brazil.}, journal = {Veterinary research communications}, volume = {49}, number = {1}, pages = {60}, pmid = {39738989}, issn = {1573-7446}, support = {2020/11755-6//Fundação de Amparo à Pesquisa do Estado de São Paulo/ ; 2020/07826-5//Fundação de Amparo à Pesquisa do Estado de São Paulo/ ; 2021/06758-9//Fundação de Amparo à Pesquisa do Estado de São Paulo/ ; 2022/05615-2//Fundação de Amparo à Pesquisa do Estado de São Paulo/ ; 2019/19853-0, 2024/01231-0//Fundação de Amparo à Pesquisa do Estado de São Paulo/ ; #303701/2021-8//Conselho Nacional de Desenvolvimento Científico e Tecnológico/ ; #303802/2021-9//Conselho Nacional de Desenvolvimento Científico e Tecnológico/ ; 402575/2021-0//Conselho Nacional de Desenvolvimento Científico e Tecnológico/ ; }, mesh = {Animals ; *Wolbachia/genetics/isolation & purification/classification ; Brazil ; *Mites/microbiology ; *Phylogeny ; *Chiroptera/microbiology/parasitology ; *RNA, Ribosomal, 16S/genetics ; }, abstract = {Wolbachia is an intracellular endosymbiont bacterium found in nematodes and arthopods. Regarding mites, the Wolbachia supergroup U has been described based on strains found in the genus Spinturnix. In this study, ten specimens of Periglischrus iheringi (Mesostigmata: Spinturnicidae), collected from Artibeus obscurus (Chiroptera: Phyllostomidae) in Santa Catarina State, were found to be infected with Wolbachia. Phylogenetic analysis based on the 16 S rRNA gene revealed that the detected Wolbachia strain belongs to Supergroup F, which has also been detected in other ectoparasitic arthropods, such as Columbicola columbae (slender pigeon lice) and Cimex lectularius (bed bug). This study presents the first molecular detection of Wolbachia in P. iheringi.}, } @article {pmid39737459, year = {2024}, author = {Ogunlade, ST and Adekunle, AI and McBryde, ES}, title = {Mitigating dengue transmission in Africa: the need for Wolbachia-infected mosquitoes' rollout.}, journal = {Frontiers in public health}, volume = {12}, number = {}, pages = {1506072}, pmid = {39737459}, issn = {2296-2565}, } @article {pmid39733938, year = {2024}, author = {Angelella, GM and Foutz, JJ and Galindo-Schuller, J}, title = {Wolbachia infection modifies phloem feeding behavior but not plant virus transmission by a hemipteran host.}, journal = {Journal of insect physiology}, volume = {}, number = {}, pages = {104746}, doi = {10.1016/j.jinsphys.2024.104746}, pmid = {39733938}, issn = {1879-1611}, abstract = {Wolbachia-infected and uninfected subpopulations of beet leafhoppers, Circulifer tenellus (Baker) (Hemiptera: Cicadellidae), co-occur in the Columbia Basin region of Washington and Oregon. While facultative endosymbionts such as Hamiltonella defensa have demonstrably altered feeding/probing behavior in hemipteran hosts, the behavioral phenotypes conferred by Wolbachia to its insect hosts, including feeding/probing, are largely understudied. We studied the feeding/probing behavior of beet leafhoppers with and without Wolbachia using electropenetrography, along with corresponding inoculation rates of beet curly top virus, a phloem-limited plant pathogen vectored by beet leafhoppers. Insects carrying the virus with and without Wolbachia were individually recorded for four hours while interacting with a potato plant, and wavelengths annotated following established conventions. Virus inoculation rates and the duration of phloem salivation events did not vary. Wolbachia-infected insects more than tripled the duration of phloem ingestion, but despite this, Wolbachia infection was linked with marginally lower, not enhanced, acquisition. Regardless, results suggest potential for Wolbachia to increase the acquisition rate of other phloem-limited plant pathogens.}, } @article {pmid39718247, year = {2024}, author = {Santos, PKF and de Souza Araujo, N and Françoso, E and Werren, JH and Kapheim, KM and Arias, MC}, title = {The genome of the solitary bee Tetrapedia diversipes (Hymenoptera, Apidae).}, journal = {G3 (Bethesda, Md.)}, volume = {}, number = {}, pages = {}, doi = {10.1093/g3journal/jkae264}, pmid = {39718247}, issn = {2160-1836}, support = {001//Coordenação de Aperfeiçoamento de Pessoal de Nível Superior/ ; 306932/2016-4//CNPq - Conselho Nacional de Desenvolvimento Científico e Tecnológico/ ; 2013/12530-4//Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)/ ; DEB1257053//USNSF/ ; }, abstract = {Tetrapedia diversipes is a Neotropical solitary bee commonly found in trap-nests, known for its morphological adaptations for floral oil collection and prepupal diapause during the cold and dry season. Here, we present the genome assembly of T. diversipes (332 Mbp), comprising 2,575 scaffolds, with 15,028 predicted protein-coding genes. Repetitive elements constitute 38.68% of the genome, notably Class II transposable elements. An investigation into lateral gene transfers identified a low frequency (0.037%) of nuclear copies of mitochondrial DNA and 18 candidate regions from bacterial origins. Furthermore, the annotation of 3 scaffolds reveals the presence of the Wolbachia endosymbiont genome, confirming the infection by 2 strains in T. diversipes populations. This genome contributes valuable insights into Neotropical bee genomics, offering a resource for comparative studies and enhancing our understanding of the molecular basis of solitary bee adaptations and interactions.}, } @article {pmid39717715, year = {2024}, author = {Lai, CT and Hsiao, YT and Wu, LH}, title = {Evidence of horizontal transmission of Wolbachia wCcep in rice moths parasitized by Trichogramma chilonis and its persistence across generations.}, journal = {Frontiers in insect science}, volume = {4}, number = {}, pages = {1519986}, pmid = {39717715}, issn = {2673-8600}, abstract = {The horizontal transmission of endosymbionts between hosts and parasitoids plays a crucial role in biological control, yet its mechanisms remain poorly understood. This study investigates the dynamics of horizontal transfer of Wolbachia (wCcep) from the rice moth, Corcyra cephalonica, to its parasitoid, Trichogramma chilonis. Through PCR detection and phylogenetic analysis, we demonstrated the presence of identical wCcep strains in both host and parasitoid populations, providing evidence for natural horizontal transmission. To investigate thoroughly, Wolbachia-free colonies were acquired through tetracycline treatment, and the initial density of wCcep in host eggs significantly influences transmission efficiency. High-density wCcep infections led to rapid transmission, with F1 parasitoid titers increasing by as much as 100-fold, while low-density infections exhibited more gradual increases. Additionally, without continuous exposure to infected hosts, wCcep density in T. chilonis diminished over generations. These findings enhance our understanding of Wolbachia's transfer dynamics and have important implications for developing effective and sustainable biological control strategies using parasitoid wasps, particularly in managing Wolbachia-related pest populations in agricultural systems.}, } @article {pmid39716303, year = {2024}, author = {da Moura, AJF and Tomaz, F and Melo, T and Seixas, G and Sousa, CA and Pinto, J}, title = {Vector competence of Culex quinquefasciatus from Santiago Island, Cape Verde, to West Nile Virus: exploring the potential effect of the vector native Wolbachia.}, journal = {Parasites & vectors}, volume = {17}, number = {1}, pages = {536}, pmid = {39716303}, issn = {1756-3305}, support = {UID/04413/2020//Fundação para a Ciência e Tecnologia (FCT)/ ; LA/P/0117/2020//LA-REAL/ ; }, mesh = {Animals ; *Culex/virology/microbiology ; *Wolbachia/physiology ; *Mosquito Vectors/microbiology/virology ; *West Nile virus/physiology ; Female ; *West Nile Fever/transmission/virology ; Cabo Verde ; }, abstract = {BACKGROUND: Culex quinquefasciatus plays a crucial role as a vector of West Nile virus (WNV). This mosquito species is widely distributed in Cape Verde, being found in all inhabited islands of the archipelago. However, no data are currently available on the susceptibility of the local mosquito population to WNV. This study aimed to assess the vector competence of Cx. quinquefasciatus mosquitoes from Santiago Island, Cape Verde, for WNV and to explore the potential impact of its native Wolbachia on virus transmission.

METHODS: Wolbachia-infected and uninfected Cx. quinquefasciatus female mosquitoes were exposed to WNV lineage 1 PT6.39 strain using a Hemotek membrane feeding system. Mosquito samples, including the body, legs, wings and saliva, were collected at days 7, 14 and 21 post-infection (dpi) to assess WNV infection through one-step quantitative real-time PCR (RT-qPCR).

RESULTS: Culex quinquefasciatus from Cape Verde exhibited high susceptibility to the tested strain of WNV. Also, treated females without their native Wolbachia exhibited significantly higher WNV load in their bodies and greater dissemination rate at 7 dpi than their wild-type counterparts carrying Wolbachia.

CONCLUSIONS: The high susceptibility to WNV of Cx. quinquefasciatus from Cape Verde poses a potential risk for virus transmission in the archipelago. However, Wolbachia infection in this mosquito species seems to confer protection against WNV dissemination in the early stages of viral infection. Additional research is required to uncover the mechanisms driving this protection and its potential impact on WNV transmission.}, } @article {pmid39714171, year = {2024}, author = {Wang, G and Hussain, M and Qi, Z and Asgari, S}, title = {Role of Vigilin and RACK1 in dengue virus-Aedes aegypti-Wolbachia interactions.}, journal = {mSphere}, volume = {}, number = {}, pages = {e0048224}, doi = {10.1128/msphere.00482-24}, pmid = {39714171}, issn = {2379-5042}, abstract = {Vigilin is a large and evolutionary conserved RNA-binding protein (RBP), which can interact with RNA through its KH domain. Vigilin is, therefore, a multifunctional protein reported to be associated with RNA transport and metabolism, sterol metabolism, chromosome segregation, carcinogenesis, and heterochromatin-mediated gene silencing. The receptor for activated C kinase 1 (RACK1) is another highly conserved protein involved in many cellular pathways. Functional studies in human cells indicated that RACK1 interacts with Vigilin to promote dengue virus (DENV) replication. Both proteins are associated with the endoplasmic reticulum. Here, we investigated the significance of Vigilin and RACK1 homologs in Aedes aegypti mosquitoes concerning DENV replication and Wolbachia infection. We identified the homologs of the two genes in Ae. aegypti (AeVigilin and AeRACK1), which were upregulated in DENV-infected Aag2 cells and mosquitoes, and silencing them in Aag2 cells resulted in reduced DENV replication. Co-immunoprecipitation showed that AeRACK1 and AeVigilin interact in mosquito cells. Interestingly, we also found upregulation of both genes in a Wolbachia-infected cell line (Aag2.wAlbB). Furthermore, silencing AeVigilin and AeRACK1 in Aag2.wAlbB cells reduced DENV replication but increased Wolbachia density. However, we did not find a significant effect on DENV replication after silencing both genes in Ae. aegypti mosquitoes. Overall, our results support the involvement and significance of AeVigilin and AeRACK1 in DENV replication in Ae. aegypti.IMPORTANCEDengue virus (DENV), transmitted mainly by Aedes aegypti mosquitoes, poses significant health risks. Identifying factors involved in the virus replication in mosquitoes and human hosts is essential for devising control measures. In this study, we show that Vigilin and the receptor for activated C kinase 1 (RACK1), two proteins shown to play a role in the replication of DENV in human cells, are induced in mosquitoes and cell lines following DENV replication. Both proteins reside in the cytoplasm, where they interact similarly to human cells. Silencing the genes in mosquito cells significantly reduced virus replication. Furthermore, we found that both genes are induced in mosquito cells transinfected with Wolbachia, a bacterium that blocks DENV replication. The results help better understand the role of the common factors supporting DENV replication in mosquitoes and human cells.}, } @article {pmid39713442, year = {2024}, author = {Njogu, AK and Logozzo, F and Conner, WR and Shropshire, JD}, title = {Counting rare Wolbachia endosymbionts using digital droplet PCR.}, journal = {bioRxiv : the preprint server for biology}, volume = {}, number = {}, pages = {}, doi = {10.1101/2024.12.10.627731}, pmid = {39713442}, issn = {2692-8205}, abstract = {Wolbachia is the most widespread animal-associated intracellular microbe, living within the cells of over half of insect species. Since they can suppress pathogen replication and spread rapidly through insect populations, Wolbachia is at the vanguard of public health initiatives to control mosquito-borne diseases. Wolbachia's abilities to block pathogens and spread quickly are closely linked to their abundance in host tissues. The most common method for counting Wolbachia is quantitative polymerase chain reaction (qPCR), yet qPCR can be insufficient to count rare Wolbachia, necessitating tissue pooling and consequently compromising individual-level resolution of Wolbachia dynamics. Digital droplet PCR (ddPCR) offers superior sensitivity, enabling the detection of rare targets and eliminating the need for sample pooling. Here, we report three ddPCR assays to measure total Wolbachia abundance, Wolbachia abundance adjusted for DNA extraction efficiency, and Wolbachia density relative to host genome copies. Using Drosophila melanogaster with wMel Wolbachia as a model, we show these ddPCR assays can reliably detect as few as 7 to 12 Wolbachia gene copies in a 20 μL reaction. The designed oligos are homologous to sequences from at least 106 Wolbachia strains across Supergroup A and 53 host species from the Drosophila, Scaptomyza, and Zaprionus genera, suggesting broad utility. These highly sensitive ddPCR assays are expected to significantly advance Wolbachia-host interactions research by enabling the collection of molecular data from individual insect tissues. Their ability to detect rare Wolbachia will be especially valuable in applied and natural field settings where pooling samples could obscure important variation.}, } @article {pmid39709001, year = {2024}, author = {Mao, B and Wang, YY and Li, SY and Fu, Y and Xiao, YL and Wang, YF}, title = {A potential role for the interaction of Wolbachia surface proteins with the Drosophila microtubulin in maintenance of endosymbiosis and affecting spermiogenesis.}, journal = {Journal of insect physiology}, volume = {}, number = {}, pages = {104743}, doi = {10.1016/j.jinsphys.2024.104743}, pmid = {39709001}, issn = {1879-1611}, abstract = {Wolbachia, as a widely infected intracellular symbiotic bacterium in Arthropoda, is able to manipulate the reproduction of insect hosts for facilitating their own transmission. Cytoplasmic incompatibility (CI) is the most common phenotype that Wolbachia induced in insect hosts where they resulted in the failure of uninfected egg hatch when fertilized with the sperm derived from Wolbachia-infected males, suggesting that the sperm are modified by Wolbachia during spermatogenesis. Although the molecular mechanisms of CI are beginning to be understood, the effects of Wolbachia on the symbiotic relationship and the proper dynamics of spermatogenesis have not yet been fully investigated. We report here that Wolbachia infection induced a significant upregulation of betaTub85D in the testis of Drosophila melanogaster. Knockdown of betaTub85D in fly testes resulted in significant decrease of expression of Wolbachia surface protein gene (wsp), indicating a notable reduction of Wolbachia density. Pull-down analyses revealed that WSP interacted with the betaTub85D of D. melanogaster. Wolbachia infection altered the interactome between betaTub85D and other proteins in the testes, and may thus change the protein synthesis and metabolic pathways. Wolbachia infection induced not only an interaction of betaTub85D with Mst77F but also increase in phosphorylated Mst77F. These results suggest that Wolbachia WSP protein might play important roles in anchoring the endosymbiont to the host's cytoskeleton and consequently interfere the interactions among key proteins involved in spermatogenesis in the insect host testes, resulting in modified sperm.}, } @article {pmid39703404, year = {2024}, author = {Huang, K and Zhang, X and Xiong, N and Sun, L and Zhao, X and Zhou, K and Wu, J}, title = {First metagenomic sequencing for the analysis of microbial community populations of adults and pupae of Melophagus ovinus in Xinjiang, China.}, journal = {Frontiers in veterinary science}, volume = {11}, number = {}, pages = {1462772}, pmid = {39703404}, issn = {2297-1769}, abstract = {INTRODUCTION: Melophagus ovinus, a parasite on the body surface of sheep, directly attacks the host through biting and sucking blood and may also transmit pathogens in the process. There are currently only a few studies on the microbial composition of M. ovinus, while there are no such studies on pupae.

METHODS: In this study, samples AT-1 to AT-4 each contained four M. ovinus individuals, while sample AT-5 comprised four M. ovinus pupae, all used for metagenomic sequencing and analysis. Melophagus ovinus adults and pupae were collected from four regions in Xinjiang, China. DNA was extracted from the samples, amplified, and sequenced using the Illumina Novaseq 6000 System; finally, the sequencing data were analyzed using molecular biology software.

RESULTS AND DISCUSSION: From all samples, a total of 32 phyla, comprising 372 genera and 1,037 species, were detected. The highest microbial diversity was observed in Kuqa City (AT-2) and Qira County (AT-4). Pupae exhibited 40 unique microbial genera (AT-5) but did not have the highest microbial diversity. Proteobacteria was the dominant phylum in all samples. The dominant genera included Bartonella, Wolbachia, Pseudomonas, and Arsenophonus. This is the first study to report most of the bacteria (e.g., Pseudomonas versuta and Arsenophonus nasoniae), fungi (e.g., Saitoella complicata), viruses (e.g., Orf virus and Wolbachia phage WO), and protozoa (e.g., Trypanosoma theileri and Babesia bigemina) in M. ovinus. This study has enriched the microbial diversity data of M. ovinus, and the pathogens it carries may pose a threat to public health safety and the economy of related industries, necessitating further research to develop effective biological control strategies.}, } @article {pmid39696847, year = {2024}, author = {Huang, M and Wang, Z and Nie, Z}, title = {A stage structured model for mosquito suppression with immigration.}, journal = {Mathematical biosciences and engineering : MBE}, volume = {21}, number = {11}, pages = {7454-7479}, doi = {10.3934/mbe.2024328}, pmid = {39696847}, issn = {1551-0018}, mesh = {Animals ; Female ; Male ; *Wolbachia/physiology ; *Mosquito Control/methods ; *Mosquito Vectors/microbiology ; Larva/microbiology ; Malaria/prevention & control/transmission ; Culicidae ; Dengue/prevention & control/transmission ; Humans ; Aedes/microbiology ; Computer Simulation ; Models, Biological ; Seasons ; Animal Migration ; Population Dynamics ; Zika Virus Infection/prevention & control/transmission ; }, abstract = {The incompatible insect technique based on Wolbachia is a promising alternative to control mosquito-borne diseases, such as dengue fever, malaria, and Zika, which drives wild female mosquitoes sterility through a mechanism cytoplasmic incompatibility. A successful control program should be able to withstand the perturbation induced by the immigration of fertilized females from surrounding uncontrolled areas. In this paper, we formulated a system of delay differential equations, including larval and adult stages, interfered by Wolbachia-infected males. We classified the release number of infected males and immigration number of fertile females, to ensure that the system displays globally asymptotically stable or bistable dynamics. The immigration of fertile females hinders the maximum possible suppression efficiency so that the wild adults cannot be reduced to a level below $ A^*_\infty $. We identified the permitted most migration number to reduce the wild adults to a target level. To reduce up to $ 90\% $ of wild adults in the peak season within two months, an economically viable strategy is to reduce the immigration number of wild females less than $ 0.21\% $ of the carrying capacity of adults in the control area.}, } @article {pmid39695811, year = {2024}, author = {Jeon, J and Kim, HC and Donnelly, MJ and Choi, KS}, title = {Genetic diversity and Wolbachia infection in the Japanese encephalitis virus vector Culex tritaeniorhynchus in the Republic of Korea.}, journal = {Parasites & vectors}, volume = {17}, number = {1}, pages = {518}, pmid = {39695811}, issn = {1756-3305}, mesh = {Animals ; *Culex/virology/microbiology ; *Wolbachia/genetics/isolation & purification/classification ; *Genetic Variation ; *Mosquito Vectors/microbiology/virology ; Republic of Korea/epidemiology ; *Encephalitis Virus, Japanese/genetics/classification/isolation & purification ; Phylogeny ; Encephalitis, Japanese/epidemiology/transmission/virology ; Electron Transport Complex IV/genetics ; DNA Barcoding, Taxonomic ; }, abstract = {BACKGROUND: Culex tritaeniorhynchus, a major vector of Japanese encephalitis virus (JEV), is found across a broad geographical range, including Africa, Asia, Australia and Europe. Understanding the population structure and genetic diversity of pathogen vectors is increasingly seen as important for effective disease control. In China and Japan, two countries in close proximity to the Republic of Korea (ROK), Cx. tritaeniorhynchus has been categorized into two clades based on the DNA barcoding region of mitochondrial cytochrome c oxidase subunit I (COI), suggesting the presence of cryptic species. No comprehensive analysis of the genetic diversity in Cx. tritaeniorhynchus has been conducted in the ROK. To address this gap, we investigated the population structure of Cx. tritaeniorhynchus in the ROK.

METHODS: In Daegu, mosquito collections were conducted over a 2-year period from 2022 to 2023. For all other regions, Cx. tritaeniorhynchus specimens collected in 2023 were used. The COI barcoding region was analyzed to determine the genetic structure of the populations, supplemented with data from the 28S ribosomal DNA region. Each population was also examined for the eventual presence of Wolbachia infection. Finally, a back trajectory analysis was conducted to assess the possibility of international introduction of Cx. tritaeniorhynchus into the ROK.

RESULTS: The analysis of the COI region revealed the presence of two distinct clades within Cx. tritaeniorhynchus; these clades were the same as Cx. tritaeniorhynchus continental type (Ct-C) and C. tritaeniorhynchus Japanese type (Ct-J) previously reported. In contrast, the nuclear 28S region showed no significant genetic differentiation between these clades. Wolbachia infection was confirmed in some populations, but there was no evidence of an association with Wolbachia in Ct-C and Ct-J. It was also confirmed that the ROK is currently dominated by the Ct-J clade, with a possible introduction of Ct-C via air currents.

CONCLUSIONS: Determining the presence of cryptic species is important for preventing vector-borne diseases. The results of this study confirm the existence of two clades of Cx. tritaeniorhynchus in the ROK, with Ct-J being the dominant clade. Our findings enhance current understanding of the genetic diversity within Cx. tritaeniorhynchus and provide valuable insights for the prevention of JEV outbreaks and the effective management of Cx. tritaeniorhynchus populations in East Asia.}, } @article {pmid39681734, year = {2024}, author = {Rajendran, D and Vinayagam, S and Sekar, K and Bhowmick, IP and Sattu, K}, title = {Symbiotic Bacteria: Wolbachia, Midgut Microbiota in Mosquitoes and Their Importance for Vector Prevention Strategies.}, journal = {Microbial ecology}, volume = {87}, number = {1}, pages = {154}, pmid = {39681734}, issn = {1432-184X}, mesh = {*Wolbachia/physiology ; Animals ; *Symbiosis ; *Gastrointestinal Microbiome ; *Mosquito Vectors/microbiology ; Culicidae/microbiology ; Mosquito Control/methods ; }, abstract = {Mosquito-borne illnesses pose a significant threat to eradication under existing vector management measures. Chemo-based vector control strategies (use of insecticides) raise a complication of resistance and environmental pollution. Biological control methods are an alternative approach to overcoming this complication arising from insecticides. The mosquito gut microbiome is essential to supporting the factors that involve metabolic regulation and metamorphic development (from juvenile to adult), as well as the induction of an immune response. The induced immune response includes the JAK-STAT, IMD, and Toll pathways due to the microbial interaction with the midgut cells (MG cells) that prevent disease transmission to humans. The aforementioned sequel to the review provides information about endosymbiont Wolbachia, which contaminates insect cells, including germline and somatic cytoplasm, and inhibits disease-causing pathogen development and transmission by competing for resources within the cell. Moreover, it reduces the host population via cytoplasmic incompatibility (CI), feminization, male killing, and parthenogenesis. Furthermore, the Cif factor in Wolbachia is responsible for CI induction that produces inviable cells with the translocating systems and the embryonic defect-causing protein factor, WalE1 (WD0830), which manipulates the host actin. This potential of Wolbachia can be used to design a paratransgenic system to control vectors in the field. An extracellular symbiotic bacterium such as Asaia, which is grown in the growth medium, is used to transfer lethal genes within itself. Besides, the genetically transferred symbiotic bacteria infect the wild mosquito population and are easily manifold. So, it might be suitable for vector control strategies in the future.}, } @article {pmid39656697, year = {2024}, author = {Rasool, B and Younis, T and Zafar, S and Parvaiz, A and Javed, Z and Rasool, I and Shakeel, M}, title = {Incidence of endosymbiont bacteria Wolbachia in cowpea weevil Callosobruchus maculatus Fabricius (Coleoptera, Chrysomelidae).}, journal = {PloS one}, volume = {19}, number = {12}, pages = {e0313449}, doi = {10.1371/journal.pone.0313449}, pmid = {39656697}, issn = {1932-6203}, mesh = {*Wolbachia/genetics/isolation & purification/classification ; Animals ; *Phylogeny ; *Weevils/microbiology ; *Symbiosis ; RNA, Ribosomal/genetics ; Coleoptera/microbiology ; Electron Transport Complex IV/genetics ; }, abstract = {This study focuses on the cowpea weevil, Callosobruchus maculatus, a globally distributed grain pest that affects cereals and pulses. Using chemicals to store grains can harm pest control and pose risks to consumers and the environment. The facultative intracellular symbiont bacteria Wolbachia can affect host's reproductive capacities in a variety of ways, which makes it useful in the management of pests such as C. maculatus. The main goal of the study was to identify Wolbachia diversity in the C. maculatus population. Phylogenetic analysis utilized mitochondrial COI and 12S rRNA genes to identify the host C. maculatus, while screening for Wolbachia was conducted using genes (wsp, coxA, and ftsZ) genes. Molecular phylogenetic analysis of the Wolbachia genes resulted in one new Wolbachia strain (wCmac1) in C. maculatus populations and contrasting already published data of other Callosobruchus strains. The study discussed the detection of Wolbachia and its phylogenetic comparison with other C. maculatus and Coleopteran populations. It is important to take these findings into account when considering host-pathogen interactions.}, } @article {pmid39630881, year = {2024}, author = {Wang, Y and Yu, J}, title = {Dynamics of a non-autonomous delay mosquito population suppression model with Wolbachia-infected male mosquitoes.}, journal = {Journal of biological dynamics}, volume = {18}, number = {1}, pages = {2437034}, doi = {10.1080/17513758.2024.2437034}, pmid = {39630881}, issn = {1751-3766}, mesh = {Animals ; *Wolbachia/physiology ; Male ; *Models, Biological ; *Culicidae/microbiology/physiology ; Population Dynamics ; }, abstract = {In this paper, we develop a non-autonomous delay differential equation model for mosquito population suppression. After establishing the positiveness and boundedness of the solutions, we study the dynamical behaviours of the model with or without Wolbachia-infected male mosquitoes. More specifically, for the model without infected male mosquitoes, we analyse the asymptotic stability of the equilibria and demonstrate that the model undergo Hopf bifurcations under certain conditions. For the model incorporating infected male mosquitoes, we derive sufficient conditions for the global asymptotic stability of the origin. Numerical examples are provided to illustrate and support our theoretical findings.}, } @article {pmid39629982, year = {2024}, author = {Ross, PA and Yeatman, E and Berran, MS and Gu, X and Hoffmann, AA and van Heerwaarden, B}, title = {Wolbachia strain wMelM disrupts egg retention by Aedes aegypti females prevented from ovipositing.}, journal = {Applied and environmental microbiology}, volume = {}, number = {}, pages = {e0149124}, doi = {10.1128/aem.01491-24}, pmid = {39629982}, issn = {1098-5336}, abstract = {Aedes aegypti mosquitoes are well adapted to dry climates and can retain their eggs for extended periods in the absence of suitable habitat. Wolbachia strains transferred from other insects to mosquitoes can be released to combat dengue transmission by blocking virus replication and spreading through populations, but host fitness costs imposed by Wolbachia, particularly under some environments, can impede spread. We, therefore, assessed the impact of two Wolbachia strains being released for dengue control (wAlbB and wMelM) on fecundity and egg viability following extended egg retention (up to 24 days) under laboratory conditions. Egg viability following retention decreased to a greater extent in females carrying wMelM compared to uninfected or wAlbB females. Fertility fully recovered in uninfected females following a second blood meal after laying retained eggs, while wMelM females experienced only partial recovery. Effects of wMelM on egg retention were similar regardless of whether females were crossed to uninfected or wMelM males, suggesting that fitness costs were triggered by Wolbachia presence in females. The fecundity and hatch proportions of eggs of wMelM females declined with age, regardless of whether females used stored sperm or were recently inseminated. Costs of some Wolbachia strains during egg retention may affect the invasion and persistence of Wolbachia in release sites where larval habitats are scarce and/or intermittent.IMPORTANCEWolbachia mosquito releases are expanding around the world with substantial impacts on dengue transmission. Releases have succeeded in many locations, but the establishment of Wolbachia has been challenging in some environments, and the factors contributing to this outcome remain unresolved. Here, we explore the effects of Wolbachia on a novel trait, egg retention, which is likely to be important for the persistence of mosquito populations in locations with intermittent rainfall. We find substantial impacts of the Wolbachia strain wMelM on the quality of retained eggs but not the wAlbB strain. This cost is driven by the Wolbachia infection status of the female and can partially recover following a second blood meal. The results of our study may help to explain the difficulty in establishing Wolbachia strains at some field release sites and emphasize the need to characterize Wolbachia phenotypes across a variety of traits and strains.}, } @article {pmid39627857, year = {2024}, author = {Rahul, A and Reegan, AD and Shriram, AN and Fouque, F and Rahi, M}, title = {Innovative sterile male release strategies for Aedes mosquito control: progress and challenges in integrating evidence of mosquito population suppression with epidemiological impact.}, journal = {Infectious diseases of poverty}, volume = {13}, number = {1}, pages = {91}, pmid = {39627857}, issn = {2049-9957}, abstract = {BACKGROUND: Aedes mosquitoes pose a significant global threat as vectors for several debilitating arboviruses, including dengue, Zika, yellow fever, and chikungunya. Their unique breeding habits, behavior, and daytime activity complicate control efforts, prompting the search for innovative solutions. The sterile insect technique (SIT) and incompatible insect technique (IIT) are promising new techniques under investigation. This review synthesizes findings from field trials on SIT and/or IIT for Aedes mosquito control.

METHODS: A scoping review was conducted through comprehensive searches on Scopus, Web of Science, MEDLINE, PubMed, and preprint repositories up to April 25, 2024. Studies were initially screened for relevance based on their titles and abstracts, followed by a full-text review conducted by two independent extractors. Only field trials with control groups were included, with the final assessment focusing on trials reporting epidemiological outcomes. Data were abstracted into templates, emphasizing study design, intervention details, and outcomes. The review adhered to the PRISMA-ScR guidelines.

RESULTS: The search identified 21 field trials in various countries against Aedes mosquitoes. These trials employed diverse methodologies and mosquito release strategies, achieving varying levels of mosquito population suppression. Notably, two SIT and two Wolbachia-based IIT trials reported epidemiological outcomes, including reductions in dengue incidence and associated risk ratios. However, the reliance on national surveillance data for assessing dengue incidence suggests caution due to the potential underreporting of subclinical cases.

CONCLUSIONS: The review underscores the promise of SIT and IIT for controlling Aedes mosquito populations, citing successful reductions in mosquito densities and dengue transmission. However, it calls for more rigorous study designs and standardized methodologies, as well as the adoption of comprehensive frameworks to accurately assess the effectiveness of these interventions. Future research should focus on bridging gaps in real-world effectiveness by addressing factors such as feasibility, acceptability, scalability, and cost, which are crucial for guiding their successful large-scale deployment in any country.}, } @article {pmid39615475, year = {2024}, author = {Řezáč, M and Řezáčová, V and Heneberg, P}, title = {Differences in the abundance and diversity of endosymbiotic bacteria drive host resistance of Philodromus cespitum, a dominant spider of central European orchards, to selected insecticides.}, journal = {Journal of environmental management}, volume = {373}, number = {}, pages = {123486}, doi = {10.1016/j.jenvman.2024.123486}, pmid = {39615475}, issn = {1095-8630}, abstract = {The ability of tissue endosymbionts to degrade and detoxify agrochemicals is increasingly recognized as a mechanism supporting the survival of arthropods in agroecosystems. Therefore, tissue endosymbionts have the potential to drive insecticide resistance in agrobiont spiders, i.e., in major generalist predators and pest control agents within agroecosystems. We hypothesized that the abundance and diversity of the endosymbiotic bacteria of Philodromus cespitum, a philodromid spider dominating central European apple orchards, vary with regard to differences in predation capacity and drive host insecticide resistance. We provisioned P. cespitum with diets of varying protein and lipid content and topically exposed them to field-relevant doses of commonly used insecticides, namely Mospilan (acetamiprid), Movento (spirotetramat), Gondola (sulfoxaflor), Decis (deltamethrin), Coragen (chlorantraniliprole), and Benevia (cyantraniliprole). The analyses were based on 16S rDNA profiles from lysates of the cephalothorax and legs of the tested spiders. The application of Benevia, Mospilan, and Movento was partially lethal. The spiders that were resistant to the treatments with Benevia, Mospilan, or Movento were associated with the increased relative abundance of Mycoplasmatota by more than one order of magnitude. Additionally, the abundance of other bacteria differed in Mospilan-resistant and Mospilan-sensitive individuals. In contrast, the diet regimens were not associated with any major differences in the microbiome diversity nor the diversity of endosymbionts. Philodromus cespitum hosts assemblages with unexpectedly high beta diversity of endosymbionts. The OTU identified as the alpha proteobacterium endosymbiont of Coelostomidia zealandica was an obligate endosymbiont of the analyzed P. cespitum population. Wolbachia, Rickettsia, and Spiroplasma endosymbionts were also highly prevalent and differed in their responses to the applied treatments. In conclusion, differences in the abundance and diversity of endosymbiotic bacteria drove the resistance of the spider host to selected insecticides.}, } @article {pmid39615459, year = {2024}, author = {Chow, JY and Bansal, S and Dickens, BSL and Ma, P and Hoffmann, A and Cheong, YL and Ahmad, NW and Lim, JT}, title = {Assessing the direct and spillover protective effectiveness of Wolbachia-mediated introgression to combat dengue.}, journal = {EBioMedicine}, volume = {110}, number = {}, pages = {105456}, doi = {10.1016/j.ebiom.2024.105456}, pmid = {39615459}, issn = {2352-3964}, abstract = {BACKGROUND: Dengue remains a global health challenge with limited treatment options, highlighting the need for effective vector control strategies. The introduction of Wolbachia pipientis into Aedes aegypti populations has shown success in reducing dengue transmission across global field trials. However, the spillover effectiveness of the technology on untreated areas is not well-known. This study estimates the spillover protective effectiveness (PE) of Wolbachia-mediated introgression on dengue.

METHODS: We used the synthetic control method (SCM) under assumption of partial interference to evaluate the direct and spillover PEs of Wolbachia-mediated introgression in a long-running operational trial of the intervention in Malaysia. Synthetic controls (SCs), which comprise of a weighted sum of non-spillover controls, were constructed for each directly-treated and spillover site in the pre-intervention period to account for historical imbalances in dengue risk and risk trajectories. SCs were compared to directly/spillover-treated sites to estimate the impact of Wolbachia-introgression on dengue incidence across each site, calendar year and intervention time. Robustness checks, including visual inspections, root-mean-square error (RMSE) calculations, in-space and in-time placebo checks, and permutation tests, were used to inspect the model's ability in attributing dengue incidence reductions to the Wolbachia interventions.

FINDINGS: The direct and spillover PEs of Wolbachia on dengue incidence were expressed as a percentage reduction of dengue incidence, or the absolute case reductions, by comparing SCs to actual intervention/spillover sites. Findings indicate a direct reduction in dengue incidence by 64.35% (95% CI: 63.50-66.71, p < 0.05 using permutation tests) in directly treated areas, corresponding to 1802 (95% CI: 1768-1932) cases averted. Meanwhile, spillover effects contributed to a 37.69% (95% CI: 36.45-38.49, p < 0.05) reduction in adjacent non-intervention areas, accounting for 115 (95% CI: 104-132) absolute cases averted. Tracking PEs by intervention time revealed a dose-response relationship, where PEs increased concomitantly with Wolbachia frequency. Model checks confirmed the robustness of these results, and ascertained that these PEs were not an artefact of poor control selection, pre-trends in dengue incidence or poor predictive ability of the fitted SCs.

INTERPRETATION: Wolbachia-introgression effectively diminished dengue incidence in directly-treated and surrounding spillover regions. This dual effectiveness highlights the potential of Wolbachia-infected mosquitoes as a sustainable, cost-effective strategy against dengue.

FUNDING: This research is hosted by CNRS@CREATE and supported by the National Research Foundation, Prime Minister's Office, Singapore, under its Campus for Research Excellence and Technological Enterprise (CREATE) program, and is funded by the Lee Kong Chian School of Medicine-Ministry of Education Start-Up Grant. The original Hoffmann et al. (2024) study was funded by the Wellcome Trust Awards 226166, 108508, 202888 and the Ministry of Health Malaysia NMRR-16-297-28898.}, } @article {pmid39614636, year = {2024}, author = {Jiménez-Florido, P and Aquilino, M and Buckley, D and Bella, JL and Planelló, R}, title = {Differential gene expression in Chorthippus parallelus (Zetterstedt, 1821) (Orthoptera: Acrididae: Gomphocerinae) induced by Wolbachia infection.}, journal = {Insect science}, volume = {}, number = {}, pages = {}, doi = {10.1111/1744-7917.13481}, pmid = {39614636}, issn = {1744-7917}, support = {PID2019-104952GBI00//Ministerio de Economía y Competitividad/ ; FPU contract 22/02220//Ministerioa de Ciencia, Innovación y Universidades/ ; PEJD-2019-POST/AMB-16425//Comunidada de Madrid/European Social Fund/ ; María Zambrano contract//Ministerio de Universidades/Europeana Union-NextGeneration Programme/ ; }, abstract = {Distinct lineages of the grasshopper Chorthippus parallelus (Orthoptera: Acrididae) form well-known hybrid zones (HZs) both in the Pyrenees and the Alps mountain ranges in South Europe. These HZs represent unique experimental systems to identify "key genes" that maintain genetic boundaries between emerging species. The Iberian endemism C. p. erythropus (Cpe) and the subspecies C. p. parallelus (Cpp), widely distributed throughout the rest of Europe, overlap and form the Pyrenean HZ. Both subspecies differ morphologically, as well as in behavioral, mitochondrial, nuclear, and chromosomal traits, and in the strains of the maternally transmitted bacterial endosymbiont Wolbachia infecting them. This results in either unidirectional and bidirectional cytoplasmic incompatibility between both grasshopper subspecies, pointing out that Wolbachia clearly affects gene expression in the infected individuals. Here we explore how Wolbachia may modify the expression of some major genes involved in relevant pathways in Cpp in the Pyrenean HZ. We have analyzed, through molecular biomarkers, the physiological responses in C. parallelus individuals infected by Wolbachia, with particular attention to the energy metabolism, the immune system response, and the reproduction. qPCR was used to evaluate the expression of selected genes in the gonads of infected and uninfected adults of both sexes, since this tissue constitutes the main target of Wolbachia infection. Transcriptional analyses also showed differential sex-dependent responses in most of the analyzed biomarkers in infected and noninfected individuals. We identified for the first time new sensitive biomarkers that might be involved in the reproductive barrier induced by Wolbachia in the hybrid zone.}, } @article {pmid39605481, year = {2024}, author = {Culotta, J and Lindsey, AR}, title = {A reference genome for Trichogramma kaykai : A tiny desert-dwelling parasitoid wasp with competing sex-ratio distorters.}, journal = {bioRxiv : the preprint server for biology}, volume = {}, number = {}, pages = {}, doi = {10.1101/2024.11.22.624848}, pmid = {39605481}, issn = {2692-8205}, abstract = {The tiny parasitoid wasp Trichogramma kaykai inhabits the Mojave Desert of the southwest United States. Populations of this tiny insect variably host up to two different sex-distorting genetic elements: (1) the endosymbiotic bacterium Wolbachia which induces the parthenogenetic reproduction of females, and (2) a B-chromosome, "Paternal Sex Ratio" (PSR), which converts would-be female offspring to PSR-transmitting males. We report here the genome of a Wolbachia -infected Trichogramma kaykai isofemale colony KSX58. Using Oxford Nanopore sequencing we produced a final genome assembly of 203 Mbp with 45x coverage, consisting of 213 contigs with an N50 of 1.9 Mbp. The assembly is quite complete, with 91.41% complete BUSCOs recovered: a very high score for Trichogrammatids that have been previously characterized for having high levels of core gene losses. We also report a complete mitochondrial genome for T. kaykai, and an assembly of the associated Wolbachia , strain w Tkk. We identified copies of the parthenogenesis-inducing genes pifA and pifB in a remnant prophage region of the w Tkk genome. The Trichogramma kaykai assembly is the highest quality genome assembly for the genus to-date and will serve as a great resource for understanding the evolution of sex and selfish genetic elements.}, } @article {pmid39597595, year = {2024}, author = {da Silva, LMI and da Silva, JI and da Silva, AF and Dezordi, FZ and Machado, LC and Qin, S and Fan, H and Tong, Y and Campos, TL and Paiva, MHS and Wallau, GL}, title = {Sequencing and Analysis of Wolbachia Strains from A and B Supergroups Detected in Sylvatic Mosquitoes from Brazil.}, journal = {Microorganisms}, volume = {12}, number = {11}, pages = {}, doi = {10.3390/microorganisms12112206}, pmid = {39597595}, issn = {2076-2607}, abstract = {Wolbachia are endosymbiotic bacteria that infect a wide range of arthropods and filarial nematodes, often manipulating host reproduction. The efficacy of Wolbachia-based interventions for dengue and chikungunya control has been validated through numerous field studies in recent years. This study aimed to investigate the diversity and prevalence of Wolbachia infections in sylvatic mosquitoes from two locations in Recife, Brazil. Multiple mosquito species were screened for Wolbachia using both target marker gene amplification coupled with Sanger sequencing and whole-genome sequencing (WGS) approaches. Phylogenetic analyses were conducted to classify Wolbachia strains into supergroups and assess their evolutionary relationships. Results revealed the presence of Wolbachia in eleven mosquito species examined, with different infection rates. Both supergroups A and B of Wolbachia strains were identified, with Aedes albopictus showing co-infection by both supergroups through the WGS approach. We also detected indirect evidence of Wolbachia horizontal transmission among mosquitoes and other distant host orders. This study provides valuable insights into the distribution and diversity of Wolbachia in sylvatic mosquitoes from Brazil and adds new important data about Wolbachia detection through target marker gene amplicon coupled with Sanger sequencing and WGS methods, highlighting its complementarity to ascertain the presence of Wolbachia in mosquito samples.}, } @article {pmid39588098, year = {2024}, author = {Fukatsu, T and Gottlieb, Y and Tsiamis, G and McGraw, E and Perlman, S and Bouchon, D and Johnson, K and Taylor, MJ}, title = {Editorial: Frontiers in Wolbachia biology 2023.}, journal = {Frontiers in microbiology}, volume = {15}, number = {}, pages = {1513314}, pmid = {39588098}, issn = {1664-302X}, } @article {pmid39581403, year = {2024}, author = {Gao, M and Li, B and Zhang, K and Li, D and Chen, R and Elumalai, P and Gao, X and Wang, L and Zhu, X and Luo, J and Ji, J and Cui, J}, title = {Worldwide used bio-insecticides Cry1Ac toxin have no detrimental effects on E. balteatus but alter the symbiotic microbial communities.}, journal = {International journal of biological macromolecules}, volume = {}, number = {}, pages = {137995}, doi = {10.1016/j.ijbiomac.2024.137995}, pmid = {39581403}, issn = {1879-0003}, abstract = {Hoverflies, capable of abilities providing dual ecosystem services including pest control and pollination, are exposed to insecticidal proteins from transgenic plants via pollen and prey aphids. However, the effects of such exposures on hoverflies have never been adequately assessed. Here, we investigated impacts of the most widely used biotoxin Cry1Ac on a representative hoverfly species Episyrphus balteatus through food chain transmission and active toxin exposure. The results showed Cry1Ac can be transmitted into E. balteatus through feeding on Aphis gossypii reared by Bt insect-resistant cotton variety expressing the Cry1Ac toxin, but the biological parameters of E. balteatus including survival rate, growth, development, reproductive capacity, and detoxification-related gene expression, were not significantly affected. Furthermore, the exposure to high-dose Cry1Ac toxin (500 μg/mL) resulted in slight increase of 16.67 % in the activity of detoxification and antioxidant enzyme catalase in E. balteatus and inhibited the egg hatching, partially inducing stress responses. Notably, the exposure to Cry1Ac toxin disrupted the microbiota homeostasis in E. balteatus, and the relative abundances of three dominant symbiotic bacterial genera (Cosenzaea, Wolbachia, and Commensalibacter) in E. balteatus exhibited a 10 % ~ 40 % fluctuation under Cry1Ac toxin stress. Taken together, these results suggest Cry1Ac toxin is not lethal to E. balteatus, but it poses a potential threat to its endosymbiotic bacteria.}, } @article {pmid39576110, year = {2024}, author = {Hussain, M and Qi, Z and Asgari, S}, title = {Interaction of the Wolbachia surface protein with a novel pro-viral protein from Aedes aegypti.}, journal = {mBio}, volume = {}, number = {}, pages = {e0148624}, doi = {10.1128/mbio.01486-24}, pmid = {39576110}, issn = {2150-7511}, abstract = {UNLABELLED: Dengue virus (DENV) and other flaviviruses are prevented from replicating in mosquitoes by Wolbachia. To date, several reports have appeared that highlight multiple molecular and cellular pathways involved in the blocking mechanism, which underlines the complicated nature of the mechanism. Here, we developed a hypothesis on whether Wolbachia proteins interact with pro-viral host proteins by using a unique approach to study the antiviral mechanism based on Wolbachia-host protein-protein interaction. We selected Wolbachia surface protein (WSP) for co-immunoprecipitation because of its abundance and possible secretion. We first confirmed WSP's secretion in mosquito cells and found two host proteins, Ae. aegypti serine-threonine kinase (STK) and synaptic vesicle membrane (SVM) protein VAT-1, and one Wolbachia protein (wGroEL) interacting with WSP. We examined the role of STK and SVM genes in relation to DENV replication in Ae. aegypti mosquitoes and mosquito cell lines with and without Wolbachia. In DENV-infected Aag2 cells, the expression of SVM and STK was significantly increased. However, although these genes were induced in Wolbachia-infected Aag2 cells, they were downregulated after DENV infection. Silencing of STK, but not SVM, reduced DENV replication in Aag2 cells and mosquitoes. Conversely, RNA activation of STK, by utilizing promoter induction via short activating oligos, resulted in higher DENV replication in Wolbachia-infected and uninfected cell lines. Overall, our findings suggest that STK is a pro-viral gene, and Wolbachia WSP binds to STK, possibly making it less accessible for DENV replication.

IMPORTANCE: Wolbachia is an endosymbiotic bacterium that blocks the replication of arboviruses in transinfected Aedes aegypti mosquitoes. In this study, we focused on identifying the potential interaction of Wolbachia proteins with the host pro-viral proteins. For this, we embarked on identifying the interacting proteins with a major Wolbachia protein, WSP, which is both structural and also secreted into the host cells. An Ae. aegypti STK was identified, which is induced in DENV and Wolbachia-infected cells. Silencing or induction of the gene led to reduced and increased DENV replication in vitro. Consistently, knocking down the gene in mosquitoes resulted in decreased virus replication. We hypothesize that WSP may sequester STK, which is pro-viral, contributing to Wolbachia virus blocking.}, } @article {pmid39575185, year = {2024}, author = {Brischetto, C and Rossi, V and Fedele, G}, title = {The microbiome analysis of ripen grape berries supports the complex etiology of sour rot.}, journal = {Frontiers in microbiology}, volume = {15}, number = {}, pages = {1450443}, pmid = {39575185}, issn = {1664-302X}, abstract = {Sour rot (SR) is a grapevine disease complex that is not completely understood in its etiology and epidemiology. Recently, SR has received special attention due to its increasing economic importance due to crop losses and reduced wine quality. In this study, the fungal and bacterial microbiota of healthy (i.e., without rot symptoms) and rotten (i.e., exhibiting visual and olfactory SR symptoms) ripe bunches were characterized across 47 epidemics (39 vineyards in six Italian grape-growing areas) over three years. The 16S rRNA gene, ITS high-throughput amplicon sequencing, and quantitative PCR were used to assess the relative abundance and dynamic changes of microorganisms associated with SR. The estimators of genera richness of fungal communities within samples indicated a significantly different diversity between healthy and rotten bunches. For bacterial communities, the healthy and rotten bunches significantly differed in the total number of species, but not in abundance distribution across species. The bunch status (i.e., healthy and rotten) was a significant source of diversity (p < 0.01) when the community composition between samples was evaluated, indicating that microbiome composition varied between healthy and rotten bunches. In particular, healthy and rotten bunches shared 43.1 and 54.8% of fungal and bacterial genera, respectively; 31.3% (fungal) and 26.2% (bacterial) genera were associated with rotten bunches only. The yeast genera Zygosaccharomyces, Zygoascus, Saccharomycopsis, Issatchenkia, and Pichia and the bacterial genera Orbus, Gluconobacter, Komagataeibacter, Gluconacetobacter, and Wolbachia were strongly associated with bunches showing SR symptoms based on a linear discriminant analysis. These microorganisms have been associated with Drosophila insects in literature. The relationships between the microflora associated with SR-affected bunches and the roles of Drosophila in SR development need further investigation, which may open perspectives for more effective disease control.}, } @article {pmid39574078, year = {2024}, author = {Lim, PL and Cook, AR and Bansal, S and Chow, JY and Lim, JT}, title = {Wolbachia incompatible insect technique program optimization over large spatial scales using a process-based model of mosquito metapopulation dynamics.}, journal = {BMC biology}, volume = {22}, number = {1}, pages = {269}, pmid = {39574078}, issn = {1741-7007}, mesh = {*Wolbachia/physiology ; Animals ; *Mosquito Control/methods ; *Population Dynamics ; Singapore ; Mosquito Vectors/microbiology/physiology ; Male ; Models, Biological ; Aedes/microbiology/physiology ; Pest Control, Biological/methods ; Culicidae/physiology/microbiology ; }, abstract = {BACKGROUND: Wolbachia incompatible insect technique (IIT) programs have been shown in field trials to be highly effective in suppressing populations of mosquitoes that carry diseases such as dengue, chikungunya, and Zika. However, the frequent and repeated release of Wolbachia-infected male mosquitoes makes such programs resource-intensive. While the need for optimization is recognized, potential strategies to optimize releases and reduce resource utilization have not been fully explored.

RESULTS: We developed a process-based model to study the spatio-temporal metapopulation dynamics of mosquitoes in a Wolbachia IIT program, which explicitly incorporates climatic influence in mosquito life-history traits. We then used the model to simulate various scale-down and redistribution strategies to optimize the existing program in Singapore. Specifically, the model was used to study the trade-offs between the intervention efficacy outcomes and resource requirements of various release program strategies, such as the total number of release events and the number of mosquitoes released. We found that scaling down releases in existing sites from twice a week to only once a week yielded small changes in suppression efficacy (from 87 to 80%), while requiring 44% fewer mosquitoes and release events. Additionally, redistributing mosquitoes from already suppressed areas and releasing them in new areas once a week led to a greater total suppressive efficacy (83% compared to 61%) while also yielding a 16% and 14% reduction in the number of mosquitoes and release events required, respectively.

CONCLUSIONS: Both scale-down and redistribution strategies can be implemented to significantly reduce program resource requirements without compromising the suppressive efficacy of IIT. These findings will inform planners on ways to optimize existing and future IIT programs, potentially allowing for the wider adoption of this method for mosquito-borne disease control.}, } @article {pmid39562330, year = {2024}, author = {Higashi, CHV and Patel, V and Kamalaker, B and Inaganti, R and Bressan, A and Russell, JA and Oliver, KM}, title = {Another tool in the toolbox: Aphid-specific Wolbachia protect against fungal pathogens.}, journal = {Environmental microbiology}, volume = {26}, number = {11}, pages = {e70005}, doi = {10.1111/1462-2920.70005}, pmid = {39562330}, issn = {1462-2920}, support = {1754302//National Science Foundation/ ; 2109582//National Science Foundation/ ; 2240392//National Science Foundation/ ; }, mesh = {*Aphids/microbiology ; Animals ; *Wolbachia/physiology ; *Symbiosis ; Musa/microbiology ; }, abstract = {Aphids harbor nine common facultative symbionts, most mediating one or more ecological interactions. Wolbachia pipientis, well-studied in other arthropods, remains poorly characterized in aphids. In Pentalonia nigronervosa and P. caladii, global pests of banana, Wolbachia was initially hypothesized to function as a co-obligate nutritional symbiont alongside the traditional obligate Buchnera. However, genomic analyses failed to support this role. Our sampling across numerous populations revealed that more than 80% of Pentalonia aphids carried an M-supergroup strain of Wolbachia (wPni). The lack of fixation further supports a facultative status for Wolbachia, while high infection frequencies in these entirely asexual aphids strongly suggest Wolbachia confers net fitness benefits. Finding no correlation between Wolbachia presence and food plant use, we challenged Wolbachia-infected aphids with common natural enemies. Bioassays revealed that Wolbachia conferred significant protection against a specialized fungal pathogen (Pandora neoaphidis) but not against generalist pathogens or parasitoids. Wolbachia also improved aphid fitness in the absence of enemy challenge. Thus, we identified the first clear benefits for aphid-associated Wolbachia and M-supergroup strains specifically. Aphid-Wolbachia systems provide unique opportunities to merge key models of symbiosis to better understand infection dynamics and mechanisms underpinning symbiont-mediated phenotypes.}, } @article {pmid39556610, year = {2024}, author = {Garrigós, M and Garrido, M and Ruiz-López, MJ and García-López, MJ and Veiga, J and Magallanes, S and Soriguer, R and Moreno-Indias, I and Figuerola, J and Martínez-de la Puente, J}, title = {Microbiota composition of Culex perexiguus mosquitoes during the West Nile virus outbreak in southern Spain.}, journal = {PloS one}, volume = {19}, number = {11}, pages = {e0314001}, doi = {10.1371/journal.pone.0314001}, pmid = {39556610}, issn = {1932-6203}, mesh = {Animals ; Spain/epidemiology ; *West Nile Fever/epidemiology/virology ; *Culex/microbiology/virology ; *Disease Outbreaks ; *West Nile virus/isolation & purification/genetics ; *Microbiota ; *Mosquito Vectors/microbiology/virology ; Humans ; RNA, Ribosomal, 16S/genetics ; Female ; }, abstract = {West Nile virus (WNV) is a flavivirus naturally circulating between mosquito vectors and birds, occasionally infecting horses and humans and causing epidemiologically relevant outbreaks. In Spain, the first big WNV outbreak was recorded in 2020, resulting in 77 people infected and 8 fatalities, most of them in southern Spain. Culex perexiguus was identified as the primary vector of WNV maintaining its enzootic circulation of the virus. Growing evidence highlights the role of mosquito microbiota as a key component determining the vectorial capacity of mosquitoes, largely contributing to disease epidemiology. Here, we develop, to our knowledge, the first identification of the microbiota composition of this mosquito vector under natural conditions and test for the potential relationship between mosquito microbiota composition and WNV infection. To do so, we collected mosquitoes in a natural area of southern Spain during the 2020 WNV outbreak and identified the microbiota composition of mosquitoes using a 16S rRNA gene metabarcoding approach. The microbiota of Cx. perexiguus was dominated by the phylum Proteobacteria. The most abundant families were Burkholderiaceae and Erwiniaceae, including the genera Burkholderia, Erwinia, and Pantoea. The genus Wolbachia, which use to dominate the microbiota of Cx. pipiens and negatively interact with WNV according to the literature, had a low prevalence and relative abundance in Cx. perexiguus and its abundance did not differ between WNV-positive and WNV-negative mosquito pools. The microbiota diversity and composition of Cx. perexiguus were not significantly related to the WNV infection status. These results provide the first identification of the mosquito microbiota in an endemic area of WNV circulation in Spain.}, } @article {pmid39548000, year = {2024}, author = {Visser, B and Scheifler, M}, title = {Insect Lipid Metabolism in the Presence of Symbiotic and Pathogenic Viruses and Bacteria.}, journal = {Advances in experimental medicine and biology}, volume = {}, number = {}, pages = {}, pmid = {39548000}, issn = {0065-2598}, abstract = {Insects, like most animals, have intimate interactions with microorganisms that can influence the insect host's lipid metabolism. In this chapter, we describe what is known so far about the role prokaryotic microorganisms play in insect lipid metabolism. We start exploring microbe-insect lipid interactions focusing on endosymbionts, and more specifically the gut microbiota that has been predominantly studied in Drosophila melanogaster. We then move on to an overview of the work done on the common and well-studied endosymbiont Wolbachia pipientis, also in interaction with other microbes. Taking a slightly different angle, we then look at the effect of human pathogens, including dengue and other viruses, on the lipids of mosquito vectors. We extend the work on human pathogens and include interactions with the endosymbiont Wolbachia that was identified as a natural tool to reduce the spread of mosquito-borne diseases. Research on lipid metabolism of plant disease vectors is up and coming and we end this chapter by highlighting current knowledge in that field.}, } @article {pmid39547593, year = {2024}, author = {Hu, H and Lu, Z and Ma, Y and Song, X and Wang, D and Wu, C and Ma, X and Shan, Y and Ren, X and Ma, Y}, title = {Impact of transinfection of Wolbachia from the planthopper Laodelphax striatellus on reproductive fitness and transcriptome of the whitefly Bemisia tabaci.}, journal = {Journal of invertebrate pathology}, volume = {}, number = {}, pages = {108230}, doi = {10.1016/j.jip.2024.108230}, pmid = {39547593}, issn = {1096-0805}, abstract = {The whitefly Bemisia tabaci is critical global pest threatening crops and leading to agricultural losses. Wolbachia is an intracellular symbiotic bacterium in insects, which can regulate the growth and development of the host through various ways. In a prior study, Wolbachia was found to be transferred to whitefly and induce fitness changes. However, little is known about the underlying mechanisms of host-Wolbachia interactions in B. tabaci. In this study, a Wolbachia strain wStri was isolated from the small brown planthopper, Laodelphex striatellus, and transferred to B. tabaci. The distribution of Wolbachia in whiteflies was determined using fluorescence in situ hybridization. Reciprocal crossing experiments demonstrated that wStri did not induce cytoplasmic incompatibility phenotypes in B. tabaci, but prolonged the developmental duration of the offspring. We performed transcriptomic analysis of Wolbachia-infected female and male adults using Illumina-based RNA-Seq. A total of 843 differentially expressed genes (DEGs) were identified in infected females, among them 141 were significantly up-regulated and 702 were down-regulated by Wolbachia infection. In infected males, of 511 gene sets, 279 host genes were significantly up-regulated, and 232 were down-regulated by Wolbachia infection. KEGG analysis of DEGs demonstrated significant differences in gene pathway distribution between up-regulated and down-regulated genes. These genes are involved in various biological processes, including, but not limited to, detoxification, oxidation-reduction, metabolic processes, and immunity. The transcriptomic profiling of this study offers valuable information on the differential expression of genes in whiteflies following Wolbachia infection, and enhances our understanding of this host-symbiotic interaction.}, } @article {pmid39543157, year = {2024}, author = {Edenborough, K and Supriyati, E and Dufault, S and Arguni, E and Indriani, C and Denton, J and Sasmono, RT and Ahmad, RA and Anders, KL and Simmons, CP}, title = {Dengue virus genomic surveillance in the applying Wolbachia to eliminate dengue trial reveals genotypic efficacy and disruption of focal transmission.}, journal = {Scientific reports}, volume = {14}, number = {1}, pages = {28004}, pmid = {39543157}, issn = {2045-2322}, support = {1173928//National Health and Medical Research Council/ ; 1173928//National Health and Medical Research Council/ ; }, abstract = {Release of Aedes aegypti mosquitoes infected with Wolbachia pipientis (wMel strain) is a biocontrol approach against Ae. aegypti-transmitted arboviruses. The Applying Wolbachia to Eliminate Dengue (AWED) cluster-randomised trial was conducted in Yogyakarta, Indonesia in 2018-2020 and provided pivotal evidence for the efficacy of wMel-Ae. aegypti mosquito population replacement in significantly reducing the incidence of virologically-confirmed dengue (VCD) across all four dengue virus (DENV) serotypes. Here, we sequenced the DENV genomes from 318 dengue cases detected in the AWED trial, with the aim of characterising DENV genetic diversity, measuring genotype-specific intervention effects, and inferring DENV transmission dynamics in wMel-treated and untreated areas of Yogyakarta. Phylogenomic analysis of all DENV sequences revealed the co-circulation of five endemic DENV genotypes: DENV-1 genotype I (12.5%) and IV (4.7%), DENV-2 Cosmopolitan (47%), DENV-3 genotype I (8.5%), and DENV-4 genotype II (25.7%), and one recently imported genotype, DENV-4 genotype I (1.6%). The diversity of genotypes detected among AWED trial participants enabled estimation of the genotype-specific protective efficacies of wMel, which were similar (± 10%) to the point estimates of the respective serotype-specific efficacies reported previously. This indicates that wMel afforded protection to all of the six genotypes detected in Yogyakarta. We show that within this substantial overall viral diversity, there was a strong spatial and temporal structure to the DENV genomic relationships, consistent with highly focal DENV transmission around the home in wMel-untreated areas and a near-total disruption of transmission by wMel. These findings can inform long-term monitoring of DENV transmission dynamics in Wolbachia-treated areas including Yogyakarta.}, } @article {pmid39533947, year = {2024}, author = {Serga, S and Kovalenko, PA and Maistrenko, OM and Deconninck, G and Shevchenko, O and Iakovenko, N and Protsenko, Y and Susulovsky, A and Kaczmarek, Ł and Pavlovska, M and Convey, P and Kozeretska, I}, title = {Wolbachia in Antarctic terrestrial invertebrates: Absent or undiscovered?.}, journal = {Environmental microbiology reports}, volume = {16}, number = {6}, pages = {e70040}, doi = {10.1111/1758-2229.70040}, pmid = {39533947}, issn = {1758-2229}, support = {//Ukrainian State Special-Purpose Research Program in Antarctica for 2011-2025/ ; //Natural Environment Research Council (NERC)/ ; 22-28778S//Czech Science Foundation/ ; RVO 67985904//Czech Science Foundation/ ; CZ.02.1.01/0.0/0.0/16_019/0000803//OP RDE/ ; EVA4.0//OP RDE/ ; ANR-20-CE02-0011-01//ANR (Agence Nationale de la Recherche Française)/ ; //Scholarship from the National Academy of Sciences of Ukraine for Young Scientists/ ; //PAUSE-ANR Ukraine Program/ ; }, mesh = {*Wolbachia/genetics/classification/isolation & purification/physiology ; Animals ; Antarctic Regions ; *Invertebrates/microbiology ; *Symbiosis ; Phylogeny ; }, abstract = {Interactions between a host organism and its associated microbiota, including symbiotic bacteria, play a crucial role in host adaptation to changing environmental conditions. Antarctica provides a unique environment for the establishment and maintenance of symbiotic relationships. One of the most extensively studied symbiotic bacteria in invertebrates is Wolbachia pipientis, which is associated with a wide variety of invertebrates. Wolbachia is known for manipulating host reproduction and having obligate or facultative mutualistic relationships with various hosts. However, there is a lack of clear understanding of the prevalence of Wolbachia in terrestrial invertebrates in Antarctica. We present the outcomes of a literature search for information on the occurrence of Wolbachia in each of the major taxonomic groups of terrestrial invertebrates (Acari, Collembola, Diptera, Rotifera, Nematoda, Tardigrada). We also performed profiling of prokaryotes based on three marker genes and Kraken2 in available whole genome sequence data obtained from Antarctic invertebrate samples. We found no reports or molecular evidence of Wolbachia in these invertebrate groups in Antarctica. We discuss possible reasons underlying this apparent absence and suggest opportunities for more targeted future research to confirm bacteria's presence or absence.}, } @article {pmid39532731, year = {2024}, author = {Almeida, L and Bellver-Arnau, J and Privat, Y and Rebelo, C}, title = {Vector-borne disease outbreak control via instant releases.}, journal = {Journal of mathematical biology}, volume = {89}, number = {6}, pages = {63}, pmid = {39532731}, issn = {1432-1416}, support = {754362//H2020 Marie Sklodowska-Curie Actions/ ; 44567YH-5460//PHC/FCT Pessoa Program 2020/ ; UIDB/04621/2020//FCT projects/ ; UIDP/04621/2020//FCT projects/ ; }, mesh = {Animals ; *Mathematical Concepts ; *Disease Outbreaks/prevention & control/statistics & numerical data ; *Vector Borne Diseases/prevention & control/epidemiology/transmission ; *Mosquito Vectors/virology/microbiology ; Humans ; *Wolbachia/physiology ; *Models, Biological ; Male ; *Mosquito Control/methods/statistics & numerical data ; Pest Control, Biological/methods/statistics & numerical data ; Algorithms ; Dengue/prevention & control/epidemiology/transmission ; Computer Simulation ; Female ; Malaria/prevention & control/transmission/epidemiology ; Zika Virus Infection/epidemiology/prevention & control/transmission ; }, abstract = {This paper is devoted to the study of optimal release strategies to control vector-borne diseases, such as dengue, Zika, chikungunya and malaria. Two techniques are considered: the sterile insect one (SIT), which consists in releasing sterilized males among wild vectors in order to perturb their reproduction, and the Wolbachia one (presently used mainly for mosquitoes), which consists in releasing vectors, that are infected with a bacterium limiting their vectorial capacity, in order to replace the wild population by one with reduced vectorial capacity. In each case, the time dynamics of the vector population is modeled by a system of ordinary differential equations in which the releases are represented by linear combinations of Dirac measures with positive coefficients determining their intensity. We introduce optimal control problems that we solve numerically using ad-hoc algorithms, based on writing first-order optimality conditions characterizing the best combination of Dirac measures. We then discuss the results obtained, focusing in particular on the complexity and efficiency of optimal controls and comparing the strategies obtained. Mathematical modeling can help testing a great number of scenarios that are potentially interesting in future interventions (even those that are orthogonal to the present strategies) but that would be hard, costly or even impossible to test in the field in present conditions.}, } @article {pmid39525364, year = {2023}, author = {Tiley, K and Entwistle, J and Thomas, B and Yakob, L and Brady, O}, title = {Using models and maps to inform Target Product Profiles and Preferred Product Characteristics: the example of Wolbachia replacement.}, journal = {Gates open research}, volume = {7}, number = {}, pages = {68}, pmid = {39525364}, issn = {2572-4754}, mesh = {*Wolbachia ; Animals ; *Aedes/microbiology ; *Dengue/prevention & control/transmission/epidemiology ; Humans ; *Mosquito Control/methods/economics ; Mosquito Vectors/microbiology ; }, abstract = {BACKGROUND: The global prevalence of diseases transmitted by Aedes aegypti mosquitoes, such as dengue, Zika and Yellow Fever, is increasing, but development of promising new mosquito control technologies could reverse this trend. Target Product Profiles (TPPs) and Preferred Product Characteristics (PPCs) documents issued by the World Health Organization can guide the research and development pathways of new products and product combinations transitioning from proof of concept to operational use.

METHODS: We used high resolution global maps of the case and economic burden of dengue to derive programmatic cost targets to support a TPP for Wolbachia replacement. A compartmental entomological model was used to explore how release size, spacing and timing affect replacement speed and acceptability. To support a PPC for a hybrid suppress-then-replace approach we tested whether Wolbachia replacement could be achieved faster, more acceptably or at a lower cost if preceded by a mosquito suppression programme.

RESULTS: We show how models can reveal trade-offs, identify quantitative thresholds and prioritise areas and intervention strategies for further development. We estimate that for Wolbachia replacement to be deployable in enough areas to make major contributions to reducing global dengue burden by 25% (in line with 2030 WHO targets), it must have the potential for cost to be reduced to between $7.63 and $0.24 (USD) per person protected or less. Suppression can reduce the number of Wolbachia mosquitoes necessary to achieve replacement fixation by up to 80%. A hybrid approach can also achieve fixation faster and potentially improve acceptability, but may not justify their cost if they require major new investments in suppression technologies.

CONCLUSIONS: Here we demonstrate the value dedicated modelling can provide for interdisciplinary groups of experts when developing TPPs and PPCs. These models could be used by product developers to prioritise and shape development decisions for new Wolbachia replacement products.}, } @article {pmid39506857, year = {2024}, author = {Bernardini, I and Poggi, C and Porretta, D and Máca, J and Perugini, E and Manzi, S and Gabrielli, S and Pichler, V and Latrofa, MS and Fourie, J and Lia, RP and Beugnet, F and Otranto, D and Pombi, M}, title = {Population dynamics of sympatric Phortica spp. and first record of stable presence of Phortica oldenbergi in a Thelazia callipaeda-endemic area of Italy.}, journal = {Parasites & vectors}, volume = {17}, number = {1}, pages = {455}, pmid = {39506857}, issn = {1756-3305}, mesh = {Animals ; *Drosophilidae/parasitology ; *Population Dynamics ; Italy/epidemiology ; *Thelazioidea/isolation & purification/genetics/physiology ; *Insect Vectors/parasitology ; Seasons ; Male ; Female ; Wolbachia/isolation & purification/genetics ; }, abstract = {BACKGROUND: Five species of the Phortica genus (Diptera: Drosophilidae) are known in Europe and the Middle East. Among these, Phortica variegata and Phortica okadai are better known for their role as vectors of the zoonotic eyeworm Thelazia callipaeda. Other species, such as Phortica semivirgo and Phortica oldenbergi, have been studied less. Given the paucity of data about these Phortica spp. vectors, we explored the population dynamics and ecology of Phortica spp. in an area highly endemic for T. callipeada (Manziana, Rome, Central Italy).

METHODS: Phortica spp. flies were collected over a 3-year period (2018-2020) during their active season (April-October) with a sweep net while hovering around fermenting fruits or a human operator acting as baits. Collected flies were morphologically identified and tested for a T. callipeada infection and for the presence of Wolbachia, by polymerase chain reaction (PCR). Population dynamics of species collected was associated to environmental drivers through generalized additive models.

RESULTS: Of the 5564 flies collected, 90.8% were P. variegata, 9.1% were P. oldenbergi, 0.05% were P. semivirgo, and one specimen was P. okadai. Only P. variegata scored molecularly infected with T. callipeada throughout the 3-year sampling period (1.8%). Phortica oldenbergi, observed consistently during the entire sampling period, exhibited a marked preference for fruit traps, contrasting with the lachryphagous activity of P. variegata. Analysis of environmental drivers of P. oldenbergi and P. variegata population dynamics indicated temperature, wind speed, and pressure as significant factors. In addition, Wolbachia pipientis endosymbiont was detected in P. oldenbergi and P. okadai.

CONCLUSIONS: For the first time, this study analysed several ecological aspects of Phortica species coexisting in a T. callipeada endemic area, highlighting different behaviors in the same environment and their vectorial role. Notably, this is also the first report of the presence of P. oldenbergi in Italy and P. okadai in Europe, underscoring the importance of extensive sampling for detecting potential vectors and alien species with direct implications for vector-borne disease epidemiology.}, } @article {pmid39498151, year = {2024}, author = {Dias, YJM and Dezordi, FZ and Wallau, GDL}, title = {EEfinder, a general purpose tool for identification of bacterial and viral endogenized elements in eukaryotic genomes.}, journal = {Computational and structural biotechnology journal}, volume = {23}, number = {}, pages = {3662-3668}, pmid = {39498151}, issn = {2001-0370}, abstract = {Horizontal gene transfer is a phenomenon of genetic material transmission between species with no parental relationship. It has been characterized among several major branches of life, including among prokaryotes, viruses and eukaryotes. The characterization of endogenous elements derived from viruses or bacteria provides a snapshot of past host-pathogen interactions and coevolution as well as reference information to remove false positive results from metagenomic studies. Currently there is a lack of general purpose standardized tools for endogenous elements screening which limits reproducibility and hinder comparative analysis between studies. Here we describe EEfinder, a new general purpose tool for identification and classification of endogenous elements derived from viruses or bacteria found in eukaryotic genomes. The tool was developed to include six common steps performed in this type of analysis: data cleaning, similarity search through sequence alignment, filtering candidate elements, taxonomy assignment, merging of truncated elements and flanks extraction. We evaluated the sensitivity of EEfinder to identify endogenous elements through comparative analysis using data from the literature and showed that EEfinder automatically detected 97 % of the EVEs compared to published results obtained by manual curation and detected an almost exact full integration of a Wolbachia genome described using wet-lab experiments. Therefore, EEfinder can effectively and systematically identify endogenous elements with bacterial/viral origin integrated in eukaryotic genomes. EEfinder is publicly available on https://github.com/WallauBioinfo/EEfinder.}, } @article {pmid39495807, year = {2024}, author = {Dodson, BL and Pujhari, S and Brustolin, M and Metz, HC and Rasgon, JL}, title = {Variable effects of transient Wolbachia infections on alphaviruses in Aedes aegypti.}, journal = {PLoS neglected tropical diseases}, volume = {18}, number = {11}, pages = {e0012633}, doi = {10.1371/journal.pntd.0012633}, pmid = {39495807}, issn = {1935-2735}, abstract = {Wolbachia pipientis (= Wolbachia) has promise as a tool to suppress virus transmission by Aedes aegypti mosquitoes. However, Wolbachia can have variable effects on mosquito-borne viruses. This variation remains poorly characterized, yet the multimodal effects of Wolbachia on diverse pathogens could have important implications for public health. Here, we examine the effects of transient somatic infection with two strains of Wolbachia (wAlbB and wMel) on the alphaviruses Sindbis virus (SINV), O'nyong-nyong virus (ONNV), and Mayaro virus (MAYV) in Ae. aegypti. We found variable effects of Wolbachia including enhancement and suppression of viral infections, with some effects depending on Wolbachia strain. Both wAlbB- and wMel-infected mosquitoes showed enhancement of SINV infection rates one week post-infection, with wAlbB-infected mosquitoes also having higher viral titers than controls. Infection rates with ONNV were low across all treatments and no significant effects of Wolbachia were observed. The effects of Wolbachia on MAYV infections were strikingly strain-specific; wMel strongly blocked MAYV infections and suppressed viral titers, while wAlbB had more modest effects. The variable effects of Wolbachia on vector competence underscore the importance of further research into how this bacterium impacts the virome of wild mosquitoes including the emergent human pathogens they transmit.}, } @article {pmid39495046, year = {2024}, author = {Yang, SY and Lin, YY and Hao, Z and Li, ZJ and Peng, ZQ and Jin, T}, title = {Bacterial communities in Asecodes hispinarum (Hymenoptera: Eulophidae) and its host Brontispa longissima (Coleoptera: Chrysomelidae), with comparison of Wolbachia dominance.}, journal = {Journal of economic entomology}, volume = {}, number = {}, pages = {}, doi = {10.1093/jee/toae234}, pmid = {39495046}, issn = {1938-291X}, support = {2021YFD2600405//National Key R&D Program of China/ ; CATASCXTD202311//Chinese Academy of Tropical Agricultural Sciences for Science and Technology Innovation Team/ ; }, abstract = {The endoparasitoid Asecodes hispinarum (Bouček) (Hymenoptera: Eulophidae) serves as an effective biological control agent against Brontispa longissima (Gestro) (Coleoptera: Chrysomelidae), a notorious palm pest. Endosymbionts found in parasitoids and their hosts have attracted significant attention due to their substantial influence on biocontrol efficacy. In this study, we employed 16S rRNA sequencing, polymerase chain reaction, and fluorescence in situ hybridization to assess the symbiotic bacteria composition, diversity, phylogeny, and localization in A. hispinarum and its host B. longissima. Our findings showed significant differences in the richness, diversity, and composition of symbiotic bacteria among different life stages of B. longissima. Notably, the bacterial richness, diversity, and composition of A. hispinarum was similar to that of B. longissima. Firmicutes and Proteobacteria were the dominant phyla, while Wolbachia was the dominant genera across the parasitoid and host. It was discovered for the first time that Wolbachia was present in A. hispinarum with a high infection rate at ≥ 96.67%. Notably, the Wolbachia strain in A. hispinarum was placed in supergroup A, whereas it was categorized under supergroup B in B. longissima. Furthermore, Wolbachia is concentrated in the abdomen of A. hispinarum, with particularly high levels observed in the ovipositors of female adults. These findings highlight the composition and diversity of symbiotic bacteria in both A. hispinarum and its host B. longissima, providing a foundation for the development of population regulation strategies targeting B. longissima.}, } @article {pmid39471850, year = {2024}, author = {Tourani, AH and Katlav, A and Cook, JM and Riegler, M}, title = {Mating receptivity mediated by endosymbiont interactions in a haplodiploid thrips species.}, journal = {Proceedings. Biological sciences}, volume = {291}, number = {2033}, pages = {20241564}, doi = {10.1098/rspb.2024.1564}, pmid = {39471850}, issn = {1471-2954}, support = {//University of Western Sydney/ ; }, mesh = {Animals ; *Symbiosis ; *Thysanoptera/physiology ; *Wolbachia/physiology ; Female ; Male ; *Sexual Behavior, Animal ; Bacteroidetes/physiology ; Reproduction ; }, abstract = {Many arthropods carry maternally inherited endosymbionts that cause cytoplasmic incompatibility (CI), manifested as embryonic mortality in matings of infected males with uninfected females. Infected females, however, do not suffer this cost. Therefore, in populations with mixed endosymbiont infections, selection is expected to favour mechanisms that enable hosts to avoid or mitigate CI. This may include changes in mating behaviour, such as reduced female receptivity to mating and/or remating when approached by incompatible males. Here, we investigated mating behavioural traits in haplodiploid thrips naturally associated with two CI-inducing endosymbionts, Cardinium and Wolbachia. Compared with females with both endosymbionts, those with only Cardinium showed reduced receptivity to males carrying both. However, surprisingly, females without endosymbionts were not less receptive to incompatible males. Furthermore, in contrast to females without endosymbionts, females with Cardinium were far less likely to remate with incompatible than compatible males irrespective of the compatibility type of the first mating. Our results suggest that endosymbiont-specific sexual selection processes occur, whereby females carrying only Cardinium recognize Wolbachia in coinfected males to avoid CI. This may hinder a CI-driven Wolbachia spread. Endosymbiont-mediated mating behaviours may be crucial for the dynamics of CI-inducing endosymbionts and their application in pest management strategies.}, } @article {pmid39458303, year = {2024}, author = {Clervil, E and Guidez, A and Talaga, S and Carinci, R and Gaborit, P and Lavergne, A and Tirera, S and Duchemin, JB}, title = {Wolbachia Natural Infection of Mosquitoes in French Guiana: Prevalence, Distribution, and Genotyping.}, journal = {Microorganisms}, volume = {12}, number = {10}, pages = {}, doi = {10.3390/microorganisms12101994}, pmid = {39458303}, issn = {2076-2607}, abstract = {Wolbachia are the most spread bacterial endosymbionts in the world. These bacteria can manipulate host reproduction or block virus transmission in mosquitoes. For this reason, Wolbachia-based strategies for vector control are seriously considered or have already been applied in several countries around the world. In South America, Wolbachia have been studied in human pathogen vectors such as sand flies and mosquitoes. In French Guiana, the diversity and distribution of Wolbachia are not well known in mosquitoes. In this study, we screened for Wolbachia natural infection in mosquitoes in French Guiana by using 16S rRNA, Wolbachia surface protein (WSP), and multi-locus sequence typing (MLST) molecular assays. A total of 29 out of 44 (65.9%) mosquito species were positive for natural Wolbachia infection according to the PCR results, and two Wolbachia strains co-infected three specimens of Mansonia titillans. Then, we analyzed the phylogenetic relationships among the Wolbachia detected. All of the tested specimens of Aedes aegypti, the major dengue vector of French Guiana, were negative. These results regarding Wolbachia strain, distribution, and prevalence in mosquitoes from French Guiana highlight Wolbachia-mosquito associations and pave the way for a future Wolbachia-based strategy for vector control in this Amazonian territory.}, } @article {pmid39455905, year = {2024}, author = {Aželytė, J and Maitre, A and Abuin-Denis, L and Wu-Chuang, A and Žiegytė, R and Mateos-Hernandez, L and Obregon, D and Palinauskas, V and Cabezas-Cruz, A}, title = {Nested patterns of commensals and endosymbionts in microbial communities of mosquito vectors.}, journal = {BMC microbiology}, volume = {24}, number = {1}, pages = {434}, pmid = {39455905}, issn = {1471-2180}, support = {SGCE-RAPPORT No. 0300//Collectivité de Corse/ ; S-MIP-22-52//Lietuvos Mokslo Taryba/ ; ANR-10-LABX-62-IBEID//Agence Nationale de la Recherche/ ; }, mesh = {Animals ; *Symbiosis ; *Mosquito Vectors/microbiology/physiology ; *Culex/microbiology ; *Wolbachia/physiology/genetics ; *Microbiota ; Bacteria/classification/genetics/isolation & purification ; }, abstract = {BACKGROUND: Mosquitoes serve as vectors for numerous pathogens, posing significant health risks to humans and animals. Understanding the complex interactions within mosquito microbiota is crucial for deciphering vector-pathogen dynamics and developing effective disease management strategies. Here, we investigated the nested patterns of Wolbachia endosymbionts and Escherichia-Shigella within the microbiota of laboratory-reared Culex pipiens f. molestus and Culex quinquefasciatus mosquitoes. We hypothesized that Wolbachia would exhibit a structured pattern reflective of its co-evolved relationship with both mosquito species, while Escherichia-Shigella would display a more dynamic pattern influenced by environmental factors.

RESULTS: Our analysis revealed different microbial compositions between the two mosquito species, although some microorganisms were common to both. Network analysis revealed distinct community structures and interaction patterns for these bacteria in the microbiota of each mosquito species. Escherichia-Shigella appeared prominently within major network modules in both mosquito species, particularly in module P4 of Cx. pipiens f. molestus, interacting with 93 nodes, and in module Q3 of Cx. quinquefasciatus, interacting with 161 nodes, sharing 55 nodes across both species. On the other hand, Wolbachia appeared in disparate modules: module P3 in Cx. pipiens f. molestus and a distinct module with a single additional taxon in Cx. quinquefasciatus, showing species-specific interactions and no shared taxa. Through computer simulations, we evaluated how the removal of Wolbachia or Escherichia-Shigella affects network robustness. In Cx. pipiens f. molestus, removal of Wolbachia led to a decrease in network connectivity, while Escherichia-Shigella removal had a minimal impact. Conversely, in Cx. quinquefasciatus, removal of Escherichia-Shigella resulted in decreased network stability, whereas Wolbachia removal had minimal effect.

CONCLUSIONS: Contrary to our hypothesis, the findings indicate that Wolbachia displays a more dynamic pattern of associations within the microbiota of Culex pipiens f. molestus and Culex quinquefasciatus mosquitoes, than Escherichia-Shigella. The differential effects on network robustness upon Wolbachia or Escherichia-Shigella removal suggest that these bacteria play distinct roles in maintaining community stability within the microbiota of the two mosquito species.}, } @article {pmid39453968, year = {2024}, author = {Szentiványi, T and Takács, N and Sándor, AD and Péter, Á and Boldogh, SA and Kováts, D and Foster, JT and Estók, P and Hornok, S}, title = {Bat-associated ticks as a potential link for vector-borne pathogen transmission between bats and other animals.}, journal = {PLoS neglected tropical diseases}, volume = {18}, number = {10}, pages = {e0012584}, doi = {10.1371/journal.pntd.0012584}, pmid = {39453968}, issn = {1935-2735}, abstract = {BACKGROUND: Potentially zoonotic pathogens have been previously detected in bat-associated ticks, but their role in disease transmission and their frequency of feeding on non-bat hosts is poorly known.

We used molecular blood meal analysis to reveal feeding patterns of the bat-associated tick species Ixodes ariadnae, I. simplex, and I. vespertilionis collected from cave and mine walls in Central and Southeastern Europe. Vertebrate DNA, predominantly from bats, was detected in 43.5% of the samples (70 of 161 ticks) but in these ticks we also detected the DNA of non-chiropteran hosts, such as dog, Canis lupus familiaris, wild boar, Sus scrofa, and horse, Equus caballus, suggesting that bat-associated ticks may exhibit a much broader host range than previously thought, including domestic and wild mammals. Furthermore, we detected the zoonotic bacteria Neoehrlichia mikurensis in bat ticks for the first time, and other bacteria, such as Bartonella and Wolbachia.

CONCLUSIONS/SIGNIFICANCE: In the light of these findings, the role of bat ticks as disease vectors should be urgently re-evaluated in more diverse host systems, as they may contribute to pathogen transmission between bats and non-chiropteran hosts.}, } @article {pmid39452360, year = {2024}, author = {Zhu, Y and Wang, X and Wang, S and Song, Z and Du, Y}, title = {No Evidence for Wolbachia Effects on the Thermal Preference of the Invasive Pest Liriomyza huidobrensis.}, journal = {Insects}, volume = {15}, number = {10}, pages = {}, doi = {10.3390/insects15100784}, pmid = {39452360}, issn = {2075-4450}, support = {BK20231330//Natural Science Foundation of Jiangsu Province/ ; }, abstract = {Heritable endosymbiont Wolbachia is prevalent among arthropods, serving multiple functions for their hosts. However, the role of Wolbachia in mediating thermal preference selection remains largely unexplored. In this study, we utilized a custom-built thermal gradient to evaluate the thermal preference (Tp) of 1367 individuals of the invasive leaf-miner Liriomyza huidobrensis with or without Wolbachia wLhui from Yunnan and Xinjiang populations. Under meticulously controlled conditions and with a vast sample size, we found no significant difference in the mean Tp between wLhui-infected and uninfected leaf miners from either population when host age and sex were not considered. Furthermore, generalized linear model (GLM) analysis revealed no significant correlation between average Tp and age, sex, or Wolbachia infection, nor interactions among these factors, except in the Xinjiang population, where Tp was strongly associated with host age. Finally, we discuss the ecological implications of these findings and propose future research directions on Wolbachia-mediated host Tp in the leaf miner. Overall, our findings do not provide evidence that Wolbachia significantly affects the thermal preference of L. huidobrensis. Further studies across different systems are needed to investigate the complex interactions between Wolbachia and insect thermal behavior.}, } @article {pmid39447296, year = {2024}, author = {Antonelli, P and Grizard, S and Tran, FH and Lejon, D and Bellemain, A and Van, and Mavingui, P and Roiz, D and Simard, F and Martin, E and Abrouk, D and Vigneron, A and Minard, G and Valiente Moro, C}, title = {Bioaccumulation of polycyclic aromatic hydrocarbons and microbiota dynamics across developmental stages of the Asian tiger mosquito, Aedes albopictus exposed to urban pollutants.}, journal = {Ecotoxicology and environmental safety}, volume = {286}, number = {}, pages = {117214}, doi = {10.1016/j.ecoenv.2024.117214}, pmid = {39447296}, issn = {1090-2414}, abstract = {Aedes albopictus mosquitoes face numerous anthropic stressors in urban areas. These xenobiotics not only impact mosquito physiology but also shape the composition of their microbiota, which play important roles in host physiological traits. Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous environmental pollutants known to alter mosquito metabolism, but no studies have yet investigated their impact on microbiota. Using a bespoke indoor mesocosm tailored for Ae. albopictus mosquitoes, we investigated the dynamics of bacterial communities in both mosquitoes and their larval breeding sites following chronic exposure to a cocktail of PAHs consisting of benzo[a]pyrene, benz[a]anthracene, chrysene and benzo[b]fluoranthene. Our findings showed that PAHs have a stage-specific effect on mosquito microbiota, with a higher impact in larvae than in adults, contributing to 12.5 % and 4.5 % of the PAHs-induced variations, respectively. The presence of PAHs in the treated mesocosm led to the enrichment of bacterial families and genera known for their ability to catabolize PAHs, such as Comamonadaceae and Raoultella (increasing from 19 % to 30 % and from 1.2 % to 5.6 %, respectively). Conversely, prevalent taxa found in mosquito microbiota like Wolbachia and Cedecea exhibited a reduction (decreasing from 4 % to 0.8 % and from 12.8 % to 6.4 %, respectively). This reduction could be attributed to the competitive advantage gained by PAH-degrading taxa, or it could reflect a direct sensitivity to PAH exposure. Overall, this indicates a shift in microbiota composition favoring bacteria that can thrive in a PAH-contaminated environment. PAHs persisted in the water of breeding sites only the first 45 days of the experiment. Benzo[a]pyrene and benzo[b]fluoranthene were more susceptible to bioaccumulation in larval tissues over time. Overall, this study enhances our understanding of the impact of pollution on mosquitoes and could facilitate future research on the importance of symbiosis in urban-dwelling insect disease vectors. Given the recent advancements in the generation of axenic (microbe-free) and gnotobiotic (mosquitoes with a defined or specific microbiota) mosquitoes, further studies are needed to explore how changes in microbiota composition could influence mosquito responses to pollution, particularly in relation to host fitness, immunity, and vector competence.}, } @article {pmid39442642, year = {2024}, author = {Efon-Ekangouo, A and Nana-Djeunga, HC and Nwane, PB and Nzune-Toche, N and Sondi-Dissake, JC and Sempere, G and Domche, A and Njiokou, F and Kamgno, J and Moundipa-Fewou, P and Geiger, A}, title = {Spatial and temporal diversity of Simulium damnosum s.l. gut microbiota and association with Onchocerca volvulus infection in Cameroon.}, journal = {Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases}, volume = {}, number = {}, pages = {105683}, doi = {10.1016/j.meegid.2024.105683}, pmid = {39442642}, issn = {1567-7257}, abstract = {Arthropod microbiota plays an important role in host physiology, and there is growing interest in using vector symbionts to modify vector competence and control parasite transmission. This study aims to characterise the blackfly Simulium damnosum s.l. gut microbiota and to explore possible associations with various bio-ecological determinants of the Onchocerca volvulus establishment and the transmission in blackfly. Adult female blackflies were caught in three Cameroonian health districts belonging to different bioecological zones endemic for onchocerciasis. Flies were dissected and qPCR screened for Onchocerca volvulus infection. The diversity of the blackflies gut microbiota was assessed by high-throughput sequencing of the V3-V4 hypervariable region of the bacterial 16S ribosomal RNA. Subsequent metataxo-genomic, multivariate, and association analysis were used to investigate the variables that influence the microbiota diversity. Transmission index rates ranging from 20.7 to 6.0 % and 6.2-2.0 % for infection and infectivity rates, respectively, indicate ongoing transmission of onchocerciasis across all surveyed health districts. The identified bacterial taxa were clustered into four phyla, five classes, and 23 genera. The S. damnosum s.l. gut microbiota was dominated by Wolbachia and by Rosenbergiella in Wolbachia-free Simulium. Significant differences were observed in the diversity of S. damnosum s.l. microbiota concerning parity status (P = 0.007), health district of origin (P = 0.001), and the presence of the Onchocerca volvulus. Simulium from the Bafia health district also showed increased bacterial diversity between two consecutive years (P = 0.001). Four bacterial taxa, including Serratia, were associated with the absence of the O. volvulus infection. These results indicate that S. damnosum s.l. from different onchocerciasis foci in Cameroon, exhibit distinguishable gut microbial compositions which are dynamic over time. Some bacterial species are associated with the O. volvulus infection and could be further investigated as biological target/tool for vector modified-based onchocerciasis control.}, } @article {pmid39432669, year = {2024}, author = {Cruz, MA and Magalhães, S and Bakırdöven, M and Zélé, F}, title = {Wolbachia strengthens the match between pre-mating and early post-mating isolation in spider mites.}, journal = {Evolution; international journal of organic evolution}, volume = {}, number = {}, pages = {}, doi = {10.1093/evolut/qpae149}, pmid = {39432669}, issn = {1558-5646}, abstract = {Endosymbiotic reproductive manipulators are widely studied as sources of post-zygotic isolation in arthropods, but their effect on pre-zygotic isolation between genetically differentiated populations has garnered less attention. We tested this using two partially isolated populations of the red and green colour forms of Tetranychus urticae, either uninfected or infected with different Wolbachia strains, one inducing cytoplasmic incompatibility and the other not. We first investigated male and female preferences, and found that, in absence of infection, females were not choosy, but all males preferred red-form females. Wolbachia effects were more subtle, with only the CI-inducing strain slightly strengthening colour-form based preferences. We then performed a double-mating experiment to test how incompatible matings affect subsequent mating behaviour and offspring production, as compared to compatible matings. Females mated with an incompatible male (infected and/or heterotypic) were more attractive and/or receptive to subsequent (compatible) matings, although analyses of offspring production revealed no clear benefit for this re-mating behaviour (i.e., apparently unaltered first male sperm precedence). Finally, by computing the relative contributions of each reproductive barrier to total isolation, we showed that pre-mating isolation matches both host-associated and Wolbachia-induced post-mating isolation, suggesting that Wolbachia could contribute to reproductive isolation in this system.}, } @article {pmid39431789, year = {2024}, author = {Bickerstaff, JRM and Walsh, T and Court, L and Pandey, G and Ireland, K and Cousins, D and Caron, V and Wallenius, T and Slipinski, A and Rane, R and Escalona, HE}, title = {Chromosome structural rearrangements in invasive haplodiploid ambrosia beetles revealed by the genomes of Euwallacea fornicatus (Eichhoff) and Euwallacea similis (Ferrari) (Coleoptera, Curculionidae, Scolytinae).}, journal = {Genome biology and evolution}, volume = {}, number = {}, pages = {}, doi = {10.1093/gbe/evae226}, pmid = {39431789}, issn = {1759-6653}, abstract = {Bark and ambrosia beetles are among the most ecologically and economically damaging introduced plant pests worldwide. Life history traits including polyphagy, haplodiploidy, inbreeding polygyny and symbiosis with fungi contribute to their dispersal and impact. Species vary in their interactions with host trees, with many attacking stressed or recently dead trees, such as the globally distributed E. similis (Ferrari). Other species, like the Polyphagous Shot Hole Borer (PSHB) Euwallacea fornicatus (Eichhoff), can attack over 680 host plants and is causing considerable economic damage in several countries. Despite their notoriety, publicly accessible genomic resources for Euwallacea Hopkins species are scarce, hampering our understanding of their invasive capabilities as well as modern control measures, surveillance and management. Using a combination of long and short read sequencing platforms we assembled and annotated high quality (BUSCO > 98% complete) pseudo-chromosome level genomes for these species. Comparative macro-synteny analysis identified an increased number of pseudo-chromosome scaffolds in the haplodiploid inbreeding species of Euwallacea compared to diploid outbred species, due to fission events. This suggests that life history traits can impact chromosome structure. Further, the genome of E. fornicatus had a higher relative proportion of repetitive elements, up to 17% more, than E. similis. Metagenomic assembly pipelines identified microbiota associated with both species including Fusarium fungal symbionts and a novel Wolbachia strain. These novel genomes of haplodiploid inbreeding species will contribute to the understanding of how life history traits are related to their evolution and to the management of these invasive pests.}, } @article {pmid39411028, year = {2024}, author = {Angotti, AA and da Silva, JGST and Yonekura, T and Figueiró, MF}, title = {[Rapid review: effectiveness of the Wolbachia strategy for arbovirus controlRevisión sistemática rápida: efectividad de la estrategia de control con Wolbachia en la lucha contra las arbovirosis].}, journal = {Revista panamericana de salud publica = Pan American journal of public health}, volume = {48}, number = {}, pages = {e98}, pmid = {39411028}, issn = {1680-5348}, abstract = {OBJECTIVE: To identify and synthesize evidence of the effectiveness and cost-effectiveness of the Wolbachia-Aedes methodology for reducing cases of arboviral infection.

METHODOLOGY: Rapid systematic review. A search of five online portals and bibliographic databases was conducted in February 2022. Two reviewers conducted the selection stages, with any disagreements resolved by a third investigator. Data were extracted by one reviewer and subsequently checked by another. The inclusion criteria were studies describing the cost-effectiveness and effectiveness of the Wolbachia-Aedes methodology, based on field interventions, published in Portuguese, English, and Spanish, with no restrictions on date of publication.

RESULTS: Four studies were included: one cluster randomized trial, two quasi-experimental studies, and one pre-post study, published between 2019 and 2021 in four countries. The included studies reported effectiveness for some of the outcomes of interest, such as a reduction in the incidence of dengue, Zika, and chikungunya cases. Although no experimental studies addressing the cost-effectiveness of the Wolbachia-Aedes strategy were identified, some modeling studies have demonstrated potential cost-benefit of this methodology.

CONCLUSION: Considering the results of the included studies, Wolbachia has the potential to be an economically effective strategy that leads to reductions in the incidence of dengue, Zika, and chikungunya. Nevertheless, this strategy cannot yet be recommended as a public policy; additional large-scale studies with high methodological quality are still needed to inform political decision-making.}, } @article {pmid39407097, year = {2024}, author = {Chow, JY and Geng, L and Bansal, S and Dickens, BSL and Ng, LC and Hoffmann, AA and Lim, JT}, title = {Correction: Evaluating quasi-experimental approaches for estimating epidemiological efficacy of non-randomised field trials: applications in Wolbachia interventions for dengue.}, journal = {BMC medical research methodology}, volume = {24}, number = {1}, pages = {237}, pmid = {39407097}, issn = {1471-2288}, } @article {pmid39405448, year = {2024}, author = {}, title = {Correction to: Life history parameters of Ectropis grisescens (Lepidoptera: Geometridae) in different Wolbachia infection states.}, journal = {Journal of economic entomology}, volume = {}, number = {}, pages = {}, doi = {10.1093/jee/toae238}, pmid = {39405448}, issn = {1938-291X}, support = {32371581//National Natural Science Foundation of China/ ; 2021YFD1601105//National Key Research and Development Programs/ ; 2023n06020060//Anhui Key Research and Development Program/ ; }, } @article {pmid39403558, year = {2024}, author = {Fisher, AM and McKenzie, AV and Price, TAR and Bonsall, MB and Knell, RJ}, title = {Do Sex Ratio Distorting Microbes Inhibit the Evolution of Pesticide Resistance? An Experimental Test.}, journal = {Evolutionary applications}, volume = {17}, number = {10}, pages = {e70003}, pmid = {39403558}, issn = {1752-4571}, abstract = {We are still largely reliant on pesticides for the suppression of arthropod pests which threaten human health and food production, but the recent rise of evolved resistance among important pest species has reduced pesticide efficacy. Despite this, our understanding of strategies that effectively limit the evolution of resistance remains weak. Male-killing sex ratio distorting microbes (SRDMs), such as Wolbachia and Spiroplasma, are common among arthropod species. Previous theoretical work has suggested that they could limit adaptive potential in two ways: first, because by distorting sex ratios they reduce the effective population size, and second, because infected females produce no male offspring which restricts gene flow. Here we present the results of a novel experiment in which we test the extent by which these two mechanisms limit the adaptive response of arthropods to pesticide. Using a fully factorial design, we manipulated the adult sex ratio of laboratory populations of Drosophila melanogaster, both in the presence and absence of SRDMs, and exposed these populations to six generations of pesticide poisoning. This design allows the effects of SRDMs on sex ratio and their effects on gene flow to be estimated separately. After six generations, individuals from populations with even sex ratios displayed a higher resistance to pesticide relative to individuals from female-biased populations. By contrast, we found no effect of the presence of SRDMs in host populations on pesticide resistance independent of sex ratio. In addition, males were more susceptible to pesticide than females-this was true of flies from both naïve and previously exposed populations. These findings provide the first empirical proof of concept that sex ratio distortion arising from SRDMs can limit adaptation to pesticides, but cast doubt on the theoretical effect of male-killers limiting adaptation by disrupting gene flow.}, } @article {pmid39402267, year = {2024}, author = {Kaczmarczyk-Ziemba, A and Wagner, GK and Staniec, B and Zagaja, M and Pietrykowska-Tudruj, E and Iorgu, EI and Iorgu, IŞ}, title = {Intraspecific diversity of Myrmecophilus acervorum (Orthoptera: Myrmecophilidae) indicating an ongoing cryptic speciation.}, journal = {Scientific reports}, volume = {14}, number = {1}, pages = {23984}, pmid = {39402267}, issn = {2045-2322}, mesh = {Animals ; *Wolbachia/genetics/classification/isolation & purification ; *Phylogeny ; *Genetic Variation ; Genetic Speciation ; Male ; Female ; Orthoptera/genetics/classification ; DNA, Mitochondrial/genetics ; Poland ; Symbiosis ; RNA, Ribosomal, 16S/genetics ; Europe ; Gryllidae ; }, abstract = {Myrmecophilus acervorum, previously considered a parthenogenetic species widely-distributed in Europe, has been observed to have both sexes in populations inhabiting the central part of the distribution range. Specimens from those heterosexual populations have been found being infected with Wolbachia. New mitochondrial data (COI and 16S markers) revealed the well-supported differentiation of M. acervorum populations inhabiting western Polesie (Poland) and southern Europe. In turn, analyses of EF1α marker support the hypothesis on the unfinished lineage sorting at the nuclear DNA level. Interestingly, we found that parthenogenetic populations inhabiting western Polesie are infected with Wolbachia belonging to supergroup A, while endosymbionts occurring in sexual populations of M. acervorum observed in Romania belong to supergroup B. Furthermore, new and potentially diagnostic characteristics in the external structures of the eyes of M. acervorum were identified. The surface of ommatidia in specimens occurring in southern Europe was smooth. In contrast, the ommatidia surface of individuals collected in Poland was visibly sculptured. To sum up, the significant genetic variability found in the present case, and the differentiating morphological character, are almost certainly effects of cryptic species being present within M. acervorum. This is indicative of ongoing speciation within the populations of this insect, and of simultaneous unfinished lineage sorting at the nuclear DNA level.}, } @article {pmid39400605, year = {2024}, author = {Ma, X and Su, Y}, title = {Wolbachia Invasion in Mosquitoes with Incomplete CI, Imperfect Maternal Transmission and Maturation Delay.}, journal = {Bulletin of mathematical biology}, volume = {86}, number = {12}, pages = {137}, pmid = {39400605}, issn = {1522-9602}, support = {12371164//National Natural Science Foundation of China/ ; 11971129//National Natural Science Foundation of China/ ; }, mesh = {*Wolbachia/physiology/pathogenicity ; Animals ; *Mathematical Concepts ; *Models, Biological ; Female ; *Computer Simulation ; *Mosquito Vectors/microbiology ; Cytoplasm/microbiology ; Male ; Culicidae/microbiology ; Maternal Inheritance ; }, abstract = {The mechanism of cytoplasmic incompatibility (CI) is important in the study of Wolbachia invasion in wild mosquitoes. Su et al. (Bull Math Biol 84(9):95, 2022) proposed a delay differential equation model by relating the CI effect to maturation delay. In this paper, we investigate the dynamics of this model by allowing the same density-dependent death rate and distinct density-independent death rates. Through analyzing the existence and stability of equilibria, we obtain the parameter conditions for Wolbachia successful invasion if the maternal transmission is perfect. While if the maternal transmission is imperfect, we give the ranges of parameters to ensure failure invasion, successful invasion and partially suppressing, respectively. Meanwhile, numerical simulations indicate that the system may exhibit monostable and bistable dynamics when parameters vary. Particularly, in the bistable situation an unstable separatrix, like a line, exists when choosing constant functions as initial values; and the maturation delay affects this separatrix in an interesting way.}, } @article {pmid39386366, year = {2024}, author = {Kaur, T and Brown, AMV}, title = {Discovery of a novel Wolbachia in Heterodera expands nematode host distribution.}, journal = {Frontiers in microbiology}, volume = {15}, number = {}, pages = {1446506}, pmid = {39386366}, issn = {1664-302X}, abstract = {Bioinformatics sequence data mining can reveal hidden microbial symbionts that might normally be filtered and removed as contaminants. Data mining can be helpful to detect Wolbachia, a widespread bacterial endosymbiont in insects and filarial nematodes whose distribution in plant-parasitic nematodes (PPNs) remains underexplored. To date, Wolbachia has only been reported a few PPNs, yet nematode-infecting Wolbachia may have been widespread in the evolutionary history of the phylum based on evidence of horizontal gene transfers, suggesting there may be undiscovered Wolbachia infections in PPNs. The goal of this study was to more broadly sample PPN Wolbachia strains in tylenchid nematodes to enable further comparative genomic analyses that may reveal Wolbachia's role and identify targets for biocontrol. Published whole-genome shotgun assemblies and their raw sequence data from 33 Meloidogyne spp. assemblies, seven Globodera spp. assemblies, and seven Heterodera spp. assemblies were analyzed to look for Wolbachia. No Wolbachia was found in Meloidogyne spp. and Globodera spp., but among seven genome assemblies for Heterodera spp., an H. schachtii assembly from the Netherlands was found to have a large Wolbachia-like sequence that, when re-assembled from reads, formed a complete, circular genome. Detailed analyses comparing read coverage, GC content, pseudogenes, and phylogenomic patterns clearly demonstrated that the H. schachtii Wolbachia represented a novel strain (hereafter, denoted wHet). Phylogenomic tree construction with PhyloBayes showed wHet was most closely related to another PPN Wolbachia, wTex, while 16S rRNA gene analysis showed it clustered with other Heterodera Wolbachia assembled from sequence databases. Pseudogenes in wHet suggested relatedness to the PPN clade, as did the lack of significantly enriched GO terms compared to PPN Wolbachia strains. It remains unclear whether the lack of Wolbachia in other published H. schachtii isolates represents the true absence of the endosymbiont from some hosts.}, } @article {pmid39382727, year = {2024}, author = {Chuchuy, A and Rodriguero, MS and Alonso, AC and Stein, M and Micieli, MV}, title = {Wolbachia infection in natural mosquito populations from Argentina.}, journal = {Parasitology research}, volume = {123}, number = {10}, pages = {343}, pmid = {39382727}, issn = {1432-1955}, support = {2015-0665//Agencia Nacional de Promoción Científica y Tecnológica/ ; 2015-0665//Agencia Nacional de Promoción Científica y Tecnológica/ ; 2015-0665//Agencia Nacional de Promoción Científica y Tecnológica/ ; 2015-0665//Agencia Nacional de Promoción Científica y Tecnológica/ ; 2015-0665//Agencia Nacional de Promoción Científica y Tecnológica/ ; 584//Consejo Nacional de Investigaciones Científicas y Técnicas/ ; 584//Consejo Nacional de Investigaciones Científicas y Técnicas/ ; 584//Consejo Nacional de Investigaciones Científicas y Técnicas/ ; 584//Consejo Nacional de Investigaciones Científicas y Técnicas/ ; 584//Consejo Nacional de Investigaciones Científicas y Técnicas/ ; }, mesh = {*Wolbachia/isolation & purification/genetics ; Animals ; Argentina ; *Aedes/microbiology/virology ; Culex/microbiology ; Mosquito Vectors/microbiology ; Female ; }, abstract = {The increasing spread of mosquito vectors has made mosquito-borne arboviral diseases a global threat to public health, leading to the urgent need for effective population control methods. Strategies based in the intracellular bacterium Wolbachia Hertig, 1936 are considered environmentally friendly, safe for humans, and potentially cost-effective for controlling arboviral diseases. To minimize undesirable side effects, it is relevant to assess whether Wolbachia is present in the area and understand the diversity associated with native infections before implementing these strategies. With this purpose, we investigated Wolbachia infection status, diversity, and prevalence in populations of Aedes albifasciatus (Macquart, 1838), Aedes fluviatilis (Lutz, 1904), and hybrids of the Culex pipiens (Linnaeus, 1758) complex from Argentina. Aedes albifasciatus and C. pipiens complex samples were collected in the province of Buenos Aires, and A. fluviatilis in the province of Misiones. Aedes albifasciatus was found to be uninfected, while infections with strains wFlu and wPip were detected in A. fluviatilis and hybrids of the C. pipiens complex, respectively. All strains were fixed or close to fixation and clustered within supergroup B. These finding provides valuable information on Wolbachia strains found in natural mosquito populations in Argentina that might be used in heterologous infections in the future or be considered when designing control strategies based on Wolbachia infection.}, } @article {pmid39366500, year = {2024}, author = {Tobón, AC and Rojo-Ospina, R and Zuluaga, S and Giraldo-Muñoz, JF and Cadavid, JM}, title = {Evaluation of Wolbachia Infection in Aedes aegypti suggests low prevalence and highly heterogeneous distribution in Medellín, Colombia.}, journal = {Acta tropica}, volume = {}, number = {}, pages = {107423}, doi = {10.1016/j.actatropica.2024.107423}, pmid = {39366500}, issn = {1873-6254}, abstract = {Dengue virus, transmitted mainly by Aedes aegypti mosquitoes, is a significant public health challenge in tropical and subtropical countries, with an incidence that is growing at an alarming rate. The release of Wolbachia-carrying mosquitoes has been suggested as a strategy to reduce the incidence of multiple arboviruses. In Medellín, Colombia, large-scale releases of Wolbachia-infected Ae. aegypti mosquitoes were performed between 2017 and 2022 by the World Mosquito Program to facilitate population replacement. In this study, we evaluated the prevalence and distribution of Wolbachia-infected Ae. aegypti two years after completion of these releases. We conducted the sampling across 19 communes in Medellín, using 416 ovitraps to collect Ae. aegypti eggs from epidemiological weeks 26 to 41 in 2023. Upon hatching the collected eggs, we identified and pooled adult female Ae. aegypti for DNA extraction. Subsequently, we conducted PCR assays for the detection of Wolbachia infection in these mosquitoes. We used maximum likelihood estimation (MLE) and Bayesian methods to estimate the prevalence of Wolbachia infection, while using QGIS to analyze spatial distribution of infection in the region. A total of 774 female Ae. aegypti mosquitoes from 182 pools were evaluated. We detected Wolbachia in 33.5% of pools, with an estimated individual minimum infection rate of 9.5% and a maximum of 33.2%. The prevalence varied significantly across communes, with the highest rates observed in the northeastern and southwestern areas. Spatial analysis revealed a highly heterogeneous island-like distribution of Wolbachia across Medellín with a few hotspots. The observed Wolbachia prevalence in this work was lower than previously reported. We suspect a decline in the prevalence of Wolbachia-infected Ae. aegypti mosquitoes in Medellín following the completion of their release.}, } @article {pmid39353088, year = {2024}, author = {Wang, GH and Hoffmann, A and Champer, J}, title = {Gene Drive and Symbiont Technologies for Control of Mosquito-Borne Diseases.}, journal = {Annual review of entomology}, volume = {}, number = {}, pages = {}, doi = {10.1146/annurev-ento-012424-011039}, pmid = {39353088}, issn = {1545-4487}, abstract = {Mosquito-borne diseases, such as dengue and malaria, pose a significant burden to global health. Current control strategies with insecticides are only moderately effective. Scalable solutions are needed to reduce the transmission risk of these diseases. Symbionts and genome engineering-based mosquito control strategies have been proposed to address these problems. Bacterial, fungal, and viral symbionts affect mosquito reproduction, reduce mosquito lifespan, and block pathogen transmission. Field tests of endosymbiont Wolbachia-based methods have yielded promising results, but there are hurdles to overcome due to the large-scale rearing and accurate sex sorting required for Wolbachia-based suppression approaches and the ecological impediments to Wolbachia invasion in replacement approaches. Genome engineering-based methods, in which mosquitoes are genetically altered for the modification or suppression of wild populations, offer an additional approach for control of mosquito-borne diseases. In particular, the use of gene drive alleles that bias inheritance in their favor is a potentially powerful approach. Several drives are frequency dependent, potentially giving them broadly similar population dynamics to Wolbachia. However, public acceptance and the behavior of released drives in natural mosquito populations remain challenges. We summarize the latest developments and discuss the knowledge gaps in both symbiont- and gene drive-based methods.}, } @article {pmid39356896, year = {2024}, author = {Lima-Camara, TN}, title = {Dengue is a product of the environment: an approach to the impacts of the environment on the Aedes aegypti mosquito and disease cases.}, journal = {Revista brasileira de epidemiologia = Brazilian journal of epidemiology}, volume = {27}, number = {}, pages = {e240048}, doi = {10.1590/1980-549720240048}, pmid = {39356896}, issn = {1980-5497}, mesh = {*Dengue/transmission/epidemiology/prevention & control ; *Aedes/virology ; Animals ; Humans ; Brazil/epidemiology ; *Climate Change ; *Mosquito Vectors ; Mosquito Control/methods ; Environment ; Socioeconomic Factors ; }, abstract = {Dengue is an arbovirus infection whose etiologic agent is transmitted by the Aedes aegypti mosquito. Since the early 1980s, when the circulation of the dengue virus (DENV) was confirmed in Brazil, the disease has become a growing multifactorial public health problem. This article presented the main factors that have contributed to the frequent dengue epidemics in recent years, such as the behavior of the vector, climate change, and social, political, and economic aspects. The intersection between these different factors in the dynamics of the disease is highlighted, including the increase in the mosquito population due to higher temperatures and rainy periods, as well as the influence of socioeconomic conditions on the incidence of dengue. Some mosquito control strategies are also addressed, including the use of innovative technologies such as drones and the Wolbachia bacterium, as well as the hope represented by the dengue vaccine. Nevertheless, the need for integrated and effective public policies to reduce social inequalities and the impacts of climate change on the spread of dengue is emphasized.}, } @article {pmid39349234, year = {2024}, author = {Montenegro-López, D and Cortés-Cortés, G and Balbuena-Alonso, MG and Warner, C and Camps, M}, title = {Wolbachia-Based Emerging Strategies for Control of Vector-Transmitted Disease.}, journal = {Acta tropica}, volume = {}, number = {}, pages = {107410}, doi = {10.1016/j.actatropica.2024.107410}, pmid = {39349234}, issn = {1873-6254}, abstract = {Dengue fever is a mosquito-transmitted disease of great public health importance. Dengue lacks adequate vaccine protection and insecticide-based methods of mosquito control are proving increasingly ineffective. Here we review the emerging use of mosquitoes transinfected with the obligate intracellular bacterium Wolbachia pipientis for vector control. Wolbachia often induces cytoplasmic incompatibility in its mosquito hosts, resulting in infertile progeny between an infected male and an uninfected female. Wolbachia infection also suppresses the replication of pathogens in the mosquito, a process known as "pathogen blocking". Two strategies have emerged. The first one releases Wolbachia-carriers (both male and female) to replace the wild mosquito population, a process driven by cytoplasmic incompatibility and that becomes irreversible once a threshold is reached. This suppresses disease transmission mainly by pathogen blocking and frequently requires a single intervention. The second strategy floods the field population with an exclusively male population of Wolbachia-carrying mosquitoes to generate infertile hybrid progeny. In this case, transmission suppression depends largely on decreasing the population density of mosquitoes caused by infertility and requires continued mosquito release. The efficacy of both Wolbachia-based approaches has been conclusively demonstrated by randomized and non-randomized studies of deployments across the world. However, results conducted in one setting cannot be directly or easily extrapolated to other settings because dengue incidence is highly affected by the conditions into which the mosquitoes are released. Compared to traditional methods, Wolbachia-based approaches are much more environmentally friendly and can be effective in the medium/long term. On the flip side, they are much more complex and cost-intensive operations, requiring a substantial investment, infrastructure, trained personnel, coordination between agencies, and community engagement. Finally, we discuss recent evidence suggesting that transinfected Wolbachia in released mosquitoes has a moderate potential risk of spreading genes in the environment.}, } @article {pmid39342132, year = {2024}, author = {Mfopit, YM and Bilgo, E and Boma, S and Somda, MB and Gnambani, JE and Konkobo, M and Diabate, A and Dayo, GK and Mamman, M and Kelm, S and Balogun, EO and Shuaibu, MN and Kabir, J}, title = {Symbiotic bacteria Sodalis glossinidius, Spiroplasma sp and Wolbachia do not favour Trypanosoma grayi coexistence in wild population of tsetse flies collected in Bobo-Dioulasso, Burkina Faso.}, journal = {BMC microbiology}, volume = {24}, number = {1}, pages = {373}, pmid = {39342132}, issn = {1471-2180}, abstract = {BACKGROUND: Tsetse flies, the biological vectors of African trypanosomes, have established symbiotic associations with different bacteria. Their vector competence is suggested to be affected by bacterial endosymbionts. The current study provided the prevalence of three tsetse symbiotic bacteria and trypanosomes in Glossina species from Burkina Faso.

RESULTS: A total of 430 tsetse flies were captured using biconical traps in four different collection sites around Bobo-Dioulasso (Bama, Bana, Nasso, and Peni), and their guts were removed. Two hundred tsetse were randomly selected and their guts were screened by PCR for the presence of Sodalis glossinidius, Spiroplasma sp., Wolbachia and trypanosomes. Of the 200 tsetse, 196 (98.0%) were Glossina palpalis gambiensis and 4 (2.0%) Glossina tachinoides. The overall symbiont prevalence was 49.0%, 96.5%, and 45.0%, respectively for S. glossinidius, Spiroplasma and Wolbachia. Prevalence varied between sampling locations: S. glossinidius (54.7%, 38.5%, 31.6%, 70.8%); Spiroplasma (100%, 100%, 87.7%, 100%); and Wolbachia (43.4%, 38.5%, 38.6%, 70.8%), respectively in Bama, Bana, Nasso and Peni. Noteworthy, no G. tachnoides was infected by S. glossinidius and Wolbachia, but they were all infected by Spiroplasma sp. A total of 196 (98.0%) harbored at least one endosymbionts. Fifty-five (27.5%) carried single endosymbiont. Trypanosomes were found only in G. p. gambiensis, but not G. tachinoides. Trypanosomes were present in flies from all study locations with an overall prevalence of 29.5%. In Bama, Bana, Nasso, and Peni, the trypanosome infection rate was respectively 39.6%, 23.1%, 8.8%, and 37.5%. Remarkably, only Trypanosoma grayi was present. Of all trypanosome-infected flies, 55.9%, 98.3%, and 33.9% hosted S. glossinidius, Spiroplasma sp and Wolbachia, respectively. There was no association between Sodalis, Spiroplasma and trypanosome presence, but there was a negative association with Wolbachia presence. We reported 1.9 times likelihood of trypanosome absence when Wolbachia was present.

CONCLUSION: This is the first survey reporting the presence of Trypanosoma grayi in tsetse from Burkina Faso. Tsetse from these localities were highly positive for symbiotic bacteria, more predominantly with Spiroplasma sp. Modifications of symbiotic interactions may pave way for disease control.}, } @article {pmid39341970, year = {2024}, author = {Shirozu, T and Regilme, MAF and Ote, M and Sasaki, M and Soga, A and Bochimoto, H and Kawabata, H and Umemiya-Shirafuji, R and Kanuka, H and Fukumoto, S}, title = {Wolbachia infection in Aedes aegypti does not affect its vectorial capacity for Dirofilaria immitis.}, journal = {Scientific reports}, volume = {14}, number = {1}, pages = {22528}, pmid = {39341970}, issn = {2045-2322}, support = {22H02510//Japan Society for the Promotion of Science/ ; }, abstract = {Mosquito-borne diseases such as dengue and filariasis are a growing public health concern in endemic countries. Biological approaches, such as the trans-infection of Wolbachia pipientis in mosquitoes, are an alternative vector control strategy, especially for arthropod-borne viruses such as dengue. In the present study, the effect of Wolbachia (wMel strain) on the vectorial capacity of Aedes aegypti for Dirofilaria immitis was studied. Our results showed that Wolbachia does not affect the phenotype of mosquito survival or the prevalence, number, and molting rate of third-stage larvae in both susceptible and resistant strains of Ae. aegypti. RNA-seq analysis of Malpighian tubules at 2 days post-infection with D. immitis showed the differentially expressed genes (DEGs) with and without wMel infection. No characteristic immune-related gene expression patterns were observed among the DEGs. No significant change in the amount of Wolbachia was observed in the Ae. aegypti after D. immitis infection. Our results suggest that infection of D. immitis in Ae. aegypti populations will not interfere with Wolbachia-based vector control strategies in dengue-endemic areas where cases of D. immitis are present. This study demonstrated the veterinary medical validity of a dengue control program using Wolbachia.}, } @article {pmid39338567, year = {2024}, author = {Salvioni Recalde, OD and Rolón, MS and Velázquez, MC and Kowalewski, MM and Alfonso Ruiz Diaz, JJ and Rojas de Arias, A and Moraes, MO and Magdinier Gomes, H and de Azevedo Baêta, B and Dias Cordeiro, M and Vega Gómez, MC}, title = {Diversity of Anaplasmataceae Transmitted by Ticks (Ixodidae) and the First Molecular Evidence of Anaplasma phagocytophilum and Candidatus Anaplasma boleense in Paraguay.}, journal = {Microorganisms}, volume = {12}, number = {9}, pages = {}, doi = {10.3390/microorganisms12091893}, pmid = {39338567}, issn = {2076-2607}, support = {COF N°03/11//Organization for the Structural Convergence in the Mercosur Region FOCEM/ ; 14-INV-052 and PINV18-178//National Council of Science and Technology CONACYT-Paraguay/ ; }, abstract = {Anaplasmataceae bacteria are emerging infectious agents transmitted by ticks. The aim of this study was to identify the molecular diversity of this bacterial family in ticks and hosts, both domestic and wild, as well as blood meal sources of free-living ticks in northeastern Paraguay. The bacteria were identified using PCR-HRM, a method optimized for this purpose, while the identification of ticks and their blood meal was performed using conventional PCR. All amplified products were subsequently sequenced. The bacteria detected in the blood hosts included Ehrlichia canis, Anaplasma platys, and Anaplasma phagocytophilum, Candidatus Anaplasma boleense, and Wolbachia spp., which had not been previously reported in the country. Free-living and parasitic ticks on dogs (Canis lupus familiaris) and wild armadillos (Dasypus novemcinctus) were collected and identified as Rhipicephalus sanguineus and Amblyomma spp. The species E. canis, A. platys, A. phagocytophilum, and Ca. A. boleense were detected in domestic dog ticks, and E. canis and A. platys were found for the first time in armadillos and free-living ticks. Blood feeding sources detected in free-living ticks were rodents, humans, armadillos and dogs. Results show a high diversity of tick-borne pathogens circulating among domestic and wild animals in the northeastern region of Paraguay.}, } @article {pmid39336695, year = {2024}, author = {Li, J and Tian, X and Hsiang, T and Yang, Y and Shi, C and Wang, H and Li, W}, title = {Microbial Community Structure and Metabolic Function in the Venom Glands of the Predatory Stink Bug, Picromerus lewisi (Hemiptera: Pentatomidae).}, journal = {Insects}, volume = {15}, number = {9}, pages = {}, doi = {10.3390/insects15090727}, pmid = {39336695}, issn = {2075-4450}, support = {(ZK [2023] 023)//Key Project of Guizhou Provincial Science and Technology Foundation/ ; (ZK-2021-Key036, Qiankehe Talent Platform - CXTD [2023]021)//Guizhou Science Technology Foundation/ ; (GCC[2022]028-1)//Hundred' Level Innovative Talent Foundation of Guizhou Province/ ; }, abstract = {The predatory stink bug, Picromerus lewisi (Hemiptera: Pentatomidae), is an important and valuable natural enemy of insect pests in their ecosystems. While insects are known to harbor symbiotic microorganisms, and these microbial symbionts play a crucial role in various aspects of the host's biology, there is a paucity of knowledge regarding the microbiota present in the venom glands of P. lewisi. This study investigated the venom glands of adult bugs using both traditional in vitro isolation and cultural methods, as well as Illumina high-throughput sequencing technology. Additionally, the carbon metabolism of the venom gland's microorganisms was analyzed using Biolog ECO metabolic phenotyping technology. The results showed 10 different culturable bacteria where the dominant ones were Enterococcus spp. and Lactococcus lactis. With high-throughput sequencing, the main bacterial phyla in the microbial community of the venom glands of P. lewisi were Proteobacteria (78.1%) and Firmicutes (20.3%), with the dominant bacterial genera being Wolbachia, Enterococcus, Serratia, and Lactococcus. At the fungal community level, Ascomycota accounted for the largest proportion (64.1%), followed by Basidiomycota (27.6%), with Vishniacozyma, Cladosporium, Papiliotrema, Penicillium, Fusarium, and Aspergillus as the most highly represented fungal genera. The bacterial and fungal community structure of the venom glands of P. lewisi exhibited high species richness and diversity, along with a strong metabolism of 22 carbon sources. Functional prediction indicated that the primary dominant function of P. lewisi venom-gland bacteria was metabolism. The dominant eco-functional groups of the fungal community included undefined saprotroph, fungal parasite-undefined saprotroph, unassigned, endophyte-plant pathogen, plant pathogen-soil saprotroph-wood saprotroph, animal pathogen-endophyte-plant pathogen-wood saprotroph, plant pathogen, and animal pathogen-endophyte-epiphyte-plant pathogen-undefined saprotroph. These results provide a comprehensive characterization of the venom-gland microbiota of P. lewisi and demonstrate the stability (over one week) of the microbial community within the venom glands. This study represents the first report on the characterization of microbial composition from the venom glands of captive-reared P. lewisi individuals. The insights gained from this study are invaluable for future investigations into P. lewisi's development and the possible interactions between P. lewisi's microbiota and some Lepidopteran pests.}, } @article {pmid39336625, year = {2024}, author = {Kepngop, LRK and Wosula, EN and Amour, M and Ghomsi, PGT and Wakam, LN and Kansci, G and Legg, JP}, title = {Genetic Diversity of Whiteflies Colonizing Crops and Their Associated Endosymbionts in Three Agroecological Zones of Cameroon.}, journal = {Insects}, volume = {15}, number = {9}, pages = {}, doi = {10.3390/insects15090657}, pmid = {39336625}, issn = {2075-4450}, support = {S/CMR21-10//Arturo Falaschi ICGEB fellowships program/ ; N/A//The CGIAR Trust Fund: https://www.cgiar.org/funders through the CGIAR Initiative on Plant Health and Rapid Response to Protect Food Security and Livelihoods (Plant Health Initiative)./ ; }, abstract = {Bemisia tabaci (Gennadius) is as a major pest of vegetable crops in Cameroon. These sap-sucking insects are the main vector of many viruses infecting plants, and several cryptic species have developed resistance against insecticides. Nevertheless, there is very little information about whitefly species on vegetable crops and the endosymbionts that infect them in Cameroon. Here, we investigated the genetic diversity of whiteflies and their frequency of infection by endosymbionts in Cameroon. Ninety-two whitefly samples were collected and characterized using mitochondrial cytochrome oxidase I (mtCOI) markers and Kompetitive Allele Specific PCR (KASP). The analysis of mtCOI sequences of whiteflies indicated the presence of six cryptic species (mitotypes) of Bemisia tabaci, and two distinct clades of Bemisia afer and Trialeurodes vaporariorum. Bemisia tabaci mitotypes identified included: MED on tomato, pepper, okra, and melon; and SSA1-SG1, SSA1-SG2, SSA1-SG5, SSA3, and SSA4 on cassava. The MED mitotype predominated in all regions on the solanaceous crops, suggesting that MED is probably the main phytovirus vector in Cameroonian vegetable cropping systems. The more diverse cassava-colonizing B. tabaci were split into three haplogroups (SNP-based grouping) including SSA-WA, SSA4, and SSA-ECA using KASP genotyping. This is the first time that SSA-ECA has been reported in Cameroon. This haplogroup is predominant in regions currently affected by the severe cassava mosaic virus disease (CMD) and cassava brown streak virus disease (CBSD) pandemics. Three endosymbionts including Arsenophonus, Rickettsia, and Wolbachia were present in female whiteflies tested in this study with varying frequency. Arsenophonus, which has been shown to influence the adaptability of whiteflies, was more frequent in the MED mitotype (75%). Cardinium and Hamiltonella were absent in all whitefly samples. These findings add to the knowledge on the diversity of whiteflies and their associated endosymbionts, which, when combined, influence virus epidemics and responses to whitefly control measures, especially insecticides.}, } @article {pmid39336607, year = {2024}, author = {Lilja, T and Lindström, A and Hernández-Triana, LM and Di Luca, M and Lwande, OW}, title = {European Culex pipiens Populations Carry Different Strains of Wolbachia pipientis.}, journal = {Insects}, volume = {15}, number = {9}, pages = {}, doi = {10.3390/insects15090639}, pmid = {39336607}, issn = {2075-4450}, support = {2020-01056//Formas, Sweden/ ; }, abstract = {The mosquito Culex pipiens occurs in two ecotypes differing in their mating and overwintering behavior: pipiens mate in open environments and diapause, and molestus also mate in small spaces and is active throughout the year. Cx. pipiens carry Wolbachia endosymbionts of the wPip strain, but the frequency of infection differs between studied populations. Wolbachia infection affects the host reproductive success through cytoplasmic incompatibility. wPip Wolbachia is divided into five types, wPip I-V. The type of wPip carried varies among Cx. pipiens populations. In northern European locations different wPip types are found in the two ecotypes, whereas in southern locations, they often carry the same type, indicating differences in hybridization between ecotypes. In this study, Cx. pipiens specimens of both ecotypes were collected from Sweden and compared to specimens from Norway, England, Italy, and the Netherlands, as well as Cx. quinquefasciatus from Mali and Thailand. The abundance varied, but all specimens were infected by Wolbachia, while the tested specimens of other mosquito species were often uninfected. The wPip strains were determined through the sequence analysis of Wolbachia genes ank2 and pk1, showing that Cx. pipiens ecotypes in Scandinavia carry different wPip strains. The observed differences in wPip strains indicate that hybridization is not frequent and may contribute to barriers against hybridization of the ecotypes in Sweden and Norway.}, } @article {pmid39325838, year = {2024}, author = {Liang, Y and Liu, J and Wu, Y and Wu, Y and Xi, Z}, title = {Stable introduction of Wolbachia wPip into invasive Anopheles stephensi for potential malaria control.}, journal = {PLoS neglected tropical diseases}, volume = {18}, number = {9}, pages = {e0012523}, doi = {10.1371/journal.pntd.0012523}, pmid = {39325838}, issn = {1935-2735}, abstract = {The spread and invasion of the urban malaria vector Anopheles stephensi has emerged as a significant threat to ongoing malaria control and elimination efforts, particularly in Africa. The successful use of the maternally inherited endosymbiotic bacterium Wolbachia for arbovirus control has inspired the exploration of similar strategies for managing malaria vectors, necessitating the establishment of a stable Wolbachia-Anopheles symbiosis. In this study, we successfully transferred Wolbachia wPip into An. stephensi, resulting in the establishment of a stable transinfected HP1 line with 100% maternal transmission efficiency. We demonstrate that wPip in the HP1 line induces nearly complete unidirectional cytoplasmic incompatibility (CI) and maintains high densities in both somatic and germline tissues. Despite a modest reduction in lifespan and female reproductive capacity, our results suggest the Wolbachia infection in the HP1 line has little impact on life history traits, body size, and male mating competitiveness, as well as the ability of its larvae to tolerate rearing temperatures up to 38°C, although wPip densities moderately decrease when larvae are exposed to a constant 33°C and diurnal cyclic temperatures of 27-36°C and 27-38°C. These findings highlight the potential of the HP1 line as a robust candidate for further development in malaria control.}, } @article {pmid39325836, year = {2024}, author = {Holt, CI and Dunning Hotopp, JC}, title = {Updated annotation and meta-analysis of Brugia malayi transcriptomics data reveals consistent transcriptional profiles across time and space with some study-specific differences in adult female worm transcriptional profiles.}, journal = {PLoS neglected tropical diseases}, volume = {18}, number = {9}, pages = {e0012511}, doi = {10.1371/journal.pntd.0012511}, pmid = {39325836}, issn = {1935-2735}, abstract = {Genomics, transcriptomics, and proteomics have significantly advanced our understanding of obligately host-associated microbes, where interrogation of the biology is often limited by the complexity of the biological system and limited tools. This includes the causative agents of many neglected tropical diseases, including filarial nematodes. Therefore, numerous transcriptomics studies have been undertaken on filarial nematodes. Most of these transcriptomics studies focus on Brugia malayi, which causes lymphatic filariasis and is a laboratory model for human filarial disease. Here, we undertook a meta-analysis of the publicly available B. malayi transcriptomics data enabling the direct cross comparison of samples from almost a dozen studies. This reanalysis highlights the consistency of transcriptomics results across many different studies and experimental designs from across the globe for over a decade of research, across many different generations of a sequencing technology, library preparation protocols, and differential expression tools. Males and microfilariae across samples had similar expression profiles. However, female samples were clustered into two differential expression patterns that were significantly different from one another. Largely, we confirm previous results for all studies reanalyzed including tissue-specific gene expression and anti-Wolbachia doxycycline treatment of microfilaria. However, we did not detect previously reported differential expression upon in vitro or in vivo treatment with ivermectin, albendazole, and DEC, instead identifying a consistent lack of transcriptomic change upon exposure to these anthelminthic drugs. Updated annotation has been provided that denotes poorly supported genes including those overlapping rRNAs.}, } @article {pmid39321893, year = {2024}, author = {Behrmann, LV and Lämmer, C and Schiefer, A and Neufeld, H and Grosse, M and Stadler, M and Bierbaum, G and Hoerauf, A and Pfarr, K}, title = {No resistance development against corallopyronin A in Wolbachia in C6/36 cell culture.}, journal = {International journal of antimicrobial agents}, volume = {}, number = {}, pages = {107344}, doi = {10.1016/j.ijantimicag.2024.107344}, pmid = {39321893}, issn = {1872-7913}, } @article {pmid39320534, year = {2024}, author = {Elsawy, BSM and Abdel-Ghany, HSM and Alzan, HF and Abdel-Shafy, S and Shahein, YE}, title = {Molecular screening of piroplasms and Anaplasmataceae agents in Hyalomma dromedarii ticks from camels over different seasons in Egypt.}, journal = {Experimental & applied acarology}, volume = {}, number = {}, pages = {}, pmid = {39320534}, issn = {1572-9702}, abstract = {Piroplasmosis, a disease of domestic and wild animals, is caused by tick-borne protozoa of the genera Babesia and Theileria, while anaplasmosis is caused by tick-borne bacteria of genera Anaplasma. Hyalomma dromedarii is the most dominant tick species infesting camels in Egypt and act as a vector of piroplasms, Anaplasma, Rickettsia and Ehrlichia spp. The available information concerning the detection of these pathogens in H. dromedarii infesting camels is limited. The present study aimed to evaluate the status of these pathogens in H. dromedarii ticks over four seasons of a year, in addition to investigate the infections of piroplasms and Anaplasmataceae besides their genetic diversity starting from June 2021 till April 2022. A total of 275 semi-engorged females of H. dromedarii were collected from different slaughtered camels, Toukh city slaughterhouse then investigated by Polymerase Chain Reaction (PCR) to detect piroplasms (Babesia spp., Theileria spp.) and Anaplasmataceae DNA targeting 18 S rRNA and 16 S rRNA genes, respectively followed by sequencing and phylogenetic analyses. Overall, piroplasms were detected in 38 ticks (13.8%), Babesia spp. was detected in 35 ticks (12.7%), while Theileria spp. was detected in one tick (0.4%). Anaplasmataceae was detected in 57 ticks (20.7%). Mixed infections of piroplasms and Anaplasmataceae were detected in 13 ticks (5%). Single infection either with piroplasms or Anaplasmataceae was detected in 25 (9%) and 44 (16%) ticks, respectively. The highest monthly rate of piroplasms was in April (spring) and Anaplasmataceae was in July (summer). Sequence analysis revealed that Babesia bigemina, Wolbachia spp. and Anaplasma marginale are the most dominant species in the examined tick samples. To the best of our knowledge, this study confirms the presence of B. bigemina, Wolbachia spp. and A. marginale in H. dromedarii in Egypt by sequencing.}, } @article {pmid39319953, year = {2024}, author = {Gurgel-Gonçalves, R and Oliveira, WK and Croda, J}, title = {The greatest Dengue epidemic in Brazil: Surveillance, Prevention, and Control.}, journal = {Revista da Sociedade Brasileira de Medicina Tropical}, volume = {57}, number = {}, pages = {e002032024}, doi = {10.1590/0037-8682-0113-2024}, pmid = {39319953}, issn = {1678-9849}, mesh = {*Dengue/prevention & control/epidemiology ; Humans ; Brazil/epidemiology ; *Mosquito Control/methods ; Animals ; *Aedes/virology ; *Epidemics/prevention & control ; *Mosquito Vectors ; Dengue Vaccines/administration & dosage ; Population Surveillance ; }, abstract = {In this review, we discuss dengue surveillance, prevention, and control measures in Brazil. Data on dengue epidemics between 2000 and 2024 indicates an increase in the number of dengue cases and deaths. Global climate change is a key driver of this growth. Over the past 25 years, nearly 18 million Brazilians have been infected with the dengue virus, and the highest number of dengue cases in Brazil's history is projected to reach 2024. Dengue mortality in Brazil increased geographically over time. As of June, there were approximately 6 million probable cases and 4,000 confirmed deaths in Brazil, which represents the greatest dengue epidemic to date. Several technologies have been developed to control Aedes aegypti, including the deployment of Wolbachia-infected mosquitoes, indoor residual spraying, sterile insect techniques, and mosquito-disseminated insecticides. The Ministry of Health recommends integrating these technologies into health services. Brazil is the first country to incorporate the Takeda vaccine into its public health system, and the Butantan vaccine is currently undergoing Phase 3 clinical trials. Increasing the vaccination coverage and implementing novel Ae. aegypti control technologies could reduce the number of dengue cases in Brazil in the coming years. Community activities such as home cleaning and elimination of potential mosquito breeding sites, facilitated by social media and health education initiatives, must continue to achieve this reduction. Ultimately, a multisectoral approach encompassing sanitary improvements, mosquito control, vaccination, and community mobilization is crucial in the fight against dengue epidemics.}, } @article {pmid39313916, year = {2024}, author = {Zhu, YX and Zhang, YY and Wang, XY and Yin, Y and Du, YZ}, title = {Wolbachia modify host cell metabolite profiles in response to short-term temperature stress.}, journal = {Environmental microbiology reports}, volume = {16}, number = {5}, pages = {e70013}, doi = {10.1111/1758-2229.70013}, pmid = {39313916}, issn = {1758-2229}, support = {BK20231330//The Natural Science Foundation of Jiangsu Province/ ; }, mesh = {*Wolbachia/metabolism/physiology/genetics ; Animals ; *Stress, Physiological ; *Temperature ; Cell Line ; *Metabolome ; Drosophila/microbiology ; Symbiosis ; Diptera/microbiology ; Fatty Acids/metabolism ; }, abstract = {Wolbachia are common heritable endosymbionts that influence many aspects of ecology and evolution in various insects, yet Wolbachia-mediated intracellular metabolic responses to temperature stress have been largely overlooked. Here, we introduced the Wolbachia strain wLhui from the invasive Liriomyza huidobrensis (Blanchard) into a Drosophila Schneider 2 cell line (S2) and investigated the metabolite profile of wLhui-infected (S2_wLhui) and uninfected cell lines (S2_wu) under short-term exposure to either high (37°C), moderate (27°C), or low (7 and 17°C) temperatures. We find that Wolbachia infection, temperature stress, and their interactions significantly affect cellular metabolic profiles. Most significantly, when comparing the changes in metabolites between S2_wLhui and S2_wu, glycerophospholipids, amino acids, and fatty acids associated with metabolic pathways, microbial metabolism in diverse environments, and other pathways were significantly accumulated at either low or high temperatures. Our findings suggest Wolbachia-induced cellular physiological responses to short-term temperature stress, which may in turn affect the fitness and adaptive ability of its host as an invasive species.}, } @article {pmid39310793, year = {2024}, author = {Lečić, S and Wolfe, TM and Ghosh, A and Satar, S and Souza Beraldo, C and Smith, E and Dombroskie, JJ and Jernigan, E and Hood, GR and Schuler, H and Stauffer, C}, title = {Spatially Varying Wolbachia Frequencies Reveal the Invasion Origin of an Agricultural Pest Recently Introduced From Europe to North America.}, journal = {Evolutionary applications}, volume = {17}, number = {9}, pages = {e70016}, pmid = {39310793}, issn = {1752-4571}, abstract = {The introduction of non-native species across the world represents a major global challenge. Retracing invasion origin is an important first step in understanding the invasion process, often requiring detailed sampling within the native range. Insect species frequently host Wolbachia, a widespread endosymbiotic bacterium that manipulates host reproduction to increase infected female fitness. Here, we draw on the spatial variation in infection frequencies of an actively spreading Wolbachia strain wCer2 to investigate the invasion origin of the European cherry fruit fly, Rhagoletis cerasi. This pest of cherries was introduced from Europe to North America within the last decade. First, we screen the introduced fly population for the presence of Wolbachia. The introduced populations lack the wCer2 strain and the strongly associated mitochondrial haplotype, suggesting strain absence due to founder effects with invading individuals originating from wCer2-uninfected native population(s). To narrow down geographic regions of invasion origin, we perform spatial interpolation of the wCer2 infection frequency across the native range and predict the infection frequency in unsampled regions. For this, we use an extensive dataset of R. cerasi infection covering 238 populations across Europe over 25 years, complemented with 14 additional populations analyzed for this study. We find that R. cerasi was unlikely introduced from wCer2-infected populations in Central and Western Europe. We propose wCer2-uninfected populations from Eastern Europe and the Mediterranean region as the most likely candidates for the invasion origin. This work utilizes Wolbachia as an indirect instrument to provide insights into the invasion source of R. cerasi in North America, revealing yet another application for this multifaceted heritable endosymbiont. Given the prevalence of biological invasions, rapidly uncovering invasion origins gives fundamental insights into how invasive species adapt to new environments.}, } @article {pmid39298440, year = {2024}, author = {Hafsia, S and Barbar, T and Alout, H and Baudino, F and Lebon, C and Gomard, Y and Wilkinson, DA and Fourié, T and Mavingui, P and Atyame, C}, title = {Vector competence of Aedes albopictus field populations from Reunion Island exposed to local epidemic dengue viruses.}, journal = {PloS one}, volume = {19}, number = {9}, pages = {e0310635}, doi = {10.1371/journal.pone.0310635}, pmid = {39298440}, issn = {1932-6203}, mesh = {Animals ; *Aedes/virology/microbiology ; *Dengue Virus/physiology ; *Wolbachia/physiology ; *Dengue/transmission/epidemiology/virology ; Reunion/epidemiology ; *Mosquito Vectors/virology/microbiology ; Viral Load ; Humans ; Insect Vectors/virology/microbiology ; Female ; }, abstract = {Dengue virus (DENV) is the most prevalent mosquito-borne Flavivirus that affects humans worldwide. Aedes albopictus, which is naturally infected with the bacteria Wolbachia, is considered to be a secondary vector of DENV. However, it was responsible for a recent DENV outbreak of unprecedented magnitude in Reunion Island, a French island in the South West Indian Ocean. Moreover, the distribution of the cases during this epidemic showed a spatially heterogeneous pattern across the island, leading to questions about the differential vector competence of mosquito populations from different geographic areas. The aim of this study was to gain a better understanding of the vector competence of the Ae. albopictus populations from Reunion Island for local DENV epidemic strains, while considering their infection by Wolbachia. Experimental infections were conducted using ten populations of Ae. albopictus sampled across Reunion Island and exposed to three DENV strains: one strain of DENV serotype 1 (DENV-1) and two strains of DENV serotype 2 (DENV-2). We analyzed three vector competence parameters including infection rate, dissemination efficiency and transmission efficiency, at different days post-exposition (dpe). We also assessed whether there was a correlation between the density of Wolbachia and viral load/vector competence parameters. Our results show that the Ae. albopictus populations tested were not able to transmit the two DENV-2 strains, while transmission efficiencies up to 40.79% were observed for the DENV-1 strain, probably due to difference in viral titres. Statistical analyses showed that the parameters mosquito population, generation, dpe and area of sampling significantly affect the transmission efficiencies of DENV-1. Although the density of Wolbachia varied according to mosquito population, no significant correlation was found between Wolbachia density and either viral load or vector competence parameters for DENV-1. Our results highlight the importance of using natural mosquito populations for a better understanding of transmission patterns of dengue.}, } @article {pmid39296285, year = {2024}, author = {Wang, X and Huangfu, N and Chen, L and Zhang, K and Li, D and Gao, X and Li, B and Wang, L and Zhu, X and Ji, J and Luo, J and Cui, J}, title = {Effects of developmental stages, sex difference, and diet types of the host marmalade hoverfly (Episyrphus balteatus) on symbiotic bacteria.}, journal = {Frontiers in microbiology}, volume = {15}, number = {}, pages = {1433909}, pmid = {39296285}, issn = {1664-302X}, abstract = {INTRODUCTION: Symbiotic bacteria play key roles in a variety of important life processes of insects such as development, reproduction and environmental adaptation, and the elucidation of symbiont population structure and dynamics is crucial for revealing the underlying regulatory mechanisms. The marmalade hoverfly (Episyrphus balteatus) is not only a remarkable aphid predator, but also a worldwide pollinator second to honeybees. However, its symbiont composition and dynamics remain unclear.

METHODS: Herein, we investigate the symbiotic bacterial dynamics in marmalade hoverfly throughout whole life cycle, across two sexes, and in its prey Megoura crassicauda by 16S rRNA sequencing.

RESULTS: In general, the dominant phyla were Proteobacteria and Firmicutes, and the dominant genera were Serratia and Wolbachia. Serratia mainly existed in the larval stage of hoverfly with the highest relative abundance of 86.24% in the 1st instar larvae. Wolbachia was found in adults and eggs with the highest relative abundance of 62.80% in eggs. Significant difference in species diversity was observed between the adults feeding on pollen and larvae feeding on M. crassicauda, in which the dominant symbiotic bacteria were Asaia and Serratia, respectively. However, between two sexes, the symbionts exhibited high similarity in species composition. In addition, our results suggested that E. balteatus obtainded Serratia mainly through horizontal transmission by feeding on prey aphids, whereas it acquired Wolbachia mainly through intergeneration vertical transmission. Taken together, our study revealed the effects of development stages, diet types and genders of E. balteatus on symbionts, and explored transmission modes of dominant bacteria Serratia and Wolbachia.

DISCUSSION: Our findings lay a foundation for further studying the roles of symbiotic bacteria in E. balteatus life cycle, which will benefit for revealing the co-adaptation mechanisms of insects and symbiotic bacteria.}, } @article {pmid37941933, year = {2023}, author = {Caamal-Chan, MG and Barraza, A and Loera-Muro, A and Montes-Sánchez, JJ and Castellanos, T and Rodríguez-Pagaza, Y}, title = {Bacterial communities of the psyllid pest Bactericera cockerelli (Hemiptera: Triozidae) Central haplotype of tomato crops cultivated at different locations of Mexico.}, journal = {PeerJ}, volume = {11}, number = {}, pages = {e16347}, pmid = {37941933}, issn = {2167-8359}, mesh = {Animals ; Haplotypes ; *Solanum lycopersicum ; RNA, Ribosomal, 16S/genetics ; *Hemiptera/genetics ; Phylogeny ; Mexico ; Bacteria/genetics ; Liberibacter/genetics ; Crops, Agricultural/genetics ; }, abstract = {BACKGROUND: The psyllid, Bactericera cockerelli, is an insect vector of 'Candidatus Liberibacter' causing "Zebra chip" disease that affects potato and other Solanaceae crops worldwide. In the present study, we analyzed the bacterial communities associated with the insect vector Bactericera cockerelli central haplotype of tomato crop fields in four regions from Mexico.

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

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

CONCLUSIONS: Our data support that the members with the highest representation in microbial community of B. cockerelli central haplotype, comprise their obligate symbiont, Carsonella, and facultative symbionts. We also found evidence that among the factors analyzed, the presence of the plant pathogen affects the structure and composition of the bacterial community associated with B. cockerelli.}, } @article {pmid39288812, year = {2024}, author = {Castelli, M and Nardi, T and Giovannini, M and Sassera, D}, title = {Addictive manipulation: a perspective on the role of reproductive parasitism in the evolution of bacteria-eukaryote symbioses.}, journal = {Biology letters}, volume = {20}, number = {9}, pages = {20240310}, doi = {10.1098/rsbl.2024.0310}, pmid = {39288812}, issn = {1744-957X}, mesh = {*Symbiosis ; Animals ; *Biological Evolution ; *Reproduction ; Eukaryota/physiology ; Arthropods/microbiology/physiology ; Wolbachia/physiology/genetics ; Toxin-Antitoxin Systems/genetics ; Bacteria/genetics ; }, abstract = {Wolbachia bacteria encompass noteworthy reproductive manipulators of their arthropod hosts. which influence host reproduction to favour their own transmission, also exploiting toxin-antitoxin systems. Recently, multiple other bacterial symbionts of arthropods have been shown to display comparable manipulative capabilities. Here, we wonder whether such phenomena are truly restricted to arthropod hosts. We focused on protists, primary models for evolutionary investigations on eukaryotes due to their diversity and antiquity, but still overall under-investigated. After a thorough re-examination of the literature on bacterial-protist interactions with this question in mind, we conclude that such bacterial 'addictive manipulators' of protists do exist, are probably widespread, and have been overlooked until now as a consequence of the fact that investigations are commonly host-centred, thus ineffective to detect such behaviour. Additionally, we posit that toxin-antitoxin systems are crucial in these phenomena of addictive manipulation of protists, as a result of recurrent evolutionary repurposing. This indicates intriguing functional analogy and molecular homology with plasmid-bacterial interplays. Finally, we remark that multiple addictive manipulators are affiliated with specific bacterial lineages with ancient associations with diverse eukaryotes. This suggests a possible role of addictive manipulation of protists in paving the way to the evolution of bacteria associated with multicellular organisms.}, } @article {pmid39275847, year = {2024}, author = {MacDonald, ZG and Schoville, S and Escalona, M and Marimuthu, MPA and Nguyen, O and Chumchim, N and Fairbairn, CW and Seligmann, W and Toffelmier, E and Gillespie, T and Shaffer, HB}, title = {A genome assembly for the Chryxus Arctic (Oeneis chryxus), the highest butterfly in North America.}, journal = {The Journal of heredity}, volume = {}, number = {}, pages = {}, doi = {10.1093/jhered/esae051}, pmid = {39275847}, issn = {1465-7333}, abstract = {We describe a highly contiguous and complete diploid genome assembly for the Chryxus Arctic, Oeneis chryxus (E. Doubleday, [1849]), a butterfly species complex spanning much of northern and western North America. One subspecies, the Ivallda Arctic (O. c. ivallda), is endemic to California's Sierra Nevada and of particular biogeographic interest and conservation concern. Extreme alpine habitats occupied by this subspecies include the summit of Mt. Whitney, California, representing the highest elevation butterfly population in North America. The assembly presented here consists of two haplotypes, 738.92 and 770.85 Mb in length, with contig N50 values of 10.49 and 10.13 Mb, scaffold N50 values of 25.35 and 25.69 Mb, scaffold L50 values of 13 and 14, and BUSCO completeness scores of 96.5 and 98.3%, respectively. More than 97% of the assembly is organized into 29 scaffolds, which likely represent whole chromosomes. This assembly is the first major genomic resource for Oeneis, providing a foundational reference for future genomic studies on the taxonomy, evolutionary history, and conservation of the genus. As part of the California Conservation Genomics Project, we will use this assembly in conjunction with short-read resequencing to resolve patterns of evolutionary differentiation, adaptive genomic variation, and gene flow among remaining O. c. ivallda populations. These data can and will be used to inform the subspecies' conservation as warming climatic conditions continue to lead to the loss and fragmentation of alpine habitats. We also provide genome assemblies for the O. chryxus mitochondrion and a Wolbachia endosymbiont.}, } @article {pmid39270964, year = {2024}, author = {Giannotta, MM and Smith, I and Michie, M and Blasdell, K and Dunn, M and Nicholls, J and Heath, ACG and Rodriguez, J and Gofton, AW}, title = {Molecular characterisation of Australasian Ixodiphagus (Hymenoptera; Encyrtidae; Encyrtinae) reveals unexpected diversity and a potential novel host switch.}, journal = {International journal for parasitology}, volume = {}, number = {}, pages = {}, doi = {10.1016/j.ijpara.2024.09.001}, pmid = {39270964}, issn = {1879-0135}, abstract = {Ticks are important medical and veterinary parasites that represent a substantial health threat to humans, companion animals, and livestock. Ixodiphagus wasps (Hymenoptera; Encyrtidae) are known endoparasitoids of ixodid (hard) and argasid (soft) ticks, with potential utility as natural biocontrol agents. Two species, Ixodiphagus brunneus and Ixodiphagus mysorensis, are previously recorded from Australia, however, the genus lacks formal revisionary work in Australia, and the validity and host ranges of these species remain uncertain. This work aimed to investigate the diversity of Ixodiphagus in Australasia and provide a molecular data resource for future work on these understudied endoparasitoids. We extracted DNA from archival Ixodiphagus specimens from Australian and New Zealand insect collections and performed high-throughput sequencing which resulted in complete or mostly complete mitochondrial genome sequences from 11 specimens, including I. brunneus, Ixodiphagus taiaroaensis, and a novel Ixodiphagus sp. reared from Rhipicephalus linnaei from Townsville, Australia. In addition, approximately 70% of the genome of the Wolbachia endosymbiont of I. brunneus was recovered. Finally, we screened 178 recently collected pooled tick samples from southern New South Wales, Australia, for Ixodiphagus spp. using 28S rRNA and cytochrome c oxidase subunit 1(COI) gene PCR, and recovered 14 positive samples. Phylogenetic analysis of Australasian Ixodiphagus spp. based on 28S rRNA and complete mitochondrial genome sequences determined that members of the Australasian fauna are distinct from Ixodiphagus hookeri (the only other Ixodiphagus species for which genetic data exists), and that at least two distinct species are present in Australia; I. brunneus identified from Ixodes holocyclus and Haemaphysalis bancrofti ticks, and an uncharacterised Ixodiphagus sp. found in Rhipicephalus linnaei ticks from northern Queensland. Furthermore, there was substantial genetic diversity at the 28S rRNA loci among I. brunneus samples, which may represent normal genetic variability or a secondary cryptic species. The molecular data generated here represents the first known for the genus Ixodiphagus in Australasia, doubling that of the world fauna, and provides the first known complete mitochondrial genomes for these important tick parasitoids.}, } @article {pmid39270602, year = {2024}, author = {González, MA and Ruiz-Arrondo, I and Magallanes, S and Oboňa, J and Ruiz-López, MJ and Figuerola, J}, title = {Molecular and morphological analysis revealed a new Lipoptena species (Diptera: Hippoboscidae) in southern Spain harbouring Coxiella burnetii and bacterial endosymbionts.}, journal = {Veterinary parasitology}, volume = {332}, number = {}, pages = {110300}, doi = {10.1016/j.vetpar.2024.110300}, pmid = {39270602}, issn = {1873-2550}, abstract = {Hippoboscid flies (Diptera: Hippoboscidae) are obligate bloodsucking ectoparasites of animals. In Europe, limited research has been conducted on this family until the recent introduction of the deer ked Lipoptena fortisetosa Maa, 1965. A new species of the genus Lipoptena, Lipoptena andaluciensis sp. nov., was found in southern Spain after extensive sampling with carbon-dioxide baited suction traps. A total of 52 females and 32 males were collected at 29 out of 476 sites examined over eight months in 2023. Lipoptena andaluciensis sp. nov. was characterized morphologically and molecularly. The new Lipoptena species can be differentiated from the closely related L. fortisetosa by size, chaetotaxy of the dorsal and ventral thorax, abdominal plates, and genitalia. Based on DNA-barcoding, our specimens showed the highest similarity with Melophagus ovinus (Linnaeus, 1758) (88.4 %) and with L. fortisetosa (86-88 %). Individual screening of Lipoptena specimens (n = 76) for seven important zoonotic pathogens such as bacteria (Anaplasmataceae family: Bartonella spp., Borrelia spp., Coxiella burnetii and Rickettsia spp.) and protozoans (Babesia spp. and Theileria spp.) by conventional PCR and RT-PCR was performed. DNA of C. burnetii was detected in one specimen, while two other specimens harboured Anaplasmataceae (Wolbachia spp., 100 % homology and another endosymbiont probably related to Arsenophonus sp., 95.3 % homology, respectively), all representing the first records of these bacteria in the Lipoptena spp. from Europe. Carbon dioxide traps probed its effectiveness as a reliable passive method for keds surveillance. Our study highlights the existence of a new Lipoptena species, presumably widely distributed in southern Spain. The role of this species in the transmission cycle of pathogens of medical-veterinary relevance needs to be considered in the area.}, } @article {pmid39266798, year = {2024}, author = {Kang, SF and Chen, Y and Chen, J}, title = {Wolbachia of phylogenetic supergroup K identified in oribatid mite Nothrus anauniensis (Acari: Oribatida: Nothridae).}, journal = {Experimental & applied acarology}, volume = {}, number = {}, pages = {}, pmid = {39266798}, issn = {1572-9702}, abstract = {Heritable endosymbionts widely occur in arthropod and nematode hosts. Among these endosymbionts, Wolbachia has been extensively detected in many arthropods, such as insects and crustaceans. Maternal inheritance is the most basic and dominant mode of transmission of Wolbachia, and it might regulate the reproductive system of the host in four ways: feminization, parthenogenesis, male killing, and cytoplasmic incompatibility. There is a relatively high percentage (10%) of thelytokous species in Oribatida, a suborder under the subclass Acari of arthropods, but the study of the endosymbionts in oribatid mites is almost negligible. In this paper, we detected endosymbiotic bacteria in two parthenogenetic oribatid species, Nothrus anauniensis Canestrini and Fanzago, 1877, which has never been tested for endosymbionts, and Oppiella nova, in which Wolbachia and Cardinium have been reported before. The results showed that Wolbachia was first found in N. anauniensis with an infection rate of 100% across three populations. Phylogenetic analysis showed that Wolbachia in N. anauniensis belonged to the supergroup K, marking the second supergroup of Wolbachia found in oribatid mites. Unlike previous studies, our study did not detect Wolbachia in O. nova, leading to the exclusion of Wolbachia's role in mediating thelytoky in this species.}, } @article {pmid39264945, year = {2024}, author = {Zang, C and Wang, X and Liu, Y and Wang, H and Sun, Q and Cheng, P and Zhang, Y and Gong, M and Liu, H}, title = {Wolbachia and mosquitoes: Exploring transmission modes and coevolutionary dynamics in Shandong Province, China.}, journal = {PLoS neglected tropical diseases}, volume = {18}, number = {9}, pages = {e0011944}, doi = {10.1371/journal.pntd.0011944}, pmid = {39264945}, issn = {1935-2735}, abstract = {Vector-borne diseases leave a large footprint on global health. Notable culprits include West Nile virus (WNV), St. Louis encephalitis virus (SLEV), and Japanese encephalitis virus (JEV), all transmitted by Culex mosquitoes. Chemical insecticides have been widely used to reduce the spread of mosquito-borne diseases. Still, mosquitoes are becoming more and more resistant to most chemical insecticides which cause particular harm to the ecology. Wolbachia belongs to the family Ehrlichiaceae in the order Rickettsiales and is a matrilineally inherited endosymbiont present in 60% of insects in nature. Wolbachia is capable of inducing a wide range of reproductive abnormalities in its hosts, such as cytoplasmic incompatibility, and can alter mosquito resistance to pathogen infection. Wolbachia has been proposed as a biological alternative to chemical vector control, and specific research progress and effectiveness have been achieved. Despite the importance of Wolbachia, this strategy has not been tested in Culex pipiens pallens, the most prevalent mosquito species in Shandong Province, China. Little is known about how the mass release of Wolbachia-infected mosquitoes may impact the genetic structure of Culex pipiens pallens, and how the symbiotic bacterium Wolbachia interacts with mitochondria during host mosquito transmission. Based on the population genetic structure of Culex pipiens pallens in Shandong Province, this study investigated the infection rate and infection type of Wolbachia in Shandong Province and jointly analysed the evolutionary relationship between the host mosquito and the symbiotic bacterium Wolbachia. Our study showed that Wolbachia naturally infected by Culex pipiens pallens in Shandong Province was less homologous to Wolbachia infected by Aedes albopictus released from mosquito factory in Guangzhou. Our results also show that Culex pipiens pallens is undergoing demographic expansion in Shandong Province. The overall Wolbachia infection rate of Culex pipiens pallens was 92.8%, and a total of 15 WSP haplotypes were detected. We found that the genetic diversity of Wolbachia was low in Culex pipiens pallens from Shandong Province, and the mosquitoes were infected only with type B Wolbachia. Visualizing the relationship between Culex pipiens pallens and Wolbachia using a tanglegram revealed patterns of widespread associations. A specific coevolutionary relationship exists between the host mosquito and Wolbachia. Knowledge of this mosquito-Wolbachia relationship will provide essential scientific information required for Wolbachia-based vector control approaches in Shandong Province and will lead to a better understanding of the diversity and evolution of Wolbachia for its utility as a biocontrol agent.}, } @article {pmid39264544, year = {2024}, author = {Wannassi, T and Sayadi, A and Abbes, K and Djebbi, S and Naccache, C and Khemakhem, MM and Chermiti, B}, title = {Prevalence of Wolbachia infection in field natural population of the apricot seed wasp Eurytoma samsonowi (Hymenoptera: Eurytomidae).}, journal = {International microbiology : the official journal of the Spanish Society for Microbiology}, volume = {}, number = {}, pages = {}, pmid = {39264544}, issn = {1618-1905}, abstract = {Obligate endosymbiont bacteria associated with insects are naturally providing their hosts with essential nutrients such as vitamins and amino acids and biological services including protection from pathogens. In this study, we aimed to investigate the presence of Wolbachia infection among males and females of the parasitic apricot seed wasp (ASW) Eurytoma samsonowi Vassiliev (Vassiliev Petrograd 11: 1-15, 1915) (Hymenoptera: Eurytomidae), a very harmful pest of apricot (Prunus armeniaca), in the oasis of Gafsa, Southern-West of Tunisia. The detection of Wolbachia infection was assessed based on the amplification of the Wolbachia surface protein (wsp) gene and a multilocus sequence typing (MLST) as a universal genotyping tool for Wolbachia involving the analyses of genes gatB, coxA, hcpA, fbpA, and ftsz. Confirming the screening results, Wolbachia was detected in the natural apricot wasp for the first time, with a significant difference between males (5%) and females (59%) based on wsp gene. All Wolbachia strains identified in E. samsonowi were clustered among supergroups B of Wolbachia.}, } @article {pmid39258303, year = {2024}, author = {Silva, NNP and Carvalho, VR and Silva, CB and Bomfim, JPA and Ramos, GS and Oliveira, RC}, title = {First report of the association between Wolbachia and Cotesia flavipes (Hymenoptera: Braconidae): effect on life history parameters of the parasitoid.}, journal = {Bulletin of entomological research}, volume = {}, number = {}, pages = {1-8}, doi = {10.1017/S0007485324000361}, pmid = {39258303}, issn = {1475-2670}, abstract = {The symbiosis between microorganisms and host arthropods can cause biological, physiological, and reproductive changes in the host population. The present study aimed to survey facultative symbionts of the genera Wolbachia, Arsenophonus, Cardinium, Rickettsia, and Nosema in Cotesia flavipes (Cameron) (Hymenoptera: Braconidae) and Diatraea saccharalis (Fabricius) (Lepidoptera: Crambidae) in the laboratory and evaluate the influence of infection on the fitness of these hosts. For this purpose, 16S rDNA primers were used to detect these facultative symbionts in the host species, and the hosts' biological and morphological features were evaluated for changes resulting from the infection caused by these microorganisms. The bacterial symbionts studied herein were not detected in the D. saccharalis samples analysed, but the endosymbiont Wolbachia was detected in C. flavipes and altered the biological and morphological aspects of this parasitoid insect. The results of this study may help to elucidate the role of Wolbachia in maintaining the quality of populations/lineages of C. flavipes.}, } @article {pmid39257987, year = {2024}, author = {Mfopit, YM and Bilgo, E and Boma, S and Somda, MB and Gnambani, JE and Konkobo, M and Diabate, A and Dayo, GK and Mamman, M and Kelm, S and Balogun, EO and Shuaibu, MN and Kabir, J}, title = {Symbiotic bacteria Sodalis glossinidius, Spiroplasma sp and Wolbachia do not favour Trypanosoma grayi coexistence in wild population of tsetse flies collected in Bobo-Dioulasso, Burkina Faso.}, journal = {Research square}, volume = {}, number = {}, pages = {}, pmid = {39257987}, issn = {2693-5015}, support = {K43 TW012015/TW/FIC NIH HHS/United States ; }, abstract = {BACKGROUND: Tsetse flies, the biological vectors of African trypanosomes, have established symbiotic associations with different bacteria. Their vector competence is suggested to be affected by bacterial endosymbionts. The current study provided the prevalence of three tsetse symbiotic bacteria and trypanosomes in Glossina species from Burkina Faso.

RESULTS: A total of 430 tsetse flies were captured using biconical traps in four different collection sites around Bobo-Dioulasso (Bama, Bana, Nasso, and Peni), and their guts were removed. Two hundred tsetse were randomly selected and their guts were screened byPCR for the presence of Sodalis glossinidius, Spiroplasmasp., Wolbachia and trypanosomes. Of the 200 tsetse, 196 (98.0%) were Glossina palpalis gambienseand 4 (2.0%) Glossina tachinoides. The overall symbiont prevalence was 49.0%, 96.5%, and 45.0%, respectively for S. glossinidius, Spiroplasma and Wolbachia. Prevalence varied between sampling locations: S. glossinidius(54.7%, 38.5%, 31.6%, 70.8%); Spiroplasma (100%, 100%, 87.7%, 100%); and Wolbachia(43.4%, 38.5%, 38.6%, 70.8%),respectively in Bama, Bana, Nasso and Peni. Noteworthy, no G. tachhnoideswas infected by S. glossinidius and Wolbachia, but they were all infected by Spiroplasma sp. A total of 196 (98.0 %) harbored at least one endosymbionts. Fifty-five (27.5%) carried single endosymbiont. Trypanosomes were found only in G.p. gambiense, but not G. tachinoides. Trypanosomes were present in flies from all study locations with an overall prevalence of 29.5%. In Bama, Bana, Nasso, and Peni, the trypanosome infection rate was respectively 39.6%, 23.1%, 8.8%, and 37.5%. Remarkably, only Trypanosoma grayi was present. Of all trypanosome-infected flies, 55.9%, 98.3%, and 33.9% hosted S. glossinidius, Spiroplasma sp and Wolbachia, respectively. There was no association between Sodalis, Spiroplasma and trypanosome presence, but there was a negative association with Wolbachia presence. We reported1.9 times likelihood of trypanosome absence when Wolbachia was present.

CONCLUSION: This is the first survey reporting the presence of Trypanosoma grayi in tsetse from Burkina Faso. Tsetse from these localities were highly positive for symbiotic bacteria, more predominantly with Spiroplasma sp. Modifications of symbiotic interactions may pave way for disease control.}, } @article {pmid39248324, year = {2024}, author = {Cash, EI and Escalona, M and Ward, PS and Sahasrabudhe, R and Miller, C and Toffelmier, E and Fairbairn, C and Seligmann, W and Shaffer, HB and Tsutsui, ND}, title = {"The Reference Genome Of The Kidnapper Ant, Polyergus Mexicanus".}, journal = {The Journal of heredity}, volume = {}, number = {}, pages = {}, doi = {10.1093/jhered/esae047}, pmid = {39248324}, issn = {1465-7333}, abstract = {Polyergus kidnapper ants are widely distributed, but relatively uncommon, throughout the Holarctic, spanning an elevational range from sea level to over 3000 m. These species are well known for their obligate social parasitism with various Formica ant species, which they kidnap in dramatic, highly coordinated raids. Kidnapped Formica larvae and pupae become integrated into the Polyergus colony where they develop into adults and perform nearly all of the necessary colony tasks for the benefit of their captors. In California, Polyergus mexicanus is the most widely distributed Polyergus, but recent evidence has identified substantial genetic polymorphism within this species, including genetically divergent lineages associated with the use of different Formica host species. Given its unique behavior and genetic diversity, Polyergus mexicanus plays a critical role in maintaining ecosystem balance by influencing the population dynamics and genetic diversity of its host ant species, Formica, highlighting its conservation value and importance in the context of biodiversity preservation. Here, we present a high-quality genome assembly of P. mexicanus from a sample collected in Plumas County, CA, USA, in the foothills of the central Sierra Nevada. This genome assembly consists of 364 scaffolds spanning 252.31 Mb, with contig N50 of 481,250 kb, scaffold N50 of 10.36 Mb, and BUSCO completeness of 95.4%. We also assembled the genome of the Wolbachia endosymbiont of P. mexicanus - a single, circular contig spanning 1.23 Mb. These genome sequences provide essential resources for future studies of conservation genetics, population genetics, speciation, and behavioral ecology in this charismatic social insect.}, } @article {pmid39243881, year = {2024}, author = {Depeux, C and Branger, A and Paulhac, H and Pigeault, R and Beltran-Bech, S}, title = {Deleterious effects of Wolbachia on life history and physiological traits of common pill woodlice.}, journal = {Journal of invertebrate pathology}, volume = {207}, number = {}, pages = {108187}, doi = {10.1016/j.jip.2024.108187}, pmid = {39243881}, issn = {1096-0805}, abstract = {Most of eukaryotic organisms live in close interaction with micro-organisms called symbionts. Symbiotic interactions underpin the evolution of biological complexity, the health of organisms and, ultimately, the proper functioning of ecosystems. While some symbionts confer adaptive benefits on their host (mutualistic symbionts) and others clearly induce costs (parasitic symbionts), a number of micro-organisms are difficult to classify because they have been described as conferring both benefits and costs on their host. This is particularly true of the most widespread animal endosymbiont, Wolbachia pipientis. In this study, we investigated the influence of Wolbachia infection on a broad spectrum of ecological and physiological parameters of one of its native hosts, Armadillidium vulgare. The aim was to gain as complete a picture as possible of the influence of this endosymbiont on its host. Our results showed that the presence of Wolbachia resulted in a decrease in individual reproductive success and survival. Host immune cells density decreased and β-galactosidase activity (ageing biomarker) increased with the presence of Wolbachia, suggesting a negative impact of this endosymbiont on woodlice health. While previous studies have shown that Wolbachia can have a positive impact on the immunocompetence of A. vulgare, here we shed more light on the costs of infection. Our results illustrate the complex dynamics that exist between Wolbachia and its arthropod host and therefore offer valuable insights into the intricate interplay of symbiotic relationships in ecological systems.}, } @article {pmid39243778, year = {2024}, author = {Bansal, S and Lim, JT and Chong, CS and Dickens, B and Ng, Y and Deng, L and Lee, C and Tan, LY and Kakani, EG and Yoong, Y and Du Yu, D and Chain, G and Ma, P and Sim, S and Ng, LC and Tan, CH}, title = {Effectiveness of Wolbachia-mediated sterility coupled with sterile insect technique to suppress adult Aedes aegypti populations in Singapore: a synthetic control study.}, journal = {The Lancet. Planetary health}, volume = {8}, number = {9}, pages = {e617-e628}, doi = {10.1016/S2542-5196(24)00169-4}, pmid = {39243778}, issn = {2542-5196}, mesh = {*Aedes/microbiology ; Animals ; *Wolbachia/physiology ; Singapore ; *Mosquito Control/methods ; Male ; Female ; *Mosquito Vectors/microbiology ; Pest Control, Biological/methods ; }, abstract = {BACKGROUND: Incompatible insect technique (IIT) coupled with sterile insect technique (SIT) via the release of sterile male Wolbachia-infected mosquitoes is a promising tool for Aedes-borne disease control. Yet, real-world evidence on the suppressive effectiveness of IIT-SIT on mosquito abundance remains mostly limited to small semi-rural village and suburban localities over short trial durations. However, a large proportion of Aedes-borne diseases occur in dense, urban, and high-rise locations, limiting the applicability of previous studies for these settings with high disease burden. The sustainability and use of this technology over multiple years is also unknown.

METHODS: In this synthetic control study, we conducted a large-scale, field trial of IIT-SIT targeting Aedes aegypti among high-rise public housing estates in Singapore, an equatorial city state. Routinely collected data from a large, nationwide surveillance system of 57 990 unique mosquito traps, combined with a high-dimensional set of anthropogenic and environmental confounders were collected to ascertain mosquito abundance and its key drivers. Four townships were selected as the intervention groups (approximate population size of 607 872 residents as of 2022), wherein interventions that combined ITT with SIT over the course of the study period were conducted. Townships were subject to releases of wAlbB-SG male A aegypti mosquitoes twice a week. Data were assessed over the course of epidemiological weeks (EWs), which provide the finest temporal resolution of recorded Wolbachia release schedule and mosquito abundance data. A novel synthetic control framework was then developed to account for the non-randomised and staggered adoption setting of the intervention across trial sectors to identify the direct suppressive effectiveness of IIT-SIT on female A aegypti populations, the spillover effects in non-release areas, and the effect of the intervention on other mosquito populations such as Aedes albopictus. Furthermore, we recalculated effectiveness in terms of calendar time, time since intervention, and over multiple sites to examine heterogeneities in IIT-SIT effectiveness.

FINDINGS: Between EW27 2018 and EW26 2022, Wolbachia releases were conducted across 117 sectors, of which 97 had sufficient trap data, which were collected between EW8 2019 and EW26 2022. We found that Wolbachia-based IIT-SIT reduced wild-type female A aegypti populations by a mean of 62·01% (95% CI 60·68 to 63·26) by 3 months, 78·40% (77·56 to 79·18) by 6 months, and 91·32% (90·95 to 91·66) by at least 18 months of releases. We also found a smaller but non-negligible spillover suppression effect that gradually increased over time (mean spillover intervention effectiveness 61·02% [95% CI 57·89 to 63·72] in adjacent, non-intervention sectors). Although no consistent change in A albopictus populations was seen across the four intervention townships after Wolbachia releases, the average intervention effectiveness on the A albopictus population across all release sectors was -25·80% (95% CI -30·93 to -21·05), which was driven by increases in two towns.

INTERPRETATION: Our results demonstrate the potential of IIT-SIT for strengthening long-term, large-scale vector control in tropical cities, where dengue burden is the greatest. The effect of these interventions in different geographical settings should be assessed in future work.

FUNDING: Singapore's Ministry of Finance, Ministry of Sustainability and the Environment, National Environment Agency, and National Robotics Program.}, } @article {pmid39238888, year = {2024}, author = {Grève, P and Moumen, B and Bouchon, D}, title = {Three feminizing Wolbachia strains in a single host species: comparative genomics paves the way for identifying sex reversal factors.}, journal = {Frontiers in microbiology}, volume = {15}, number = {}, pages = {1416057}, pmid = {39238888}, issn = {1664-302X}, abstract = {INTRODUCTION: Endosymbiotic bacteria in the genus Wolbachia have evolved numerous strategies for manipulating host reproduction in order to promote their own transmission. This includes the feminization of males into functional females, a well-studied phenotype in the isopod Armadillidium vulgare. Despite an early description of this phenotype in isopods and the development of an evolutionary model of host sex determination in the presence of Wolbachia, the underlying genetic mechanisms remain elusive.

METHODS: Here we present the first complete genomes of the three feminizing Wolbachia (wVulC, wVulP, and wVulM) known to date in A. vulgare. These genomes, belonging to Wolbachia B supergroup, contain a large number of mobile elements such as WO prophages with eukaryotic association modules. Taking advantage of these data and those of another Wolbachia-derived feminizing factor integrated into the host genome (f element), we used a comparative genomics approach to identify putative feminizing factors.

RESULTS: This strategy has enabled us to identify three prophage-associated genes secreted by the Type IV Secretion System: one ankyrin repeat domain-containing protein, one helix-turn-helix transcriptional regulator and one hypothetical protein. In addition, a latrotoxin-related protein, associated with phage relic genes, was shared by all three genomes and the f element.

CONCLUSION: These putative feminization-inducing proteins shared canonical interaction features with eukaryotic proteins. These results pave the way for further research into the underlying functional interactions.}, } @article {pmid39232089, year = {2024}, author = {Melo, T and Sousa, CA and Delacour-Estrella, S and Bravo-Barriga, D and Seixas, G}, title = {Characterization of the microbiome of Aedes albopictus populations in different habitats from Spain and São Tomé.}, journal = {Scientific reports}, volume = {14}, number = {1}, pages = {20545}, pmid = {39232089}, issn = {2045-2322}, mesh = {Animals ; *Aedes/microbiology ; Spain ; *Microbiota ; *Wolbachia/genetics/isolation & purification/physiology ; Female ; *RNA, Ribosomal, 16S/genetics ; Mosquito Vectors/microbiology ; Ecosystem ; Male ; }, abstract = {The mosquito microbiome significantly influences vector competence, including in Aedes albopictus, a globally invasive vector. Describing the microbiome and Wolbachia strains of Ae. albopictus from different regions can guide area-specific control strategies. Mosquito samples from Spain and São Tomé were analyzed using 16S rRNA gene sequencing and metagenomic sequencing. Wolbachia infection patterns were observed by sex and population. Female mosquitoes were blood-fed, a factor considered in analyzing their microbiota. Results revealed a dominance of dual Wolbachia infections, strains A and B, in the microbiome of both populations of Ae. albopictus, especially among females. Both populations shared a core microbiome, although 5 and 9 other genera were only present in Spain and São Tomé populations, respectively. Genera like Pelomonas and Nevskia were identified for the first time in Aedes mosquitoes. This study is the first to describe the Ae. albopictus bacteriome in Spain and São Tomé, offering insights for the development of targeted mosquito control strategies. Understanding the specific microbiome composition can help in designing more effective interventions, such as microbiome manipulation and Wolbachia-based approaches, to reduce vector competence and transmission potential of these mosquitoes.}, } @article {pmid39228812, year = {2024}, author = {Arai, H and Herran, B and Sugimoto, TN and Miyata, M and Sasaki, T and Kageyama, D}, title = {Cell-based assays and comparative genomics revealed the conserved and hidden effects of Wolbachia on insect sex determination.}, journal = {PNAS nexus}, volume = {3}, number = {9}, pages = {pgae348}, pmid = {39228812}, issn = {2752-6542}, abstract = {It is advantageous for maternally transmitted endosymbionts to skew the sex ratio of their hosts toward females. Some endosymbiotic bacteria, such as Wolbachia, cause their insect hosts to exclusively produce female offspring through male killing (MK) or feminization. In some lepidopteran insects, MK is achieved by affecting the sex-determining process in males, and a unique mechanism of MK and its functional link with feminization have been implicated. However, comparative analysis of these phenotypes is often difficult because they have been analyzed in different host-symbiont systems, and transinfection of Wolbachia across different hosts is often challenging. In this study, we demonstrated the effects of nine Wolbachia strains on the splicing of sex-determining genes in Lepidoptera by fixing the host genetic background using a cell culture system. Cell transinfection assays confirmed that three MK-inducing Wolbachia strains and one feminization-inducing Wolbachia strain increased the female-type splicing products of the core sex-determining genes doublesex, masculinizer, and zinc finger protein 2. Regarding Wolbachia strains that do not induce MK/feminization, three had no effect on these sex-determining genes, whereas two strains induced female-type splicing of masculinizer and doublesex but not zinc finger protein 2. Comparative genomics confirmed that homologs of oscar, the Wolbachia gene responsible for MK in Ostrinia, were encoded by four MK/feminizing Wolbachia strains, but not by five non-MK/nonfeminizing strains. These results support the conserved effects underlying MK and feminization induced by oscar-bearing Wolbachia and suggested other potential mechanisms that Wolbachia might employ to manipulate host sex.}, } @article {pmid39227968, year = {2024}, author = {Liu, Y and Wang, M and Yu, N and Zhao, W and Wang, P and Zhang, H and Sun, W and Jin, N and Lu, H}, title = {Trends and insights in dengue virus research globally: a bibliometric analysis (1995-2023).}, journal = {Journal of translational medicine}, volume = {22}, number = {1}, pages = {818}, pmid = {39227968}, issn = {1479-5876}, mesh = {Humans ; *Bibliometrics ; *Dengue Virus ; Dengue/epidemiology/virology ; Animals ; Biomedical Research/trends ; History, 21st Century ; History, 20th Century ; }, abstract = {BACKGROUND: Dengue virus (DENV) is the most widespread arbovirus. The World Health Organization (WHO) declared dengue one of the top 10 global health threats in 2019. However, it has been underrepresented in bibliometric analyses. This study employs bibliometric analysis to identify research hotspots and trends, offering a comprehensive overview of the current research dynamics in this field.

RESULTS: We present a report spanning from 1995 to 2023 that provides a unique longitudinal analysis of Dengue virus (DENV) research, revealing significant trends and shifts not extensively covered in previous literature. A total of 10,767 DENV-related documents were considered, with a notable increase in publications, peaking at 747 articles in 2021. Plos Neglected Tropical Diseases has become the leading journal in Dengue virus research, publishing 791 articles in this field-the highest number recorded. Our bibliometric analysis provides a comprehensive mapping of DENV research across multiple dimensions, including vector ecology, virology, and emerging therapies. The study delineates a complex network of immune response genes, including IFNA1, DDX58, IFNB1, STAT1, IRF3, and NFKB1, highlighting significant trends and emerging themes, particularly the impacts of climate change and new outbreaks on disease transmission. Our findings detail the progress and current status of key vaccine candidates, including the licensed Dengvaxia, newer vaccines such as Qdenga and TV003, and updated clinical trials. The study underscores significant advancements in antiviral therapies and vector control strategies for dengue, highlighting innovative drug candidates such as AT-752 and JNJ-1802, and the potential of drug repurposing with agents like Ribavirin, Remdesivir, and Lopinavir. Additionally, it discusses biological control methods, including the introduction of Wolbachia-infected mosquitoes and gene-editing technologies.

CONCLUSION: This bibliometric study underscores the critical role of interdisciplinary collaboration in advancing DENV research, identifying key trends and areas needing further exploration, including host-virus dynamics, the development and application of antiviral drugs and vaccines, and the use of artificial intelligence. It advocates for strengthened partnerships across various disciplines to effectively tackle the challenges posed by DENV.}, } @article {pmid39226386, year = {2024}, author = {Amaya Romero, JE and Chenal, C and Ben Chehida, Y and Miles, A and Clarkson, CS and Pedergnana, V and Wertheim, B and Fontaine, MC}, title = {Mitochondrial Variation in Anopheles gambiae and Anopheles coluzzii: Phylogeographic Legacy and Mitonuclear Associations With Metabolic Resistance to Pathogens and Insecticides.}, journal = {Genome biology and evolution}, volume = {16}, number = {9}, pages = {}, pmid = {39226386}, issn = {1759-6653}, mesh = {Animals ; *Anopheles/genetics ; *Phylogeography ; *Phylogeny ; *DNA, Mitochondrial/genetics ; *Insecticide Resistance/genetics ; Genome, Mitochondrial ; Evolution, Molecular ; Genetic Variation ; Insecticides/pharmacology ; Mitochondria/genetics ; Africa ; }, abstract = {Mitochondrial DNA has been a popular marker in phylogeography, phylogeny, and molecular ecology, but its complex evolution is increasingly recognized. Here, we investigated mitochondrial DNA variation in Anopheles gambiae and Anopheles coluzzii, in relation to other species in the Anopheles gambiae complex, by assembling the mitogenomes of 1,219 mosquitoes across Africa. The mitochondrial DNA phylogeny of the Anopheles gambiae complex was consistent with previously reported highly reticulated evolutionary history, revealing important discordances with the species tree. The three most widespread species (An. gambiae, An. coluzzii, and Anopheles arabiensis), known for extensive historical introgression, could not be discriminated based on mitogenomes. Furthermore, a monophyletic clustering of the three saltwater-tolerant species (Anopheles merus, Anopheles melas, and Anopheles bwambae) in the Anopheles gambiae complex also suggested that introgression and possibly selection shaped mitochondrial DNA evolution. Mitochondrial DNA variation in An. gambiae and An. coluzzii across Africa revealed significant partitioning among populations and species. A peculiar mitochondrial DNA lineage found predominantly in An. coluzzii and in the hybrid taxon of the African "far-west" exhibited divergence comparable to the interspecies divergence in the Anopheles gambiae complex, with a geographic distribution matching closely An. coluzzii's geographic range. This phylogeographic relict of the An. coluzzii and An. gambiae split was associated with population and species structure, but not with the rare Wolbachia occurrence. The lineage was significantly associated with single nucleotide polymorphisms in the nuclear genome, particularly in genes associated with pathogen and insecticide resistance. These findings underline potential mitonuclear coevolution history and the role played by mitochondria in shaping metabolic responses to pathogens and insecticides in Anopheles.}, } @article {pmid39223450, year = {2024}, author = {Valerio, F and Martel, C and Stefanescu, C and van Nouhuys, S and Kankare, M and Duplouy, A}, title = {Wolbachia strain diversity in a complex group of sympatric cryptic parasitoid wasp species.}, journal = {BMC microbiology}, volume = {24}, number = {1}, pages = {319}, pmid = {39223450}, issn = {1471-2180}, mesh = {Animals ; *Wolbachia/genetics/classification/isolation & purification ; *Wasps/microbiology ; *Symbiosis ; *Phylogeny ; Sympatry ; Gene Transfer, Horizontal ; Genetic Variation ; Lepidoptera/microbiology/parasitology ; }, abstract = {BACKGROUND: Maternally-inherited symbionts can induce pre-mating and/or post-mating reproductive isolation between sympatric host lineages, and speciation, by modifying host reproductive phenotypes. The large parasitoid wasp genus Cotesia (Braconidae) includes a diversity of cryptic species, each specialized in parasitizing one to few related Lepidoptera host species. Here, we characterized the infection status of an assemblage of 21 Cotesia species from 15 countries by several microbial symbionts, as a first step toward investigating whether symbionts may provide a barrier to gene flow between these parasitoid host lineages.

RESULTS: The symbiotic microbes Arsenophonus, Cardinium, Microsporidium and Spiroplasma were not detected in the Cotesia wasps. However, the endosymbiotic bacterium Wolbachia was present in at least eight Cotesia species, and hence we concentrated on it upon screening additional DNA extracts and SRAs from NCBI. Some of the closely related Cotesia species carry similar Wolbachia strains, but most Wolbachia strains showed patterns of horizontal transfer between phylogenetically distant host lineages.

CONCLUSIONS: The lack of co-phylogenetic signal between Wolbachia and Cotesia suggests that the symbiont and hosts have not coevolved to an extent that would drive species divergence between the Cotesia host lineages. However, as the most common facultative symbiont of Cotesia species, Wolbachia may still function as a key-player in the biology of the parasitoid wasps. Its precise role in the evolution of this complex clade of cryptic species remains to be experimentally investigated.}, } @article {pmid39221028, year = {2024}, author = {Wilastonegoro, NN and Andriani, S and Sebong, PH and Agarwal-Harding, P and Shepard, DS}, title = {Estimating dengue disease and economic burden to inform municipal-level policymakers: Method for a pragmatic city-level observational cohort study.}, journal = {Gates open research}, volume = {8}, number = {}, pages = {3}, pmid = {39221028}, issn = {2572-4754}, mesh = {Humans ; *Dengue/epidemiology/prevention & control/economics ; Indonesia/epidemiology ; *Cost of Illness ; Prospective Studies ; Cohort Studies ; Female ; Cities/epidemiology ; Male ; }, abstract = {BACKGROUND: Recent trials have confirmed the effectiveness of promising dengue control technologies - two vaccines and Wolbachia. These would generally be applied at the municipal level. To help local officials decide which, if any, control strategy to implement, they need affordable, timely, and accurate data on dengue burden. Building on our previous work in Mexico, Indonesia, and Thailand, we developed a streamlined prospective method to estimate dengue burden at the municipal level quickly, accurately, and efficiently.

METHODS: The method entails enrolling and repeatedly interviewing 100 patients with laboratory-confirmed dengue. They will be selected after screening and testing about 1,000 patients with clinical dengue. The method will capture both acute and chronic effects relating to disease, economic burden, and psychological impacts (presenteeism). The total time requirements are 1.5 years, comprised of 0.25 years for planning and approvals, 1 year for data collection (a full dengue cycle), and 0 .25 years for data cleaning and analysis. A collaboration with municipal and academic colleagues in the city of Semarang, Central Java, Indonesia shows how the method could be readily applied in Indonesia's eighth largest city (population 1.8 million).

CONCLUSIONS: Many surveillance studies gather only information on numbers of cases. This proposed method will provide a comprehensive picture of the dengue burden to the health system, payers, and households at the local level.}, } @article {pmid39205310, year = {2024}, author = {Lefteri, DA and Rainey, SM and Murdochy, SM and Sinkins, SP}, title = {Bunyamwera Virus Infection of Wolbachia-Carrying Aedes aegypti Mosquitoes Reduces Wolbachia Density.}, journal = {Viruses}, volume = {16}, number = {8}, pages = {}, pmid = {39205310}, issn = {1999-4915}, mesh = {*Wolbachia/physiology ; Animals ; *Aedes/microbiology/virology ; *Mosquito Vectors/microbiology/virology ; *Bunyamwera virus ; Female ; Symbiosis ; }, abstract = {Wolbachia symbionts introduced into Aedes mosquitoes provide a highly effective dengue virus transmission control strategy, increasingly utilised in many countries in an attempt to reduce disease burden. Whilst highly effective against dengue and other positive-sense RNA viruses, it remains unclear how effective Wolbachia is against negative-sense RNA viruses. Therefore, the effect of Wolbachia on Bunyamwera virus (BUNV) infection in Aedes aegypti was investigated using wMel and wAlbB, two strains currently used in Wolbachia releases for dengue control, as well as wAu, a strain that typically persists at a high density and is an extremely efficient blocker of positive-sense viruses. Wolbachia was found to reduce BUNV infection in vitro but not in vivo. Instead, BUNV caused significant impacts on density of all three Wolbachia strains following infection of Ae. aegypti mosquitoes. The ability of Wolbachia to successfully persist within the mosquito and block virus transmission is partially dependent on its intracellular density. However, reduction in Wolbachia density was not observed in offspring of infected mothers. This could be due in part to a lack of transovarial transmission of BUNV observed. The results highlight the importance of understanding the complex interactions between multiple arboviruses, mosquitoes and Wolbachia in natural environments, the impact this can have on maintaining protection against diseases, and the necessity for monitoring Wolbachia prevalence at release sites.}, } @article {pmid39204291, year = {2024}, author = {Liu, H and Yin, J and Huang, X and Zang, C and Zhang, Y and Cao, J and Gong, M}, title = {Mosquito Gut Microbiota: A Review.}, journal = {Pathogens (Basel, Switzerland)}, volume = {13}, number = {8}, pages = {}, pmid = {39204291}, issn = {2076-0817}, support = {No. tsqn202312373//the Taishan Scholars Project of Shandong Province/ ; NHCKFKT2021-02//the Open Project of NHC Key Laboratory of Parasite and Vector Biology/ ; 230726153807227//the Ministry of Education Industry-University Cooperative Education Project/ ; GWVI-11.2-XD34//the Innovation Project of Shandong Academy of Medical Sciences; and the Three-Year Initiative Plan for Strengthening Public Health System Construction in Shanghai (2023-2025) Principal Investigator Project/ ; 202201-041 to XDH//Science Foundation of Shandong First Medical University (Shandong Academy of Medical Sciences)/ ; }, abstract = {Mosquitoes are vectors of many important human diseases. The prolonged and widespread use of insecticides has led to the development of mosquito resistance to these insecticides. The gut microbiota is considered the master of host development and physiology; it influences mosquito biology, disease pathogen transmission, and resistance to insecticides. Understanding the role and mechanisms of mosquito gut microbiota in mosquito insecticide resistance is useful for developing new strategies for tackling mosquito insecticide resistance. We searched online databases, including PubMed, MEDLINE, SciELO, Web of Science, and the Chinese Science Citation Database. We searched all terms, including microbiota and mosquitoes, or any specific genera or species of mosquitoes. We reviewed the relationships between microbiota and mosquito growth, development, survival, reproduction, and disease pathogen transmission, as well as the interactions between microbiota and mosquito insecticide resistance. Overall, 429 studies were included in this review after filtering 8139 search results. Mosquito gut microbiota show a complex community structure with rich species diversity, dynamic changes in the species composition over time (season) and across space (environmental setting), and variation among mosquito species and mosquito developmental stages (larval vs. adult). The community composition of the microbiota plays profound roles in mosquito development, survival, and reproduction. There was a reciprocal interaction between the mosquito midgut microbiota and virus infection in mosquitoes. Wolbachia, Asaia, and Serratia are the three most studied bacteria that influence disease pathogen transmission. The insecticide resistance or exposure led to the enrichment or reduction in certain microorganisms in the resistant mosquitoes while enhancing the abundance of other microorganisms in insect-susceptible mosquitoes, and they involved many different species/genera/families of microorganisms. Conversely, microbiota can promote insecticide resistance in their hosts by isolating and degrading insecticidal compounds or altering the expression of host genes and metabolic detoxification enzymes. Currently, knowledge is scarce about the community structure of mosquito gut microbiota and its functionality in relation to mosquito pathogen transmission and insecticide resistance. The new multi-omics techniques should be adopted to find the links among environment, mosquito, and host and bring mosquito microbiota studies to the next level.}, } @article {pmid39194765, year = {2024}, author = {Yen, TY and Cheng, CF and Tseng, LF and Carvalho, RMCA and Tsai, KH}, title = {Nationwide Inventory of Mosquitoes and the Distribution of Invasive Aedes (Stegomyia) albopictus (Skuse, 1894) on the Islands of Sao Tome and Principe in Central Africa.}, journal = {Insects}, volume = {15}, number = {8}, pages = {}, pmid = {39194765}, issn = {2075-4450}, support = {103-2314-B-002 -036 -MY2//Taiwan Ministry of Science and Technology/ ; }, abstract = {Aedes albopictus (Skuse, 1894), a mosquito originating in Asia, has been introduced to Africa since the 2000s. The mosquito is not only a nuisance but is capable of transmitting various arboviruses. The current study summarized our entomological surveys in the Democratic Republic of Sao Tome and Principe during 2000 to 2016. Adult mosquitoes were collected by sweep nets, human landing catches, and Centers for Disease Control (CDC) light traps, and the immatures were collected from water-filled habitats at 15 sentinel sites and reared to adulthood. Species identification was performed based on morphologic characteristics. Fragments of the cytochrome C oxidase subunit I (COI) and the Wolbachia surface protein (wsp) genes were amplified for mosquitoes collected in Principe. New records of four mosquito species were reported. Aedes albopictus was identified in 2015. The larvae were found distributed over the nation and were predominately in artificial water-holding containers (488/2698, 18.1%). The highest positive rate was observed in used tires in Príncipe (114/250, 45.6%). Mitochondrial DNA analysis revealed low genetic diversity among the invasive populations, but all tested specimens were superinfected by Wolbachia. The ability of Ae. albopictus to adapt to new environments and its involvement in disease transmission make the surveillance and control of this species particularly important.}, } @article {pmid39194189, year = {2024}, author = {Miao, Y-h and Dou, W-h and Liu, J and Huang, D-w and Xiao, J-h}, title = {Single-cell transcriptome sequencing reveals that Wolbachia induces gene expression changes in Drosophila ovary cells to favor its own maternal transmission.}, journal = {mBio}, volume = {}, number = {}, pages = {e0147324}, doi = {10.1128/mbio.01473-24}, pmid = {39194189}, issn = {2150-7511}, abstract = {Wolbachia is an obligate endosymbiont that is maternally inherited and widely distributed in arthropods and nematodes. It remains in the mature eggs of female hosts over generations through multiple strategies and manipulates the reproduction system of the host to enhance its spreading efficiency. However, the transmission of Wolbachia within the host's ovaries and its effects on ovarian cells during oogenesis, have not been extensively studied. We used single-cell RNA sequencing to comparatively analyze cell-typing and gene expression in Drosophila ovaries infected and uninfected with Wolbachia. Our findings indicate that Wolbachia significantly affects the transcription of host genes involved in the extracellular matrix, cytoskeleton organization, and cytomembrane mobility in multiple cell types, which may make host ovarian cells more conducive for the transmission of Wolbachia from extracellular to intracellular. Moreover, the genes nos and orb, which are related to the synthesis of ribonucleoprotein complexes, are specifically upregulated in early germline cells of ovaries infected with Wolbachia, revealing that Wolbachia can increase the possibility of its localization to the host oocytes by enhancing the binding with host ribonucleoprotein-complex processing bodies (P-bodies). All these findings provide novel insights into the maternal transmission of Wolbachia between host ovarian cells.IMPORTANCEWolbachia, an obligate endosymbiont in arthropods, can manipulate the reproduction system of the host to enhance its maternal transmission and reside in the host's eggs for generations. Herein, we performed single-cell RNA sequencing of ovaries from Drosophila melanogaster and observed the effects of Wolbachia (strain wMel) infection on different cell types to discuss the potential mechanism associated with the transmission and retention of Wolbachia within the ovaries of female hosts. It was found that the transcriptions of multiple genes in the ovary samples infected with Wolbachia are significantly altered, which possibly favors the maternal transmission of Wolbachia. Meanwhile, we also discovered that Wolbachia may flexibly regulate the expression level of specific host genes according to their needs rather than rigidly changing the expression level in one direction to achieve a more suitable living environment in the host's ovarian cells. Our findings contribute to a further understanding of the maternal transmission and possible universal effects of Wolbachia within the host.}, } @article {pmid39181959, year = {2024}, author = {Alkathiri, B and Lee, S and Ahn, K and Cho, YS and Youn, SY and Seo, K and Umemiya-Shirafuji, R and Xuan, X and Kwak, D and Shin, S and Lee, SH}, title = {16S rRNA metabarcoding for the identification of tick-borne bacteria in ticks in the Republic of Korea.}, journal = {Scientific reports}, volume = {14}, number = {1}, pages = {19708}, pmid = {39181959}, issn = {2045-2322}, support = {Z-1543081-2021-22-02//Animal and Plant Quarantine Agency/ ; Z-1543081-2021-22-02//Animal and Plant Quarantine Agency/ ; 2021R1F1A1061795//National Research Foundation of Korea/ ; }, mesh = {Animals ; *RNA, Ribosomal, 16S/genetics ; Republic of Korea ; *DNA Barcoding, Taxonomic/methods ; *Bacteria/genetics/classification/isolation & purification ; *Ticks/microbiology ; DNA, Bacterial/genetics ; Wolbachia/genetics/isolation & purification/classification ; Phylogeny ; Rickettsia/genetics/isolation & purification/classification ; }, abstract = {Ticks are blood-sucking ectoparasites that act as vectors for transmission of various pathogens. The purpose of this study was to assess tick-borne bacteria, whether pathogenic or not, in ticks distributed in Korea using 16S rRNA metabarcoding and to confirm the results by PCR. Questing ticks were collected from four provinces in Korea in 2021 using the flagging method. After pooling the DNAs from the 61 tick pools (including 372 ticks), the bacterial 16S rRNA V3-V4 hypervariable region was amplified and sequenced using the MiSeq platform. Rickettsia, Ehrlichia, and the endosymbiont Wolbachia were confirmed by conventional PCR and molecular analysis. In total, 6907 ticks (534 pools) were collected and identified as belonging to five species (Haemaphysalis spp., H. longicornis, H. flava, I. nipponensis, and A. testudinarium). Through 16S rRNA metabarcoding, 240 amplicon sequence variants were identified. The dominant taxa were Rickettsiella and Coxiella. Additionally, pathogenic bacteria such as Rickettsia and Ehrlichia, endosymbiotic bacteria such as Wolbachia and Spiroplasma were identified. Polymerase chain reaction (PCR) was performed to confirm the presence of Rickettsia, Ehrlichia, Bartonella, and Wolbachia in individual ticks. Overall, 352 (65.92%) of 534 pools tested positive for at least one of the screened tick-borne bacteria. Rickettsia was the most prevalent (61.42%), followed by Wolbachia (5.05%). Ehrlichia was detected in 4.86% of tested samples, whereas Bartonella was not detected. In this study, 16S rRNA metabarcoding revealed the presence of Rickettsia, Wolbachia, and Ehrlichia, in that order of abundance, while showing absence of Bartonella. These results were confirmed to exhibit the same trend as that of the conventional PCR. Therefore, large-scale screening studies based on pooling, as applied in this study, will be useful for examining novel or rare pathogens present in various hosts and vectors.}, } @article {pmid39176079, year = {2024}, author = {Mushtaq, I and Sarwar, MS and Munzoor, I}, title = {A comprehensive review of Wolbachia-mediated mechanisms to control dengue virus transmission in Aedes aegypti through innate immune pathways.}, journal = {Frontiers in immunology}, volume = {15}, number = {}, pages = {1434003}, pmid = {39176079}, issn = {1664-3224}, mesh = {*Aedes/immunology/virology/microbiology ; *Wolbachia/physiology/immunology ; Animals ; *Dengue Virus/immunology/physiology ; *Immunity, Innate ; *Dengue/immunology/transmission/virology ; *Mosquito Vectors/immunology/virology/microbiology ; Host-Pathogen Interactions/immunology ; Humans ; Signal Transduction/immunology ; }, abstract = {The Dengue virus (DENV), primarily spread by Aedes aegypti and also by Aedes albopictus in some regions, poses significant global health risks. Alternative techniques are urgently needed because the current control mechanisms are insufficient to reduce the transmission of DENV. Introducing Wolbachia pipientis into Ae. aegypti inhibits DENV transmission, however, the underlying mechanisms are still poorly understood. Innate immune effector upregulation, the regulation of autophagy, and intracellular competition between Wolbachia and DENV for lipids are among the theories for the mechanism of inhibition. Furthermore, mainly three immune pathways Toll, IMD, and JAK/STAT are involved in the host for the suppression of the virus. These pathways are activated by Wolbachia and DENV in the host and are responsible for the upregulation and downregulation of many genes in mosquitoes, which ultimately reduces the titer of the DENV in the host. The functioning of these immune pathways depends upon the Wolbachia, host, and virus interaction. Here, we summarize the current understanding of DENV recognition by the Ae. aegypti's immune system, aiming to create a comprehensive picture of our knowledge. Additionally, we investigated how Wolbachia regulates the activation of multiple genes associated with immune priming for the reduction of DENV.}, } @article {pmid39165109, year = {2024}, author = {Gloder, G and Bourne, ME and Cuny, MAC and Verreth, C and Crauwels, S and Dicke, M and Poelman, EH and Jacquemyn, H and Lievens, B}, title = {Caterpillar-parasitoid interactions: species-specific influences on host microbiome composition.}, journal = {FEMS microbiology ecology}, volume = {}, number = {}, pages = {}, doi = {10.1093/femsec/fiae115}, pmid = {39165109}, issn = {1574-6941}, abstract = {There is increasing evidence that host-parasitoid interactions can have a pronounced impact on the microbiome of host insects, but it is unclear to what extent this is caused by the host and/or parasitoid. Here, we compared the internal and external microbiome of caterpillars of Pieris brassicae and Pieris rapae parasitised by Cotesia glomerata or Cotesia rubecula with non-parasitised caterpillars. Additionally, we investigated the internal and external microbiome of the parasitoid larvae. Both internal and external bacterial densities were significantly higher for P. brassicae than P. rapae, while no differences were found between parasitised and non-parasitised caterpillars. In contrast, parasitism significantly affected the composition of the internal and external microbiome of the caterpillars and the parasitoid larvae, but the effects were dependent on the host and parasitoid species. Irrespective of host species, a Wolbachia species was exclusively found inside caterpillars parasitised by C. glomerata, as well as in the corresponding developing parasitoid larvae. Similarly, a Nosema species was abundantly present inside parasitised caterpillars and the parasitoid larvae, but this was independent of the host and the parasitoid species. We conclude that parasitism has pronounced effects on host microbiomes, but the effects depend on both the host and parasitoid species.}, } @article {pmid39157806, year = {2024}, author = {Kagemann, CH and Colocho, GM and Aquadro, CF}, title = {Non-ovarian Wolbachia pipientis titer correlates with fertility rescue of a Drosophila melanogaster bag of marbles hypomorph.}, journal = {microPublication biology}, volume = {2024}, number = {}, pages = {}, pmid = {39157806}, issn = {2578-9430}, support = {R01 GM095793/GM/NIGMS NIH HHS/United States ; S10 OD018516/OD/NIH HHS/United States ; }, abstract = {Bag of marbles (bam) is an essential gene that regulates germline stem cell maintenance and germline stem cell daughter cell differentiation in Drosophila melanogaster . When bam is partially functional (hypomorphic), the introduction of Wolbachia pipientis rescues the mutant fertility phenotype that would otherwise result in partial sterility. Infection by different W. pipientis variants results in differential rescue of the bam hypomorph fertility phenotype. We were intrigued by the varying degrees of rescue exhibited in the bam hypomorph when exposed to different W. pipientis variants, prompting us to investigate whether this phenomenon is attributable to variations in the titers of W. pipientis variants. We found no significant difference in ovarian titer between two W. pipientis variant groups, w Mel-like (low bam hypomorph fertility rescue) and w MelCS-like variants (higher bam hypomorph fertility rescue), at bam hypomorph peak fertility. However, carcass (whole flies without the ovaries) titer between w Mel-like and w MelCS-like infected bam hypomorph differed during peak fertility rescue. A positive correlation emerged between the combined titers of ovarian and carcass infections and fertility, implying a more extensive influence that extends beyond ovarian infection alone.}, } @article {pmid39150623, year = {2024}, author = {Gao, S and Niu, YD and Chen, L and Chen, MF and Bing, XL and Hong, XY}, title = {Transcriptomic landscapes reveal development-related physiological processes in the two-spotted spider mite, Tetranychus urticae.}, journal = {Experimental & applied acarology}, volume = {}, number = {}, pages = {}, pmid = {39150623}, issn = {1572-9702}, support = {BK20221003//Natural Science Foundation of Jiangsu Province/ ; 202310307177Y//Jiangsu Students' Platform for innovation and entrepreneurship training program/ ; 202310307177Y//Jiangsu Students' Platform for innovation and entrepreneurship training program/ ; 32372528, 32020103011//National Natural Science Foundation of China/ ; 32372528, 32020103011//National Natural Science Foundation of China/ ; TJ-2023-038//Young Elite Scientists Sponsorship Program by Jiangsu Association for Science and Technology/ ; }, abstract = {The two-spotted spider mite (Tetranychus urticae Koch, TSSM) is recognized as one of the most problematic spider mite pests. However, the precise gene expression patterns across its key developmental stages remain elusive. Here, we performed a comprehensive transcriptome analysis of TSSM eggs, nymphs and adult females using publicly available RNA sequencing (RNA-seq) data to elucidate the overarching transcriptomic differences between these developmental stages. Principal component analysis and hierarchical clustering analysis unveiled distinct separations among samples across different developmental stages, regardless of their Wolbachia infection status. Differential expression analysis revealed 4,089,2,762, and 1,282 core genes specifically enriched in eggs, nymphs, and adults, respectively. KEGG and GO enrichment analyses showed upregulation of genes in eggs are associated with proteolysis, Wnt signaling pathway, DNA transcription, RNA biosynthetic and metabolic processes, as well as protein folding, sorting, and degradation pathways. Meanwhile, nymphs exhibited increased abundance of genes related to chitin/amino sugar metabolic processes, G protein-coupled receptor signaling pathways, monoatomic ion transport, and neurotransmitter transport pathways. Pathways involving sphingolipid and carbohydrate metabolic processes, proteolysis, lipid transport, and localization were particularly enriched in older females. Altogether, our findings suggest that the egg stage exhibits higher activity in cell differentiation processes, the nymph stage is more involved in chitin development, and the adult stage shows increased metabolic and reproductive activity. This study enhances our understanding of the molecular mechanisms underlying TSSM development and paves the way for further research into the intricate physiological processes of TSSM.}, } @article {pmid39135725, year = {2024}, author = {Rohlfing, K and Grewoldt, M and Cordellier, M and Dobler, S}, title = {Evidence for feminized genetic males in a flea beetle using newly identified X-linked markers.}, journal = {Ecology and evolution}, volume = {14}, number = {8}, pages = {e70123}, pmid = {39135725}, issn = {2045-7758}, abstract = {The equilibrium of sex ratios in sexually reproducing species is often disrupted by various environmental and genetic factors, including endosymbionts like Wolbachia. In this study, we explore the highly female-biased sex ratio observed in the flea beetle, Altica lythri, and its underlying mechanisms. Ancient hybridization events between Altica species have led to mitochondrial DNA introgression, resulting in distinct mitochondrial haplotypes that go along with different Wolbachia infections (HT1-wLytA1, HT1*- uninfected, HT2-wLytA2, and HT3-wLytB). Notably, beetles with some haplotypes exclusively produce female offspring, suggesting potential Wolbachia-induced phenomena such as feminization of genetic males. However, the observed female bias could also be a consequence of the ancient hybridization resulting in nuclear-cytoplasmic conflicts between introgressed mtDNA and nuclear genes. Through transcriptomic analysis and the program SEX-DETector, we established markers for genotypic sex differentiation for A. lythri, enabling genetic sexing via qPCR. Our findings suggest that feminization of genetic males is contributing to the skewed sex ratios, highlighting the intricate dynamics of sex determination and reproductive strategies in this flea beetle. This study provides valuable insights into the dynamics of genetic conflicts, endosymbionts, and sex ratios, revealing the novel phenomenon of genetic male feminization in the flea beetle A. lythri.}, } @article {pmid39129654, year = {2024}, author = {Mostoufi, SL and Singh, ND}, title = {Pathogen infection alters the gene expression landscape of transposable elements in Drosophila melanogaster.}, journal = {G3 (Bethesda, Md.)}, volume = {14}, number = {9}, pages = {}, pmid = {39129654}, issn = {2160-1836}, mesh = {Animals ; *DNA Transposable Elements ; *Drosophila melanogaster/genetics ; Host-Pathogen Interactions/genetics ; Gene Expression Regulation ; Male ; Female ; Gene Expression Profiling ; }, abstract = {Transposable elements make up substantial proportions of eukaryotic genomes and many are thought to be remnants of ancient viral infections. Current research has begun to highlight the role transposable elements can play in the immune system response to infections. However, most of our knowledge about transposable element expression during infection is limited by the specific host and pathogen factors from each study, making it difficult to compare studies and develop broader patterns regarding the role of transposable elements during infection. Here, we use the tools and resources available in the model, Drosophila melanogaster, to analyze multiple gene expression datasets of flies subject to bacterial, fungal, and viral infections. We analyzed differences in pathogen species, host genotype, host tissue, and sex to understand how these factors impact transposable element expression during infection. Our results highlight both shared and unique transposable element expression patterns between pathogens and suggest a larger effect of pathogen factors over host factors for influencing transposable element expression.}, } @article {pmid39119885, year = {2024}, author = {Augustijnen, H and Lucek, K}, title = {Beyond gene flow: (non)-parallelism of secondary contact in a pair of highly differentiated sibling species.}, journal = {Molecular ecology}, volume = {33}, number = {17}, pages = {e17488}, doi = {10.1111/mec.17488}, pmid = {39119885}, issn = {1365-294X}, support = {//Burckhardt-Bürgin foundation/ ; 310030_184934//Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung/ ; PCEFP3_202869//Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung/ ; //Freiwillige Akademische Gesellschaft Basel/ ; }, mesh = {*Gene Flow ; Animals ; *Butterflies/genetics ; *Wolbachia/genetics ; *Genetic Speciation ; *Hybridization, Genetic ; *Reproductive Isolation ; *Genetics, Population ; }, abstract = {Replicated secondary contact zones can provide insights into the barriers to gene flow that are important during speciation and can reveal to which degree secondary contact may result in similar evolutionary outcomes. Here, we studied two secondary contact zones between highly differentiated Alpine butterflies of the genus Erebia using whole-genome resequencing data. We assessed the genomic relationships between populations and species and found hybridization to be rare, with no to little current or historical introgression in either contact zone. There are large similarities between contact zones, consistent with an allopatric origin of interspecific differentiation, with no indications for ongoing reinforcing selection. Consistent with expected reduced effective population size, we further find that scaffolds related to the Z-chromosome show increased differentiation compared to the already high levels across the entire genome, which could also hint towards a contribution of the Z chromosome to species divergence in this system. Finally, we detected the presence of the endosymbiont Wolbachia, which can cause reproductive isolation between its hosts, in all E. cassioides, while it appears to be fully or largely absent in contact zone populations of E. tyndarus. We discuss how this rare pattern may have arisen and how it may have affected the dynamics of speciation upon secondary contact.}, } @article {pmid39119296, year = {2024}, author = {Kang, K and Wang, L and Gong, J and Tang, Y and Wei, K}, title = {Diversity analyses of bacterial symbionts in four Sclerodermus (Hymenoptera: Bethylidae) parasitic wasps, the dominant biological control agents of wood-boring beetles in China.}, journal = {Frontiers in cellular and infection microbiology}, volume = {14}, number = {}, pages = {1439476}, pmid = {39119296}, issn = {2235-2988}, mesh = {Animals ; *Symbiosis ; *Wasps/microbiology/physiology ; China ; *RNA, Ribosomal, 16S/genetics ; High-Throughput Nucleotide Sequencing ; Bacteria/classification/genetics/isolation & purification ; Biological Control Agents ; Coleoptera/microbiology ; Phylogeny ; Microbiota ; Bacteroides/genetics/isolation & purification/classification ; Firmicutes/genetics/isolation & purification/classification ; Wolbachia/genetics/isolation & purification/classification/physiology ; Biodiversity ; }, abstract = {OBJECTIVE: Sclerodermus wasps are important biocontrol agents of a class of wood borers. Bacterial symbionts influence the ecology and biology of their hosts in a variety of ways, including the formation of life-long beneficial or detrimental parasitic infections. However, only a few studies have explored the species and content of the symbionts in the Sclerodermus species.

METHODS: Here, a high-throughput sequencing study of the V3-V4 region of the 16S ribosomal RNA gene revealed a high level of microbial variety in four Sclerodermus waps, and their diversities and functions were also predicted.

RESULTS: The three most prevalent phyla of microorganisms in the sample were Firmicutes, Bacteroides, and Proteus. The KEEG pathways prediction results indicated that the three pathways with the highest relative abundances in the S. sichuanensis species were translation, membrane transport, and nucleotide metabolism. These pathways differed from those observed in S. guani, S. pupariae, and S. alternatusi, which exhibited carbohydrate metabolism, membrane transport, and amino acid metabolism, respectively. Bacteroides were found to be abundant in several species, whereas Wolbachia was the most abundant among S. sichuanensis, with a significant negative correlation between temperature and carriage rate.

CONCLUSIONS: These results offer insights into the microbial communities associated with the bethylid wasps, which is crucial for understanding how to increase the reproductive capacity of wasps, enhance their parasitic effects, and lower cost in biocontrol.}, } @article {pmid39109809, year = {2024}, author = {Li, Q and Ji, R and Zi, H and Sun, W and Zhang, Y and Wu, X and Long, Y and Yang, Y}, title = {Life history parameters of Ectropis grisescens (Lepidoptera: Geometridae) in different Wolbachia infection states.}, journal = {Journal of economic entomology}, volume = {}, number = {}, pages = {}, doi = {10.1093/jee/toae174}, pmid = {39109809}, issn = {1938-291X}, support = {32371581//National Natural Science Foundation of China/ ; 2021YFD1601105//National Key Research and Development/ ; 2023n06020060//Anhui Key Research and Development/ ; }, abstract = {Wolbachia, a prevalent intracellular symbiotic bacterium in insects, plays a significant role in insect biology. Ectropis grisescens (Warren; Lepidoptera: Geometridae) is a devastating chewing pest distributed in tea plantations throughout China. However, it is unclear how Wolbachia titers affect the fitness and reproduction of E. grisescens. In this study, the impacts of 3 different infection lines, naturally Wolbachia-infected, Wolbachia-uninfected, and Wolbachia transinfected, regarding the life history traits of E. grisescens, were evaluated using the age-stage, 2-sex life table. Wolbachia infection significantly shortened preadult duration and preoviposition periods and notably increased the fecundity, net reproductive rate, and finite rate of increase. Meanwhile, population projection indicated that E. grisescens population size with Wolbachia infection can increase faster than without. These results indicate that Wolbachia plays a regulatory role in the fitness of E. grisescens. It is also noted that the life history parameters of E. grisescens may positively correlate with Wolbachia titers. These findings could aid in pest management in tea gardens.}, } @article {pmid39107710, year = {2024}, author = {Chow, JY and Geng, L and Bansal, S and Dickens, BSL and Ng, LC and Hoffmann, AA and Lim, JT}, title = {Evaluating quasi-experimental approaches for estimating epidemiological efficacy of non-randomised field trials: applications in Wolbachia interventions for dengue.}, journal = {BMC medical research methodology}, volume = {24}, number = {1}, pages = {170}, pmid = {39107710}, issn = {1471-2288}, mesh = {*Wolbachia/physiology ; *Dengue/prevention & control/epidemiology ; Animals ; *Aedes/microbiology/virology ; Humans ; Brazil/epidemiology ; Singapore/epidemiology ; Malaysia/epidemiology ; Incidence ; Mosquito Control/methods ; Mosquito Vectors/microbiology ; Symbiosis ; Pest Control, Biological/methods/statistics & numerical data ; }, abstract = {BACKGROUND: Wolbachia symbiosis in Aedes aegypti is an emerging biocontrol measure against dengue. However, assessing its real-world efficacy is challenging due to the non-randomised, field-based nature of most intervention studies. This research re-evaluates the spatial-temporal impact of Wolbachia interventions on dengue incidence using a large battery of quasi-experimental methods and assesses each method's validity.

METHODS: A systematic search for Wolbachia intervention data was conducted via PUBMED. Efficacy was reassessed using commonly-used quasi-experimental approaches with extensive robustness checks, including geospatial placebo tests and a simulation study. Intervention efficacies across multiple study sites were computed using high-resolution aggregations to examine heterogeneities across sites and study periods. We further designed a stochastic simulation framework to assess the methods' ability to estimate intervention efficacies (IE).

RESULTS: Wolbachia interventions in Singapore, Malaysia, and Brazil significantly decreased dengue incidence, with reductions ranging from 48.17% to 69.19%. IEs varied with location and duration. Malaysia showed increasing efficacy over time, while Brazil exhibited initial success with subsequent decline, hinting at operational challenges. Singapore's strategy was highly effective despite partial saturation. Simulations identified Synthetic Control Methods (SCM) and its variant, count Synthetic Control Method (cSCM), as superior in precision, with the smallest percentage errors in efficacy estimation. These methods also demonstrated robustness in placebo tests.

CONCLUSIONS: Wolbachia interventions exhibit consistent protective effects against dengue. SCM and cSCM provided the most precise and robust estimates of IEs, validated across simulated and real-world settings.}, } @article {pmid39107546, year = {2024}, author = {Durand, S and Pigeault, R and Giraud, I and Loisier, A and Bech, N and Grandjean, F and Rigaud, T and Peccoud, J and Cordaux, R}, title = {Temporal stability of sex ratio distorter prevalence in natural populations of the isopod Armadillidium vulgare.}, journal = {Heredity}, volume = {}, number = {}, pages = {}, pmid = {39107546}, issn = {1365-2540}, support = {ANR-15-CE32-0006//Agence Nationale de la Recherche (French National Research Agency)/ ; ANR-15-CE32-0006//Agence Nationale de la Recherche (French National Research Agency)/ ; ANR-20-CE02-0004//Agence Nationale de la Recherche (French National Research Agency)/ ; }, abstract = {In the terrestrial isopod Armadillidium vulgare, many females produce progenies with female-biased sex ratios due to two feminizing sex ratio distorters (SRD): Wolbachia endosymbionts and a nuclear non-mendelian locus called the f element. To investigate the potential impact of these SRD on the evolution of host sex determination, we analyzed their temporal distribution in six A. vulgare populations sampled between 2003 and 2017, for a total of 29 time points. SRD distribution was heterogeneous among populations despite their close geographic locations, so that when one SRD was frequent in a population, the other SRD was rare. In contrast with spatial heterogeneity, our results overall did not reveal substantial temporal variability in SRD prevalence within populations, suggesting equilibria in SRD evolutionary dynamics may have been reached or nearly so. Temporal stability was also generally reflected in mitochondrial and nuclear variation. Nevertheless, in a population, a Wolbachia strain replacement coincided with changes in mitochondrial composition but no change in nuclear composition, thus constituting a typical example of mitochondrial sweep caused by endosymbiont rise in frequency. Rare incongruence between Wolbachia strains and mitochondrial haplotypes suggested the occurrence of intraspecific horizontal transmission, making it a biologically relevant parameter for Wolbachia evolutionary dynamics in A. vulgare. Overall, our results provide an empirical basis for future studies on SRD evolutionary dynamics in the context of multiple sex determination factors co-existing within a single species, to ultimately evaluate the impact of SRD on the evolution of host sex determination mechanisms and sex chromosomes.}, } @article {pmid39106844, year = {2024}, author = {Sarasombath, PT and Sitthinamsuwan, P and Wijit, S and Panyasu, K and Roongruanchai, K and Silpa-Archa, S and Suwansirikul, M and Chortrakarnkij, P and Ruenchit, P and Preativatanyou, K and Wongkamchai, S}, title = {Integrated Histological and Molecular Analysis of Filarial Species and Associated Wolbachia Endosymbionts in Human Filariasis Cases Presenting Atypically in Thailand.}, journal = {The American journal of tropical medicine and hygiene}, volume = {}, number = {}, pages = {}, doi = {10.4269/ajtmh.24-0147}, pmid = {39106844}, issn = {1476-1645}, abstract = {Atypical presentations of filariasis have posed diagnostic challenges due to the complexity of identifying the causative species and the difficulties in both diagnosis and treatment. In this study, we present the integrative histological and molecular analysis of seven atypical filariasis cases observed in regions of nonendemicity of Thailand. All filariasis cases were initially diagnosed based on histological findings. To confirm the causative species, molecular characterization based on both filarial mitochondrial (mt 12S rRNA and COI genes) and nuclear ITS1 markers was performed, together with the identification of associated Wolbachia bacterial endosymbionts. Among the cases studied, Brugia pahangi (N = 3), Brugia malayi (N = 1), Dirofilaria sp. "hongkongensis" (N = 2), and a suspected novel filarial species genetically related to Pelecitus copsychi (N = 1) were identified. By targeting the 16S rRNA gene, Wolbachia was also molecularly amplified in two cases of infection with Dirofilaria sp. "hongkongensis." Phylogenetic analysis further revealed that the detected Wolbachia could be classified into supergroups C and F, indicating the high genetic diversity of this endosymbiont in Dirofilaria sp. "hongkongensis." Furthermore, this study demonstrates the consistency between histological findings and species identification based on mitochondrial loci rather than on the nuclear ITS1. This suggests the utility of mitochondrial markers, particularly COI, as a highly sensitive and reliable diagnostic tool for the detection and differentiation of filarial species in clinical specimens. Precise identification of the causative species will facilitate accurate diagnosis and treatment and is also essential for the development of epidemiological and preventive strategies for filariasis.}, } @article {pmid39106433, year = {2024}, author = {Tan, Y and Aravind, L and Zhang, D}, title = {Genomic Underpinnings of Cytoplasmic Incompatibility: CIF Gene-Neighborhood Diversification Through Extensive Lateral Transfers and Recombination in Wolbachia.}, journal = {Genome biology and evolution}, volume = {16}, number = {8}, pages = {}, pmid = {39106433}, issn = {1759-6653}, support = {//Saint Louis University/ ; //Intramural Research Program of the NIH/ ; /LM/NLM NIH HHS/United States ; }, mesh = {*Wolbachia/genetics ; *Gene Transfer, Horizontal ; *Recombination, Genetic ; Evolution, Molecular ; Phylogeny ; Genome, Bacterial ; Cytoplasm/genetics ; Animals ; Bacterial Proteins/genetics ; }, abstract = {Cytoplasmic incompatibility (CI), a non-Mendelian genetic phenomenon, involves the manipulation of host reproduction by Wolbachia, a maternally transmitted alphaproteobacterium. The underlying mechanism is centered around the CI Factor (CIF) system governed by two genes, cifA and cifB, where cifB induces embryonic lethality, and cifA counteracts it. Recent investigations have unveiled intriguing facets of this system, including diverse cifB variants, prophage association in specific strains, copy number variation, and rapid component divergence, hinting at a complex evolutionary history. We utilized comparative genomics to systematically classify CIF systems, analyze their locus structure and domain architectures, and reconstruct their diversification and evolutionary trajectories. Our new classification identifies ten distinct CIF types, featuring not just versions present in Wolbachia, but also other intracellular bacteria, and eukaryotic hosts. Significantly, our analysis of CIF loci reveals remarkable variability in gene composition and organization, encompassing an array of diverse endonucleases, variable toxin domains, deubiquitinating peptidases (DUBs), prophages, and transposons. We present compelling evidence that the components within the loci have been diversifying their sequences and domain architectures through extensive, independent lateral transfers and interlocus recombination involving gene conversion. The association with diverse transposons and prophages, coupled with selective pressures from host immunity, likely underpins the emergence of CIF loci as recombination hotspots. Our investigation also posits the origin of CifB-REase domains from mobile elements akin to CR (Crinkler-RHS-type) effectors and Tribolium Medea1 factor, which is linked to another non-Mendelian genetic phenomenon. This comprehensive genomic analysis offers novel insights into the molecular evolution and genomic foundations of Wolbachia-mediated host reproductive control.}, } @article {pmid39105274, year = {2024}, author = {Lim, JT and Mailepessov, D and Chong, CS and Dickens, B and Lai, YL and Ng, Y and Deng, L and Lee, C and Tan, LY and Chain, G and Ho, SH and Chang, CC and Ma, P and Bansal, S and Lee, V and Sim, S and Tan, CH and Ng, LC}, title = {Assessing Wolbachia-mediated Sterility for Dengue Control: Emulation of a Cluster Randomized Target Trial in Singapore.}, journal = {Journal of travel medicine}, volume = {}, number = {}, pages = {}, doi = {10.1093/jtm/taae103}, pmid = {39105274}, issn = {1708-8305}, abstract = {BACKGROUND: Matings between male Aedes aegypti mosquitoes infected with wAlbB strain of Wolbachia and wildtype females yield non-viable eggs. We evaluated the efficacy of releasing wAlbB-infected Ae. aegypti male mosquitoes to suppress dengue.

METHODS: We specified the protocol of a two-arm cluster-randomized test-negative controlled trial (cRCT) and emulated it using a nationally representative test-negative/positive database of individuals reporting for febrile illness to any public hospital, general practitioner or polyclinic. We retrospectively built a cohort of individuals who reside in Wolbachia locations versus a comparator control group who do not reside in Wolbachia locations, using a nationally representative database of all individuals whom report for febrile illness and were tested for dengue at the Environmental Health Institute/hospital laboratories/commercial diagnostic laboratories, through general practitioner clinic, polyclinic or public/private hospital from EW1 2019-EW 262022. We emulated a constrained randomization protocol used in cRCTs to balance dengue risk between intervention and control arms in the pre-intervention period. We used the inverse-probability weighting approach to further balance the intervention and control groups using a battery of algorithmically selected sociodemographic, environmental and anthropogenic variables. Intention-to-treat analyses was conducted to estimate the risk reduction of dengue given Wolbachia exposure.

RESULTS: Intention-to-treat analyses revealed that, compared with controls, Wolbachia releases for 3, 6, 12 or more months was associated to 47%(95%CI:25-69%), 44%(33-77%) and 61%(38-78%) protective efficacy against dengue, respectively. When exposed to 12 or more months of Wolbachia releases, protective efficacies ranged from 49%(13-72%) to 77%(60-94%) across years. The proportion of virologically confirmed dengue cases was lower overall in the intervention arm. Protective efficacies were found across all years, age and sex subgroups, with higher durations of Wolbachia exposure associated to greater risk reductions of dengue.

CONCLUSION: Results demonstrated that Wolbachia-mediated sterility can strengthen dengue control in tropical cities, where dengue burden is the greatest.}, } @article {pmid39102421, year = {2024}, author = {Kilpatrick, AM and Seidl, CM and Ipsaro, IJ and Garrison, CE and Fabbri, G and Howell, PI and McGowan, AG and White, BJ and Mitchell, SN}, title = {Transinfection of Wolbachia wAlbB into Culex quinquefasciatus mosquitoes does not alter vector competence for Hawaiian avian malaria (Plasmodium relictum GRW4).}, journal = {PLoS pathogens}, volume = {20}, number = {8}, pages = {e1012052}, pmid = {39102421}, issn = {1553-7374}, mesh = {Animals ; Female ; Male ; Aedes/microbiology ; *Culex/microbiology/parasitology ; Hawaii ; *Malaria, Avian/transmission ; *Mosquito Vectors/microbiology/parasitology ; *Plasmodium ; *Wolbachia/physiology ; }, abstract = {Avian malaria is expanding upslope with warmer temperatures and driving multiple species of Hawaiian birds towards extinction. Methods to reduce malaria transmission are urgently needed to prevent further declines. Releasing Wolbachia-infected incompatible male mosquitoes could suppress mosquito populations and releasing Wolbachia-infected female mosquitoes (or both sexes) could reduce pathogen transmission if the Wolbachia strain reduced vector competence. We cleared Culex quinquefasciatus of their natural Wolbachia pipientis wPip infection and transinfected them with Wolbachia wAlbB isolated from Aedes albopictus. We show that wAlbB infection was transmitted transovarially, and demonstrate cytoplasmic incompatibility with wild-type mosquitoes infected with wPip from Oahu and Maui, Hawaii. We measured vector competence for avian malaria, Plasmodium relictum, lineage GRW4, of seven mosquito lines (two with wAlbB; three with natural wPip infection, and two cleared of Wolbachia infection) by allowing them to feed on canaries infected with recently collected field isolates of Hawaiian P. relictum. We tested 73 groups (Ntotal = 1176) of mosquitoes for P. relictum infection in abdomens and thoraxes 6-14 days after feeding on a range of parasitemias from 0.028% to 2.49%, as well as a smaller subset of salivary glands. We found no measurable effect of Wolbachia on any endpoint, but strong effects of parasitemia, days post feeding, and mosquito strain on both abdomen and thorax infection prevalence. These results suggest that releasing male wAlbB-infected C. quinquefasciatus mosquitoes could suppress wPip-infected mosquito populations, but would have little positive or negative impact on mosquito vector competence for P. relictum if wAlbB became established in local mosquito populations. More broadly, the lack of Wolbachia effects on vector competence we observed highlights the variable impacts of both native and transinfected Wolbachia infections in mosquitoes.}, } @article {pmid39097980, year = {2024}, author = {Bilgo, E and Mancini, MV and Gnambani, JE and Dokpomiwa, HAT and Murdochy, S and Lovett, B and St Leger, R and Sinkins, SP and Diabate, A}, title = {Wolbachia confers protection against the entomopathogenic fungus Metarhizium pingshaense in African Aedes aegypti.}, journal = {Environmental microbiology reports}, volume = {16}, number = {4}, pages = {e13316}, pmid = {39097980}, issn = {1758-2229}, support = {202888/Z/16/Z/WT_/Wellcome Trust/United Kingdom ; AV/PP0025/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; /WT_/Wellcome Trust/United Kingdom ; 108508/Z/15/Z/WT_/Wellcome Trust/United Kingdom ; 226166/Z/22/Z/WT_/Wellcome Trust/United Kingdom ; }, mesh = {*Wolbachia/physiology/genetics ; Animals ; *Metarhizium/physiology ; *Aedes/microbiology ; Symbiosis ; Pest Control, Biological ; Burkina Faso ; Mosquito Control/methods ; Fertility ; Mosquito Vectors/microbiology ; Female ; Longevity ; }, abstract = {Symbiotic and pathogenic microorganisms such as bacteria and fungi represent promising alternatives to chemical insecticides to respond to the rapid increase of insecticide resistance and vector-borne disease outbreaks. This study investigated the interaction of two strains of Wolbachia, wAlbB and wAu, with the natural entomopathogenic fungi from Burkina Faso Metarhizium pingshaense, known to be lethal against Anopheles mosquitoes. In addition to showing the potential of Metarhizium against African Aedes aegypti wild-type populations, our study shows that the wAlbB and wAu provide a protective advantage against entomopathogenic fungal infections. Compared to controls, fungal-infected wAu and wAlbB-carrying mosquitoes showed higher longevity, without any significant impact on fecundity and fertility phenotypes. This study provides new insights into the complex multipartite interaction among the mosquito host, the Wolbachia endosymbiont and the entomopathogenic fungus that might be employed to control mosquito populations. Future research should investigate the fitness costs of Wolbachia, as well as its spread and prevalence within mosquito populations. Additionally, evaluating the impact of Wolbachia on interventions involving Metarhizium pingshaense through laboratory and semi-field population studies will provide valuable insights into the effectiveness of this combined approach.}, } @article {pmid39097253, year = {2024}, author = {Pandey, GS and Manandhar, P and Shrestha, BK and Sadaula, A and Hayashi, N and Abdelbaset, AE and Silwal, P and Tsubota, T and Kwak, ML and Nonaka, N and Nakao, R}, title = {Detection and characterization of vector-borne parasites and Wolbachia endosymbionts in greater one-horned rhinoceros (Rhinoceros unicornis) in Nepal.}, journal = {Acta tropica}, volume = {258}, number = {}, pages = {107344}, doi = {10.1016/j.actatropica.2024.107344}, pmid = {39097253}, issn = {1873-6254}, mesh = {Animals ; *Wolbachia/isolation & purification/genetics ; Nepal ; *Perissodactyla/microbiology/parasitology ; *Symbiosis ; Male ; Theileria/isolation & purification/genetics ; Female ; Vector Borne Diseases ; Filarioidea/isolation & purification/genetics/microbiology ; Filariasis/veterinary/parasitology/transmission/epidemiology ; RNA, Ribosomal, 16S/genetics ; Phylogeny ; Prevalence ; Disease Vectors ; }, abstract = {Vector-borne parasite infections affect both domestic and wild animals. They are often asymptomatic but can result in fatal outcomes under natural and human-induced stressors. Given the limited availability of molecular data on vector-borne parasites in Rhinoceros unicornis (greater one-horned rhinoceros), this study employed molecular tools to detect and characterize the vector-borne parasites in rescued rhinoceros in Chitwan National Park, Nepal. Whole blood samples were collected from thirty-six R. unicornis during rescue and treatment operations. Piroplasmida infections were first screened using nested polymerase chain reaction (PCR) targeting 18S ribosomal RNA gene. Wolbachia was detected by amplifying 16S rRNA gene, while filarial nematodes were detected through amplification of 28S rRNA, COI, myoHC and hsp70 genes. Our results confirmed the presence of Theileria bicornis with a prevalence of 75% (27/36) having two previously unreported haplotypes (H8 and H9). Wolbachia endosymbionts were detected in 25% (9/36) of tested samples and belonged to either supergroup C or F. Filarial nematodes of the genera Mansonella and Onchocerca were also detected. There were no significant association between T. bicornis infections and the age, sex, or location from which the animals were rescued. The high prevalence of Theileria with novel haplotypes along with filarial parasites has important ecological and conservational implications and highlights the need to implement parasite surveillance programs for wildlife in Nepal. Further studies monitoring vector-borne pathogens and interspecies transmission among wild animals, livestock and human are required.}, } @article {pmid39094393, year = {2024}, author = {Turco, S and Brugneti, F and Giubilei, I and Silvestri, C and Petrović, M and Drais, MI and Cristofori, V and Speranza, S and Mazzaglia, A and Contarini, M and Rossini, L}, title = {A bud's life: Metabarcoding analysis to characterise hazelnut big buds microbiome biodiversity.}, journal = {Microbiological research}, volume = {287}, number = {}, pages = {127851}, doi = {10.1016/j.micres.2024.127851}, pmid = {39094393}, issn = {1618-0623}, mesh = {*Corylus/microbiology ; *Microbiota ; Animals ; *DNA Barcoding, Taxonomic ; *Biodiversity ; *Bacteria/classification/genetics/isolation & purification ; Fungi/classification/genetics/isolation & purification ; Mites/microbiology ; Plant Tumors/microbiology ; Fusarium/genetics/classification/isolation & purification ; }, abstract = {Despite Corylus avellana L. being an economically important shrub species known for its resilience to adverse environmental conditions, it constantly faces attacks from a plethora of biotic entities. Among these, the mite pest Phytoptus avellanae is gaining importance, causing economic losses every year. This mite colonises the new generative and vegetative buds, leading them to become swollen and reddish, and drastically reducing hazelnut production. The biology behind gall formation is still poorly understood. This study provides a qualitative and quantitative description of the microbiome in both healthy and infested buds of two economically important hazelnut cultivars through metabarcoding of fungal ITS and bacterial 16 S. Potentially pathogenic genera such as Fusarium and Pseudomonas were predominant in the infested buds, along with the obligate intracellular bacterial genus Wolbachia. Akanthomyces muscarius was instead isolated from culture-based methods only from the infested buds. These findings could improve the understanding of gall ecology, supporting the management of mite populations, and they could also serve as a milestone for further studies on low-impact, monitoring-driven, and genetically targeted control strategies.}, } @article {pmid39091298, year = {2024}, author = {Behrmann, LV and Meier, K and Vollmer, J and Chiedu, CC and Schiefer, A and Hoerauf, A and Pfarr, K}, title = {In vitro extracellular replication of Wolbachia endobacteria.}, journal = {Frontiers in microbiology}, volume = {15}, number = {}, pages = {1405287}, pmid = {39091298}, issn = {1664-302X}, abstract = {Obligate intracellular endobacteria of the genus Wolbachia are widespread in arthropods and several filarial nematodes. Control programs for vector-borne diseases (dengue, Zika, malaria) and anti-filarial therapy with antibiotics are based on this important endosymbiont. Investigating Wolbachia, however, is impeded by the need for host cells. In this study, the requirements for Wolbachia wAlbB growth in a host cell-free in vitro culture system were characterized via qPCRs. A cell lysate fraction from Aedes albopictus C6/36 insect cells containing cell membranes and medium with fetal bovine serum were identified as requisite for cell-free replication of Wolbachia. Supplementation with the membrane fraction of insect cell lysate increased extracellular Wolbachia replication by 4.2-fold. Replication rates in the insect cell-free culture were lower compared to Wolbachia grown inside insect cells. However, the endobacteria were able to replicate for up to 12 days and to infect uninfected C6/36 cells. Cell-free Wolbachia treated with the lipid II biosynthesis inhibitor fosfomycin had an enlarged phenotype, seen previously for intracellular Wolbachia in C6/36 cells, indicating that the bacteria were unable to divide. In conclusion, we have developed a cell-free culture system in which Wolbachia replicate for up to 12 days, providing an in vitro tool to elucidate the biology of these endobacteria, e.g., cell division by using compounds that may not enter the C6/36 cells. A better understanding of Wolbachia biology, and in particular host-symbiont interactions, is key to the use of Wolbachia in vector control programs and to future drug development against filarial diseases.}, } @article {pmid39090271, year = {2024}, author = {Jacobs, J and Nakamoto, A and Mastoras, M and Loucks, H and Mirchandani, C and Karim, L and Penunuri, G and Wanket, C and Russell, SL}, title = {Complete de novo assembly of Wolbachia endosymbiont of Drosophila willistoni using long-read genome sequencing.}, journal = {Scientific reports}, volume = {14}, number = {1}, pages = {17770}, pmid = {39090271}, issn = {2045-2322}, support = {R00 GM135583/GM/NIGMS NIH HHS/United States ; T32 HG012344/HG/NHGRI NIH HHS/United States ; T32HG012344/NH/NIH HHS/United States ; R00GM135583/NH/NIH HHS/United States ; }, mesh = {*Wolbachia/genetics ; Animals ; *Drosophila/microbiology/genetics ; *Symbiosis/genetics ; *Genome, Bacterial ; Phylogeny ; Whole Genome Sequencing/methods ; Genomics/methods ; }, abstract = {Wolbachia is an obligate intracellular α-proteobacterium, which commonly infects arthropods and filarial nematodes. Different strains of Wolbachia are capable of a wide range of regulatory manipulations in their diverse hosts, including the modulation of host cellular differentiation to influence host reproduction. The genetic basis for the majority of these phenotypes is unknown. The wWil strain from the neotropical fruit fly, Drosophila willistoni, exhibits a remarkably high affinity for host germline-derived cells relative to the somatic cells. This trait could be leveraged for understanding how Wolbachia influences the host germline and for controlling host populations in the field. To further the use of this strain in biological and biomedical research, we sequenced the genome of the wWil strain isolated from host cell culture cells. Here, we present the first high quality Nanopore assembly of wWil, the Wolbachia endosymbiont of D. willistoni. Our assembly resulted in a circular genome of 1.27 Mb with a BUSCO completeness score of 99.7%. Consistent with other insect-associated Wolbachia strains, comparative genomic analysis revealed that wWil has a highly mosaic genome relative to the closely related wMel and wAu strains from Drosophila melanogaster and Drosophila simulans, respectively.}, } @article {pmid39084490, year = {2024}, author = {Yadav, P and Borges, RM}, title = {Oviposition decisions under environment-induced physiological stress in parasitoids.}, journal = {Current opinion in insect science}, volume = {65}, number = {}, pages = {101240}, doi = {10.1016/j.cois.2024.101240}, pmid = {39084490}, issn = {2214-5753}, mesh = {Animals ; *Oviposition ; *Stress, Physiological ; *Host-Parasite Interactions ; Environment ; Female ; }, abstract = {Parasitoids constantly evaluate their environment to optimise oviposition host utilisation based on their life history parameters and host characteristics, including density. Any factors impairing chemosensory perception, learning and memory of oviposition decisions negatively impact fitness. In the Anthropocene, stressors, for example, elevated temperatures, Wolbachia infection (likely modulated by temperature), pesticides, light pollution and water deficits in plants that provide resources for parasitoid hosts, impact parasitoid oviposition. Such physiological stressors often induce superparasitism since parasitoids are unable to remember prior oviposition on hosts or cause impaired offspring sex ratio. While the effect of these stressors on parasitoid oviposition has been examined individually, their combined effects remain unexplored. Since parasitoids are exposed to these stressors simultaneously, future work must examine their cumulative impact.}, } @article {pmid39083579, year = {2024}, author = {Crawford, JE}, title = {Virus-blocking mosquitoes take flight in the fight against dengue.}, journal = {Science robotics}, volume = {9}, number = {92}, pages = {eadr0224}, doi = {10.1126/scirobotics.adr0224}, pmid = {39083579}, issn = {2470-9476}, mesh = {Animals ; *Dengue/prevention & control/transmission ; *Dengue Virus/physiology ; *Mosquito Vectors/virology ; *Mosquito Control/methods ; *Aedes/virology/microbiology/physiology ; Humans ; Wolbachia/physiology ; Pest Control, Biological/methods ; Aircraft ; Robotics/instrumentation ; Culicidae/virology/physiology ; }, abstract = {Drone-based mosquito releases facilitate the introduction of dengue-blocking bacteria in wild mosquito populations.}, } @article {pmid39083575, year = {2024}, author = {Lin, YH and Joubert, DA and Kaeser, S and Dowd, C and Germann, J and Khalid, A and Denton, JA and Retski, K and Tavui, A and Simmons, CP and O'Neill, SL and Gilles, JRL}, title = {Field deployment of Wolbachia-infected Aedes aegypti using uncrewed aerial vehicle.}, journal = {Science robotics}, volume = {9}, number = {92}, pages = {eadk7913}, doi = {10.1126/scirobotics.adk7913}, pmid = {39083575}, issn = {2470-9476}, mesh = {Animals ; *Aedes/microbiology ; *Wolbachia/physiology ; *Mosquito Control/methods ; *Mosquito Vectors/microbiology ; Indonesia ; Female ; Dengue/prevention & control/transmission ; Humans ; Robotics/instrumentation ; Male ; Pest Control, Biological/methods ; }, abstract = {Over the past 50 years, there has been a marked increase in diseases like dengue fever, chikungunya, and Zika. The World Mosquito Program (WMP) has developed an approach that, instead of attempting to eliminate vector species, introduces Wolbachia into native Aedes aegypti populations through the release of Wolbachia-infected mosquitoes. Using this approach, a randomized controlled study recently demonstrated a 77% reduction in dengue across a treatment area within Yogyakarta, Indonesia. Existing release methods use the ground-based release of mosquito eggs or adults that are labor-intensive, are logistically challenging to scale up, and can be restrictive in areas where staff safety is a concern. To overcome these limitations, we developed a fully automated mosquito dosing release system that released smaller cohorts of mosquitoes over a wide area and integrated it into an uncrewed aerial vehicle. We established the effectiveness of this system using an aerial mark, release, and recapture approach. We then demonstrated that using only the aerial release method, we can establish Wolbachia infection in a naive Ae. aegypti population. In both cases, the use of aerial releases demonstrated comparable outcomes to ground-based releases without the required labor or risk. These two trials demonstrated the feasibility of using an aerial release approach for large-scale mosquito releases.}, } @article {pmid39079670, year = {2024}, author = {Owashi, Y and Arai, H and Adachi-Hagimori, T and Kageyama, D}, title = {Rickettsia induces strong cytoplasmic incompatibility in a predatory insect.}, journal = {Proceedings. Biological sciences}, volume = {291}, number = {2027}, pages = {20240680}, pmid = {39079670}, issn = {1471-2954}, mesh = {Animals ; *Rickettsia/physiology ; *Symbiosis ; Female ; *Hemiptera/microbiology/physiology ; *Phylogeny ; Male ; Cytoplasm ; Wolbachia/physiology ; }, abstract = {Rickettsia, a group of intracellular bacteria found in eukaryotes, exhibits diverse lifestyles, with some acting as vertebrate pathogens transmitted by arthropod vectors and others serving as maternally transmitted arthropod endosymbionts, some of which manipulate host reproduction for their own benefit. Two phenotypes, namely male-killing and parthenogenesis induction are known as Rickettsia-induced host reproductive manipulations, but it remains unknown whether Rickettsia can induce other types of host manipulation. In this study, we discovered that Rickettsia induced strong cytoplasmic incompatibility (CI), in which uninfected females produce no offspring when mated with infected males, in the predatory insect Nesidiocoris tenuis (Hemiptera: Miridae). Molecular phylogenetic analysis revealed that the Rickettsia strain was related to Rickettsia bellii, a common insect endosymbiont. Notably, this strain carried plasmid-encoded homologues of the CI-inducing factors (namely cifA-like and cifB-like genes), typically found in Wolbachia, which are well-known CI-inducing endosymbionts. Protein domain prediction revealed that the cifB-like gene encodes PD-(D/E)XK nuclease and deubiquitinase domains, which are responsible for Wolbachia-induced CI, as well as ovarian tumour-like (OTU-like) cysteine protease and ankyrin repeat domains. These findings suggest that Rickettsia and Wolbachia endosymbionts share underlying mechanisms of CI and that CI-inducing ability was acquired by microbes through horizontal plasmid transfer.}, } @article {pmid39077840, year = {2024}, author = {Duran-Ahumada, S and Karrer, L and Cheng, C and Roeske, I and Pilchik, J and Jimenez-Vallejo, D and Smith, E and Roy, K and Kirstein, OD and Martin-Park, A and Contreras-Perera, Y and Che-Mendoza, A and Gonzalez-Olvera, G and Puerta-Guardo, HN and Uribe-Soto, SI and Manrique-Saide, P and Vazquez-Prokopec, G}, title = {Wolbachia pipientis (Rickettsiales: Rickettsiaceae) mediated effects on the fitness and performance of Aedes aegypti (Diptera: Culicidae) under variable temperatures and initial larval densities.}, journal = {Journal of medical entomology}, volume = {61}, number = {5}, pages = {1155-1167}, doi = {10.1093/jme/tjae088}, pmid = {39077840}, issn = {1938-2928}, support = {00116467//Emory University/ ; }, mesh = {Animals ; *Aedes/microbiology/growth & development/physiology ; *Wolbachia/physiology ; *Larva/growth & development/microbiology ; Female ; *Temperature ; Male ; Population Density ; Fertility ; Genetic Fitness ; }, abstract = {Wolbachia pipientis (Hertig, 1936), also referred as Wolbachia, is a bacterium present across insect taxa, certain strains of which have been demonstrated to impact the fitness and capacity to transmit viruses in mosquitoes, particularly Aedes aegypti (Linnaeus, 1762). Most studies examine these impacts in limited sets of environmental regimes. Here we seek to understand the impacts of environmentally relevant conditions such as larval density, temperature, and their interaction on wAlbB-infected A. aegypti. Using a factorial design, we measured wAlbB stability (relative density, post-emergence in females, and in progeny), the ability for wAlbB to induce cytoplasmic incompatibility, and bacterial effects on mosquito fitness (fecundity, fertility, and body mass) and performance (adult survival and time to pupation) across 2 temperature regimes (fluctuating and constant) and 2 initial larval densities (low and high). Fluctuating daily regimes of temperature (27 to 40 °C) led to decreased post-emergence wAlbB density and increased wAlbB density in eggs compared to constant temperature (27 °C). An increased fecundity was found in wAlbB-carrying females reared at fluctuating temperatures compared to uninfected wild-type females. wAlbB-carrying adult females showed significantly increased survival than wild-type females. Contrarily, wAlbB-carrying adult males exhibited a significantly lower survival than wild-type males. We found differential effects of assessed treatments (Wolbachia infection status, temperature, and larval density) across mosquito sexes and life stages. Taken together, our results indicate that realistic conditions may not impact dramatically the stability of wAlbB infection in A. aegypti. Nonetheless, understanding the ecological consequence of A. aegypti-wAlbB interaction is complex due to life history tradeoffs under conditions faced by natural populations.}, } @article {pmid39076777, year = {2024}, author = {Frost, CL and Mitchell, R and Smith, JE and Hughes, WOH}, title = {Genotypes and phenotypes in a Wolbachia-ant symbiosis.}, journal = {PeerJ}, volume = {12}, number = {}, pages = {e17781}, pmid = {39076777}, issn = {2167-8359}, mesh = {Animals ; *Wolbachia/physiology/genetics ; *Symbiosis/genetics ; *Ants/microbiology/genetics ; *Genotype ; *Phenotype ; Female ; Male ; }, abstract = {The fitness effects of overt parasites, and host resistance to them, are well documented. Most symbionts, however, are more covert and their interactions with their hosts are less well understood. Wolbachia, an intracellular symbiont of insects, is particularly interesting because it is thought to be unaffected by the host immune response and to have fitness effects mostly focussed on sex ratio manipulation. Here, we use quantitative PCR to investigate whether host genotype affects Wolbachia infection density in the leaf-cutting ant Acromyrmex echinatior, and whether Wolbachia infection density may affect host morphology or caste determination. We found significant differences between host colonies in the density of Wolbachia infections, and also smaller intracolonial differences in infection density between host patrilines. However, the density of Wolbachia infections did not appear to affect the morphology of adult queens or likelihood of ants developing as queens. The results suggest that both host genotype and environment influence the host-Wolbachia relationship, but that Wolbachia infections carry little or no physiological effect on the development of larvae in this system.}, } @article {pmid39072896, year = {2024}, author = {Li, Y and Peng, J and Li, H and Zhang, R and Chen, J and Hou, X and Yang, G}, title = {Integrating pyriproxyfen into the incompatible insect technique enhances mosquito population suppression efficiency and eliminates the risk of population replacement.}, journal = {Pest management science}, volume = {}, number = {}, pages = {}, doi = {10.1002/ps.8339}, pmid = {39072896}, issn = {1526-4998}, support = {82102430//National Natural Science Foundation of China/ ; 12171112//National Natural Science Foundation of China/ ; 2020M672573//China Postdoctoral Science Foundation/ ; }, abstract = {BACKGROUND: The incompatible insect technique (IIT) has been used for Aedes mosquito population suppression to curb the transmission of dengue. However, its wide application is limited owing to the low output of male mosquitoes and the risk of population replacement from the release of fertile Wolbachia-infected females. This study aims to improve IIT efficiency for broader adoption.

RESULTS: We assessed the impact of 10% pyriproxyfen (PPF) sticky powder exposure on Wolbachia (from Culex molestus)-transinfected Aedes albopictus Guangzhou line (GUA line) (GC) mosquitoes. We found that the exposure caused chronic toxicity in adult mosquitoes without affecting the cytoplasmic incompatibility (CI)-inducing capability of males. The PPF-contaminated GC females exhibited significant sterilization and the ability to disseminate lethal doses of PPF to breeding sites. Subsequently, we conducted a field trial combining PPF with IIT aiming to suppress the Ae. albopictus population. This combined approach, termed boosted IIT (BIIT), showed a notable enhancement in population suppression efficiency. The improved efficacy of BIIT was attributed to the dispersion of PPF particles in the field via the released PPF-contaminated male mosquitoes. During the BIIT field trial, no Wolbachia wPip-positive Ae. albopictus larvae were detected, indicating the effective elimination of the risk of Wolbachia-induced population replacement. Additionally, the field trial of BIIT against Ae. albopictus resulted in the suppression of the nontarget mosquito species Culex quinquefasciatus.

CONCLUSION: Our results highlight the remarkable efficiency and feasibility of combining IIT with PPF in suppressing mosquito populations, facilitating the widespread implementation of IIT-based management of mosquito-borne diseases. © 2024 Society of Chemical Industry.}, } @article {pmid39066296, year = {2024}, author = {Madhav, M and Blasdell, KR and Trewin, B and Paradkar, PN and López-Denman, AJ}, title = {Culex-Transmitted Diseases: Mechanisms, Impact, and Future Control Strategies using Wolbachia.}, journal = {Viruses}, volume = {16}, number = {7}, pages = {}, pmid = {39066296}, issn = {1999-4915}, mesh = {*Wolbachia/physiology ; Animals ; *Culex/microbiology/virology ; *Mosquito Vectors/microbiology/virology ; Humans ; *Mosquito Control/methods ; }, abstract = {Mosquitoes of the Culex genus are responsible for a large burden of zoonotic virus transmission globally. Collectively, they play a significant role in the transmission of medically significant diseases such as Japanese encephalitis virus and West Nile virus. Climate change, global trade, habitat transformation and increased urbanisation are leading to the establishment of Culex mosquitoes in new geographical regions. These novel mosquito incursions are intensifying concerns about the emergence of Culex-transmitted diseases and outbreaks in previously unaffected areas. New mosquito control methods are currently being developed and deployed globally. Understanding the complex interaction between pathogens and mosquitoes is essential for developing new control strategies for Culex species mosquitoes. This article reviews the role of Culex mosquitos as vectors of zoonotic disease, discussing the transmission of viruses across different species, and the potential use of Wolbachia technologies to control disease spread. By leveraging the insights gained from recent successful field trials of Wolbachia against Aedes-borne diseases, we comprehensively discuss the feasibility of using this technique to control Culex mosquitoes and the potential for the development of next generational Wolbachia-based control methods.}, } @article {pmid39057829, year = {2024}, author = {Collado-Cuadrado, M and Alarcón-Torrecillas, C and Rodríguez-Escolar, I and Balmori-de la Puente, A and Infante González-Mohino, E and Pericacho, M and Morchón, R}, title = {Wolbachia Promotes an Anti-Angiogenic Response Using an In Vitro Model of Vascular Endothelial Cells in Relation to Heartworm Disease.}, journal = {Pathogens (Basel, Switzerland)}, volume = {13}, number = {7}, pages = {}, pmid = {39057829}, issn = {2076-0817}, support = {//General Foundation of University of Salamanca/ ; //Margarita-Salas as postdoctoral scholarship/ ; //University of Salamanca-Banco Santander/ ; }, abstract = {Heartworm disease caused by Dirofilaria immitis is a vector-borne zoonotic disease responsible for the infection of mainly domestic dogs and cats, or these are those for which the most data are known. Humans are an accidental host where a benign, asymptomatic pulmonary nodule may originate. Dirofilaria immitis also harbours the endosymbiont bacteria of the genus Wolbachia, which play a role in moulting, embryogenesis, inflammatory pathology, and immune response. When Wolbachia sp. is released into the bloodstream, endothelial and pulmonary damage is exacerbated, further encouraging thrombus formation and pulmonary hypertension, facilitating congestive heart failure and death of the animal. Previous studies have shown that parasite excretory/secretory products are able to activate the pro-angiogenic pathway (formation of new vessels) to facilitate parasite survival. The aim of this study was to analyse the role of Wolbachia sp. and its relationship with the cellular processes and the angiogenic pathway in a model of human endothelial cells in vitro. The use of recombinant Wolbachia Surface Protein (rWSP) showed that its stimulation exerted an anti-angiogenic effect by detecting an increase in the production of VEGFR-1/sFlt1 and sEndoglin and did not affect the production of VEGFR-2 and mEndoglin (pro-angiogenic molecules). Furthermore, it did not stimulate cell proliferation or migration, although it did negatively stimulate the formation of pseudocapillaries, slowing down this process. These cellular processes are directly related to the angiogenic pathway so, with these results, we can conclude that Wolbachia sp. is related to the stimulation of the anti-angiogenic pathway, not facilitating the survival of D. immitis in vascular endothelium.}, } @article {pmid39057258, year = {2024}, author = {Sakamoto, JM and Shikano, I and Rasgon, JL}, title = {Microbiomes of Two Pest Fly Species of Pennsylvania Mushroom Houses.}, journal = {Insects}, volume = {15}, number = {7}, pages = {}, pmid = {39057258}, issn = {2075-4450}, support = {Project #PEN04912 (Accession #7006384) and Project #PEN04769 (Accession #1010032)//United States Department of Agriculture/ ; NA//Giorgi Mushroom Fund/ ; }, abstract = {Mushroom cultivation vastly improves the yield of mushrooms under optimized, controlled conditions, but may be susceptible to opportunistic colonization by pest species that can establish themselves, as well as the pathogens and pests they may transmit. Here, we describe our investigation into the bacterial communities of adult Lycoriella ingenua (Diptera: Sciaridae) and Megaselia halterata (Diptera: Phoridae) collected from button mushroom (Agaricus bisporus) production houses in Pennsylvania. We collected adult flies and sequenced the hypervariable v4 region of the bacterial 16S rRNA using the Illumina MiSeq. The most abundant bacterial genus detected in both species was Wolbachia, but phylogenetic analysis revealed that the infections are from different clades. Future studies include the characterization of Wolbachia infections on fly behavior and biology, comparison of microbial diversity of fly species colonizing wild mushrooms, and other microbiota that may contribute to the success of certain pest fly species.}, } @article {pmid39054929, year = {2024}, author = {Singh, R and Suresh, S and Fewell, JH and Harrison, JF and Linksvayer, TA}, title = {Wolbachia-infected pharaoh ant colonies have higher egg production, metabolic rate and worker survival.}, journal = {The Journal of experimental biology}, volume = {227}, number = {16}, pages = {}, doi = {10.1242/jeb.247168}, pmid = {39054929}, issn = {1477-9145}, support = {IOS-1452520//National Science Foundation/ ; //National Science Foundation/ ; }, mesh = {Animals ; *Wolbachia/physiology ; *Ants/microbiology/physiology ; Female ; *Longevity ; Oviposition/physiology ; Symbiosis ; Ovum/microbiology/physiology ; Reproduction ; }, abstract = {Wolbachia is a widespread maternally transmitted endosymbiotic bacteria with diverse phenotypic effects on its insect hosts, ranging from parasitic to mutualistic. Wolbachia commonly infects social insects, where it faces unique challenges associated with its host's caste-based reproductive division of labor and colony living. Here, we dissect the benefits and costs of Wolbachia infection on life-history traits of the invasive pharaoh ant, Monomorium pharaonis, which are relatively short lived and show natural variation in Wolbachia infection status between colonies. We quantified the effects of Wolbachia infection on the lifespan of queen and worker castes, the egg-laying rate of queens across queen lifespan, and the metabolic rates of whole colonies and colony members. Infected queens laid more eggs than uninfected queens but had similar metabolic rates and lifespans. Interestingly, infected workers outlived uninfected workers. At the colony level, infected colonies were more productive as a consequence of increased queen egg-laying rates and worker longevity, and infected colonies had higher metabolic rates during peak colony productivity. While some effects of infection, such as elevated colony-level metabolic rates, may be detrimental in more stressful natural conditions, we did not find any costs of infection under relatively benign laboratory conditions. Overall, our study emphasizes that Wolbachia infection can have beneficial effects on ant colony growth and worker survival in at least some environments.}, } @article {pmid39052691, year = {2024}, author = {Mirchandani, C and Wang, P and Jacobs, J and Genetti, M and Pepper-Tunick, E and Sullivan, WT and Corbett-Detig, R and Russell, SL}, title = {Mixed Wolbachia infections resolve rapidly during in vitro evolution.}, journal = {PLoS pathogens}, volume = {20}, number = {7}, pages = {e1012149}, pmid = {39052691}, issn = {1553-7374}, support = {R00 GM135583/GM/NIGMS NIH HHS/United States ; R35 GM128932/GM/NIGMS NIH HHS/United States ; R35 GM139595/GM/NIGMS NIH HHS/United States ; T32 HG012344/HG/NHGRI NIH HHS/United States ; }, mesh = {*Wolbachia/physiology ; Animals ; *Drosophila melanogaster/microbiology ; *Symbiosis ; Biological Evolution ; Drosophila simulans/microbiology ; Cell Line ; }, abstract = {The intracellular symbiont Wolbachia pipientis evolved after the divergence of arthropods and nematodes, but it reached high prevalence in many of these taxa through its abilities to infect new hosts and their germlines. Some strains exhibit long-term patterns of co-evolution with their hosts, while other strains are capable of switching hosts. This makes strain selection an important factor in symbiont-based biological control. However, little is known about the ecological and evolutionary interactions that occur when a promiscuous strain colonizes an infected host. Here, we study what occurs when two strains come into contact in host cells following horizontal transmission and infection. We focus on the faithful wMel strain from Drosophila melanogaster and the promiscuous wRi strain from Drosophila simulans using an in vitro cell culture system with multiple host cell types and combinatorial infection states. Mixing D. melanogaster cell lines stably infected with wMel and wRi revealed that wMel outcompetes wRi quickly and reproducibly. Furthermore, wMel was able to competitively exclude wRi even from minuscule starting quantities, indicating that this is a nearly deterministic outcome, independent of the starting infection frequency. This competitive advantage was not exclusive to wMel's native D. melanogaster cell background, as wMel also outgrew wRi in D. simulans cells. Overall, wRi is less adept at in vitro growth and survival than wMel and its in vivo state, revealing differences between the two strains in cellular and humoral regulation. These attributes may underlie the observed low rate of mixed infections in nature and the relatively rare rate of host-switching in most strains. Our in vitro experimental framework for estimating cellular growth dynamics of Wolbachia strains in different host species and cell types provides the first strategy for parameterizing endosymbiont and host cell biology at high resolution. This toolset will be crucial to our application of these bacteria as biological control agents in novel hosts and ecosystems.}, } @article {pmid39041013, year = {2024}, author = {Henry, LP and Fernandez, M and Wolf, S and Abhyankar, V and Ayroles, JF}, title = {Wolbachia impacts microbiome diversity and fitness-associated traits for Drosophila melanogaster in a seasonally fluctuating environment.}, journal = {Ecology and evolution}, volume = {14}, number = {7}, pages = {e70004}, pmid = {39041013}, issn = {2045-7758}, abstract = {The microbiome contributes to many different host traits, but its role in host adaptation remains enigmatic. The fitness benefits of the microbiome often depend on ecological conditions, but theory suggests that fluctuations in both the microbiome and environment modulate these fitness benefits. Moreover, vertically transmitted bacteria might constrain the ability of both the microbiome and host to respond to changing environments. Drosophila melanogaster provides an excellent system to investigate the impacts of interactions between the microbiome and the environment. To address this question, we created field mesocosms of D. melanogaster undergoing seasonal environmental change with and without the vertically transmitted bacteria, Wolbachia pipientis. Sampling temporal patterns in the microbiome revealed that Wolbachia constrained microbial diversity. Furthermore, Wolbachia and a dominant member of the microbiome, Commensalibacter, were associated with differences in two higher-order fitness traits, starvation resistance and lifespan. Our work here suggests that the interplay between the abiotic context and microbe-microbe interactions may shape key host phenotypes that underlie adaptation to changing environments. We conclude by exploring the consequences of complex interactions between Wolbachia and the microbiome for our understanding of eco-evolutionary processes that shape host-microbiome interactions.}, } @article {pmid39031863, year = {2024}, author = {Lombardi, G and Lampazzi, E and Calvitti, M}, title = {Incompatible insect technique: insights on potential outcomes of releasing contaminant females: a proof of concept under semi-field conditions.}, journal = {Pest management science}, volume = {}, number = {}, pages = {}, doi = {10.1002/ps.8263}, pmid = {39031863}, issn = {1526-4998}, abstract = {BACKGROUND: Releasing large numbers of Aedes albopictus males, carrying the artificially introduced Wolbachia 'wPip' strain, results in a decrease in the reproductive capacity of wild females due to a phenomenon known as cytoplasmic incompatibility (CI). This vector control strategy is referred to as the incompatible insect technique (IIT). However, its widespread implementation faces various challenges, including the complexity of removing fertile females from the males intended for release. Here, we present the results of semi-field experiments comparing the impact of minimal female co-release on two IIT modes: unidirectional CI-based (UnCI IIT) and bidirectional CI-based (BiCI IIT), specifically targeting Ae. albopictus.

RESULTS: The contamination of 'wPip' infected females (2%) during male releases significantly weakened the overall effectiveness of IIT, emphasizing the need for thorough sex separation. Specifically, with UnCI IIT, despite the low rate of co-released females, there was a gradual rise in 'wPip' infection frequency, resulting in more compatible mating and subsequently higher rates of egg hatching. Conversely, this pattern was effectively mitigated in BiCI IIT owing to the reciprocal sterility between the wild-type and the 'wPip' infected populations.

CONCLUSION: Through an experimental approach, conducted in a semi-field setting, we have contributed to advancing scientific understanding regarding the potential outcomes of implementing the IIT strategy in the absence of a complete sexing system. The results suggest that safety measures for mitigating the potential impacts of co-released females can be tailored according to the specific type of IIT being utilized. © 2024 Society of Chemical Industry.}, } @article {pmid39029846, year = {2024}, author = {Mathimaran, A and Nagarajan, H and Mathimaran, A and Huang, YC and Chen, CJ and Vetrivel, U and Jeyaraman, J}, title = {Deciphering the pH-dependent oligomerization of aspartate semialdehyde dehydrogenase from Wolbachia endosymbiont of Brugia malayi: An in vitro and in silico approaches.}, journal = {International journal of biological macromolecules}, volume = {276}, number = {Pt 2}, pages = {133977}, doi = {10.1016/j.ijbiomac.2024.133977}, pmid = {39029846}, issn = {1879-0003}, mesh = {*Brugia malayi/enzymology/microbiology ; Hydrogen-Ion Concentration ; Animals ; *Aspartate-Semialdehyde Dehydrogenase/metabolism/chemistry/genetics ; *Protein Multimerization ; *Wolbachia/enzymology ; Molecular Dynamics Simulation ; Computer Simulation ; Symbiosis ; NADP/metabolism ; }, abstract = {The enzyme aspartate semialdehyde dehydrogenase (ASDH) plays a pivotal role in the amino acid biosynthesis pathway, making it an attractive target for the development of new antimicrobial drugs due to its absence in humans. This study aims to investigate the presence of ASDH in the filarial parasite Wolbachia endosymbiont of Brugia malayi (WBm) using both in vitro and in silico approaches. The size exclusion chromatography (SEC) and Native-PAGE analysis demonstrate that WBm-ASDH undergoes pH-dependent oligomerization and dimerization. To gain a deeper understanding of this phenomenon, the modelled monomer and dimer structures were subjected to pH-dependent dynamics simulations in various conditions. The results reveal that residues Val240, Gln161, Thr159, Tyr160, and Trp316 form strong hydrogen bond contacts in the intersurface area to maintain the structure in the dimeric form. Furthermore, the binding of NADP[+] induces conformational changes, leading to an open or closed conformation in the structure. Importantly, the binding of NADP[+] does not disturb either the dimerization or oligomerization of the protein, a finding confirmed through both in vitro and in silico analysis. These findings shed light on the structural characteristics of WBm-ASDH and offer valuable insights for the development of new inhibitors specific to WBm, thereby contributing to the development of potential therapies for filarial parasitic infections.}, } @article {pmid39019394, year = {2024}, author = {Santana, TDD and Rodrigues, TM and Andrade, LA and Santos, ER and Ardisson-Araújo, DMP}, title = {Three picorna-like viruses found associated with the spider mite, Tetranychus truncatus (Acari: Tetranychidae).}, journal = {Journal of invertebrate pathology}, volume = {206}, number = {}, pages = {108169}, doi = {10.1016/j.jip.2024.108169}, pmid = {39019394}, issn = {1096-0805}, mesh = {Animals ; *Tetranychidae/virology ; *Picornaviridae/genetics/classification ; Phylogeny ; Genome, Viral ; }, abstract = {Herbivorous arthropods, such as mites and insects, host a variety of microorganisms that significantly influence their ecology and evolution. While insect viruses have been extensively studied, our understanding of the diversity and composition of mite viromes and the interactions with mite hosts remains limited. The Asian spider mite, Tetranychus truncatus Ehara (Acari: Tetranychidae), a major agricultural pest, has not yet been reported to harbor any viruses. Here, using publicly available RNA-Seq data, we identified and characterized three picorna-like viruses associated with T. truncatus: Tetranychus truncatus-associated iflavirus 1 (TtAIV-1), Tetranychus truncatus-associated picorna-like virus 1 (TtAV-1), and Tetranychus truncatus-associated picorna-like virus 2 (TtAV-2). TtAIV-1 has a typical Iflaviridae genome structure with a single ORF, representing the first iflavirus associated with the Tetranychus genus. TtAV-1 and TtAV-2 exhibit bicistronic arrangements similar to dicistroviruses and other picorna-like viruses, with complex secondary structures in their non-coding regions. Phylogenetic analysis places TtAIV-1 within Iflaviridae, possibly as a new species, while TtAV-1 and TtAV-2 form distinct clades within unclassified picorna-like viruses, suggesting new families within Picornavirales. We analyzed in silico the presence and abundance of these viruses in T. truncatus across four bioproject SRAs, mostly finding them co-associated, with viral reads reaching up to 30% of total reads. Their presence and abundance varied by mite treatment and origin, with no significant impact from Wolbachia infection or abamectin exposure, although TtAV-2 was absent in abamectin-treated mites. Temperature influenced virus abundance, and variations were observed among Chinese mite populations based on geography and host plant association. Our findings offer insights into picorna-like virus diversity and dynamics in T. truncatus, revealing potential roles in mite biology and suggesting applications for mite population control, thereby enhancing agricultural productivity and food security.}, } @article {pmid39009178, year = {2024}, author = {Power, RI and Doyle, SR and Šlapeta, J}, title = {Whole genome amplification and sequencing of individual Dirofilaria immitis microfilariae.}, journal = {Experimental parasitology}, volume = {263-264}, number = {}, pages = {108806}, doi = {10.1016/j.exppara.2024.108806}, pmid = {39009178}, issn = {1090-2449}, mesh = {Animals ; *Dirofilaria immitis/genetics/isolation & purification ; Dogs ; *Genome, Helminth ; *Dog Diseases/parasitology ; *Dirofilariasis/parasitology ; *Microfilariae/genetics/isolation & purification ; *Whole Genome Sequencing ; *DNA, Helminth/isolation & purification/chemistry ; Female ; Male ; }, abstract = {Dirofilaria immitis is a filarial parasitic nematode of veterinary significance. With the emergence of drug-resistant isolates in the USA, it is imperative to determine the likelihood of resistance occurring in other regions of the world. One approach is to conduct population genetic studies across an extensive geographical range, and to sequence the genomes of individual worms to understand genome-wide genetic variation associated with resistance. The immature life stages of D. immitis found in the host blood are more accessible and less invasive to sample compared to extracting adult stages from the host heart. To assess the use of immature life stages for population genetic analyses, we have performed whole genome amplification and whole-genome sequencing on nine (n = 9) individual D. immitis microfilaria samples isolated from dog blood. On average, less than 1% of mapped reads aligned to each D. immitis genome (nuclear, mitochondrial, and Wolbachia endosymbiont). For the dog genome, an average of over 99% of mapped reads aligned to the nuclear genome and less than 1% aligned to the mitochondrial genome. The average coverage for all D. immitis genomes and the dog nuclear genome was less than 1, while the dog mitochondrial genome had an average coverage of 2.87. The overwhelming proportion of sequencing reads mapping to the dog host genome can be attributed to residual dog blood cells in the microfilariae samples. These results demonstrate the challenges of conducting genome-wide studies on individual immature parasite life stages, particularly in the presence of extraneous host DNA.}, } @article {pmid39004260, year = {2024}, author = {Tischer, M and Bleidorn, C}, title = {Further evidence of low infection frequencies of Wolbachia in soil arthropod communities.}, journal = {Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases}, volume = {123}, number = {}, pages = {105641}, doi = {10.1016/j.meegid.2024.105641}, pmid = {39004260}, issn = {1567-7257}, mesh = {*Wolbachia/genetics/classification ; Animals ; *Arthropods/microbiology ; *Phylogeny ; *Soil Microbiology ; Germany/epidemiology ; Multilocus Sequence Typing ; }, abstract = {Endosymbiotic Alphaproteobacteria of the genus Wolbachia are exclusively transferred maternally from mother to offspring, but horizontal transfer across species boundaries seems to be frequent as well. However, the (ecological) mechanisms of how these bacteria are transferred between distantly related arthropod hosts remain unclear. Based on the observation that species that are part of the same ecological community often also share similar Wolbachia strains, host ecology has been hypothesized as an important factor enabling transmission and a key factor in explaining the global distribution of Wolbachia lineages. In this study, we focus on the diversity and abundance of Wolbachia strains in soil arthropods, a so far rather neglected community. We screened 82 arthropod morphotypes collected in the beech forest (dominated by Fagus sp.) soil in the area of Göttingen in central Germany for the presence of Wolbachia. By performing a PCR screen with Wolbachia-MLST markers (coxA, dnaA, fbpA, ftsZ, gatB, and hcpA), we found a rather low infection frequency of 12,2%. Additionally, we performed metagenomic screening of pooled individuals from the same sampling site and could not find evidence that this low infection frequency is an artefact due to PCR-primer bias. Phylogenetic analyses of the recovered Wolbachia strains grouped them in three known supergroups (A, B, and E), with the first report of Wolbachia in Protura (Hexapoda). Moreover, Wolbachia sequences from the pseudoscorpion Neobisium carcinoides cluster outside the currently known supergroup diversity. Our screening supports results from previous studies that the prevalence of Wolbachia infections seems to be lower in soil habitats than in above-ground terrestrial habitats. The reasons for this pattern are not completely understood but might stem from the low opportunity of physical contact and the prevalence of supergroups that are less suited for horizontal transfer.}, } @article {pmid38997103, year = {2024}, author = {Lau, MJ and Nie, S and Ross, PA and Endersby-Harshman, NM and Hoffmann, AA}, title = {Long-term impacts of egg quiescence and Wolbachia infection on lipid profiles in Aedes aegypti: Ovarian roles in lipid synthesis during reproduction.}, journal = {Journal of insect physiology}, volume = {157}, number = {}, pages = {104674}, doi = {10.1016/j.jinsphys.2024.104674}, pmid = {38997103}, issn = {1879-1611}, mesh = {Animals ; *Aedes/microbiology/metabolism/physiology ; *Wolbachia/physiology ; Female ; *Ovary/microbiology/metabolism ; Ovum/microbiology/metabolism ; Reproduction ; Lipids/biosynthesis ; Lipid Metabolism ; Lipidomics ; }, abstract = {Wolbachia, an endosymbiotic bacterium, relies on nutrients from its host to complete its life cycle. The presence of Wolbachia strain wAlbB in the mosquito Aedes aegypti during egg or larval stages affects the host's development, leading to the absence of developed and visible ovaries in adult mosquito females. In this study, we investigated the impacts of egg quiescence and Wolbachia infection on lipid profiles of adult Ae. aegypti females, and discerned the role of ovaries in lipid synthesis in the reproductive process. The lipidomes of Wolbachia infected and uninfected female individuals at various developmental stages were quantitatively analyzed by LC-MS/MS. Lipidomic change patterns were systematically further investigated in wAlbB-infected fertile females and infertile females following blood feeding. Prolonged egg quiescence induced a shortage of acyl-carnitine (CAR) and potentially impacted some molecules of diacyl-phospholipid (diacyl-PL) and sphingolipid (SL) in young adult mosquitoes. After the first gonotrophic cycle, infertile females accumulated more CAR and lyso-phospholipid (lyso-PL) than fertile females. Then in the second gonotrophic cycle, the patterns of different lipid groups remained similar between fertile and infertile females. Only a small proportion of molecules of triglyceride (TG), phospholipid (lyso-PL and diacyl-PL) and ceramide (Cer) increased exclusively in fertile females from 0 h to 16 h post blood meal, suggesting that the generation or prescence of these lipids rely on ovaries. In addition, we found cardiolipins (CL) might be impacted by Wolbachia infection at the egg stage, and infected mosquitoes also showed distinct patterns between fertile and infertile females at their second gonotrophic cycle. Our study provides new insights into the long-term influence of Wolbachia on lipid profiles throughout various life stages of mosquitoes. Additionally, it suggests a role played by ovaries in lipid synthesis during mosquito reproduction.}, } @article {pmid38992783, year = {2024}, author = {Klimov, PB and Hubert, J and Erban, T and Alejandra Perotti, M and Braig, HR and Flynt, A and He, Q and Cui, Y}, title = {Genomic and metagenomic analyses of the domestic mite Tyrophagus putrescentiae identify it as a widespread environmental contaminant and a host of a basal, mite-specific Wolbachia lineage (supergroup Q).}, journal = {International journal for parasitology}, volume = {}, number = {}, pages = {}, doi = {10.1016/j.ijpara.2024.07.001}, pmid = {38992783}, issn = {1879-0135}, abstract = {Tyrophagus putrescentiae (mould mite) is a global, microscopic trophic generalist that commonly occurs in various human-created habitats, causing allergies and damaging stored food. Its ubiquity and extraordinary ability to penetrate research samples or cultures through air currents or by active walking through tights spaces (such as treads of screw caps) may lead to sample contamination and introduction of its DNA to research materials in the laboratory. This prompts a thorough investigation into potential sequence contamination in public genomic databases. The trophic success of T. putrescentiae is primarily attributed to the symbiotic bacteria housed in specialized internal mite structures, facilitating adaptation to varied nutritional niches. However, recent work suggests that horizontal transfer of bacterial/fungal genes related to nutritional functionality may also contribute to the mite's trophic versatility. This aspect requires independent confirmation. Additionally, T. putrescentiae harbors an uncharacterized and genetically divergent bacterium, Wolbachia, displaying blocking and microbiome-modifying effects. The phylogenomic position and supergroup assignment of this bacterium are unknown. Here, we sequenced and assembled the T. putrescentiae genome, analyzed its microbiome, and performed detailed phylogenomic analyses of the mite-specific Wolbachia. We show that T. putrescentiae DNA is a substantial source of contamination of research samples. Its DNA may inadvertently be co-extracted with the DNA of the target organism, eventually leading to sequence contamination in public databases. We identified a diversity of bacterial species associated with T. putrescentiae, including those capable of rapidly developing antibiotic resistance, such as Escherichia coli. Despite the presence of diverse bacterial communities in T. putrescentiae, we did not detect any recent horizontal gene transfers in this mite species and/or in astigmatid (domestic) mites in general. Our phylogenomic analysis of Wolbachia recovered a basal, mite-specific lineage (supergroup Q) represented by two Wolbachia spp. from the mould mite and a gall-inducing plant mite. Fluorescence in situ hybridization confirmed the presence of Wolbachia inside the mould mite. The discovery of an early derivative Wolbachia lineage (supergroup Q) in two phylogenetically unrelated and ecologically dissimilar mites suggests that this endosymbiotic bacterial lineage formed a long-term association with mites. This finding provides a unique insight into the early evolution and host associations of Wolbachia. Further discoveries of Wolbachia diversity in acariform mites are anticipated.}, } @article {pmid38985862, year = {2024}, author = {Hoffmann, G and Lukarska, M and Clare, RH and Masters, EKG and Johnston, KL and Ford, L and Turner, JD and Ward, SA and Taylor, MJ and Jensen, MR and Palencia, A}, title = {Targeting a microbiota Wolbachian aminoacyl-tRNA synthetase to block its pathogenic host.}, journal = {Science advances}, volume = {10}, number = {28}, pages = {eado1453}, pmid = {38985862}, issn = {2375-2548}, mesh = {*Wolbachia/drug effects ; Humans ; *Microbiota ; Animals ; Leucine-tRNA Ligase/metabolism/antagonists & inhibitors ; Amino Acyl-tRNA Synthetases/metabolism/antagonists & inhibitors ; Crystallography, X-Ray ; Boron Compounds/pharmacology/chemistry ; Symbiosis ; Models, Molecular ; }, abstract = {The interplay between humans and their microbiome is crucial for various physiological processes, including nutrient absorption, immune defense, and maintaining homeostasis. Microbiome alterations can directly contribute to diseases or heighten their likelihood. This relationship extends beyond humans; microbiota play vital roles in other organisms, including eukaryotic pathogens causing severe diseases. Notably, Wolbachia, a bacterial microbiota, is essential for parasitic worms responsible for lymphatic filariasis and onchocerciasis, devastating human illnesses. Given the lack of rapid cures for these infections and the limitations of current treatments, new drugs are imperative. Here, we disrupt Wolbachia's symbiosis with pathogens using boron-based compounds targeting an unprecedented Wolbachia enzyme, leucyl-tRNA synthetase (LeuRS), effectively inhibiting its growth. Through a compound demonstrating anti-Wolbachia efficacy in infected cells, we use biophysical experiments and x-ray crystallography to elucidate the mechanism behind Wolbachia LeuRS inhibition. We reveal that these compounds form adenosine-based adducts inhibiting protein synthesis. Overall, our study underscores the potential of disrupting key microbiota to control infections.}, } @article {pmid38982550, year = {2024}, author = {Du, CH and Xiang, R and Bie, SS and Yang, X and Yang, JH and Yao, MG and Zhang, Y and He, ZH and Shao, ZT and Luo, CF and Pu, EN and Li, YQ and Wang, F and Luo, Z and Du, CB and Zhao, J and Li, M and Cao, WC and Sun, Y and Jiang, JF}, title = {Genetic diversity and prevalence of emerging Rickettsiales in Yunnan Province: a large-scale study.}, journal = {Infectious diseases of poverty}, volume = {13}, number = {1}, pages = {54}, pmid = {38982550}, issn = {2049-9957}, support = {U200221//National Natural Science Foundation of China/ ; 2019YFC1200501//Key Technologies Research and Development Program/ ; 2022YFC2305001//Key Technologies Research and Development Program/ ; }, mesh = {China/epidemiology ; Animals ; *Genetic Variation ; *Phylogeny ; Prevalence ; *Rickettsiales/genetics/isolation & purification/classification ; *Ticks/microbiology ; RNA, Ribosomal, 16S/genetics/analysis ; Livestock/microbiology ; Rickettsia Infections/epidemiology/microbiology/veterinary ; Rickettsia/isolation & purification/genetics/classification ; Mammals/microbiology ; Humans ; }, abstract = {BACKGROUND: Rickettsia and related diseases have been identified as significant global public health threats. This study involved comprehensive field and systematic investigations of various rickettsial organisms in Yunnan Province.

METHODS: Between May 18, 2011 and November 23, 2020, field investigations were conducted across 42 counties in Yunnan Province, China, encompassing small mammals, livestock, and ticks. Preliminary screenings for Rickettsiales involved amplifying the 16S rRNA genes, along with additional genus- or species-specific genes, which were subsequently confirmed through sequencing results. Sequence comparisons were carried out using the Basic Local Alignment Search Tool (BLAST). Phylogenetic relationships were analyzed using the default parameters in the Molecular Evolutionary Genetics Analysis (MEGA) program. The chi-squared test was used to assess the diversities and component ratios of rickettsial agents across various parameters.

RESULTS: A total of 7964 samples were collected from small mammals, livestock, and ticks through Yunnan Province and submitted for screening for rickettsial organisms. Sixteen rickettsial species from the genera Rickettsia, Anaplasma, Ehrlichia, Neoehrlichia, and Wolbachia were detected, with an overall prevalence of 14.72%. Among these, 11 species were identified as pathogens or potential pathogens to humans and livestock. Specifically, 10 rickettsial organisms were widely found in 42.11% (24 out of 57) of small mammal species. High prevalence was observed in Dremomys samples at 5.60%, in samples from regions with latitudes above 4000 m or alpine meadows, and in those obtained from Yuanmou County. Anaplasma phagocytophilum and Candidatus Neoehrlichia mikurensis were broadly infecting multiple genera of animal hosts. In contrast, the small mammal genera Neodon, Dremomys, Ochotona, Anourosorex, and Mus were carrying individually specific rickettsial agents, indicating host tropism. There were 13 rickettsial species detected in 57.14% (8 out of 14) of tick species, with the highest prevalence (37.07%) observed in the genus Rhipicephalus. Eight rickettsial species were identified in 2375 livestock samples. Notably, six new Rickettsiales variants/strains were discovered, and Candidatus Rickettsia longicornii was unambiguously identified.

CONCLUSIONS: This large-scale survey provided further insight into the high genetic diversity and overall prevalence of emerging Rickettsiales within endemic hotspots in Yunnan Province. The potential threats posed by these emerging tick-borne Rickettsiales to public health warrant attention, underscoring the need for effective strategies to guide the prevention and control of emerging zoonotic diseases in China.}, } @article {pmid38976758, year = {2024}, author = {Lau, MJ and Valdez, AR and Jones, MJ and Aranson, I and Hoffmann, AA and McGraw, EA}, title = {The effect of repeat feeding on dengue virus transmission potential in Wolbachia-infected Aedes aegypti following extended egg quiescence.}, journal = {PLoS neglected tropical diseases}, volume = {18}, number = {7}, pages = {e0012305}, pmid = {38976758}, issn = {1935-2735}, support = {R56 AI155573/AI/NIAID NIH HHS/United States ; }, mesh = {*Aedes/microbiology/virology/physiology ; Animals ; *Wolbachia/physiology ; *Dengue Virus/physiology ; *Mosquito Vectors/microbiology/virology/physiology ; *Dengue/transmission ; Female ; *Feeding Behavior ; Viral Load ; Ovum/virology/microbiology ; }, abstract = {As Wolbachia pipientis is more widely being released into field populations of Aedes aegypti for disease control, the ability to select the appropriate strain for differing environments is increasingly important. A previous study revealed that longer-term quiescence in the egg phase reduced the fertility of mosquitoes, especially those harboring the wAlbB Wolbachia strain. This infertility was also associated with a greater biting rate. Here, we attempt to quantify the effect of this heightened biting behavior on the transmission potential of the dengue virus using a combination of assays for fitness, probing behavior, and vector competence, allowing repeat feeding, and incorporate these effects in a model of R0. We show that Wolbachia-infected infertile mosquitoes are more interested in feeding almost immediately after an initial blood meal relative to wild type and Wolbachia-infected fertile mosquitoes and that these differences continue for up to 8 days over the period we measured. As a result, the infertile Wolbachia mosquitoes have higher virus prevalence and loads than Wolbachia-fertile mosquitoes. We saw limited evidence of Wolbachia-mediated blocking in the disseminated tissue (legs) in terms of prevalence but did see reduced viral loads. Using a previously published estimate of the extrinsic incubation period, we demonstrate that the effect of repeat feeding/infertility is insufficient to overcome the effects of Wolbachia-mediated blocking on R0. These estimates are very conservative, however, and we posit that future studies should empirically measure EIP under a repeat feeding model. Our findings echo previous work where periods of extensive egg quiescence affected the reproductive success of Wolbachia-infected Ae. aegypti. Additionally, we show that increased biting behavior in association with this infertility in Wolbachia-infected mosquitoes may drive greater vector competence. These relationships require further exploration, given their ability to affect the success of field releases of Wolbachia for human disease reduction in drier climates where longer egg quiescence periods are expected.}, } @article {pmid38971773, year = {2024}, author = {Mosi, FA and Rutha, I and Velez, R and Swai, JK and Mlacha, YP and Marques, J and Silveira, H and Tarimo, BB}, title = {Effects of a blood-free mosquito diet on fitness and gonotrophic cycle parameters of laboratory reared Anopheles gambiae sensu stricto.}, journal = {Parasites & vectors}, volume = {17}, number = {1}, pages = {289}, pmid = {38971773}, issn = {1756-3305}, mesh = {Animals ; *Anopheles/physiology ; Female ; *Fertility ; *Diet/veterinary ; Male ; Cattle ; Mosquito Control/methods ; Genetic Fitness ; Blood ; Mosquito Vectors/physiology/genetics ; Reproduction ; }, abstract = {BACKGROUND: The current rise of new innovative tools for mosquito control, such as the release of transgenic mosquitoes carrying a dominant lethal gene and Wolbachia-based strategies, necessitates a massive production of mosquitoes in the insectary. However, currently laboratory rearing depends on vertebrate blood for egg production and maintenance. This practice raises ethical concerns, incurs logistical and cost limitations, and entails potential risk associated with pathogen transmission and blood storage. Consequently, an artificial blood-free diet emerges as a desirable alternative to address these challenges. This study aims to evaluate the effects of a previously formulated artificial blood-free diet (herein referred to as BLOODless) on Anopheles gambiae (An. gambiae s.s.; IFAKARA) gonotrophic parameters and fitness compared with bovine blood.

METHODS: The study was a laboratory-based comparative evaluation of the fitness, fecundity and fertility of An. gambiae s.s. (IFAKARA) reared on BLOODless versus vertebrate blood from founder generation (F0) to eighth generation (F8). A total of 1000 female mosquitoes were randomly selected from F0, of which 500 mosquitoes were fed with bovine blood (control group) and the other 500 mosquitoes were fed with BLOODless diet (experimental group). The feeding success, number of eggs per female, hatching rate and pupation rate were examined post-feeding. Longevity and wing length were determined as fitness parameters for adult male and female mosquitoes for both populations.

RESULTS: While blood-fed and BLOODless-fed mosquitoes showed similar feeding success, 92.3% [95% confidence interval (CI) 89.7-94.9] versus 93.6% (95% CI 90.6-96.6), respectively, significant differences emerged in their reproductive parameters. The mean number of eggs laid per female was significantly higher for blood-fed mosquitoes (P < 0.001) whereas BLOODless-fed mosquitoes had significantly lower hatching rates [odds ratio (OR) 0.17, 95% CI 0.14-0.22, P < 0.001]. Wing length and longevity were similar between both groups.

CONCLUSIONS: This study demonstrates the potential of the BLOODless diet as a viable and ethical alternative to vertebrate blood feeding for rearing An. gambiae s.s. This breakthrough paves the way for more efficient and ethical studies aimed at combating malaria and other mosquito-borne diseases.}, } @article {pmid38971524, year = {2024}, author = {Shannon, DM and Richardson, N and Lahondère, C and Peach, D}, title = {Mosquito floral visitation and pollination.}, journal = {Current opinion in insect science}, volume = {65}, number = {}, pages = {101230}, doi = {10.1016/j.cois.2024.101230}, pmid = {38971524}, issn = {2214-5753}, mesh = {Animals ; *Pollination ; *Flowers/physiology ; *Culicidae/physiology ; Cues ; }, abstract = {We often consider mosquitoes through an 'anthropocentric lens' that disregards their interactions with nonhuman and nonpathogenic organisms, even though these interactions can be harnessed for mosquito control. Mosquitoes have been recognized as floral visitors, and pollinators, for more than a century. However, we know relatively little about mosquito-plant interactions, excepting some nutrition and chemical ecology-related topics, compared with mosquito-host interactions, and frequently use flawed methodology when investigating them. Recent work demonstrates mosquitoes use multimodal sensory cues to locate flowers, including ultraviolet visual cues, and we may underestimate mosquito pollination. This review focuses on current knowledge of how mosquitoes locate flowers, floral visitation assay methodology, mosquito pollination, and implications for technologies such as sterile male mosquito release through genetic control programs or Wolbachia infection.}, } @article {pmid38969975, year = {2024}, author = {Zhang, Y and Wang, H and Du, J and Wang, Y and Zang, C and Cheng, P and Liu, L and Zhang, C and Lou, Z and Lei, J and Wu, J and Gong, M and Liu, H}, title = {Population genetic structure of Culex tritaeniorhynchus in different types of climatic zones in China.}, journal = {BMC genomics}, volume = {25}, number = {1}, pages = {673}, pmid = {38969975}, issn = {1471-2164}, support = {81871685//National Natural Science Foundation of China/ ; 81871685//National Natural Science Foundation of China/ ; 81871685//National Natural Science Foundation of China/ ; 81871685//National Natural Science Foundation of China/ ; tsqn202312373//Taishan Scholars Project of Shandong Province/ ; tsqn202312373//Taishan Scholars Project of Shandong Province/ ; tsqn202312373//Taishan Scholars Project of Shandong Province/ ; No.230726153807227//Ministry of Education industry-university cooperative education project/ ; No.230726153807227//Ministry of Education industry-university cooperative education project/ ; No.230726153807227//Ministry of Education industry-university cooperative education project/ ; No.230726153807227//Ministry of Education industry-university cooperative education project/ ; No.230726153807227//Ministry of Education industry-university cooperative education project/ ; No.230726153807227//Ministry of Education industry-university cooperative education project/ ; No.230726153807227//Ministry of Education industry-university cooperative education project/ ; No.230726153807227//Ministry of Education industry-university cooperative education project/ ; No.230726153807227//Ministry of Education industry-university cooperative education project/ ; No.230726153807227//Ministry of Education industry-university cooperative education project/ ; No.230726153807227//Ministry of Education industry-university cooperative education project/ ; 2019QL005//Academic promotion programme of Shandong First Medical University/ ; 2019QL005//Academic promotion programme of Shandong First Medical University/ ; 2019QL005//Academic promotion programme of Shandong First Medical University/ ; 2019QL005//Academic promotion programme of Shandong First Medical University/ ; 2019QL005//Academic promotion programme of Shandong First Medical University/ ; 2019QL005//Academic promotion programme of Shandong First Medical University/ ; 2019QL005//Academic promotion programme of Shandong First Medical University/ ; NHCKFKT2021-02//NHC Key Laboratory of Parasite and Vector Biology/ ; NHCKFKT2021-02//NHC Key Laboratory of Parasite and Vector Biology/ ; NHCKFKT2021-02//NHC Key Laboratory of Parasite and Vector Biology/ ; NHCKFKT2021-02//NHC Key Laboratory of Parasite and Vector Biology/ ; NHCKFKT2021-02//NHC Key Laboratory of Parasite and Vector Biology/ ; NHCKFKT2021-02//NHC Key Laboratory of Parasite and Vector Biology/ ; NHCKFKT2021-02//NHC Key Laboratory of Parasite and Vector Biology/ ; NHCKFKT2021-02//NHC Key Laboratory of Parasite and Vector Biology/ ; }, mesh = {*Culex/genetics/virology/microbiology ; Animals ; China ; *Phylogeny ; *Haplotypes ; Climate ; Genetic Variation ; Genetics, Population ; Wolbachia/genetics ; Mosquito Vectors/genetics/virology/microbiology ; Electron Transport Complex IV/genetics ; }, abstract = {BACKGROUND: Culex tritaeniorhynchus is widely distributed in China, from Hainan Island in the south to Heilongjiang in the north, covering tropical, subtropical, and temperate climate zones. Culex tritaeniorhynchus carries 19 types of arboviruses. It is the main vector of the Japanese encephalitis virus (JEV), posing a serious threat to human health. Understanding the effects of environmental factors on Culex tritaeniorhynchus can provide important insights into its population structure or isolation patterns, which is currently unclear.

RESULTS: In total, 138 COI haplotypes were detected in the 552 amplified sequences, and the haplotype diversity (Hd) value increased from temperate (0.534) to tropical (0.979) regions. The haplotype phylogeny analysis revealed that the haplotypes were divided into two high-support evolutionary branches. Temperate populations were predominantly distributed in evolutionary branch II, showing some genetic isolation from tropical/subtropical populations and less gene flow between groups. The neutral test results of HNQH (Qionghai) and HNHK(Haikou) populations were negative (P < 0.05), indicating many low-frequency mutations in the populations and that the populations might be in the process of expansion. Moreover, Wolbachia infection was detected only in SDJN (Jining) (2.24%), and all Wolbachia genotypes belonged to supergroup B. To understand the influence of environmental factors on mosquito-borne viruses, we examined the prevalence of Culex tritaeniorhynchus infection in three ecological environments in Shandong Province. We discovered that the incidence of JEV infection was notably greater in Culex tritaeniorhynchus from lotus ponds compared to those from irrigation canal regions. In this study, the overall JEV infection rate was 15.27 per 1000, suggesting the current risk of Japanese encephalitis outbreaks in Shandong Province.

CONCLUSIONS: Tropical and subtropical populations of Culex tritaeniorhynchus showed higher genetic diversity and those climatic conditions provide great advantages for the establishment and expansion of Culex tritaeniorhynchus. There are differences in JEV infection rates in wild populations of Culex tritaeniorhynchus under different ecological conditions. Our results suggest a complex interplay of genetic differentiation, population structure, and environmental factors in shaping the dynamics of Culex tritaeniorhynchus. The low prevalence of Wolbachia in wild populations may reflect the recent presence of Wolbachia invasion in Culex tritaeniorhynchus.}, } @article {pmid38969648, year = {2024}, author = {Jia, H and Tan, S and Cai, Y and Guo, Y and Shen, J and Zhang, Y and Ma, H and Zhang, Q and Chen, J and Qiao, G and Ruan, J and Zhang, YE}, title = {Low-input PacBio sequencing generates high-quality individual fly genomes and characterizes mutational processes.}, journal = {Nature communications}, volume = {15}, number = {1}, pages = {5644}, pmid = {38969648}, issn = {2041-1723}, support = {32325014//National Natural Science Foundation of China (National Science Foundation of China)/ ; }, mesh = {Animals ; *Drosophila melanogaster/genetics ; *DNA Transposable Elements/genetics ; *Wolbachia/genetics ; *Genome, Insect/genetics ; *Mutation ; High-Throughput Nucleotide Sequencing/methods ; Sequence Analysis, DNA/methods ; Genomics/methods ; Gene Conversion ; }, abstract = {Long-read sequencing, exemplified by PacBio, revolutionizes genomics, overcoming challenges like repetitive sequences. However, the high DNA requirement (> 1 µg) is prohibitive for small organisms. We develop a low-input (100 ng), low-cost, and amplification-free library-generation method for PacBio sequencing (LILAP) using Tn5-based tagmentation and DNA circularization within one tube. We test LILAP with two Drosophila melanogaster individuals, and generate near-complete genomes, surpassing preexisting single-fly genomes. By analyzing variations in these two genomes, we characterize mutational processes: complex transpositions (transposon insertions together with extra duplications and/or deletions) prefer regions characterized by non-B DNA structures, and gene conversion of transposons occurs on both DNA and RNA levels. Concurrently, we generate two complete assemblies for the endosymbiotic bacterium Wolbachia in these flies and similarly detect transposon conversion. Thus, LILAP promises a broad PacBio sequencing adoption for not only mutational studies of flies and their symbionts but also explorations of other small organisms or precious samples.}, } @article {pmid38957696, year = {2024}, author = {Ross, PA and Hoffmann, AA}, title = {Revisiting Wolbachia detections: Old and new issues in Aedes aegypti mosquitoes and other insects.}, journal = {Ecology and evolution}, volume = {14}, number = {7}, pages = {e11670}, pmid = {38957696}, issn = {2045-7758}, abstract = {Wolbachia continue to be reported in species previously thought to lack them, particularly Aedes aegypti mosquitoes. The presence of Wolbachia in this arbovirus vector is considered important because releases of mosquitoes with transinfected Wolbachia are being used around the world to suppress pathogen transmission and these efforts depend on a lack of Wolbachia in natural populations of this species. We previously assessed papers reporting Wolbachia in natural populations of Ae. aegypti and found little evidence that seemed convincing. However, since our review, more and more papers are emerging on Wolbachia detections in this species. Our purpose here is to evaluate these papers within the context of criteria we previously established but also new criteria that include the absence of releases of transinfections within the local areas being sampled which has contaminated natural populations in at least one case where novel detections have been reported. We also address the broader issue of Wolbachia detection in other insects where similar issues may arise which can affect overall estimates of this endosymbiont more generally. We note continuing shortcomings in papers purporting to find natural Wolbachia in Ae. aegypti which are applicable to other insects as well.}, } @article {pmid38952690, year = {2024}, author = {de Mello, VVC and de Oliveira, LB and Coelho, TFSB and Lee, DAB and das Neves, LF and Franco, EO and Mongruel, ACB and Machado, RZ and André, MR}, title = {Diversity of Ehrlichia spp., Anaplasma spp. and Neorickettsia spp. in vampire bats.}, journal = {Current research in parasitology & vector-borne diseases}, volume = {5}, number = {}, pages = {100182}, pmid = {38952690}, issn = {2667-114X}, abstract = {Although bats (Mammalia: Chiroptera) act as natural reservoirs for many zoonotic pathogens around the world, few studies have investigated the occurrence of Anaplasmataceae agents in bats, especially vampire bats. The family Anaplasmataceae (order Rickettsiales) encompasses obligate intracellular bacteria of the genera Anaplasma, Ehrlichia, Neorickettsia, Neoehrlichia, Wolbachia, and Allocryptoplasma. The present study aimed to investigate, using molecular techniques, the presence of species of Anaplasma, Ehrlichia, and Neorickettsia in vampire bats sampled in northern Brazil. Between 2017 and 2019, spleen samples were collected from vampire bats belonging to two species, Desmodus rotundus (n = 228) from the states of Pará (n = 207), Amazonas (n = 1), Roraima (n = 18) and Amapá (n = 3), and Diaemus youngii (n = 1) from Pará. Positivity rates of 5.2% (12/229), 3% (7/229), and 10.9% (25/229) were found in PCR assays for Anaplasma spp. (16S rRNA gene), Ehrlichia spp. (dsb gene) and Neorickettsia spp. (16S rRNA gene), respectively. The present study revealed, for the first time, the occurrence of Anaplasma spp. and different genotypes of Ehrlichia spp. in vampire bats from Brazil. While phylogenetic analyses based on the dsb and ftsZ genes of Ehrlichia and 16S rRNA of Anaplasma spp. revealed phylogenetic proximity of the genotypes detected in vampire bats with Anaplasmataceae agents associated with domestic ruminants, phylogenetic inferences based on the gltA and groEL genes evidenced the occurrence of genotypes apparently exclusive to bats. Neorickettsia sp. phylogenetically associated with N. risticii was also detected in vampire bats sampled in northern Brazil.}, } @article {pmid38946980, year = {2024}, author = {Jacobs, J and Nakamoto, A and Mastoras, M and Loucks, H and Mirchandani, C and Karim, L and Penunuri, G and Wanket, C and Russell, SL}, title = {Complete de novo assembly of Wolbachia endosymbiont of Drosophila willistoni using long-read genome sequencing.}, journal = {Research square}, volume = {}, number = {}, pages = {}, pmid = {38946980}, issn = {2693-5015}, support = {R00 GM135583/GM/NIGMS NIH HHS/United States ; T32 HG012344/HG/NHGRI NIH HHS/United States ; }, abstract = {Wolbachia is an obligate intracellular α-proteobacterium which commonly infects arthropods and filarial nematodes. Different strains of Wolbachia are capable of a wide range of regulatory manipulations in many hosts and modulate host cellular differentiation to influence host reproduction. The genetic basis for the majority of these phenotypes is unknown. The wWil strain from the neotropical fruit fly, Drosophila willistoni, exhibits a remarkably high affinity for host germline-derived cells relative to the soma. This trait could be leveraged for understanding how Wolbachia influences the host germline and for controlling host populations in the field. To further the use of this strain in biological and biomedical research, we sequenced the genome of the wWil strain isolated from host cell culture cells. Here, we present the first high quality nanopore assembly of wWil, the Wolbachia endosymbiont of D. willistoni. Our assembly resulted in a circular genome of 1.27 Mb with a BUSCO completeness score of 99.7%. Consistent with other insect-associated Wolbachia strains, comparative genomic analysis revealed that wWil has a highly mosaic genome relative to the closely related wMel strain from Drosophila melanogaster.}, } @article {pmid38940615, year = {2024}, author = {Hrdina, A and Serra Canales, M and Arias-Rojas, A and Frahm, D and Iatsenko, I}, title = {The endosymbiont Spiroplasma poulsonii increases Drosophila melanogaster resistance to pathogens by enhancing iron sequestration and melanization.}, journal = {mBio}, volume = {15}, number = {8}, pages = {e0093624}, pmid = {38940615}, issn = {2150-7511}, support = {IA 81/2-1//Deutsche Forschungsgemeinschaft (DFG)/ ; }, mesh = {Animals ; *Spiroplasma/physiology ; *Symbiosis ; *Drosophila melanogaster/microbiology/immunology ; *Iron/metabolism ; Melanins/metabolism ; Staphylococcus aureus/physiology/immunology ; Providencia/metabolism/physiology/genetics ; Disease Resistance ; }, abstract = {UNLABELLED: Facultative endosymbiotic bacteria, such as Wolbachia and Spiroplasma species, are commonly found in association with insects and can dramatically alter their host physiology. Many endosymbionts are defensive and protect their hosts against parasites or pathogens. Despite the widespread nature of defensive insect symbioses and their importance for the ecology and evolution of insects, the mechanisms of symbiont-mediated host protection remain poorly characterized. Here, we utilized the fruit fly Drosophila melanogaster and its facultative endosymbiont Spiroplasma poulsonii to characterize the mechanisms underlying symbiont-mediated host protection against bacterial and fungal pathogens. Our results indicate a variable effect of S. poulsonii on infection outcome, with endosymbiont-harboring flies being more resistant to Rhyzopus oryzae, Staphylococcus aureus, and Providencia alcalifaciens but more sensitive or as sensitive as endosymbiont-free flies to the infections with Pseudomonas species. Further focusing on the protective effect, we identified Transferrin-mediated iron sequestration induced by Spiroplasma as being crucial for the defense against R. oryzae and P. alcalifaciens. In the case of S. aureus, enhanced melanization in Spiroplasma-harboring flies plays a major role in protection. Both iron sequestration and melanization induced by Spiroplasma require the host immune sensor protease Persephone, suggesting a role of proteases secreted by the symbiont in the activation of host defense reactions. Hence, our work reveals a broader defensive range of Spiroplasma than previously appreciated and adds nutritional immunity and melanization to the defensive arsenal of symbionts.

IMPORTANCE: Defensive endosymbiotic bacteria conferring protection to their hosts against parasites and pathogens are widespread in insect populations. However, the mechanisms by which most symbionts confer protection are not fully understood. Here, we studied the mechanisms of protection against bacterial and fungal pathogens mediated by the Drosophila melanogaster endosymbiont Spiroplasma poulsonii. We demonstrate that besides the previously described protection against wasps and nematodes, Spiroplasma also confers increased resistance to pathogenic bacteria and fungi. We identified Spiroplasma-induced iron sequestration and melanization as key defense mechanisms. Our work broadens the known defense spectrum of Spiroplasma and reveals a previously unappreciated role of melanization and iron sequestration in endosymbiont-mediated host protection. We propose that the mechanisms we have identified here may be of broader significance and could apply to other endosymbionts, particularly to Wolbachia, and potentially explain their protective properties.}, } @article {pmid38938126, year = {2024}, author = {Pan, Q and Yu, SJ and Lei, S and Zhang, SH and Ding, LL and Liu, L and Li, SC and Wang, XF and Lou, BH and Ran, C}, title = {Bacterial Symbionts Contribute to Insecticide Susceptibility of Diaphorina citri via Changing the Expression Level of Host Detoxifying Genes.}, journal = {Journal of agricultural and food chemistry}, volume = {72}, number = {27}, pages = {15164-15175}, doi = {10.1021/acs.jafc.4c03049}, pmid = {38938126}, issn = {1520-5118}, mesh = {*Symbiosis ; Animals ; *Insecticides/pharmacology ; *Hemiptera/microbiology/genetics/drug effects ; Insect Proteins/genetics/metabolism ; Insecticide Resistance/genetics ; Wolbachia/drug effects/genetics ; Pyrethrins/pharmacology ; Bacteria/genetics/drug effects/classification/isolation & purification/metabolism ; Inactivation, Metabolic/genetics ; }, abstract = {Insecticide susceptibility is mainly determined by the insect host, but symbiotic bacteria are also an important affecting factor. In this study, we investigate the relationship between the structure of gut bacterial symbionts and insecticide susceptibility in Diaphorina citri, the important carrier of Candidatus Liberibacter asiaticus (CLas), the causal agent of Huanglongbing (HLB). Our results indicated that antibiotic treatment significantly increased the susceptibility of D. citri to bifenthrin and thiamethoxam, and significantly decreased the relative abundance of Wolbachia and Profftella, enzyme activities of CarEs, and expression level of multiple CarE genes. The relative loads of Wolbachia and Profftella were positively correlated with DcitCCE13, DcitCCE14, DcitCCE15, and DcitCCE16. RNAi and prokaryotic expression revealed that DcitCCE15 is associated with bifenthrin metabolism. These results revealed that bacterial symbionts might regulate DcitCCE15 expression, which is involved in the susceptibility of D. citri to bifenthrin.}, } @article {pmid38934782, year = {2024}, author = {Amini, S and Fathipour, Y and Hoffmann, A and Mehrabadi, M}, title = {Wolbachia affect female mate preference and offspring fitness in a parasitoid wasp.}, journal = {Pest management science}, volume = {}, number = {}, pages = {}, doi = {10.1002/ps.8272}, pmid = {38934782}, issn = {1526-4998}, abstract = {BACKGROUND: Wolbachia are widespread intracellular bacteria in insects that often have high rates of spread due to their impact on insect reproduction. These bacteria may also affect the mating behavior of their host with impacts on the fitness of host progeny. In this study, we investigated the impact of Wolbachia on a preference for mating with young or old males in the parasitoid wasp Habrobracon hebetor.

RESULTS: Our results showed that uninfected females from a tetracycline-treated line preferred to mate with young males, whereas Wolbachia-infected females had no preference. Time to mating was relatively shorter in the infected lines. Regardless of Wolbachia infection status, progeny resulting from matings with young males showed higher fitness than those from crosses with old males, and infected females crossed with infected young males showed the highest performance.

CONCLUSION: These results suggest an impact of Wolbachia on female mate preference and offspring fitness although it is unclear how this phenomenon increases Wolbachia transmission of infected wasps. © 2024 Society of Chemical Industry.}, } @article {pmid38934580, year = {2024}, author = {Mendel, BM and Asselin, AK and Johnson, KN and McGuigan, K}, title = {Effects of spontaneous mutations on survival and reproduction of Drosophila serrata infected with Drosophila C virus.}, journal = {Evolution; international journal of organic evolution}, volume = {}, number = {}, pages = {}, doi = {10.1093/evolut/qpae101}, pmid = {38934580}, issn = {1558-5646}, abstract = {The impact of selection on host immune function genes has been widely documented. However, it remains essentially unknown how mutation influences the quantitative immune traits that selection acts on. Applying a classical mutation accumulation (MA) experimental design in Drosophila serrata, we found the mutational variation in susceptibility (median time of death, LT50) to Drosophila C virus (DCV) was of similar magnitude to that reported for intrinsic survival traits. Mean LT50 did not change as mutations accumulated, suggesting no directional bias in mutational effects. Maintenance of genetic variance in immune function is hypothesised to be influenced by pleiotropic effects on immunity and other traits that contribute to fitness. To investigate this, we assayed female reproductive output for a subset of MA lines with relatively long or short survival times under DCV infection. Longer survival time tended to be associated with lower reproductive output, suggesting that mutations affecting susceptibility to DCV had pleiotropic effects on investment in reproductive fitness. Further studies are needed to uncover the general patterns of mutational effect on immune responses and other fitness traits, and to determine how selection might typically act on new mutations via their direct and pleiotropic effects.}, } @article {pmid38924387, year = {2024}, author = {Devereux, G and Bula, M and Tripp, K and Fitzgerald, R and Eraut, N and Alam, MS and Moriyama, T and Shinkyo, R and Walker, L and Wang, D and Gusovsky, F and van der Velde, J and Turner, JD and Hong, WD and O'Neill, PM and Taylor, MJ and Ward, SA}, title = {A Phase 1, Randomized, Double-Blind, Placebo-Controlled, Single Ascending Dose Trial of AWZ1066S, an Anti-Wolbachia Candidate Macrofilaricide.}, journal = {Clinical pharmacology in drug development}, volume = {13}, number = {9}, pages = {1071-1081}, doi = {10.1002/cpdd.1441}, pmid = {38924387}, issn = {2160-7648}, mesh = {Humans ; Double-Blind Method ; Male ; Female ; Adult ; Middle Aged ; Young Adult ; *Wolbachia/drug effects ; Dose-Response Relationship, Drug ; Elephantiasis, Filarial/drug therapy ; Healthy Volunteers ; Anti-Bacterial Agents/pharmacokinetics/administration & dosage/adverse effects ; Administration, Oral ; Onchocerciasis/drug therapy ; }, abstract = {AWZ1066S has been developed as a potential treatment for the neglected tropical diseases lymphatic filariasis and onchocerciasis. AWZ1066S targets the Wolbachia bacterial endosymbiont present in the causative nematode parasites. This phase 1, first-in-human study aimed to assess the safety and pharmacokinetics of AWZ1066S in healthy human participants. In a randomized double-blind, placebo-controlled, single ascending dose study, healthy adults received a single oral dose of AWZ1066S (or placebo) and were followed up for 10 days. The planned single doses of AWZ1066S ranged from 100 to 1600 mg, and each dose was administered to a cohort of 8 participants (6 AWZ1066S and 2 placebo). In total 30 people participated, 18 (60%) female, median age 30.0 years (minimum 20, maximum 61). The cohorts administered 100, 200, 300, and 400 mg of AWZ1066S progressed unremarkably. After single 700-mg doses all 4 participants developed symptoms of acute gastritis and transient increases in liver enzymes. The severity of these adverse events ranged from mild to severe, with 1 participant needing hospital admission. Pharmacokinetic analysis indicated that AWZ1066S is rapidly absorbed with predictable pharmacokinetics. In conclusion, safety concerns prevented this study from reaching the human exposures needed for AWZ1066S to be clinically effective against lymphatic filariasis and onchocerciasis.}, } @article {pmid38923204, year = {2024}, author = {Pratelli, A and Riparbelli, MG and Callaini, G}, title = {Axonemal tubules in the distal sperm tail of Wolbachia-infected Drosophila simulans males contain ring-like intraluminal structures that persist after axoneme fragmentation.}, journal = {Cytoskeleton (Hoboken, N.J.)}, volume = {}, number = {}, pages = {}, doi = {10.1002/cm.21891}, pmid = {38923204}, issn = {1949-3592}, support = {2020CLZ5XW//Ministero dell'Istruzione, dell'Università e della Ricerca/ ; 2022TLYYPS_003//Ministero dell'Istruzione, dell'Università e della Ricerca/ ; }, abstract = {Wolbachia are obligate intracellular alphaproteobacteria that enhance their spreading by altering the reproductive mechanisms of several invertebrates. Among the reproductive alterations, Wolbachia also causes cytoplasmic incompatibility that leads to embryo death when infected males are crossed with uninfected females, thus selecting infected females. However, the presence of Wolbachia has important fitness costs and infected Drosophila simulans males produce less sperm than their uninfected counterparts. Such sperm suffer, indeed, of some structural alterations that hinder their proper function. We took advantage of the fact that several sperm have abnormal distal regions of the tail, in which the plasma membrane is broken and the axonemal components splayed, making the ultrastructural aspects clearly observable. We found that axoneme reduction in the distal region of the sperm does not follow a unique pattern as observed in other insects, but occurs by losing accessory tubules or peripheral doublets. The axonemal tubules contain distinct coaxial ring-like structures that are still observed after axoneme fragmentation and form large clusters of several units.}, } @article {pmid38915450, year = {2024}, author = {Trouche, B and Schrieke, H and Duron, O and Eren, AM and Reveillaud, J}, title = {Wolbachia populations across organs of individual Culex pipiens: highly conserved intra-individual core pangenome with inter-individual polymorphisms.}, journal = {ISME communications}, volume = {4}, number = {1}, pages = {ycae078}, pmid = {38915450}, issn = {2730-6151}, abstract = {Wolbachia is a maternally inherited intracellular bacterium that infects a wide range of arthropods including mosquitoes. The endosymbiont is widely used in biocontrol strategies due to its capacity to modulate arthropod reproduction and limit pathogen transmission. Wolbachia infections in Culex spp. are generally assumed to be monoclonal but the potential presence of genetically distinct Wolbachia subpopulations within and between individual organs has not been investigated using whole genome sequencing. Here we reconstructed Wolbachia genomes from ovary and midgut metagenomes of single naturally infected Culex pipiens mosquitoes from Southern France to investigate patterns of intra- and inter-individual differences across mosquito organs. Our analyses revealed a remarkable degree of intra-individual conservancy among Wolbachia genomes from distinct organs of the same mosquito both at the level of gene presence-absence signal and single-nucleotide polymorphisms (SNPs). Yet, we identified several synonymous and non-synonymous substitutions between individuals, demonstrating the presence of some level of genomic heterogeneity among Wolbachia that infect the same C. pipiens field population. Overall, the absence of genetic heterogeneity within Wolbachia populations in a single individual confirms the presence of a dominant Wolbachia that is maintained under strong purifying forces of evolution.}, } @article {pmid38911346, year = {2024}, author = {Zimmermann, IR and Alves Fernandes, RR and Santos da Costa, MG and Pinto, M and Peixoto, HM}, title = {Simulation-based economic evaluation of the Wolbachia method in Brazil: a cost-effective strategy for dengue control.}, journal = {Lancet regional health. Americas}, volume = {35}, number = {}, pages = {100783}, pmid = {38911346}, issn = {2667-193X}, abstract = {BACKGROUND: Dengue virus (DENV) is an arbovirus transmitted by Aedes aegypti mosquitoes, which can cause severe conditions such as hemorrhagic fever and dengue shock syndrome. These conditions are associated with adverse social, clinical, and economic consequences in Brazil. Herein, the Wolbachia mosquito replacement method is a promising dengue control strategy.

METHODS: We estimated the economic impact of implementing the Wolbachia mosquito replacement method in seven Brazilian cities. A mathematical microsimulation model tracked nearly 23 million inhabitants over a 20-year period, considering the transitions between five different health states (susceptible, inapparent, outpatient, hospitalised and death). Direct costs included local dengue control programs, Wolbachia implementation and dengue care. Indirect costs related to death and productivity loss, as well as disability-adjusted life-years (DALY) averted were also considered.

FINDINGS: Without Wolbachia, the model projected 1,762,688 reported dengue cases over 20 years. Implementing the Wolbachia method would avert at least 1,295,566 dengue cases, resulting in lower costs and greater effectiveness in all simulated cities. On average, for every 1000 inhabitants followed for 20 years, the Wolbachia method yielded a cost difference of USD 538,233.68 (BRL 2,691,168.40) and averted 5.56 DALYs. Net monetary benefits (NMB) were positive in all seven cities, ranging from USD 110.72 (BRL 553.59) to USD 1399.19 (BRL 6995.95) per inhabitant. Alternative scenarios have also shown a favourable return on investment with a positive benefit-cost ratio (BCR).

INTERPRETATION: Wolbachia is likely a cost-effective strategy in the Brazilian context, consistent with international studies. Sensitivity analysis and alternative scenarios confirmed the robustness of the results.

FUNDING: This study was funded by the Wellcome Trust under a grant (224459/Z/21/Z).}, } @article {pmid38908822, year = {2024}, author = {Schinkel, M and Bousema, T and van Rij, RP}, title = {Tripartite interactions between viruses, parasites, and mosquitoes.}, journal = {Current opinion in insect science}, volume = {64}, number = {}, pages = {101222}, doi = {10.1016/j.cois.2024.101222}, pmid = {38908822}, issn = {2214-5753}, mesh = {Animals ; *Mosquito Vectors/virology/physiology ; *Anopheles/virology/parasitology ; Insect Viruses/physiology ; Malaria/transmission ; Plasmodium/physiology ; }, abstract = {Mosquito-borne diseases have a major impact on global human health. Biological agents that colonize the mosquito vector are increasingly explored as an intervention strategy to prevent vector-borne disease transmission. For instance, the release of mosquitoes carrying the endosymbiotic bacterium Wolbachia effectively reduced dengue virus incidence and disease. Insect-specific viruses are likewise considered as biocontrol agents against vector-borne diseases. While most studies focused on insect-specific viruses as an intervention against arthropod-borne viruses, we here consider whether mosquito-specific viruses may affect the transmission of the malaria-causing Plasmodium parasite by Anopheles mosquitoes. Although there is no direct experimental evidence addressing this question, we found that viral infections in dipteran insects activate some of the immune pathways that are antiparasitic in Anopheles. These findings suggest that indirect virus-parasite interactions could occur and that insect-specific viruses may modulate malaria transmission. Tripartite interactions between viruses, parasites, and Anopheles mosquitoes thus merit further investigation.}, } @article {pmid38903791, year = {2024}, author = {Tafesh-Edwards, G and Kyza Karavioti, M and Markollari, K and Bunnell, D and Chtarbanova, S and Eleftherianos, I}, title = {Wolbachia endosymbionts in Drosophila regulate the resistance to Zika virus infection in a sex dependent manner.}, journal = {Frontiers in microbiology}, volume = {15}, number = {}, pages = {1380647}, pmid = {38903791}, issn = {1664-302X}, abstract = {Drosophila melanogaster has been used extensively for dissecting the genetic and functional bases of host innate antiviral immunity and virus-induced pathology. Previous studies have shown that the presence of Wolbachia endosymbionts in D. melanogaster confers resistance to infection by certain viral pathogens. Zika virus is an important vector-borne pathogen that has recently expanded its range due to the wide geographical distribution of the mosquito vector. Here, we describe the effect of Wolbachia on the immune response of D. melanogaster adult flies following Zika virus infection. First, we show that the presence of Wolbachia endosymbionts promotes the longevity of uninfected D. melanogaster wild type adults and increases the survival response of flies following Zika virus injection. We find that the latter effect is more pronounced in females rather than in males. Then, we show that the presence of Wolbachia regulates Zika virus replication during Zika virus infection of female flies. In addition, we demonstrate that the antimicrobial peptide-encoding gene Drosocin and the sole Jun N-terminal kinase-specific MAPK phosphatase Puckered are upregulated in female adult flies, whereas the immune and stress response gene TotM is upregulated in male individuals. Finally, we find that the activity of RNA interference and Toll signaling remain unaffected in Zika virus-infected female and male adults containing Wolbachia compared to flies lacking the endosymbionts. Our results reveal that Wolbachia endosymbionts in D. melanogaster affect innate immune signaling activity in a sex-specific manner, which in turn influences host resistance to Zika virus infection. This information contributes to a better understanding of the complex interrelationship between insects, their endosymbiotic bacteria, and viral infection. Interpreting these processes will help us design more effective approaches for controlling insect vectors of infectious disease.}, } @article {pmid38902790, year = {2024}, author = {Lim, JT and Mailepessov, D and Chong, CS and Chang, CC and Dickens, B and Lai, YL and Deng, L and Lee, C and Tan, LY and Chain, G and Ho, SH and Zulkifli, MF and Liew, J and Vasquez, K and Lee, V and Wong, JCC and Sim, S and Tan, CH and Ng, LC}, title = {Update to: Assessing the efficacy of male Wolbachia-infected mosquito deployments to reduce dengue incidence in Singapore.}, journal = {Trials}, volume = {25}, number = {1}, pages = {400}, pmid = {38902790}, issn = {1745-6215}, mesh = {*Dengue/prevention & control/epidemiology/transmission ; Animals ; *Wolbachia ; Singapore/epidemiology ; Male ; *Aedes/microbiology/virology ; Humans ; Incidence ; *Mosquito Vectors/microbiology/virology ; *Randomized Controlled Trials as Topic ; Mosquito Control/methods ; Female ; Pest Control, Biological/methods ; }, abstract = {BACKGROUND: This trial is a parallel, two-arm, non-blinded cluster randomised controlled trial that is under way in Singapore, with the aim of measuring the efficacy of male Wolbachia-infected Aedes aegypti deployments in reducing dengue incidence in an endemic setting with all four dengue serotypes in circulation. The trial commenced in July 2022 and is expected to conclude in September 2024. The original study protocol was published in December 2022. Here, we describe amendments that have been made to the study protocol since commencement of the trial.

METHODS: The key protocol amendments are (1) addition of an explicit definition of Wolbachia exposure for residents residing in intervention sites based on the duration of Wolbachia exposure at point of testing, (2) incorporation of a high-dimensional set of anthropogenic and environmental characteristics in the analysis plan to adjust for baseline risk factors of dengue transmission, and (3) addition of alternative statistical analyses for endpoints to control for post hoc imbalance in cluster-based environmental and anthropogenic characteristics.

DISCUSSION: The findings from this study will provide the first experimental evidence for the efficacy of releasing male-Wolbachia infected mosquitoes to reduce dengue incidence in a cluster-randomised controlled trial. The trial will conclude in 2024 and results will be reported shortly thereafter.

TRIAL REGISTRATION: ClinicalTrials.gov, identifier: NCT05505682. Registered on 16 August 2022. Retrospectively registered. Last updated 11 November 2023.}, } @article {pmid38899428, year = {2024}, author = {Laifi-Necibi, N and Amor, N and Merella, P and Mohammed, OB and Medini, L}, title = {DNA barcoding reveals cryptic species in the sea slater Ligia italica (Crustacea, Isopoda) from Tunisia.}, journal = {Mitochondrial DNA. Part A, DNA mapping, sequencing, and analysis}, volume = {}, number = {}, pages = {1-11}, doi = {10.1080/24701394.2024.2363350}, pmid = {38899428}, issn = {2470-1408}, abstract = {Barcoding studies have provided significant insights into phylogenetic relationships among species belonging to the genus Ligia (Crustacea, Isopoda). Herein the diversity of the Italian sea slater Ligia italica from Tunisia is studied for the first time. Samples were collected from 18 localities in Tunisia, and the analysis included previously published sequences from Italy and Greece available in GenBank. Bayesian and Maximum Likelihood phylogenetic analyses were carried out using a fragment of the mitochondrial COI gene. Putative cryptic species were explored using the 'barcode gap' approach in the software ASAP. A genetic landscape shape analysis was carried out using the program Alleles in Space. The analyses revealed highly divergent and well-supported clades of L. italica dispersed across Tunisia (Clades A1 and A2), Greece (Clade B) and Italy (Clades C1 and C2). High genetic dissimilarity among clades suggested that L. italica constitute a cryptic species complex. Divergence among different L. italica lineages (Clades A, B and C) occurred around 7-4.5 Ma. The detected high genetic distances among clades did not result from atypical mitochondrial DNAs or intracellular infection by Wolbachia bacteria. The complex history of the Mediterranean Sea appears to have played a significant role in shaping the phylogeographic pattern of Ligia italica. Additional morphological and molecular studies are needed to confirm the existence of cryptic species in Ligia italica in Mediterranean.}, } @article {pmid38885278, year = {2024}, author = {She, L and Shi, M and Cao, T and Yuan, H and Wang, R and Wang, W and She, Y and Wang, C and Zeng, Q and Mao, W and Zhang, Y and Wang, Y and Xi, Z and Pan, X}, title = {Wolbachia mediates crosstalk between miRNA and Toll pathways to enhance resistance to dengue virus in Aedes aegypti.}, journal = {PLoS pathogens}, volume = {20}, number = {6}, pages = {e1012296}, pmid = {38885278}, issn = {1553-7374}, support = {R01 AI080597/AI/NIAID NIH HHS/United States ; }, mesh = {*Wolbachia/physiology ; *Aedes/microbiology/virology/immunology ; Animals ; *MicroRNAs/genetics/metabolism ; *Dengue Virus/immunology ; *Dengue/immunology/virology ; Toll-Like Receptors/metabolism/immunology ; Mosquito Vectors/virology/microbiology/immunology ; Signal Transduction ; RNA, Long Noncoding/genetics/immunology ; Immunity, Innate ; Symbiosis ; }, abstract = {The obligate endosymbiont Wolbachia induces pathogen interference in the primary disease vector Aedes aegypti, facilitating the utilization of Wolbachia-based mosquito control for arbovirus prevention, particularly against dengue virus (DENV). However, the mechanisms underlying Wolbachia-mediated virus blockade have not been fully elucidated. Here, we report that Wolbachia activates the host cytoplasmic miRNA biogenesis pathway to suppress DENV infection. Through the suppression of the long noncoding RNA aae-lnc-2268 by Wolbachia wAlbB, aae-miR-34-3p, a miRNA upregulated by the Wolbachia strains wAlbB and wMelPop, promoted the expression of the antiviral effector defensin and cecropin genes through the Toll pathway regulator MyD88. Notably, anti-DENV resistance induced by Wolbachia can be further enhanced, with the potential to achieve complete virus blockade by increasing the expression of aae-miR-34-3p in Ae. aegypti. Furthermore, the downregulation of aae-miR-34-3p compromised Wolbachia-mediated virus blockade. These findings reveal a novel mechanism by which Wolbachia establishes crosstalk between the cytoplasmic miRNA pathway and the Toll pathway via aae-miR-34-3p to strengthen antiviral immune responses against DENV. Our results will aid in the advancement of Wolbachia for arbovirus control by enhancing its virus-blocking efficiency.}, } @article {pmid38877196, year = {2024}, author = {Hague, MTJ and Wheeler, TB and Cooper, BS}, title = {Comparative analysis of Wolbachia maternal transmission and localization in host ovaries.}, journal = {Communications biology}, volume = {7}, number = {1}, pages = {727}, pmid = {38877196}, issn = {2399-3642}, support = {R35 GM124701/GM/NIGMS NIH HHS/United States ; 2145195//National Science Foundation (NSF)/ ; R35GM124701//U.S. Department of Health & Human Services | NIH | National Institute of General Medical Sciences (NIGMS)/ ; }, mesh = {*Wolbachia/physiology/genetics ; Animals ; Female ; *Ovary/microbiology ; *Drosophila melanogaster/microbiology ; *Aedes/microbiology ; Symbiosis ; Temperature ; Oocytes/microbiology ; }, abstract = {Many insects and other animals carry microbial endosymbionts that influence their reproduction and fitness. These relationships only persist if endosymbionts are reliably transmitted from one host generation to the next. Wolbachia are maternally transmitted endosymbionts found in most insect species, but transmission rates can vary across environments. Maternal transmission of wMel Wolbachia depends on temperature in natural Drosophila melanogaster hosts and in transinfected Aedes aegypti, where wMel is used to block pathogens that cause human disease. In D. melanogaster, wMel transmission declines in the cold as Wolbachia become less abundant in host ovaries and at the posterior pole plasm (the site of germline formation) in mature oocytes. Here, we assess how temperature affects maternal transmission and underlying patterns of Wolbachia localization across 10 Wolbachia strains diverged up to 50 million years-including strains closely related to wMel-and their natural Drosophila hosts. Many Wolbachia maintain high transmission rates across temperatures, despite highly variable (and sometimes low) levels of Wolbachia in the ovaries and at the developing germline in late-stage oocytes. Identifying strains like closely related wMel-like Wolbachia with stable transmission across variable environmental conditions may improve the efficacy of Wolbachia-based biocontrol efforts as they expand into globally diverse environments.}, } @article {pmid38869236, year = {2024}, author = {Taprogge, M and Grath, S}, title = {Modelling suggests Wolbachia-induced cytoplasmic incompatibility in oak gall wasps with cyclical parthenogenesis.}, journal = {Journal of evolutionary biology}, volume = {37}, number = {8}, pages = {926-934}, doi = {10.1093/jeb/voae077}, pmid = {38869236}, issn = {1420-9101}, mesh = {*Wolbachia/physiology ; Animals ; *Wasps/microbiology/physiology ; Female ; *Parthenogenesis ; Male ; Quercus/microbiology ; Models, Biological ; Symbiosis ; Cytoplasm ; }, abstract = {Oak gall wasps typically exhibit a life cycle with one sexual and one asexual generation each year. These wasps can carry various endosymbionts, one of which is the maternally inherited bacterium Wolbachia that can induce several reproductive manipulations on its host. Cytoplasmic incompatibility (CI) has been described as the most prominent of these manipulations. CI leads to embryonic mortality in the hosts' offspring when infected males mate with either uninfected females or with females that harbour different Wolbachia strains. It has been hypothesized that Wolbachia can induce CI in oak gall wasps. To address this hypothesis, we derived a mathematical model to investigate the spread of a bacterial infection in naive populations and to determine the plausibility of CI occurrence. To validate our model, we used published data from Wolbachia-infected Belonocnema kinseyi populations in two approaches. Our first approach uses measurements of infection frequencies and maternal transmission in the sexual generation. For the second approach, we extended the model to compare predictions to estimates of mtDNA-haplotypes, which, like Wolbachia, are maternally inherited, and can therefore be associated with the infection. Both approaches indicate that CI is present in these populations. Our model can be generalized to investigate the occurrence of CI not only for oak gall wasps but also for other species.}, } @article {pmid38860195, year = {2023}, author = {Naseri-Karimi, N and Vatandoost, H and Mehdi Sedaghat, M and Moosa-Kazemi, SH and Amidi, F and Ali Oshaghi, M}, title = {Drosophila melanogaster Laboratory Rearing for Wolbachia-Based Control Programs, a Component of Dengue Control.}, journal = {Journal of arthropod-borne diseases}, volume = {17}, number = {3}, pages = {214-228}, pmid = {38860195}, issn = {2322-1984}, abstract = {BACKGROUND: Drosophila melanogaster flies are smooth, low upkeep and safe model organisms, they can be effortlessly used in different fields of life sciences like genomics, biotechnology, genetics, disease model, and Wolbachia-based approaches to fight vectors and the pathogens they transmit.

METHODS: Fruit fly specimens were collected in 25 districts (14 provinces) of Iran and their morphological recognition was proven by molecular analysis based on sequence homology of mitochondrial COI barcode region. Essential information and specific requirements were provided for laboratory rearing of D. melanogaster.

RESULTS: Drosophila melanogaster colonies were found in 23 out of 25 districts. Also, five related species coincident with D. melanogaster were reported in this study including D. ananassae/D. parapallidosa, D. hydei, D. repleta, Zaprionus indianus (Diptera: Drosophilidae), and Megaselia scalaris (Diptera: Phoridae). The Iranian D. melanogaster molecular signature and their rearing techniques have been described here. The complete life cycle, from (egg to adult), takes approximately 8 days at 25 °C. Some biological points have been presented with highlighting capturing, rearing, culturing, and embryo collection along with primitive recognition and segregation between females and males have been presented. A recipe for culture media and the quantity of various ingredients have been provided.

CONCLUSION: This is the first report on the D. repleta and D. ananassae/D. parapallidosa species for the country. Results of this study provide efficient and effective rearing procedures which are requirement for both small-scale for facilitating entomological research and large-scale use in justifiable vector control management such as disease model or Dengue control.}, } @article {pmid38857820, year = {2024}, author = {Kumar, T and Maitra, S and Rai, R and Priyanka, and Maitra, S and Tirkey, NN and Kumari, R}, title = {The dichotomy between probiotic lactic acid bacteria and Plasmodium: A promising therapeutic avenue.}, journal = {Acta tropica}, volume = {257}, number = {}, pages = {107284}, doi = {10.1016/j.actatropica.2024.107284}, pmid = {38857820}, issn = {1873-6254}, mesh = {*Probiotics/administration & dosage/therapeutic use ; Humans ; *Gastrointestinal Microbiome/drug effects ; *Malaria/prevention & control/drug therapy/parasitology ; Animals ; *Lactobacillales/physiology ; *Plasmodium/physiology/drug effects ; }, abstract = {Our understanding of gut microbial populations and their immense influence on host immunity, health, and diseases has increased deeply in recent years. Numerous reports have identified the role of mosquito and mammalian gut microbiota in the modulation of host susceptibility to Plasmodium infection. Artemisinin resistance in malaria-endemic regions necessitates the development of new, safer, and more affordable treatments to supplement existing therapies. In this review, we compiled a colossal amount of data from numerous studies that have assessed the roles played by gut microbial communities in Plasmodium infection, progression, transmission, and severity. Most interestingly, our study points to the overwhelming evidence from experimental studies in mural malaria to human trials, suggesting that the presence of lactic acid bacteria in the gut microbiota of mammalian hosts provides a great degree of protection against malaria. Therefore, our study provides a compelling narrative for probiotic administration as an adjunct therapy for combatting malaria.}, } @article {pmid38855918, year = {2024}, author = {Mulio, SÅ and Zwolińska, A and Klejdysz, T and Prus-Frankowska, M and Michalik, A and Kolasa, M and Łukasik, P}, title = {Limited variation in microbial communities across populations of Macrosteles leafhoppers (Hemiptera: Cicadellidae).}, journal = {Environmental microbiology reports}, volume = {16}, number = {3}, pages = {e13279}, pmid = {38855918}, issn = {1758-2229}, support = {PPN/PPO/2018/1/00015//Narodowa Agencja Wymiany Akademickiej/ ; RGP 0024/2015//Human Frontier Science Program/ ; 2018/30/E/NZ8/00880//Narodowe Centrum Nauki/ ; 2021/41/B/NZ8/04526//Narodowe Centrum Nauki/ ; }, mesh = {Animals ; *Hemiptera/microbiology ; *Symbiosis ; *Microbiota ; *RNA, Ribosomal, 16S/genetics ; *Bacteria/classification/genetics/isolation & purification ; Phylogeny ; Electron Transport Complex IV/genetics ; }, abstract = {Microbial symbionts play crucial roles in insect biology, yet their diversity, distribution, and temporal dynamics across host populations remain poorly understood. In this study, we investigated the spatio-temporal distribution of bacterial symbionts within the widely distributed and economically significant leafhopper genus Macrosteles, with a focus on Macrosteles laevis. Using host and symbiont marker gene amplicon sequencing, we explored the intricate relationships between these insects and their microbial partners. Our analysis of the cytochrome oxidase subunit I (COI) gene data revealed several intriguing findings. First, there was no strong genetic differentiation across M. laevis populations, suggesting gene flow among them. Second, we observed significant levels of heteroplasmy, indicating the presence of multiple mitochondrial haplotypes within individuals. Third, parasitoid infections were prevalent, highlighting the complex ecological interactions involving leafhoppers. The 16S rRNA data confirmed the universal presence of ancient nutritional endosymbionts-Sulcia and Nasuia-in M. laevis. Additionally, we found a high prevalence of Arsenophonus, another common symbiont. Interestingly, unlike most previously studied species, M. laevis exhibited only occasional cases of infection with known facultative endosymbionts and other bacteria. Notably, there was no significant variation in symbiont prevalence across different populations or among sampling years within the same population. Comparatively, facultative endosymbionts such as Rickettsia, Wolbachia, Cardinium and Lariskella were more common in other Macrosteles species. These findings underscore the importance of considering both host and symbiont dynamics when studying microbial associations. By simultaneously characterizing host and symbiont marker gene amplicons in large insect collections, we gain valuable insights into the intricate interplay between insects and their microbial partners. Understanding these dynamics contributes to our broader comprehension of host-microbe interactions in natural ecosystems.}, } @article {pmid38834927, year = {2024}, author = {Guo, W and Zhu, W and Jia, L and Tao, Y}, title = {Unique microbial communities of parasitic fleas on wild animals from the Qinghai-Tibet Plateau.}, journal = {Environmental science and pollution research international}, volume = {31}, number = {28}, pages = {40916-40924}, pmid = {38834927}, issn = {1614-7499}, support = {2020-ZJ-788//Applied Basic Research Project of Science and Technology Department of Qinghai Province/ ; }, mesh = {Animals ; *Siphonaptera/microbiology ; Tibet ; Animals, Wild ; Microbiota ; China ; Phylogeny ; }, abstract = {Fleas, one of the most significant ectoparasites, play a crucial role as vectors in spreading zoonotic diseases globally. The Qinghai Province, as part of the Qinghai-Tibet Plateau, is one of the provinces in China with the largest number of flea species. In this study, we characterized the microbial communities of eighty-five adult fleas, belonging to nineteen species within four families (Ceratophyllidae, Ctenophthalmidae, Leptopsyllidae, and Pulicidae). We identified a total of 1162 unique operational taxonomic units at the genus level, with flea-borne pathogens such as Wolbachia, Bartonella, Rickettsia being the members of top abundant taxa. Except for comparison between Ctenophthalmidae and Leptopsyllidae families, the analyses of both alpha- and beta- diversity indicators suggested that bacterial diversity varied among flea families. This could be attributed to flea phylogeny, which also influenced by their geographical sites and animal hosts. Results of Linear discriminant analysis effect size (LEfSe) indicated that 29 genera in Ceratophylloidea, 11 genera in Ctenophthalmidae, 15 genera in Leptopsyllidae, and 22 genera in Pulicidae were significantly responsible for explaining the differences among the four flea families (linear discriminant analysis score > 2, P < 0.05). Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt2) analyses showed that the functional pathways varied significantly across flea families, which was supported by the significant correlation between the functional pathways and the microbial communities.}, } @article {pmid38834030, year = {2024}, author = {Engl, T}, title = {Reproductive manipulation: Wolbachia induce host parthenogenesis using a stolen transformer.}, journal = {Current biology : CB}, volume = {34}, number = {11}, pages = {R547-R549}, doi = {10.1016/j.cub.2024.04.076}, pmid = {38834030}, issn = {1879-0445}, mesh = {*Wolbachia/physiology/genetics ; *Parthenogenesis ; Animals ; *Wasps/microbiology/physiology ; Female ; Reproduction ; }, abstract = {The Wolbachia strain that infects the parasitoid wasp Encarsia formosa induces female-producing parthenogenesis. A new study shows that a Wolbachia-encoded gene has replaced the use of the ancestral wasp homologue that normally controls sexual reproduction, resulting in parthenogenesis.}, } @article {pmid38833496, year = {2024}, author = {Chen, J and Lin, G and Ma, K and Li, Z and Liégeois, S and Ferrandon, D}, title = {A specific innate immune response silences the virulence of Pseudomonas aeruginosa in a latent infection model in the Drosophila melanogaster host.}, journal = {PLoS pathogens}, volume = {20}, number = {6}, pages = {e1012252}, pmid = {38833496}, issn = {1553-7374}, mesh = {Animals ; *Drosophila melanogaster/microbiology/immunology ; *Pseudomonas aeruginosa/pathogenicity/immunology ; *Immunity, Innate ; *Pseudomonas Infections/immunology/microbiology ; Virulence ; Disease Models, Animal ; Host-Pathogen Interactions/immunology ; }, abstract = {Microbial pathogenicity often depends on the route of infection. For instance, P. aeruginosa or S. marcescens cause acute systemic infections when low numbers of bacteria are injected into D. melanogaster flies whereas flies succumb much slower to the continuous ingestion of these pathogens, even though both manage to escape from the gut compartment and reach the hemocoel. Here, we have developed a latent P. aeruginosa infection model by feeding flies on the bacteria for a short period. The bacteria stably colonize internal tissues yet hardly cause any damage since latently-infected flies live almost as long as noninfected control flies. The apparently dormant bacteria display particular characteristics in terms of bacterial colony morphology, composition of the outer cell wall, and motility. The virulence of these bacteria can however be reactivated upon wounding the host. We show that melanization but not the cellular or the systemic humoral response is the predominant host defense that establishes latency and may coerce the bacteria to a dormant state. In addition, the lasting activation of the melanization responses in latently-infected flies provides a degree of protection to the host against a secondary fungal infection. Latent infection by an ingested pathogen protects against a variety of homologous or heterologous systemic secondary infectious challenges, a situation previously described for the endosymbiotic Wolbachia bacteria, a guard against viral infections.}, } @article {pmid38832111, year = {2024}, author = {Cantin, LJ and Gregory, V and Blum, LN and Foster, JM}, title = {Dual RNA-seq in filarial nematodes and Wolbachia endosymbionts using RNase H based ribosomal RNA depletion.}, journal = {Frontiers in microbiology}, volume = {15}, number = {}, pages = {1418032}, pmid = {38832111}, issn = {1664-302X}, abstract = {Lymphatic filariasis is caused by parasitic nematodes and is a leading cause of disability worldwide. Many filarial worms contain the bacterium Wolbachia as an obligate endosymbiont. RNA sequencing is a common technique used to study their molecular relationships and to identify potential drug targets against the nematode and bacteria. Ribosomal RNA (rRNA) is the most abundant RNA species, accounting for 80-90% of the RNA in a sample. To reduce sequencing costs, it is necessary to remove ribosomal reads through poly-A enrichment or ribosomal depletion. Bacterial RNA does not contain a poly-A tail, making it difficult to sequence both the nematode and Wolbachia from the same library preparation using standard poly-A selection. Ribosomal depletion can utilize species-specific oligonucleotide probes to remove rRNA through pull-down or degradation methods. While species-specific probes are commercially available for many commonly studied model organisms, there are currently limited depletion options for filarial parasites. Here, we performed total RNA sequencing from Brugia malayi containing the Wolbachia symbiont (wBm) and designed ssDNA depletion probes against their rRNA sequences. We compared the total RNA library to poly-A enriched, Terminator 5'-Phosphate-Dependent Exonuclease treated, NEBNext Human/Bacteria rRNA depleted and our custom nematode probe depleted libraries. The custom nematode depletion library had the lowest percentage of ribosomal reads across all methods, with a 300-fold decrease in rRNA when compared to the total RNA library. The nematode depletion libraries also contained the highest percentage of Wolbachia mRNA reads, resulting in a 16-1,000-fold increase in bacterial reads compared to the other enrichment and depletion methods. Finally, we found that the Brugia malayi depletion probes can remove rRNA from the filarial worm Dirofilaria immitis and the majority of rRNA from the more distantly related free living nematode Caenorhabditis elegans. These custom filarial probes will allow for future dual RNA-seq experiments between nematodes and their bacterial symbionts from a single sequencing library.}, } @article {pmid38828283, year = {2024}, author = {Turner, HC}, title = {Cost-effectiveness of a Wolbachia-based replacement strategy for dengue control in Brazil.}, journal = {Lancet regional health. Americas}, volume = {35}, number = {}, pages = {100789}, pmid = {38828283}, issn = {2667-193X}, } @article {pmid38800519, year = {2024}, author = {Jeffries, CL and Tantely, LM and Kadriaj, P and Blagrove, MSC and Lytra, I and Orsborne, J and Al-Amin, HM and Mohammed, AR and Alam, MS and Girod, R and Afrane, YA and Bino, S and Robert, V and Boyer, S and Baylis, M and Velo, E and Hughes, GL and Walker, T}, title = {Mitochondrial and microbial diversity of the invasive mosquito vector species Culex tritaeniorhynchus across its extensive inter-continental geographic range.}, journal = {Wellcome open research}, volume = {9}, number = {}, pages = {18}, pmid = {38800519}, issn = {2398-502X}, abstract = {BACKGROUND: Culex (Cx.) tritaeniorhynchus is an invasive mosquito species with an extensive and expanding inter-continental distribution, currently reported across Asia, Africa, the Middle East, Europe and now Australia. It is an important vector of medical and veterinary pathogens which cause significant morbidity and mortality in human and animal populations. Across regions endemic for Japanese encephalitis virus (JEV), Cx. tritaeniorhynchus is considered the major vector and has also been shown to contribute to the transmission of several other zoonotic arboviruses including Rift Valley fever virus (RVFV) and West Nile virus (WNV).

METHODS: In this study, we used laboratory vector competence experiments to determine if Cx. tritaeniorhynchus from a Southern European population were competent JEV vectors. We also obtained samples from multiple geographically dispersed Cx. tritaeniorhynchus populations from countries within Europe, Africa, Eurasia and Asia to perform phylogenetic analysis to measure the level of mitochondrial divergence using the cytochrome oxidase subunit 1 (CO1) gene. We also undertook bacterial 16S rRNA gene amplicon sequencing to determine microbial diversity and used multi-locus sequence typing (MLST) to determine any evidence for the presence of strains of the naturally occurring endosymbiotic bacterium Wolbachia.

RESULTS: Cx. tritaeniorhynchus from a Greek population were shown be be competent vectors of JEV with high levels of virus present in saliva. We found a signficant level of mitochondrial genetic diversity using the mosquito CO1 gene between geographically dispersed populations. Furthermore, we report diverse microbiomes identified by 16S rRNA gene amplicon sequencing within and between geographical populations. Evidence for the detection of the endosymbiotic bacteria Wolbachia was confirmed using Wolbachia-specific PCR and MLST.

CONCLUSIONS: This study enhances our understanding of the diversity of Cx. tritaeniorhynchus and the associated microbiome across its inter-continental range and highlights the need for greater surveillance of this invasive vector species in Europe.}, } @article {pmid38796552, year = {2024}, author = {Wijegunawardana, NDAD and Gunawardene, YINS and Abeyewickreme, W and Chandrasena, TGAN and Thayanukul, P and Kittayapong, P}, title = {Diversity of Wolbachia infections in Sri Lankan mosquitoes with a new record of Wolbachia Supergroup B infecting Aedes aegypti vector populations.}, journal = {Scientific reports}, volume = {14}, number = {1}, pages = {11966}, pmid = {38796552}, issn = {2045-2322}, mesh = {Animals ; *Wolbachia/genetics/isolation & purification ; *Aedes/microbiology/virology ; Sri Lanka ; *Mosquito Vectors/microbiology ; *Phylogeny ; Female ; Male ; RNA, Ribosomal, 16S/genetics ; Multilocus Sequence Typing/methods ; }, abstract = {Wolbachia bacteria are common endosymbionts of insects and have recently been applied for controlling arboviral vectors, especially Aedes aegypti mosquito populations. However, several medically important mosquito species in Sri Lanka were present with limited information for the Wolbachia infection status. Therefore, the screening of Wolbachia in indigenous mosquitoes is required prior to a successful application of Wolbachia-based vector control strategy. In this study, screening of 78 mosquito species collected from various parts of the country revealed that 13 species were positive for Wolbachia infection, giving ~ 17% infection frequency of Wolbachia among the Sri Lankan mosquitoes. Twelve Wolbachia-positive mosquito species were selected for downstream Wolbachia strain genotyping using Multi Locus Sequencing Type (MLST), wsp gene, and 16S rRNA gene-based approaches. Results showed that these Wolbachia strains clustered together with the present Wolbachia phylogeny of world mosquito populations with some variations. Almost 90% of the mosquito populations were infected with supergroup B while the remaining were infected with supergroup A. A new record of Wolbachia supergroup B infection in Ae. aegypti, the main vectors of dengue, was highlighted. This finding was further confirmed by real-time qPCR, revealing Wolbachia density variations between Ae. aegypti and Ae. albopictus (p = 0.001), and between males and females (p < 0.05). The evidence of natural Wolbachia infections in Ae. aegypti populations in Sri Lanka is an extremely rare incident that has the potential to be used for arboviral vector control.}, } @article {pmid38792834, year = {2024}, author = {Moerbeck, L and Parreira, R and Szczotko, M and Seixas, G and Velez, R and Dmitryjuk, M and Santos, AS and Domingos, A and Antunes, S}, title = {Ticks and Tick-Borne Pathogens Circulating in Peri-Domestic Areas in Mainland Portugal.}, journal = {Microorganisms}, volume = {12}, number = {5}, pages = {}, pmid = {38792834}, issn = {2076-2607}, support = {2022.14376.BD//Fundação para a Ciência e Tecnologia/ ; }, abstract = {Over the years, tick-borne pathogens (TBPs) have garnered significant interest due to their medical, veterinary and economic importance. Additionally, TBPs have drawn attention to how these microorganisms interact with their own vectors, increasing the risk to human and animal infection of emerging and reemerging zoonoses. In this sense, ticks, which are obligate hematophagous ectoparasites, have a key role in maintaining and transmitting TBPs among humans and animals. The aim of this study was to assess the prevalence of neglected TBPs in mainland Portugal, namely Anaplasma spp., Babesia spp., Ehrlichia spp. and Neoehrlichia mikurensis. DNA fragments were detected in questing ticks collected from five different ecological areas under investigation. To the best of the authors' knowledge, this study reports new worldwide findings, including B. bigemina infecting Ixodes frontalis, Ixodes ricinus and Rhipicephalus sanguineus sensu lato. Additionally, it presents new findings in Portugal of N. mikurensis infecting I. ricinus and of presumably Wolbachia endosymbionts being detected in I. ricinus. Overall, there were 208 tick samples that were negative for all screened TBPs. The results herein obtained raise concerns about the circulation of neglected TBPs in mainland Portugal, especially in anthropophilic ticks, highlighting the importance of adopting a One Health perspective.}, } @article {pmid38786149, year = {2024}, author = {Löckener, I and Behrmann, LV and Reuter, J and Schiefer, A and Klöckner, A and Krannich, S and Otten, C and Mölleken, K and Ichikawa, S and Hoerauf, A and Schneider, T and Pfarr, KM and Henrichfreise, B}, title = {The MraY Inhibitor Muraymycin D2 and Its Derivatives Induce Enlarged Cells in Obligate Intracellular Chlamydia and Wolbachia and Break the Persistence Phenotype in Chlamydia.}, journal = {Antibiotics (Basel, Switzerland)}, volume = {13}, number = {5}, pages = {}, pmid = {38786149}, issn = {2079-6382}, support = {398967434 - TRR261//Deutsche Forschungsgemeinschaft (DFG, German Research Foundation)/ ; N.a.//BONFOR intramural funding program, Medical Faculty of Bonn University/ ; N.a.//Jürgen Manchot foundation/ ; N.a.//FEMHABIL, Medical Faculty, University of Bonn/ ; JP22H02738//JSPS KAKENHI Grant-in-Aid for Scientific Research (B)/ ; JP22ama121039//AMED/ ; N.a.//Studienstiftung des deutschen Volkes/ ; }, abstract = {Chlamydial infections and diseases caused by filarial nematodes are global health concerns. However, treatment presents challenges due to treatment failures potentially caused by persisting Chlamydia and long regimens against filarial infections accompanied by low compliance. A new treatment strategy could be the targeting of the reduced peptidoglycan structures involved in cell division in the obligate intracellular bacteria Chlamydia and Wolbachia, the latter being obligate endosymbionts supporting filarial development, growth, and survival. Here, cell culture experiments with C. trachomatis and Wolbachia showed that the nucleoside antibiotics muraymycin and carbacaprazamycin interfere with bacterial cell division and induce enlarged, aberrant cells resembling the penicillin-induced persistence phenotype in Chlamydia. Enzymatic inhibition experiments with purified C. pneumoniae MraY revealed that muraymycin derivatives abolish the synthesis of the peptidoglycan precursor lipid I. Comparative in silico analyses of chlamydial and wolbachial MraY with the corresponding well-characterized enzyme in Aquifex aeolicus revealed a high degree of conservation, providing evidence for a similar mode of inhibition. Muraymycin D2 treatment eradicated persisting non-dividing C. trachomatis cells from an established penicillin-induced persistent infection. This finding indicates that nucleoside antibiotics may have additional properties that can break bacterial persistence.}, } @article {pmid38784793, year = {2024}, author = {, }, title = {Erratum: Genetic diversity of endosymbiotic bacteria Wolbachia infecting two mosquito species of the genus Eretmapodites occurring in sympatry in the Comoros archipelago.}, journal = {Frontiers in microbiology}, volume = {15}, number = {}, pages = {1425304}, doi = {10.3389/fmicb.2024.1425304}, pmid = {38784793}, issn = {1664-302X}, abstract = {[This corrects the article DOI: 10.3389/fmicb.2024.1343917.].}, } @article {pmid38776328, year = {2024}, author = {Khosravi, G and Akbarzadeh, K and Karimian, F and Koosha, M and Saeedi, S and Oshaghi, MA}, title = {A survey of Wolbachia infection in brachyceran flies from Iran.}, journal = {PloS one}, volume = {19}, number = {5}, pages = {e0301274}, pmid = {38776328}, issn = {1932-6203}, mesh = {*Wolbachia/genetics/isolation & purification ; Animals ; Iran ; *Diptera/microbiology ; *Phylogeny ; RNA, Ribosomal, 16S/genetics ; }, abstract = {Wolbachia is a maternally inherited intracellular bacterium that is considered to be the most plentiful endosymbiont found in arthropods. It reproductively manipulates its host to increase the chances of being transmitted to the insect progeny; and it is currently used as a means of suppressing disease vector populations or controlling vector-borne diseases. Studies of the dissemination and prevalence of Wolbachia among its arthropod hosts are important for its possible use as a biological control agent. The molecular identification of Wolbachia relies on different primers sets due to Wolbachia strain variation. Here, we screened for the presence of Wolbachia in a broad range of Brachycera fly species (Diptera), collected from different regions of Iran, using nine genetic markers (wsp, ftsZ, fbpA, gatB, CoxA, gltA, GroEL dnaA, and 16s rRNA), for detecting, assessing the sensitivity of primers for detection, and phylogeny of this bacterium. The overall incidence of Wolbachia among 22 species from six families was 27.3%. The most commonly positive fly species were Pollenia sp. and Hydrotaea armipes. However, the bacterium was not found in the most medically important flies or in potential human disease vectors, including Musca domestica, Sarcophaga spp., Calliphora vicinia, Lucilia sericata, and Chrysomya albiceps. The primer sets of 16s rRNA with 53.0% and gatB with 52.0% were the most sensitive primers for detecting Wolbachia. Blast search, phylogenetic, and MLST analysis of the different locus sequences of Wolbachia show that all the six distantly related fly species likely belonging to supergroup A. Our study showed some primer sets generated false negatives in many of the samples, emphasizing the importance of using different loci in detecting Wolbachia. The study provides the groundwork for future studies of a Wolbachia-based program for control of flies.}, } @article {pmid38774500, year = {2024}, author = {Poulain, M and Rosinski, E and Henri, H and Balmand, S and Delignette-Muller, ML and Heddi, A and Lasseur, R and Vavre, F and Zaidman-Rémy, A and Kremer, N}, title = {Development, feeding, and sex shape the relative quantity of the nutritional obligatory symbiont Wolbachia in bed bugs.}, journal = {Frontiers in microbiology}, volume = {15}, number = {}, pages = {1386458}, pmid = {38774500}, issn = {1664-302X}, abstract = {The common bed bug, Cimex lectularius, is a hemipteran insect that feeds only on blood, and whose bites cause public health issues. Due to globalization and resistance to insecticides, this pest has undergone a significant and global resurgence in recent decades. Blood is an unbalanced diet, lacking notably sufficient B vitamins. Like all strict hematophagous arthropods, bed bugs host a nutritional symbiont supplying B vitamins. In C. lectularius, this nutritional symbiont is the intracellular bacterium Wolbachia (wCle). It is located in specific symbiotic organs, the bacteriomes, as well as in ovaries. Experimental depletion of wCle has been shown to result in longer nymphal development and lower fecundity. These phenotypes were rescued by B vitamin supplementation. Understanding the interaction between wCle and the bed bug may help to develop new pest control methods targeting the disruption of this symbiotic interaction. The objective of this work was thus to quantify accurately the density of wCle over the life cycle of the host and to describe potential associated morphological changes in the bacteriome. We also sought to determine the impact of sex, feeding status, and aging on the bacterial population dynamics. We showed that the relative quantity of wCle continuously increases during bed bug development, while the relative size of the bacteriome remains stable. We also showed that adult females harbor more wCle than males and that wCle relative quantity decreases slightly in adults with age, except in weekly-fed males. These results are discussed in the context of bed bug ecology and will help to define critical points of the symbiotic interaction during the bed bug life cycle.}, } @article {pmid38773487, year = {2024}, author = {Connolly, JB and Burt, A and Christophides, G and Diabate, A and Habtewold, T and Hancock, PA and James, AA and Kayondo, JK and Lwetoijera, DW and Manjurano, A and McKemey, AR and Santos, MR and Windbichler, N and Randazzo, F}, title = {Considerations for first field trials of low-threshold gene drive for malaria vector control.}, journal = {Malaria journal}, volume = {23}, number = {1}, pages = {156}, pmid = {38773487}, issn = {1475-2875}, mesh = {*Mosquito Control/methods ; *Mosquito Vectors/genetics ; *Malaria/prevention & control/transmission ; Animals ; *Anopheles/genetics ; *Gene Drive Technology/methods ; }, abstract = {Sustainable reductions in African malaria transmission require innovative tools for mosquito control. One proposal involves the use of low-threshold gene drive in Anopheles vector species, where a 'causal pathway' would be initiated by (i) the release of a gene drive system in target mosquito vector species, leading to (ii) its transmission to subsequent generations, (iii) its increase in frequency and spread in target mosquito populations, (iv) its simultaneous propagation of a linked genetic trait aimed at reducing vectorial capacity for Plasmodium, and (v) reduced vectorial capacity for parasites in target mosquito populations as the gene drive system reaches fixation in target mosquito populations, causing (vi) decreased malaria incidence and prevalence. Here the scope, objectives, trial design elements, and approaches to monitoring for initial field releases of such gene dive systems are considered, informed by the successful implementation of field trials of biological control agents, as well as other vector control tools, including insecticides, Wolbachia, larvicides, and attractive-toxic sugar bait systems. Specific research questions to be addressed in initial gene drive field trials are identified, and adaptive trial design is explored as a potentially constructive and flexible approach to facilitate testing of the causal pathway. A fundamental question for decision-makers for the first field trials will be whether there should be a selective focus on earlier points of the pathway, such as genetic efficacy via measurement of the increase in frequency and spread of the gene drive system in target populations, or on wider interrogation of the entire pathway including entomological and epidemiological efficacy. How and when epidemiological efficacy will eventually be assessed will be an essential consideration before decisions on any field trial protocols are finalized and implemented, regardless of whether initial field trials focus exclusively on the measurement of genetic efficacy, or on broader aspects of the causal pathway. Statistical and modelling tools are currently under active development and will inform such decisions on initial trial design, locations, and endpoints. Collectively, the considerations here advance the realization of developer ambitions for the first field trials of low-threshold gene drive for malaria vector control within the next 5 years.}, } @article {pmid38772333, year = {2024}, author = {Miyata, M and Nomura, M and Kageyama, D}, title = {Rapid spread of a vertically transmitted symbiont induces drastic shifts in butterfly sex ratio.}, journal = {Current biology : CB}, volume = {34}, number = {10}, pages = {R490-R492}, doi = {10.1016/j.cub.2024.04.027}, pmid = {38772333}, issn = {1879-0445}, mesh = {Animals ; *Sex Ratio ; *Wolbachia/physiology/genetics ; *Symbiosis ; *Butterflies/microbiology/physiology/genetics ; Female ; Male ; }, abstract = {The causes and consequences of sex-ratio dynamics constitutes a pivotal subject in evolutionary biology[1]. Under conditions of evolutionary equilibrium, the male-to-female ratio tends to be approximately 1:1; however, this equilibrium is susceptible to distortion by selfish genetic elements exemplified by driving sex chromosomes and cytoplasmic elements[2][,][3]. Although previous studies have documented instances of these genetic elements distorting the sex ratio, studies specifically tracking the process with which these distorters spread within populations, leading to a transition from balanced parity to a skewed, female-biased state, are notably lacking. Herein, we present compelling evidence documenting the rapid spread of the cytoplasmic endosymbiont Wolbachia within a localized population of the pierid butterfly Eurema hecabe (Figure 1A). This spread resulted in a shift in the sex ratio from near parity to an exceedingly skewed state overwhelmingly biased toward females, reaching 93.1% within a remarkably brief period of 4 years.}, } @article {pmid38771858, year = {2024}, author = {Maleki-Ravasan, N and Ghafari, SM and Najafzadeh, N and Karimian, F and Darzi, F and Davoudian, R and Farshbaf Pourabad, R and Parvizi, P}, title = {Characterization of bacteria expectorated during forced salivation of the Phlebotomus papatasi: A neglected component of sand fly infectious inoculums.}, journal = {PLoS neglected tropical diseases}, volume = {18}, number = {5}, pages = {e0012165}, pmid = {38771858}, issn = {1935-2735}, mesh = {Animals ; *Phlebotomus/microbiology ; *RNA, Ribosomal, 16S/genetics ; *Phylogeny ; *Saliva/microbiology ; *Bacteria/classification/genetics/isolation & purification ; Iran ; Insect Vectors/microbiology/physiology ; Female ; Microbiota ; Leishmaniasis, Cutaneous/transmission/microbiology/parasitology ; Male ; }, abstract = {The infectious inoculum of a sand fly, apart from its metacyclic promastigotes, is composed of factors derived from both the parasite and the vector. Vector-derived factors, including salivary proteins and the gut microbiota, are essential for the establishment and enhancement of infection. However, the type and the number of bacteria egested during salivation is unclear. In the present study, sand flies of Phlebotomus papatasi were gathered from three locations in hyperendemic focus of zoonotic cutaneous leishmaniasis (ZCL) in Isfahan Province, Iran. By using the forced salivation assay and targeting the 16S rRNA barcode gene, egested bacteria were characterized in 99 (44%) out of 224 sand flies. Culture-dependent and culture-independent methods identified the members of Enterobacter cloacae and Spiroplasma species as dominant taxa, respectively. Ten top genera of Spiroplasma, Ralstonia, Acinetobacter, Reyranella, Undibacterium, Bryobacter, Corynebacterium, Cutibacterium, Psychrobacter, and Wolbachia constituted >80% of the saliva microbiome. Phylogenetic analysis displayed the presence of only one bacterial species for the Spiroplasma, Ralstonia, Reyranella, Bryobacter and Wolbachia, two distinct species for Cutibacterium, three for Undibacterium and Psychrobacter, 16 for Acinetobacter, and 27 for Corynebacterium, in the saliva. The abundance of microbes in P. papatasi saliva was determined by incorporating the data on the read counts and the copy number of 16S rRNA gene, about 9,000 bacterial cells, per sand fly. Both microbiological and metagenomic data indicate that bacteria are constant companions of Leishmania, from the intestine of the vector to the vertebrate host. This is the first forced salivation experiment in a sand fly, addressing key questions on infectious bite and competent vectors.}, } @article {pmid38769579, year = {2024}, author = {Muharromah, AF and Carvajal, TM and Regilme, MAF and Watanabe, K}, title = {Fine-scale adaptive divergence and population genetic structure of Aedes aegypti in Metropolitan Manila, Philippines.}, journal = {Parasites & vectors}, volume = {17}, number = {1}, pages = {233}, pmid = {38769579}, issn = {1756-3305}, mesh = {Animals ; *Aedes/genetics/classification/physiology ; Philippines ; Female ; Male ; *Polymorphism, Single Nucleotide ; *Microsatellite Repeats/genetics ; *Genetics, Population ; *Mosquito Vectors/genetics/physiology ; Ecosystem ; Genetic Variation ; Dengue/transmission ; Adaptation, Physiological/genetics ; }, abstract = {BACKGROUND: The adaptive divergence of Aedes aegypti populations to heterogeneous environments can be a driving force behind the recent expansion of their habitat distribution and outbreaks of dengue disease in urbanized areas. In this study, we investigated the population genomics of Ae. aegypti at a regional scale in Metropolitan Manila, Philippines.

METHODS: We used the Pool-Seq double digestion restriction-site association DNA sequencing (ddRAD-Seq) approach to generate a high number of single nucleotide polymorphisms (SNPs), with the aim to determine local adaptation and compare the population structure with 11 microsatellite markers. A total of 217 Ae. aegypti individuals from seven female and seven male populations collected from Metropolitan Manila were used in the assays.

RESULTS: We detected 65,473 SNPs across the populations, of which 76 were non-neutral SNPs. Of these non-neutral SNPs, the multivariate regression test associated 50 with eight landscape variables (e.g. open space, forest, etc.) and 29 with five climate variables (e.g. air temperature, humidity, etc.) (P-value range 0.005-0.045) in female and male populations separately. Male and female populations exhibited contrasting spatial divergence, with males exhibiting greater divergence than females, most likely reflecting the different dispersal abilities of male and female mosquitoes. In the comparative analysis of the same Ae. aegypti individuals, the pairwise FST values of 11 microsatellite markers were lower than those of the neutral SNPs, indicating that the neutral SNPs generated via pool ddRAD-Seq were more sensitive in terms of detecting genetic differences between populations at fine-spatial scales.

CONCLUSIONS: Overall, our study demonstrates the utility of pool ddRAD-Seq for examining genetic differences in Ae. aegypti populations in areas at fine-spatial scales that could inform vector control programs such as Wolbachia-infected mosquito mass-release programs. This in turn would provide information on mosquito population dispersal patterns and the potential barriers to mosquito movement within and around the release area. In addition, the potential of environmental adaptability observed in Ae. aegypti could help population control efforts.}, } @article {pmid38760668, year = {2024}, author = {Bell-Sakyi, L and Haines, LR and Petrucci, G and Beliavskaia, A and Hartley, C and Khoo, JJ and Makepeace, BL and Abd-Alla, AMM and Darby, AC}, title = {Establishment and partial characterisation of a new cell line derived from adult tissues of the tsetse fly Glossina morsitans morsitans.}, journal = {Parasites & vectors}, volume = {17}, number = {1}, pages = {231}, pmid = {38760668}, issn = {1756-3305}, support = {BB/P024270/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; AV/PP0021/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; BB/P024270/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; BB/P024270/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; BB/P024270/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; BB/P024270/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; 223743/Z/21/Z/WT_/Wellcome Trust/United Kingdom ; 204806/Z/16/Z/WT_/Wellcome Trust/United Kingdom ; 223743/Z/21/Z/WT_/Wellcome Trust/United Kingdom ; 223743/Z/21/Z/WT_/Wellcome Trust/United Kingdom ; 223743/Z/21/Z/WT_/Wellcome Trust/United Kingdom ; 223743/Z/21/Z/WT_/Wellcome Trust/United Kingdom ; 223743/Z/21/Z/WT_/Wellcome Trust/United Kingdom ; Joint FAO/IAEA Insect Pest Control Subprogramme//FAO/IAEA/ ; Joint FAO/IAEA Insect Pest Control Subprogramme//FAO/IAEA/ ; }, mesh = {*Tsetse Flies/parasitology ; Animals ; Cell Line ; Female ; RNA, Ribosomal, 16S/genetics ; Karyotyping ; Insect Vectors/virology ; }, abstract = {BACKGROUND: Insect cell lines play a vital role in many aspects of research on disease vectors and agricultural pests. The tsetse fly Glossina morsitans morsitans is an important vector of salivarian trypanosomes in sub-Saharan Africa and, as such, is a major constraint on human health and agricultural development in the region.

METHODS: Here, we report establishment and partial characterisation of a cell line, GMA/LULS61, derived from tissues of adult female G. m. morsitans. GMA/LULS61 cells, grown at 28 °C in L-15 (Leibovitz) medium supplemented with foetal bovine serum and tryptose phosphate broth, have been taken through 23 passages to date and can be split 1:1 at 2-week intervals. Karyotyping at passage 17 revealed a predominantly haploid chromosome complement. Species origin and absence of contaminating bacteria were confirmed by PCR amplification and sequencing of fragments of the COI gene and pan-bacterial 16S rRNA gene respectively. However, PCR screening of RNA extracted from GMA/LULS61 cells confirmed presence of the recently described Glossina morsitans morsitans iflavirus and Glossina morsitans morsitans negevirus, but absence of Glossina pallipides salivary gland hypertrophy virus. GMA/LULS61 cells supported infection and growth of 6/7 different insect-derived strains of the intracellular bacterial symbiont Wolbachia.

CONCLUSIONS: The GMA/LULS61 cell line has potential for application in a variety of studies investigating the biology of G. m. morsitans and its associated pathogenic and symbiotic microorganisms.}, } @article {pmid38755197, year = {2024}, author = {Dufault, SM and Tanamas, SK and Indriani, C and Ahmad, RA and Utarini, A and Jewell, NP and Simmons, CP and Anders, KL}, title = {Reanalysis of cluster randomised trial data to account for exposure misclassification using a per-protocol and complier-restricted approach.}, journal = {Scientific reports}, volume = {14}, number = {1}, pages = {11207}, pmid = {38755197}, issn = {2045-2322}, support = {/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Humans ; *Wolbachia ; *Aedes/microbiology ; Animals ; *Dengue/prevention & control/transmission ; Mosquito Vectors/microbiology ; Randomized Controlled Trials as Topic ; Cluster Analysis ; Mosquito Control/methods ; Female ; }, abstract = {The intention-to-treat (ITT) analysis of the Applying Wolbachia to Eliminate Dengue (AWED) trial estimated a protective efficacy of 77.1% for participants resident in areas randomised to receive releases of wMel-infected Aedes aegypti mosquitoes, an emerging dengue preventive intervention. The limiting assumptions of ITT analyses in cluster randomised trials and the mobility of mosquitoes and humans across cluster boundaries indicate the primary analysis is likely to underestimate the full public health benefit. Using spatiotemporally-resolved data on the distribution of Wolbachia mosquitoes and on the mobility of AWED participants (n = 6306), we perform complier-restricted and per-protocol re-examinations of the efficacy of the Wolbachia intervention. Increased intervention efficacy was estimated in all analyses by the refined exposure measures. The complier-restricted analysis returned an estimated efficacy of 80.7% (95% CI 65.9, 89.0) and the per-protocol analysis estimated 82.7% (71.7, 88.4) efficacy when comparing participants with an estimated wMel exposure of ≥ 80% compared to those with <20%. These reanalyses demonstrate how human and mosquito movement can lead to underestimation of intervention effects in trials of vector interventions and indicate that the protective efficacy of Wolbachia is even higher than reported in the primary trial results.}, } @article {pmid38754459, year = {2024}, author = {Idro, R and Ogwang, R and Anguzu, R and Akun, P and Ningwa, A and Abbo, C and Giannoccaro, MP and Kubofcik, J and Mwaka, AD and Nakamya, P and Opar, B and Taylor, M and Nutman, TB and Elliott, A and Vincent, A and Newton, CR and Marsh, K}, title = {Doxycycline for the treatment of nodding syndrome: a randomised, placebo-controlled, phase 2 trial.}, journal = {The Lancet. Global health}, volume = {12}, number = {7}, pages = {e1149-e1158}, pmid = {38754459}, issn = {2214-109X}, support = {Z01 AI000439/ImNIH/Intramural NIH HHS/United States ; }, mesh = {Humans ; Child ; Adolescent ; Female ; Male ; *Doxycycline/therapeutic use ; *Nodding Syndrome/drug therapy ; Double-Blind Method ; Uganda ; Treatment Outcome ; Anti-Bacterial Agents/therapeutic use ; Onchocerca volvulus/drug effects ; }, abstract = {BACKGROUND: Nodding syndrome is a poorly understood neurological disorder that predominantly occurs in Africa. We hypothesised that nodding syndrome is a neuroinflammatory disorder, induced by antibodies to Onchocerca volvulus or its Wolbachia symbiont, cross-reacting with host neuronal proteins (HNPs), and that doxycycline can be used as treatment.

METHODS: In this randomised, double-blind, placebo-controlled, phase 2 trial, we recruited participants from districts affected by nodding syndrome in northern Uganda. We included children and adolescents aged 8-18 years with nodding syndrome, as defined by WHO consensus criteria. Participants were randomly assigned (1:1) to receive either 100 mg doxycycline daily or placebo for 6 weeks via a computer-generated schedule stratified by skin microscopy results, and all parties were masked to group assignment. Diagnoses of O volvulus and antibodies to HNPs were made using luciferase immunoprecipitation system assays and immunohistochemistry. The primary outcome was change in the proportion with antibodies to HNPs, assessed at 24 months. All participants were included in safety analyses, and surviving participants (those with samples at 24 months) were included in primary analyses. Secondary outcomes were: change in concentrations of antibodies to HNPs at 24 months compared with baseline; proportion of participants testing positive for antibodies to O volvulus-specific proteins and concentrations of Ov16 or OVOC3261 antibodies at 24 months compared with baseline; change in seizure burden, proportion achieving seizure freedom, and the proportions with interictal epileptiform discharges on the diagnostic EEG; overall quality of life; disease severity at 24 months; and incidence of all-cause adverse events, serious adverse events, and seizure-related mortality by 24 months. This trial is registered with ClinicalTrials.gov, NCT02850913.

FINDINGS: Between Sept 1, 2016, and Aug 31, 2018, 329 children and adolescents were screened, of whom 240 were included in the study. 140 (58%) participants were boys and 100 (42%) were girls. 120 (50%) participants were allocated to receive doxycycline and 120 (50%) to receive placebo. At recruitment, the median duration of symptoms was 9 years (IQR 6-10); 232 (97%) participants had O volvulus-specific antibodies and 157 (65%) had autoantibodies to HNPs. The most common plasma autoantibodies were to human protein deglycase DJ-1 (85 [35%] participants) and leiomodin-1 (77 [32%] participants) and, in cerebrospinal fluid (CSF), to human DJ-1 (27 [11%] participants) and leiomodin-1 (14 [6%] participants). On immunohistochemistry, 46 (19%) participants had CSF autoantibodies to HNPs, including leiomodin-1 (26 [11%]), γ-aminobutyric acid B receptors (two [<1%]), CASPR2 (one [<1%]), or unknown targets (28 [12%]). At 24 months, 161 (72%) of 225 participants had antibodies to HNPs compared with 157 (65%) of 240 at baseline. 6 weeks of doxycycline did not affect the concentration of autoantibodies to HNPs, seizure control, disease severity, or quality of life at the 24-month follow-up but substantially decreased Ov16 antibody concentrations; the median plasma signal-to-noise Ov16 ratio was 16·4 (95% CI 6·4-38·4), compared with 27·9 (8·2-65·8; p=0·033) for placebo. 14 (6%) participants died and, other than one traffic death, all deaths were seizure-related. Acute seizure-related hospitalisations (rate ratio [RR] 0·43 [95% CI 0·20-0·94], p=0·028) and deaths (RR 0·46 [0·24-0·89], p=0·028) were significantly lower in the doxycycline group. At 24 months, 96 (84%) of 114 participants who received doxycycline tested positive for antibodies to Ov16, compared with 97 (87%) of 111 on placebo (p=0·50), and 74 (65%) participants on doxycycline tested positive for antibodies to OVOC3261, compared with 57 (51%) on placebo (p=0·039). Doxycycline was safe; there was no difference in the incidence of grade 3-5 adverse events across the two groups.

INTERPRETATION: Nodding syndrome is strongly associated with O volvulus and the pathogenesis is probably mediated through an O volvulus induced autoantibody response to multiple proteins. Although it did not reverse disease symptoms, doxycycline or another prophylactic antibiotic could be considered as adjunct therapy to antiseizure medication, as it might reduce fatal complications from acute seizures and status epilepticus induced by febrile infections.

FUNDING: Medical Research Council (UK).

TRANSLATION: For the Luo translation of the abstract see Supplementary Materials section.}, } @article {pmid38742876, year = {2024}, author = {De Coninck, L and Soto, A and Wang, L and De Wolf, K and Smitz, N and Deblauwe, I and Mbigha Donfack, KC and Müller, R and Delang, L and Matthijnssens, J}, title = {Lack of abundant core virome in Culex mosquitoes from a temperate climate region despite a mosquito species-specific virome.}, journal = {mSystems}, volume = {9}, number = {6}, pages = {e0001224}, pmid = {38742876}, issn = {2379-5077}, support = {11L1323N//Fonds Wetenschappelijk Onderzoek (FWO)/ ; C14/20/108//KU Leuven/ ; }, mesh = {Animals ; *Culex/virology/microbiology ; *Virome/genetics ; *Wolbachia/genetics/isolation & purification ; Belgium ; Species Specificity ; Mosquito Vectors/virology/microbiology ; Metagenomics ; Insect Viruses/genetics/isolation & purification ; Climate ; }, abstract = {In arthropod-associated microbial communities, insect-specific viruses (ISVs) are prevalent yet understudied due to limited infectivity outside their natural hosts. However, ISVs might play a crucial role in regulating mosquito populations and influencing arthropod-borne virus transmission. Some studies have indicated a core virome in mosquitoes consisting of mostly ISVs. Employing single mosquito metagenomics, we comprehensively profiled the virome of native and invasive mosquito species in Belgium. This approach allowed for accurate host species determination, prevalence assessment of viruses and Wolbachia, and the identification of novel viruses. Contrary to our expectations, no abundant core virome was observed in Culex mosquitoes from Belgium. In that regard, we caution against rigidly defining mosquito core viromes and encourage nuanced interpretations of other studies. Nonetheless, our study identified 45 viruses of which 28 were novel, enriching our understanding of the mosquito virome and ISVs. We showed that the mosquito virome in this study is species-specific and less dependent on the location where mosquitoes from the same species reside. In addition, because Wolbachia has previously been observed to influence arbovirus transmission, we report the prevalence of Wolbachia in Belgian mosquitoes and the detection of several Wolbachia mobile genetic elements. The observed prevalence ranged from 83% to 92% in members from the Culex pipiens complex.IMPORTANCECulex pipiens mosquitoes are important vectors for arboviruses like West Nile virus and Usutu virus. Virome studies on individual Culex pipiens, and on individual mosquitoes in general, have been lacking. To mitigate this, we sequenced the virome of 190 individual Culex and 8 individual Aedes japonicus mosquitoes. We report the lack of a core virome in these mosquitoes from Belgium and caution the interpretation of other studies in this light. The discovery of new viruses in this study will aid our comprehension of insect-specific viruses and the mosquito virome in general in relation to mosquito physiology and mosquito population dynamics.}, } @article {pmid38732070, year = {2024}, author = {Zhang, J and Liu, Q and Dai, L and Zhang, Z and Wang, Y}, title = {Pan-Genome Analysis of Wolbachia, Endosymbiont of Diaphorina citri, Reveals Independent Origin in Asia and North America.}, journal = {International journal of molecular sciences}, volume = {25}, number = {9}, pages = {}, pmid = {38732070}, issn = {1422-0067}, support = {2021YFD1400805//Nation Key R & D Program of China/ ; 31672031//National Natural Science Foundation of China/ ; 32272537//National Natural Science Foundation of China/ ; }, mesh = {*Wolbachia/genetics/classification ; *Symbiosis/genetics ; *Genome, Bacterial ; Animals ; *Phylogeny ; Asia ; North America ; Hemiptera/microbiology/genetics ; Diptera/microbiology/genetics ; Polymorphism, Single Nucleotide ; }, abstract = {Wolbachia, a group of Gram-negative symbiotic bacteria, infects nematodes and a wide range of arthropods. Diaphorina citri Kuwayama, the vector of Candidatus Liberibacter asiaticus (CLas) that causes citrus greening disease, is naturally infected with Wolbachia (wDi). However, the interaction between wDi and D. citri remains poorly understood. In this study, we performed a pan-genome analysis using 65 wDi genomes to gain a comprehensive understanding of wDi. Based on average nucleotide identity (ANI) analysis, we classified the wDi strains into Asia and North America strains. The ANI analysis, principal coordinates analysis (PCoA), and phylogenetic tree analysis supported that the D. citri in Florida did not originate from China. Furthermore, we found that a significant number of core genes were associated with metabolic pathways. Pathways such as thiamine metabolism, type I secretion system, biotin transport, and phospholipid transport were highly conserved across all analyzed wDi genomes. The variation analysis between Asia and North America wDi showed that there were 39,625 single-nucleotide polymorphisms (SNPs), 2153 indels, 10 inversions, 29 translocations, 65 duplications, 10 SV-based insertions, and 4 SV-based deletions. The SV-based insertions and deletions involved genes encoding transposase, phage tail tube protein, ankyrin repeat (ANK) protein, and group II intron-encoded protein. Pan-genome analysis of wDi contributes to our understanding of the geographical population of wDi, the origin of hosts of D. citri, and the interaction between wDi and its host, thus facilitating the development of strategies to control the insects and huanglongbing (HLB).}, } @article {pmid38730102, year = {2024}, author = {Taglialegna, A}, title = {A mosquito symbiont controls flaviviruses.}, journal = {Nature reviews. Microbiology}, volume = {22}, number = {7}, pages = {387}, pmid = {38730102}, issn = {1740-1534}, mesh = {Animals ; *Symbiosis ; *Flavivirus/genetics/physiology/classification ; Mosquito Vectors/virology ; Culicidae/virology ; Humans ; Aedes/virology ; Wolbachia/physiology ; Flavivirus Infections/virology/transmission ; }, } @article {pmid38725798, year = {2024}, author = {Setegn, A and Amare, GA and Mihret, Y}, title = {Wolbachia and Lymphatic Filarial Nematodes and Their Implications in the Pathogenesis of the Disease.}, journal = {Journal of parasitology research}, volume = {2024}, number = {}, pages = {3476951}, pmid = {38725798}, issn = {2090-0023}, abstract = {Lymphatic filariasis (LF) is an infection of three closely related filarial worms such as Wuchereria bancrofti, Brugia malayi, and Brugia timori. These worms can cause a devastating disease that involves acute and chronic lymphoedema of the extremities, which can cause elephantiasis in both sexes and hydroceles in males. These important public health nematodes were found to have a mutualistic relationship with intracellular bacteria of the genus Wolbachia, which is essential for the development and survival of the nematode. The host's inflammatory response to parasites and possibly also to the Wolbachia endosymbiont is the cause of lymphatic damage and disease pathogenesis. This review tried to describe and highlight the mutualistic associations between Wolbachia and lymphatic filarial nematodes and the role of bacteria in the pathogenesis of lymphatic filariasis. Articles for this review were searched from PubMed, Google Scholar, and other databases. Article searching was not restricted by publication year; however, only English version full-text articles were included.}, } @article {pmid38716193, year = {2024}, author = {Mushtaq, I and Sarwar, MS and Chaudhry, A and Shah, SAH and Ahmad, MM}, title = {Updates on traditional methods for combating malaria and emerging Wolbachia-based interventions.}, journal = {Frontiers in cellular and infection microbiology}, volume = {14}, number = {}, pages = {1330475}, pmid = {38716193}, issn = {2235-2988}, mesh = {*Wolbachia ; *Malaria/prevention & control ; Animals ; Humans ; *Mosquito Vectors/microbiology ; Mosquito Control/methods ; Pakistan ; }, abstract = {The escalating challenge of malaria control necessitates innovative approaches that extend beyond traditional control strategies. This review explores the incorporation of traditional vector control techniques with emerging Wolbachia-based interventions. Wolbachia, a naturally occurring bacteria, offers a novel approach for combatting vector-borne diseases, including malaria, by reducing the mosquitoes' ability to transmit these diseases. The study explores the rationale for this integration, presenting various case studies and pilot projects that have exhibited significant success. Employing a multi-dimensional approach that includes community mobilization, environmental modifications, and new biological methods, the paper posits that integrated efforts could mark a turning point in the struggle against malaria. Our findings indicate that incorporating Wolbachia-based strategies into existing vector management programs not only is feasible but also heightens the efficacy of malaria control initiatives in different countries especially in Pakistan. The paper concludes that continued research and international collaboration are imperative for translating these promising methods from the laboratory to the field, thereby offering a more sustainable and effective malaria control strategy.}, } @article {pmid38705185, year = {2024}, author = {Łukasik, P and Kolasa, MR}, title = {With a little help from my friends: the roles of microbial symbionts in insect populations and communities.}, journal = {Philosophical transactions of the Royal Society of London. Series B, Biological sciences}, volume = {379}, number = {1904}, pages = {20230122}, pmid = {38705185}, issn = {1471-2970}, mesh = {Animals ; *Insecta/microbiology/physiology ; *Symbiosis ; *Microbiota/physiology ; Biodiversity ; }, abstract = {To understand insect abundance, distribution and dynamics, we need to understand the relevant drivers of their populations and communities. While microbial symbionts are known to strongly affect many aspects of insect biology, we lack data on their effects on populations or community processes, or on insects' evolutionary responses at different timescales. How these effects change as the anthropogenic effects on ecosystems intensify is an area of intense research. Recent developments in sequencing and bioinformatics permit cost-effective microbial diversity surveys, tracking symbiont transmission, and identification of functions across insect populations and multi-species communities. In this review, we explore how different functional categories of symbionts can influence insect life-history traits, how these effects could affect insect populations and their interactions with other species, and how they may affect processes and patterns at the level of entire communities. We argue that insect-associated microbes should be considered important drivers of insect response and adaptation to environmental challenges and opportunities. We also outline the emerging approaches for surveying and characterizing insect-associated microbiota at population and community scales. This article is part of the theme issue 'Towards a toolkit for global insect biodiversity monitoring'.}, } @article {pmid38701242, year = {2024}, author = {Roldán, EL and Stelinski, LL and Pelz-Stelinski, KS}, title = {Reduction of Wolbachia in Diaphorina citri (Hemiptera: Liviidae) increases phytopathogen acquisition and decreases fitness.}, journal = {Journal of economic entomology}, volume = {117}, number = {3}, pages = {733-749}, doi = {10.1093/jee/toae089}, pmid = {38701242}, issn = {1938-291X}, support = {2021-70029-36053//United States Department of Agriculture National Institute of Food and Agriculture/ ; 2021-70029-36053//U.S. Department of Agriculture/ ; 2021-70029-36053//National Institute of Food and Agriculture/ ; }, mesh = {Animals ; *Wolbachia/physiology ; *Hemiptera/microbiology ; Female ; *Doxycycline/pharmacology ; Male ; *Genetic Fitness ; Nymph/microbiology/growth & development ; Liberibacter ; Plant Diseases/microbiology/prevention & control ; Symbiosis ; Anti-Bacterial Agents/pharmacology ; Citrus/microbiology ; }, abstract = {Wolbachia pipientis is a maternally inherited intracellular bacterium that infects a wide range of arthropods. Wolbachia can have a significant impact on host biology and development, often due to its effects on reproduction. We investigated Wolbachia-mediated effects in the Asian citrus psyllid, Diaphorina citri Kuwayama, which transmits Candidatus Liberibacter asiaticus (CLas), the causal agent of citrus greening disease. Diaphorina citri are naturally infected with Wolbachia; therefore, investigating Wolbachia-mediated effects on D. citri fitness and CLas transmission required artificial reduction of this endosymbiont with the application of doxycycline. Doxycycline treatment of psyllids reduced Wolbachia infection by approximately 60% in both male and female D. citri. Psyllids treated with doxycycline exhibited higher CLas acquisition in both adults and nymphs as compared with negative controls. In addition, doxycycline-treated psyllids exhibited decreased fitness as measured by reduced egg and nymph production as well as adult emergence as compared with control lines without the doxycycline treatment. Our results indicate that Wolbachia benefits D. citri by improving fitness and potentially competes with CLas by interfering with phytopathogen acquisition. Targeted manipulation of endosymbionts in this phytopathogen vector may yield disease management tools.}, } @article {pmid38692276, year = {2024}, author = {Li, C and Li, CQ and Chen, ZB and Liu, BQ and Sun, X and Wei, KH and Li, CY and Luan, JB}, title = {Wolbachia symbionts control sex in a parasitoid wasp using a horizontally acquired gene.}, journal = {Current biology : CB}, volume = {34}, number = {11}, pages = {2359-2372.e9}, doi = {10.1016/j.cub.2024.04.035}, pmid = {38692276}, issn = {1879-0445}, mesh = {Animals ; *Wolbachia/physiology/genetics ; *Wasps/physiology/microbiology/genetics ; *Symbiosis/genetics ; Female ; Male ; *Gene Transfer, Horizontal ; Parthenogenesis/genetics ; Insect Proteins/genetics/metabolism ; Sex Determination Processes/genetics ; }, abstract = {Host reproduction can be manipulated by bacterial symbionts in various ways. Parthenogenesis induction is the most effective type of reproduction manipulation by symbionts for their transmission. Insect sex is determined by regulation of doublesex (dsx) splicing through transformer2 (tra2) and transformer (tra) interaction. Although parthenogenesis induction by symbionts has been studied since the 1970s, its underlying molecular mechanism is unknown. Here we identify a Wolbachia parthenogenesis-induction feminization factor gene (piff) that targets sex-determining genes and causes female-producing parthenogenesis in the haplodiploid parasitoid Encarsia formosa. We found that Wolbachia elimination repressed expression of female-specific dsx and enhanced expression of male-specific dsx, which led to the production of wasp haploid male offspring. Furthermore, we found that E. formosa tra is truncated and non-functional, and Wolbachia has a functional tra homolog, termed piff, with an insect origin. Wolbachia PIFF can colocalize and interact with wasp TRA2. Moreover, Wolbachia piff has coordinated expression with tra2 and dsx of E. formosa. Our results demonstrate the bacterial symbiont Wolbachia has acquired an insect gene to manipulate the host sex determination cascade and induce parthenogenesis in wasps. This study reveals insect-to-bacteria horizontal gene transfer drives the evolution of animal sex determination systems, elucidating a striking mechanism of insect-microbe symbiosis.}, } @article {pmid38691215, year = {2024}, author = {Tokash-Peters, AG and Niyonzima, JD and Kayirangwa, M and Muhayimana, S and Tokash, IW and Jabon, JD and Lopez, SG and Kearns, PJ and Woodhams, DC}, title = {Mosquito Microbiomes of Rwanda: Characterizing Mosquito Host and Microbial Communities in the Land of a Thousand Hills.}, journal = {Microbial ecology}, volume = {87}, number = {1}, pages = {64}, pmid = {38691215}, issn = {1432-184X}, support = {DGE 1249946//Directorate for Biological Sciences/ ; 1950051//Directorate for Biological Sciences/ ; 1947684//National Science Foundation/ ; 1947684//National Science Foundation/ ; }, mesh = {Rwanda ; Animals ; *Microbiota ; *Culicidae/microbiology ; *Wolbachia/genetics/isolation & purification/classification ; *Bacteria/classification/genetics/isolation & purification ; Mosquito Vectors/microbiology ; Female ; Male ; RNA, Ribosomal, 16S/genetics ; Serratia/genetics/isolation & purification/classification ; Electron Transport Complex IV/genetics ; High-Throughput Nucleotide Sequencing ; }, abstract = {Mosquitoes are a complex nuisance around the world and tropical countries bear the brunt of the burden of mosquito-borne diseases. Rwanda has had success in reducing malaria and some arboviral diseases over the last few years, but still faces challenges to elimination. By building our understanding of in situ mosquito communities in Rwanda at a disturbed, human-occupied site and at a natural, preserved site, we can build our understanding of natural mosquito microbiomes toward the goal of implementing novel microbial control methods. Here, we examined the composition of collected mosquitoes and their microbiomes at two diverse sites using Cytochrome c Oxidase I sequencing and 16S V4 high-throughput sequencing. The majority (36 of 40 species) of mosquitoes captured and characterized in this study are the first-known record of their species for Rwanda but have been characterized in other nations in East Africa. We found significant differences among mosquito genera and among species, but not between mosquito sexes or catch method. Bacteria of interest for arbovirus control, Asaia, Serratia, and Wolbachia, were found in abundance at both sites and varied greatly by species.}, } @article {pmid38683843, year = {2024}, author = {Hernández, AM and Alcaraz, LD and Hernández-Álvarez, C and Romero, MF and Jara-Servín, A and Barajas, H and Ramírez, CM and Peimbert, M}, title = {Revealing the microbiome diversity and biocontrol potential of field Aedes ssp.: Implications for disease vector management.}, journal = {PloS one}, volume = {19}, number = {4}, pages = {e0302328}, pmid = {38683843}, issn = {1932-6203}, mesh = {*Aedes/microbiology ; Animals ; *Microbiota ; *Mosquito Vectors/microbiology ; *RNA, Ribosomal, 16S/genetics ; Wolbachia/genetics/physiology/isolation & purification ; Larva/microbiology ; Metagenomics/methods ; Mexico ; Mosquito Control/methods ; }, abstract = {The mosquito Aedes spp. holds important relevance for human and animal health, as it serves as a vector for transmitting multiple diseases, including dengue and Zika virus. The microbiome's impact on its host's health and fitness is well known. However, most studies on mosquito microbiomes have been conducted in laboratory settings. We explored the mixed microbial communities within Aedes spp., utilizing the 16S rRNA gene for diversity analysis and shotgun metagenomics for functional genomics. Our samples, which included Ae. aegypti and Ae. albopictus, spanned various developmental stages-eggs, larvae, and adults-gathered from five semiurban areas in Mexico. Our findings revealed a substantial diversity of 8,346 operational taxonomic units (OTUs), representing 967 bacterial genera and 126,366 annotated proteins. The host developmental stage was identified as the primary factor associated with variations in the microbiome composition. Subsequently, we searched for genes and species involved in mosquito biocontrol. Wolbachia accounted for 9.6% of the 16S gene sequences. We observed a high diversity (203 OTUs) of Wolbachia strains commonly associated with mosquitoes, such as wAlb, with a noticeable increase in abundance during the adult stages. Notably, we detected the presence of the cifA and cifB genes, which are associated with Wolbachia's cytoplasmic incompatibility, a biocontrol mechanism. Additionally, we identified 221 OTUs related to Bacillus, including strains linked to B. thuringiensis. Furthermore, we discovered multiple genes encoding insecticidal toxins, such as Cry, Mcf, Vip, and Vpp. Overall, our study contributes to the understanding of mosquito microbiome biodiversity and metabolic capabilities, which are essential for developing effective biocontrol strategies against this disease vector.}, } @article {pmid38681850, year = {2024}, author = {Junsiri, W and Kamkong, P and Phojun, A and Taweethavonsawat, P}, title = {Unveiling zoonotic threats: molecular identification of Brugia sp. infection in a lion.}, journal = {Frontiers in veterinary science}, volume = {11}, number = {}, pages = {1376208}, pmid = {38681850}, issn = {2297-1769}, abstract = {Brugia malayi and B. pahangi, potential zoonotic pathogens transmitted by mosquitoes, are believed to primarily infect dogs and cats as reservoir hosts. Although previous studies have indicated nematode infections in lions, particularly in zoo environments where human contact with these reservoirs is possible, limited documentation exists regarding Brugia sp. infections in lions in Thailand. This study aims to investigate a case of Brugia infection in a lion from a zoo in Thailand. The blood sample was collected and examined from a female lion, using staining methods to morphologically identify microfilaria at the genus level. Subsequently, the PCR was employed targeting specific genes, including mitochondrial 12S rDNA, 18S rDNA, cytochrome oxidase I (COI) and Wolbachia surface protein (wsp), to confirm the species of the filarial nematode parasite. The genetic sequencing results revealed a high similarity (99-100%) to B. malayi for the 12S rDNA, 18S rDNA, COI and wsp genes. Phylogenetic analysis based on nucleotide sequences from the 12S rDNA, 18S rDNA, COI and wsp genes showed that the sequences from this study belong to different clusters. This marks the inaugural documentation of molecular identification of Brugia infection in a lion, signifying that lions could function as reservoirs for this parasite and present a potential public health risk in the region. Our research underscores the effectiveness of molecular techniques and phylogenetic analysis in discerning and comprehending the evolution of filarial parasites. Additionally, it emphasizes the significance of these methods in enhancing the diagnosis, control, and prevention of zoonotic filarial nematode infections.}, } @article {pmid38680182, year = {2024}, author = {Andreenkova, OV and Adonyeva, NV and Efimov, VM and Gruntenko, NE}, title = {Fertility differences between two wild-type Drosophila melanogaster lines correlate with differences in the expression of the Jheh1 gene, which codes for an enzyme degrading juvenile hormone.}, journal = {Vavilovskii zhurnal genetiki i selektsii}, volume = {28}, number = {2}, pages = {185-189}, doi = {10.18699/vjgb-24-22}, pmid = {38680182}, issn = {2500-0462}, abstract = {Juvenile hormone plays a "status quo" role in Drosophila melanogaster larvae, preventing the untimely metamorphosis, and performs a gonadotropic function in imagoes, ensuring the ovaries' preparedness for vitellogenesis. The decreased level of juvenile hormone results in reproductive disorders in D. melanogaster females including a delay in the oviposition onset and a fertility decrease. Another factor that can affect the insect reproduction is an infection with the maternally inherited symbiotic α-proteobacterium Wolbachia. The present study is devoted to the analysis of the expression of two juvenile hormone metabolism genes encoding enzymes of its synthesis and degradation, juvenile hormone acid O-methyltransferase (jhamt) and juvenile hormone epoxide hydrase (Jheh1), respectively, in four wild-type D. melanogaster lines, two of them being infected with Wolbachia. Lines w153 and Bi90 were both derived from an individual wild-caught females infected with Wolbachia, while lines w153T and Bi90T were derived from them by tetracycline treatment and are free of infection. Line Bi90 is known to be infected with the Wolbachia strain wMel, and line w153, with the Wolbachia strain wMelPlus belonging to the wMelCS genotype. It was found that infection with either Wolbachia strain does not affect the expression of the studied genes. At the same time, it was shown that the w153 and w153T lines differ from the Bi90 and Bi90T lines by an increased level of the Jheh1 gene expression and do not differ in the jhamt gene expression level. Analysis of the fertility of these four lines showed that it does not depend on Wolbachia infection either, but differs between lines with different nuclear genotypes: in w153 and w153T, it is significantly lower than in lines Bi90 and Bi90T. The data obtained allow us to reasonably propose that the inter-line D. melanogaster polymorphism in the metabolism of the juvenile hormone is determined by its degradation (not by its synthesis) and correlates with the fertility level.}, } @article {pmid38678110, year = {2024}, author = {Wan, H and Wu, Y and Fan, G and Li, D}, title = {Wolbachia invasion dynamics of a random mosquito population model with imperfect maternal transmission and incomplete CI.}, journal = {Journal of mathematical biology}, volume = {88}, number = {6}, pages = {72}, doi = {10.1007/s00285-024-02094-9}, pmid = {38678110}, issn = {1432-1416}, support = {11971240//national natural science foundation of china/ ; 12171003; 11801209//national natural science foundation of china/ ; }, mesh = {*Wolbachia/physiology/pathogenicity ; Animals ; *Mathematical Concepts ; *Models, Biological ; Female ; *Mosquito Vectors/microbiology ; Population Dynamics/statistics & numerical data ; Cytoplasm/microbiology ; Culicidae/microbiology ; Male ; Computer Simulation ; Maternal Inheritance ; }, abstract = {In this work, we formulate a random Wolbachia invasion model incorporating the effects of imperfect maternal transmission and incomplete cytoplasmic incompatibility (CI). Under constant environments, we obtain the following results: Firstly, the complete invasion equilibrium of Wolbachia does not exist, and thus the population replacement is not achievable in the case of imperfect maternal transmission; Secondly, imperfect maternal transmission or incomplete CI may obliterate bistability and backward bifurcation, which leads to the failure of Wolbachia invasion, no matter how many infected mosquitoes would be released; Thirdly, the threshold number of the infected mosquitoes to be released would increase with the decrease of the maternal transmission rate or the intensity of CI effect. In random environments, we investigate in detail the Wolbachia invasion dynamics of the random mosquito population model and establish the initial release threshold of infected mosquitoes for successful invasion of Wolbachia into the wild mosquito population. In particular, the existence and stability of invariant probability measures for the establishment and extinction of Wolbachia are determined.}, } @article {pmid38677361, year = {2024}, author = {Tuñon, A and García, J and Carrera, LC and Chaves, LF and Lenhart, AE and Loaiza, JR}, title = {Chemical control of medically important arthropods in Panama: A systematic literature review of historical efforts.}, journal = {Acta tropica}, volume = {255}, number = {}, pages = {107217}, doi = {10.1016/j.actatropica.2024.107217}, pmid = {38677361}, issn = {1873-6254}, mesh = {Animals ; Humans ; Aedes/drug effects ; Anopheles/drug effects ; History, 20th Century ; History, 21st Century ; Insecticide Resistance ; *Insecticides/pharmacology ; *Mosquito Control/methods ; *Mosquito Vectors/drug effects ; Panama ; *Vector Borne Diseases/prevention & control ; }, abstract = {Vector-borne diseases are a major source of morbidity in Panama. Herein, we describe historical usage patterns of synthetic insecticides to control arthropod disease vectors in this country. We examine the influence of interventions by vector control programs on the emergence of insecticide resistance. Chemical control has traditionally focused on two mosquito species: Anopheles albimanus, a major regional malaria vector, and Aedes aegypti, a historical vector of yellow fever, and current vector of dengue, chikungunya, and Zika. Countrywide populations of An. albimanus depict hyperirritability to organochlorine insecticides administered by indoor residual spraying, although they appear susceptible to these insecticides in bioassays settings, as well as to organophosphate and carbamate insecticides in field tests. Populations of Ae. aegypti show resistance to pyrethroids, particularly in areas near Panama City, but the spread of resistance remains unknown in Ae. aegypti and Aedes albopictus. A One Health approach is needed in Panama to pinpoint the insecticide resistance mechanisms including the frequency of knockdown mutations and behavioral plasticity in populations of Anopheles and Aedes mosquitoes. This information is necessary to guide the sustainable implementation of chemical control strategies and the use of modern vector control technologies such as genetically modified mosquitoes, and endosymbiont Wolbachia-based biological control.}, } @article {pmid38667347, year = {2024}, author = {Liu, Y and Zhang, L and Cai, X and Rutikanga, A and Qiu, B and Hou, Y}, title = {The Diversity of Wolbachia and Other Bacterial Symbionts in Spodoptera frugiperda.}, journal = {Insects}, volume = {15}, number = {4}, pages = {}, pmid = {38667347}, issn = {2075-4450}, support = {2022YFC2601400//the National Key R & D Program of China/ ; NT2021003//the project of Guangdong Laboratory for Lingnan Modern Agriculture/ ; 2017YFC1200605//the National Key Research and Development Programme of China/ ; 2017NZ0003-1-6//the Science and Technology Major Project of Fujian/ ; 202102021290//the Guangzhou Basic and Applied Basic Research Program, China/ ; }, abstract = {Bacterial symbionts associated with insects can be crucial in insect nutrition, metabolism, immune responses, development, and reproduction. However, the bacterial symbionts of the fall armyworm Spodoptera frugiperda remain unclear. S. frugiperda is an invasive polyphagous pest that severely damages many crops, particularly maize and wheat. Here, we investigated the infection, composition, abundance, and diversity of bacterial symbionts, especially Wolbachia, in different tissues of S. frugiperda female adults. The infection prevalence frequencies of Wolbachia in five provinces of China, namely Pu'er, Yunnan; Nanning, Guangxi; Sanya, Hainan; Yunfu, Guangdong; and Nanping, Fujian, were assessed. The results indicated that Proteobacteria, Firmicutes, and Bacteroidetes were the three most dominant bacterial phyla in S. frugiperda adults. At the genus level, the abundant microbiota, which included Enterobacter and Enterococcus, varied in abundance between tissues of S. frugiperda. Wolbachia was found in the ovaries and salivary glands of S. frugiperda adults, and was present in 33.33% of the Pu'er, Yunnan, 23.33% of the Nanning, Guangxi, and 13.33% of the Sanya, Hainan populations, but Wolbachia was absent in the Yunfu, Guangdong and Nanping, Fujian populations. Further phylogenetic analyses revealed that all of the Wolbachia strains from the different S. frugiperda populations belonged to the supergroup B and were named the wFru strain. Since there were Wolbachia strains inducing cytoplasmic incompatibility in supergroup B, these findings may provide a foundation for developing potential biocontrol techniques against S. frugiperda.}, } @article {pmid38646734, year = {2024}, author = {Ng, IK}, title = {Wastewater surveillance and Wolbachia-aedes method to combat Zika virus outbreaks in tropical countries.}, journal = {Tropical doctor}, volume = {54}, number = {3}, pages = {295}, doi = {10.1177/00494755241246019}, pmid = {38646734}, issn = {1758-1133}, mesh = {*Zika Virus Infection/epidemiology/prevention & control ; Humans ; *Wolbachia ; *Disease Outbreaks/prevention & control ; *Aedes/virology/microbiology ; *Zika Virus ; *Wastewater/microbiology/virology ; Animals ; Tropical Climate ; Mosquito Vectors/virology/microbiology ; }, } @article {pmid38645127, year = {2024}, author = {Penunuri, G and Wang, P and Corbett-Detig, R and Russell, SL}, title = {A Structural Proteome Screen Identifies Protein Mimicry in Host-Microbe Systems.}, journal = {bioRxiv : the preprint server for biology}, volume = {}, number = {}, pages = {}, doi = {10.1101/2024.04.10.588793}, pmid = {38645127}, issn = {2692-8205}, support = {R00 GM135583/GM/NIGMS NIH HHS/United States ; T32 HG012344/HG/NHGRI NIH HHS/United States ; }, abstract = {Host-microbe systems are evolutionary niches that produce coevolved biological interactions and are a key component of global health. However, these systems have historically been a difficult field of biological research due to their experimental intractability. Impactful advances in global health will be obtained by leveraging in silico screens to identify genes involved in mediating interspecific interactions. These predictions will progress our understanding of these systems and lay the groundwork for future in vitro and in vivo experiments and bioengineering projects. A driver of host-manipulation and intracellular survival utilized by host-associated microbes is molecular mimicry, a critical mechanism that can occur at any level from DNA to protein structures. We applied protein structure prediction and alignment tools to explore host-associated bacterial structural proteomes for examples of protein structure mimicry. By leveraging the Legionella pneumophila proteome and its many known structural mimics, we developed and validated a screen that can be applied to virtually any host-microbe system to uncover signals of protein mimicry. These mimics represent candidate proteins that mediate host interactions in microbial proteomes. We successfully applied this screen to other microbes with demonstrated effects on global health, Helicobacter pylori and Wolbachia , identifying protein mimic candidates in each proteome. We discuss the roles these candidates may play in important Wolbachia -induced phenotypes and show that Wobachia infection can partially rescue the loss of one of these factors. This work demonstrates how a genome-wide screen for candidates of host-manipulation and intracellular survival offers an opportunity to identify functionally important genes in host-microbe systems.}, } @article {pmid38644035, year = {2024}, author = {Pardinilla, LM and Aljaberi, S and Procter, M and Hamdan, L and Pasha, SK and Al Aiyan, A and Qablan, MA}, title = {The prevalence of selected vector-borne diseases in dromedary camels (Camelus dromedarius) in the United Arab Emirates.}, journal = {Veterinary parasitology, regional studies and reports}, volume = {50}, number = {}, pages = {101006}, doi = {10.1016/j.vprsr.2024.101006}, pmid = {38644035}, issn = {2405-9390}, mesh = {Animals ; United Arab Emirates/epidemiology ; *Camelus/parasitology ; Prevalence ; *Vector Borne Diseases/epidemiology/parasitology/veterinary/microbiology ; Female ; Male ; Babesia/isolation & purification/genetics ; Phylogeny ; Trypanosoma/isolation & purification/genetics/classification ; Anaplasmataceae/isolation & purification/genetics ; Babesiosis/epidemiology/parasitology ; Risk Factors ; }, abstract = {Vector-borne diseases (VBDs) affecting dromedary camels (Camelus dromedarius) have considerable importance in the United Arab Emirates (UAE) because of the consequences associated with production decline and economic losses. Our study aimed to determine the prevalence of selected VBDs in camels in the UAE and identify risk factors. This research is currently affected by the low number of epidemiological molecular surveys addressing this issue. Blood samples were obtained from 425 dromedary camels from different locations across the UAE. Whole genomic DNA was isolated, and PCR screening was done to detect piroplasmids (Babesia/Theileria spp.), Trypanosoma spp., and Anaplasmataceae spp. (Anaplasma, Ehrlichia, Neorickettsia and Wolbachia spp.). Amplicons were sequenced, and phylogenetic trees were constructed. Trypanosoma sequences were identified as T. brucei evansi, whereas Anaplasmataceae sequences were identified as A. platys-like. All camels were negative for Babesia/Theileria spp. (0%); however, 18 camels were positive for T. b. evansi (4%) and 52 were positive for A. platys-like (12%). Mixed infection with T. b. evansi and A. platys-like was found in one camel. Statistical analyses revealed that camels with a brown coat colour were significantly more prone to acquire the A. platys-like strain compared with those having a clearer coat. A similar finding was observed when comparing urban moving camels with desert indoor and urban indoor camels. Continuous disease surveillance is required to ensure and maintain the good health status of the camels in the UAE. Nonetheless, the risk of disease outbreak remains if the misuse of drugs continues.}, } @article {pmid38642566, year = {2024}, author = {Salje, H and Jiggins, FM}, title = {Risks of releasing imperfect Wolbachia strains for arbovirus control.}, journal = {The Lancet. Microbe}, volume = {5}, number = {7}, pages = {622-623}, doi = {10.1016/S2666-5247(24)00072-7}, pmid = {38642566}, issn = {2666-5247}, mesh = {*Wolbachia/genetics ; Animals ; *Arboviruses ; *Arbovirus Infections/transmission/prevention & control ; Humans ; Mosquito Vectors/microbiology/virology ; }, } @article {pmid38633519, year = {2024}, author = {Wei, X and Zhu, J and Hoffmann, AA and Jia, J and Xiao, M and Duan, F and Zhang, Y and Zhong, H and Ge, J and Yu, W and Zhang, L and Jiang, W}, title = {Wolbachia infection status and molecular diversity in the species of tribe Tagiadini Mabille, 1878 (Lepidoptera: Hesperiidae) collected in China.}, journal = {Ecology and evolution}, volume = {14}, number = {4}, pages = {e11279}, pmid = {38633519}, issn = {2045-7758}, abstract = {Wolbachia, one of the most ubiquitous heritable symbionts in lepidopteran insects, can cause mitochondrial introgression in related host species. We recently found mito-nuclear discordance in the Lepidopteran tribe Tagiadini Mabille 1878 from which Wolbachia has not been reported. In this study, we found that 13 of the 46 species of Tagiadini species tested were positive for Wolbachia. Overall, 14% (15/110) of Tagiadini specimens were infected with Wolbachia and nine new STs were found from 15 isolates. A co-phylogenetic comparison, divergence time estimation and Wolbachia recombination analysis revealed that mito-nuclear discordance in Tagiadini species is not mediated by Wolbachia, but Wolbachia acquisition in Tagiadini appears to have occurred mainly through horizontal transmission rather than codivergence.}, } @article {pmid38632506, year = {2024}, author = {Alkathiry, HA and Alghamdi, SQ and Sinha, A and Margos, G and Stekolnikov, AA and Alagaili, AN and Darby, AC and Makepeace, BL and Khoo, JJ}, title = {Microbiome and mitogenomics of the chigger mite Pentidionis agamae: potential role as an Orientia vector and associations with divergent clades of Wolbachia and Borrelia.}, journal = {BMC genomics}, volume = {25}, number = {1}, pages = {380}, pmid = {38632506}, issn = {1471-2164}, mesh = {Animals ; *Borrelia/genetics ; DNA ; *Microbiota ; Multilocus Sequence Typing ; Orientia ; *Orientia tsutsugamushi/genetics ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Rodentia/genetics ; Saudi Arabia ; *Scrub Typhus/epidemiology/microbiology ; *Trombiculidae/genetics/microbiology ; *Wolbachia/genetics ; }, abstract = {BACKGROUND: Trombiculid mites are globally distributed, highly diverse arachnids that largely lack molecular resources such as whole mitogenomes for the elucidation of taxonomic relationships. Trombiculid larvae (chiggers) parasitise vertebrates and can transmit bacteria (Orientia spp.) responsible for scrub typhus, a zoonotic febrile illness. Orientia tsutsugamushi causes most cases of scrub typhus and is endemic to the Asia-Pacific Region, where it is transmitted by Leptotrombidium spp. chiggers. However, in Dubai, Candidatus Orientia chuto was isolated from a case of scrub typhus and is also known to circulate among rodents in Saudi Arabia and Kenya, although its vectors remain poorly defined. In addition to Orientia, chiggers are often infected with other potential pathogens or arthropod-specific endosymbionts, but their significance for trombiculid biology and public health is unclear.

RESULTS: Ten chigger species were collected from rodents in southwestern Saudi Arabia. Chiggers were pooled according to species and screened for Orientia DNA by PCR. Two species (Microtrombicula muhaylensis and Pentidionis agamae) produced positive results for the htrA gene, although Ca. Orientia chuto DNA was confirmed by Sanger sequencing only in P. agamae. Metagenomic sequencing of three pools of P. agamae provided evidence for two other bacterial associates: a spirochaete and a Wolbachia symbiont. Phylogenetic analysis of 16S rRNA and multi-locus sequence typing genes placed the spirochaete in a clade of micromammal-associated Borrelia spp. that are widely-distributed globally with no known vector. For the Wolbachia symbiont, a genome assembly was obtained that allowed phylogenetic localisation in a novel, divergent clade. Cytochrome c oxidase I (COI) barcodes for Saudi Arabian chiggers enabled comparisons with global chigger diversity, revealing several cases of discordance with classical taxonomy. Complete mitogenome assemblies were obtained for the three P. agamae pools and almost 50 SNPs were identified, despite a common geographic origin.

CONCLUSIONS: P. agamae was identified as a potential vector of Ca. Orientia chuto on the Arabian Peninsula. The detection of an unusual Borrelia sp. and a divergent Wolbachia symbiont in P. agamae indicated links with chigger microbiomes in other parts of the world, while COI barcoding and mitogenomic analyses greatly extended our understanding of inter- and intraspecific relationships in trombiculid mites.}, } @article {pmid38631561, year = {2024}, author = {Orozco-Gonzales, JL and Dos Santos Benedito, A and Cardona-Salgado, D and Ferreira, CP and de Oliveira Florentino, H and Sepulveda-Salcedo, LS and Vasilieva, O}, title = {Comparing the long-term persistence of different Wolbachia strains after the release of bacteria-carrying mosquitoes.}, journal = {Mathematical biosciences}, volume = {372}, number = {}, pages = {109190}, doi = {10.1016/j.mbs.2024.109190}, pmid = {38631561}, issn = {1879-3134}, mesh = {*Wolbachia/physiology/pathogenicity ; Animals ; *Mosquito Vectors/microbiology ; Culicidae/microbiology ; Models, Biological ; Mathematical Concepts ; }, abstract = {This paper proposes a bidimensional modeling framework for Wolbachia invasion, assuming imperfect maternal transmission, incomplete cytoplasmic incompatibility, and direct infection loss due to thermal stress. Our model adapts to various Wolbachia strains and retains all properties of higher-dimensional models. The conditions for the durable coexistence of Wolbachia-carrying and wild mosquitoes are expressed using the model's parameters in a compact closed form. When the Wolbachia bacterium is locally established, the size of the remanent wild population can be assessed by a direct formula derived from the model. The model was tested for four Wolbachia strains undergoing laboratory and field trials to control mosquito-borne diseases: wMel, wMelPop, wAlbB, and wAu. As all these bacterial strains affect the individual fitness of mosquito hosts differently and exhibit different levels of resistance to temperature variations, the model helped to conclude that: (1) the wMel strain spreads faster in wild mosquito populations; (2) the wMelPop exhibits lower resilience but also guarantees the smallest size of the remanent wild population; (3) the wAlbB strain performs better at higher ambient temperatures than others; (4) the wAu strain is not sustainable and cannot persist in the wild mosquito population despite its resistance to high temperatures.}, } @article {pmid38630673, year = {2024}, author = {Durrance-Bagale, A and Hoe, N and Lai, J and Liew, JWK and Clapham, H and Howard, N}, title = {Dengue vector control in high-income, city settings: A scoping review of approaches and methods.}, journal = {PLoS neglected tropical diseases}, volume = {18}, number = {4}, pages = {e0012081}, pmid = {38630673}, issn = {1935-2735}, mesh = {Adult ; Animals ; Humans ; *Dengue Virus ; *Dengue/epidemiology ; Mosquito Vectors ; Mosquito Control/methods ; Cities ; *Aedes ; }, abstract = {BACKGROUND: Dengue virus (DENV) is endemic to many parts of the world and has serious health and socioeconomic effects even in high-income countries, especially with rapid changes in the climate globally. We explored the literature on dengue vector control methods used in high-income, city settings and associations with dengue incidence, dengue prevalence, or mosquito vector densities.

METHODS: Studies of any design or year were included if they reported effects on human DENV infection or Aedes vector indices of dengue-specific vector control interventions in high-income, city settings.

RESULTS: Of 24 eligible sources, most reported research in the United States (n = 8) or Australia (n = 5). Biocontrol (n = 12) and chemical control (n = 13) were the most frequently discussed vector control methods. Only 6 sources reported data on the effectiveness of a given method in reducing human DENV incidence or prevalence, 2 described effects of larval and adult control on Aedes DENV positivity, 20 reported effectiveness in reducing vector density, using insecticide, larvicide, source reduction, auto-dissemination of pyriproxyfen and Wolbachia, and only 1 described effects on human-vector contact.

CONCLUSIONS: As most studies reported reductions in vector densities, rather than any effects on human DENV incidence or prevalence, we can draw no clear conclusions on which interventions might be most effective in reducing dengue in high-income, city areas. More research is needed linking evidence on the effects of different DENV vector control methods with dengue incidence/prevalence or mosquito vector densities in high-income, city settings as this is likely to differ from low-income settings. This is a significant evidence gap as climate changes increase the global reach of DENV. The importance of community involvement was clear in several studies, although it is impossible to tease out the relative contributions of this from other control methods used.}, } @article {pmid38628128, year = {2024}, author = {Scussel, S and Gaudillat, B and Esnault, J and Lejarre, Q and Duployer, M and Lebon, C and Benlali, A and Mavingui, P and Tortosa, P and Cattel, J}, title = {Combining transinfected Wolbachia and a genetic sexing strain to control Aedes albopictus in laboratory-controlled conditions.}, journal = {Proceedings. Biological sciences}, volume = {291}, number = {2021}, pages = {20240429}, pmid = {38628128}, issn = {1471-2954}, mesh = {Animals ; Female ; Male ; *Wolbachia/genetics ; Artificial Intelligence ; *Aedes/genetics ; *Culex ; }, abstract = {The global expansion of Aedes albopictus has stimulated the development of environmentally friendly methods aiming to control disease transmission through the suppression of natural vector populations. Sterile male release programmes are currently being deployed worldwide, and are challenged by the availability of an efficient sex separation which can be achieved mechanically at the pupal stage and/or by artificial intelligence at the adult stage, or through genetic sexing, which allows separating males and females at an early development stage. In this study, we combined the genetic sexing strain previously established based on the linkage of dieldrin resistance to the male locus with a Wolbachia transinfected line. For this, we introduced either the wPip-I or the wPip-IV strain from Culex pipiens in an asymbiotic Wolbachia-free Ae. albopictus line. We then measured the penetrance of cytoplasmic incompatibility and life-history traits of both transinfected lines, selected the wPip-IV line and combined it with the genetic sexing strain. Population suppression experiments demonstrated a 90% reduction in population size and a 50% decrease in hatching rate. Presented results showed that such a combination has a high potential in terms of vector control but also highlighted associated fitness costs, which should be reduced before large-scale field assay.}, } @article {pmid38609398, year = {2024}, author = {Mowery, MA and Rosenwald, LC and Chapman, E and Lubin, Y and Segoli, M and Khoza, T and Lyle, R and White, JA}, title = {Endosymbiont diversity across native and invasive brown widow spider populations.}, journal = {Scientific reports}, volume = {14}, number = {1}, pages = {8556}, pmid = {38609398}, issn = {2045-2322}, support = {1953223//National Science Foundation/ ; 1020740//National Institute of Food and Agriculture/ ; }, mesh = {Humans ; Adult ; Animals ; Female ; *Animals, Poisonous ; *Chlamydiales ; Eggs ; *Spiders ; *Wolbachia ; }, abstract = {The invasive brown widow spider, Latrodectus geometricus (Araneae: Theridiidae), has spread in multiple locations around the world and, along with it, brought associated organisms such as endosymbionts. We investigated endosymbiont diversity and prevalence across putative native and invasive populations of this spider, predicting lower endosymbiont diversity across the invasive range compared to the native range. First, we characterized the microbial community in the putative native (South Africa) and invasive (Israel and the United States) ranges via high throughput 16S sequencing of 103 adult females. All specimens were dominated by reads from only 1-3 amplicon sequence variants (ASV), and most individuals were infected with an apparently uniform strain of Rhabdochlamydia. We also found Rhabdochlamydia in spider eggs, indicating that it is a maternally-inherited endosymbiont. Relatively few other ASV were detected, but included two variant Rhabdochlamydia strains and several Wolbachia, Spiroplasma and Enterobacteriaceae strains. We then diagnostically screened 118 adult female spiders from native and invasive populations specifically for Rhabdochlamydia and Wolbachia. We found Rhabdochlamydia in 86% of individuals and represented in all populations, which suggests that it is a consistent and potentially important associate of L. geometricus. Wolbachia was found at lower overall prevalence (14%) and was represented in all countries, but not all populations. In addition, we found evidence for geographic variation in endosymbiont prevalence: spiders from Israel were more likely to carry Rhabdochlamydia than those from the US and South Africa, and Wolbachia was geographically clustered in both Israel and South Africa. Characterizing endosymbiont prevalence and diversity is a first step in understanding their function inside the host and may shed light on the process of spread and population variability in cosmopolitan invasive species.}, } @article {pmid38608996, year = {2024}, author = {Akintola, AA and Hwang, UW}, title = {Microbiome profile of South Korean vector mosquitoes.}, journal = {Acta tropica}, volume = {255}, number = {}, pages = {107213}, doi = {10.1016/j.actatropica.2024.107213}, pmid = {38608996}, issn = {1873-6254}, mesh = {Animals ; Republic of Korea ; *RNA, Ribosomal, 16S/genetics ; *Aedes/microbiology ; *Mosquito Vectors/microbiology ; *Bacteria/classification/genetics/isolation & purification ; Female ; *Culex/microbiology ; Anopheles/microbiology ; Gastrointestinal Microbiome ; Microbiota/genetics ; DNA, Bacterial/genetics ; Phylogeny ; Sequence Analysis, DNA ; Wolbachia/genetics/isolation & purification/classification ; High-Throughput Nucleotide Sequencing ; }, abstract = {This research offers a comprehensive exploration of the microbial communities associated with vector mosquitoes from South Korea. Aedes albopictus, Anopheles sinensis, and Culex molestus are vectors of pathogens, and understanding the intricacies of their microbiome profile is paramount for unraveling their roles in disease transmission dynamics. In this study, we characterized the microbiome of the midguts of adult female vector mosquitoes collected from different locations in South Korea. After DNA extraction from dissected mosquito midguts, we used the Illumina MiSeq next-generation sequencing to obtain sequences spanning the V4 hypervariable region of the bacteria 16S rRNA. Morphological and molecular characterization using 506-bp mitochondrial 16S rRNA was used to identify the mosquito species before amplicon sequencing. Across the three vector mosquitoes surveyed, 21 bacteria genera belonging to 20 families and 5 phyla were discovered. Proteobacteria and Bacteriodota were the major phyla of bacteria associated with the three mosquito species. There were significant differences in the gut microbiome genera composition between the species and little variation in the gut microbiome between individuals of the same mosquito species. Wolbachia is the most dominant genus in Aedes while Aeromonas, Acinetobacter, and unassigned taxa are the most common in An. sinensis. In addition to that, Chromobacterium, Chryseobacterium, and Aeromonas are dominant in Cx. molestus. This study sheds light on the complex interactions between mosquitoes and their microbiome, revealing potential implications for vector competence, disease transmission, and vector control strategies.}, } @article {pmid38604685, year = {2024}, author = {Lei, J and Lü, W and Wang, W and Wang, H and Guo, X and Cheng, P and Gong, M and Liu, L}, title = {[Comparison of the microbiota diversity between autogenous and anautogenous Culex pipiens pallens].}, journal = {Zhongguo xue xi chong bing fang zhi za zhi = Chinese journal of schistosomiasis control}, volume = {36}, number = {1}, pages = {52-58}, doi = {10.16250/j.32.1374.2023204}, pmid = {38604685}, issn = {1005-6661}, support = {81902096//National Natural Science Foundation of China/ ; 81871685//National Natural Science Foundation of China/ ; //Medical Science and Technology Innovation Project of Shandong Academy of Medical Sciences/ ; }, mesh = {Humans ; Animals ; RNA, Ribosomal, 16S/genetics ; *Culex/genetics ; *Culicidae/genetics ; *Microbiota/genetics ; }, abstract = {OBJECTIVE: To investigate the microbiota composition and diversity between autogenous and anautogenous Culex pipiens pallens, so as to provide insights into unraveling the pathogenesis of autogeny in Cx. pipiens pallens.

METHODS: Autogenous and anautogenous adult Cx. pipiens pallens samples were collected at 25 ℃, and the hypervariable regions of the microbial 16S ribosomal RNA (16S rRNA) gene was sequenced on the Illumina NovaSeq 6000 sequencing platform. The microbiota abundance and diversity were evaluated using the alpha diversity index, and the difference in the microbiota structure was examined using the beta diversity index. The microbiota with significant differences in the abundance between autogenous and anautogenous adult Cx. pipiens pallens samples was identified using the linear discriminant analysis effect size (LEfSe).

RESULTS: The microbiota in autogenous and anautogenous Cx. pipiens pallens samples belonged to 18 phyla, 28 classes, 70 orders, 113 families, and 170 genera, and the dominant phyla included Proteobacteria, Bacteroidetes, and so on. At the genus level, Wolbachia was a common dominant genus, and the relative abundance was (77.6 ± 11.3)% in autogenous Cx. pipiens pallens samples and (47.5 ± 8.5)% in anautogenous mosquito samples, while Faecalibaculum (0.4% ± 0.1%), Dubosiella (0.5% ± 0.0%) and Massilia (0.5% ± 0.1%) were specific species in autogenous Cx. pipiens pallens samples. Alpha diversity analysis showed that higher Chao1 index and ACE index in autogenous Cx. pipiens pallens samples than in anautogenous samples (both P values > 0.05), and lower Shannon index (P > 0.05) and Simpson index (P < 0.05) in autogenous Cx. pipiens pallens samples than in anautogenous samples. LEfSe analysis showed a total of 48 significantly different taxa between autogenous and anautogenous Cx. pipiens pallens samples (all P values < 0.05).

CONCLUSIONS: There is a significant difference in the microbiota diversity between autogenous and anautogenous Cx. pipiens pallens.}, } @article {pmid38604562, year = {2024}, author = {Song, G and Shin, D and Kim, JS}, title = {Microbiome changes in Akanthomyces attenuatus JEF-147-infected two-spotted spider mites.}, journal = {Journal of invertebrate pathology}, volume = {204}, number = {}, pages = {108102}, doi = {10.1016/j.jip.2024.108102}, pmid = {38604562}, issn = {1096-0805}, mesh = {Animals ; *Tetranychidae/microbiology/physiology ; *Microbiota ; Female ; Pest Control, Biological ; }, abstract = {The two-spotted spider mite (Tetranychus urticae Koch) is an agriculturally serious polyphagous pest that has acquired strong resistance against acaricides because of its short life cycle and continuous exposure to acaricides. As an alternative, mite-pathogenic fungi with different modes of action could be used to control the mites. The spider mite has symbiotic microorganisms that could be involved in the physiological and ecological adaptations to biotic stresses. In this study, mite-pathogenic fungi were used to control female adults, and the microbiomes changes in the fungus-infected mites were analyzed. The acaricidal activity of 77 fungal isolates was tested, and Akanthomyces attenuatus JEF-147 exhibited the highest acaricidal activity. Subsequently a dose-response assay and morphological characterization was undertaken For microbiome analysis in female adults infected with A. attenuatus JEF-147, 16S rDNA and ITS1 were sequenced using Illumina Miseq. Infected mite showed a higher Shannon index in bacterial diversity but lower index in fungal diversity. In beta diversity using principal component analysis, JEF-147-treated mites were significantly different from non-treated controls in both bacteria and fungi. Particularly in bacterial abundance, arthropod defense-related Rickettsia increased, but arthropod reproduction-associated Wolbachia decreased. The change in major bacterial abundance in the infected mites could be explained by a trade-off between reproduction and immunity against the early stage of fungal attack. In fungal abundance, Akanthomyces showed up as expected. Foremost, this work reports microbiome changes in a fungus-infected mite and suggests a possible trade-off in mites against fungal pathogens. Future studies will focus on gene-based investigations related to this topic.}, } @article {pmid38601925, year = {2024}, author = {Gomard, Y and Hafsia, S and Lebon, C and Rabarison, P and Idaroussi, AB and Yssouf, A and Boussès, P and Mavingui, P and Atyame, C}, title = {Genetic diversity of endosymbiotic bacteria Wolbachia infecting two mosquito species of the genus Eretmapodites occurring in sympatry in the Comoros archipelago.}, journal = {Frontiers in microbiology}, volume = {15}, number = {}, pages = {1343917}, pmid = {38601925}, issn = {1664-302X}, abstract = {INTRODUCTION: The influence of Wolbachia on mosquito reproduction and vector competence has led to renewed interest in studying the genetic diversity of these bacteria and the phenotypes they induced in mosquito vectors. In this study, we focused on two species of Eretmapodites, namely Eretmapodites quinquevittatus and Eretmapodites subsimplicipes, from three islands in the Comoros archipelago (in the Southwestern Indian Ocean).

METHODS: Using the COI gene, we examined the mitochondrial genetic diversity of 879 Eretmapodites individuals from 54 sites. Additionally, we investigated the presence and genetic diversity of Wolbachia using the wsp marker and the diversity of five housekeeping genes commonly used for genotyping through Multiple Locus Sequence Typing (MLST).

RESULTS AND DISCUSSION: Overall, Er. quinquevittatus was the most abundant species in the three surveyed islands and both mosquito species occurred in sympatry in most of the investigated sites. We detected a higher mitochondrial genetic diversity in Er. quinquevittatus with 35 reported haplotypes (N = 615 specimens, Hd = 0.481 and π = 0.002) while 13 haplotypes were found in Er. subsimplicipes (N = 205 specimens, Hd = 0.338 and π = 0.001), this difference is likely due to the bias in sampling size between the two species. We report for the first time the presence of Wolbachia in these two Eretmapodites species. The prevalence of Wolbachia infection varied significantly between species, with a low prevalence recorded in Er. quinquevittatus (0.8%, N = 5/627) while infection was close to fixation in Er. subsimplicipes (87.7%, N = 221/252). Both male and female individuals of the two mosquito species appeared to be infected. The analysis of MLST genes revealed the presence of two Wolbachia strains corresponding to two new strain types (STs) within the supergroups A and B, which have been named wEretA and wEretB. These strains were found as mono-infections and are closely related, phylogenetically, to Wolbachia strains previously reported in Drosophila species. Finally, we demonstrate that maternal transmission of Wolbachia is imperfect in Er. subsimplicipes, which could explain the presence of a minority of uninfected individuals in the field.}, } @article {pmid38598552, year = {2024}, author = {Vandana, V and Dong, S and Sheth, T and Sun, Q and Wen, H and Maldonado, A and Xi, Z and Dimopoulos, G}, title = {Wolbachia infection-responsive immune genes suppress Plasmodium falciparum infection in Anopheles stephensi.}, journal = {PLoS pathogens}, volume = {20}, number = {4}, pages = {e1012145}, pmid = {38598552}, issn = {1553-7374}, support = {R01 AI158615/AI/NIAID NIH HHS/United States ; R01 AI182096/AI/NIAID NIH HHS/United States ; R21 AI168873/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; *Anopheles/parasitology/microbiology/immunology ; *Wolbachia/immunology ; *Plasmodium falciparum/immunology ; *Malaria, Falciparum/immunology/parasitology ; Mosquito Vectors/parasitology/microbiology/immunology ; Insect Proteins/genetics/metabolism/immunology ; Transcriptome ; Female ; }, abstract = {Wolbachia, a maternally transmitted symbiotic bacterium of insects, can suppress a variety of human pathogens in mosquitoes, including malaria-causing Plasmodium in the Anopheles vector. However, the mechanistic basis of Wolbachia-mediated Plasmodium suppression in mosquitoes is not well understood. In this study, we compared the midgut and carcass transcriptomes of stably infected Anopheles stephensi with Wolbachia wAlbB to uninfected mosquitoes in order to discover Wolbachia infection-responsive immune genes that may play a role in Wolbachia-mediated anti-Plasmodium activity. We show that wAlbB infection upregulates 10 putative immune genes and downregulates 14 in midguts, while it upregulates 31 putative immune genes and downregulates 15 in carcasses at 24 h after blood-fed feeding, the time at which the Plasmodium ookinetes are traversing the midgut tissue. Only a few of these regulated immune genes were also significantly differentially expressed between Wolbachia-infected and non-infected midguts and carcasses of sugar-fed mosquitoes. Silencing of the Wolbachia infection-responsive immune genes TEP 4, TEP 15, lysozyme C2, CLIPB2, CLIPB4, PGRP-LD and two novel genes (a peritrophin-44-like gene and a macro domain-encoding gene) resulted in a significantly greater permissiveness to P. falciparum infection. These results indicate that Wolbachia infection modulates mosquito immunity and other processes that are likely to decrease Anopheles permissiveness to Plasmodium infection.}, } @article {pmid38597256, year = {2024}, author = {Fox, T and Sguassero, Y and Chaplin, M and Rose, W and Doum, D and Arevalo-Rodriguez, I and Villanueva, G}, title = {Wolbachia-carrying Aedes mosquitoes for preventing dengue infection.}, journal = {The Cochrane database of systematic reviews}, volume = {4}, number = {4}, pages = {CD015636}, pmid = {38597256}, issn = {1469-493X}, mesh = {Animals ; Humans ; *Aedes/microbiology ; *Wolbachia ; *Dengue Virus ; Mosquito Vectors/microbiology ; *Dengue/prevention & control ; }, abstract = {BACKGROUND: Dengue is a global health problem of high significance, with 3.9 billion people at risk of infection. The geographic expansion of dengue virus (DENV) infection has resulted in increased frequency and severity of the disease, and the number of deaths has increased in recent years. Wolbachia,an intracellular bacterial endosymbiont, has been under investigation for several years as a novel dengue-control strategy. Some dengue vectors (Aedes mosquitoes) can be transinfected with specific strains of Wolbachia, which decreases their fitness (ability to survive and mate) and their ability to reproduce, inhibiting the replication of dengue. Both laboratory and field studies have demonstrated the potential effect of Wolbachia deployments on reducing dengue transmission, and modelling studies have suggested that this may be a self-sustaining strategy for dengue prevention, although long-term effects are yet to be elucidated.

OBJECTIVES: To assess the efficacy of Wolbachia-carrying Aedes speciesdeployments (specifically wMel-, wMelPop-, and wAlbB- strains of Wolbachia) for preventing dengue virus infection.

SEARCH METHODS: We searched CENTRAL, MEDLINE, Embase, four other databases, and two trial registries up to 24 January 2024.

SELECTION CRITERIA: Randomized controlled trials (RCTs), including cluster-randomized controlled trials (cRCTs), conducted in dengue endemic or epidemic-prone settings were eligible. We sought studies that investigated the impact of Wolbachia-carrying Aedes deployments on epidemiological or entomological dengue-related outcomes, utilizing either the population replacement or population suppression strategy.

DATA COLLECTION AND ANALYSIS: Two review authors independently selected eligible studies, extracted data, and assessed the risk of bias using the Cochrane RoB 2 tool. We used odds ratios (OR) with the corresponding 95% confidence intervals (CI) as the effect measure for dichotomous outcomes. For count/rate outcomes, we planned to use the rate ratio with 95% CI as the effect measure. We used adjusted measures of effect for cRCTs. We assessed the certainty of evidence using GRADE.

MAIN RESULTS: One completed cRCT met our inclusion criteria, and we identified two further ongoing cRCTs. The included trial was conducted in an urban setting in Yogyakarta, Indonesia. It utilized a nested test-negative study design, whereby all participants aged three to 45 years who presented at healthcare centres with a fever were enrolled in the study provided they had resided in the study area for the previous 10 nights. The trial showed that wMel-Wolbachia infected Ae aegypti deployments probably reduce the odds of contracting virologically confirmed dengue by 77% (OR 0.23, 95% CI 0.15 to 0.35; 1 trial, 6306 participants; moderate-certainty evidence). The cluster-level prevalence of wMel Wolbachia-carrying mosquitoes remained high over two years in the intervention arm of the trial, reported as 95.8% (interquartile range 91.5 to 97.8) across 27 months in clusters receiving wMel-Wolbachia Ae aegypti deployments, but there were no reliable comparative data for this outcome. Other primary outcomes were the incidence of virologically confirmed dengue, the prevalence of dengue ribonucleic acid in the mosquito population, and mosquito density, but there were no data for these outcomes. Additionally, there were no data on adverse events.

AUTHORS' CONCLUSIONS: The included trial demonstrates the potential significant impact of wMel-Wolbachia-carrying Ae aegypti mosquitoes on preventing dengue infection in an endemic setting, and supports evidence reported in non-randomized and uncontrolled studies. Further trials across a greater diversity of settings are required to confirm whether these findings apply to other locations and country settings, and greater reporting of acceptability and cost are important.}, } @article {pmid38594795, year = {2024}, author = {Machado, FN and Draper, ADK and Monteiro Fernandes, A and Dos Santos, FBA and Mali, MA and Pereira Tilman, AJ and Soares da Silva, E and Hornay, E and Salles de Sousa, A and Oakley, T and Cruz, ED and Sarmento, N and Niha, MAV and Soares, AF and Cardoso Gomes, EE and de Deus Alves, J and Soares, JP and Francis, JR and Yan, J and Monteiro, MA}, title = {The first confirmed outbreak of chikungunya reported in Timor-Leste, 2024.}, journal = {Communicable diseases intelligence (2018)}, volume = {48}, number = {}, pages = {}, doi = {10.33321/cdi.2024.48.17}, pmid = {38594795}, issn = {2209-6051}, support = {001/WHO_/World Health Organization/International ; }, mesh = {Male ; Animals ; Humans ; Infant ; Child, Preschool ; Child ; Adolescent ; Young Adult ; Adult ; Middle Aged ; Aged ; Female ; *Chikungunya Fever/epidemiology ; Timor-Leste/epidemiology ; Australia/epidemiology ; *Chikungunya virus/genetics ; *Zika Virus ; Disease Outbreaks ; *Zika Virus Infection/epidemiology/prevention & control ; }, abstract = {Timor-Leste is a mountainous, half-island nation with a population of 1.3 million, which shares a land border with Indonesia and is 550 km from Darwin, Australia. Since independence in 2002, Timor-Leste has achieved significant development; however, high levels of poverty remain. Chikungunya virus (CHIKV) is endemic in over 100 countries in Africa, Asia, Europe and in the Americas. It is transmitted by the bite of infected Aedes aegypti or Ae. albopictus mosquitoes, which are present in Timor-Leste and which contribute to annual rainy-season dengue virus (DENV) outbreaks. Symptomatic people typically suffer from acute onset of fever, usually accompanied by severe arthritis or arthralgia. Joint pain can be debilitating for several days, and may sometimes last for weeks, months or years. Unlike DENV infection which has significant mortality, most people recover completely. Between 2002 and 2023, there were 26 cases of CHIKV notified in Australia who acquired their infection in Timor-Leste; however, laboratory testing capability for CHIKV in Timor-Leste only became available in 2021 using polymerase chain reaction (PCR). The first locally diagnosed case was notified in November 2023. In January 2024, an outbreak of CHIKV was recognised in Timor-Leste for the first time, with 195 outbreak cases reported during 1-31 January 2024; all were PCR positive. There were no cases hospitalised, and no deaths. The median age of cases was 17 years (range 1-76 years); 51% were males. Cases were reported across the country; most (88/195) were from Dili, although the highest incidence was seen in the neighbouring municipality of Ermera (monthly incidence rate of 58.8 cases per 100,000 population). This first reported outbreak of CHIKV in Timor-Leste highlights the need for improved mosquito-borne illness control and response strategies, including minimising breeding sites and promoting early presentation for treatment and differential diagnosis from DENV, and consideration of the deployment of Wolbachia-infected mosquitoes, particularly as they have shown to reduce the transmission of CHIKV, DENV and Zika virus, all of which pose threats in Timor-Leste.}, } @article {pmid38594794, year = {2024}, author = {Machado, FN and Draper, ADK and Dos Santos, FBA and Mali, MA and Pereira Tilman, AJ and Soares da Silva, E and Soares, NG and Sarmento, N and Niha, MAV and Soares, AF and Taal, A and Francis, JR and Yan, J and Miller, M and Flint, J}, title = {A brief description of the epidemiology of dengue in Dili, Timor-Leste, 2018-2022.}, journal = {Communicable diseases intelligence (2018)}, volume = {48}, number = {}, pages = {}, doi = {10.33321/cdi.2024.48.16}, pmid = {38594794}, issn = {2209-6051}, support = {001/WHO_/World Health Organization/International ; }, mesh = {Animals ; Humans ; *Dengue Virus ; Timor-Leste/epidemiology ; Pandemics ; Australia/epidemiology ; *Dengue/epidemiology ; }, abstract = {Dengue virus (DENV) infection causes 390 million infections per year and 40,000 deaths globally. It is endemic in many countries in Asia, Africa, the Americas, the Caribbean, and Oceania. Dengue is endemic in Timor-Leste year-round, but peak transmission occurs during the rainy season. We briefly describe the epidemiology of DENV in the Municipality of Dili between 2018 and 2022. There were 6,234 cases notified, with a mean annual incidence rate of 330 cases per 100,000 population. There were 55 deaths (case fatality rate 0.9%). The peak annual incidence (3,904 cases) occurred in 2022 after an outbreak was declared in January of that year; this outbreak included 760 cases of dengue haemorrhagic fever and 35 deaths. The number of outbreak cases requiring hospital treatment exceeded the usual capacity, but facilities established for coronavirus disease 2019 (COVID-19) isolation and treatment were repurposed to meet this demand. Existing strategies of vector control, minimising breeding sites and promoting early presentation for treatment should continue, as should the utilisation of surveillance systems and treatment facilities established during the COVID-19 pandemic. However, dengue incidence remains high, and other dengue control strategies-including the deployment of Wolbachia-infected mosquitoes-should be considered in Timor-Leste.}, } @article {pmid38591028, year = {2024}, author = {Sharmin, Z and Samarah, H and Aldaya Bourricaudy, R and Ochoa, L and Serbus, LR}, title = {Cross-validation of chemical and genetic disruption approaches to inform host cellular effects on Wolbachia abundance in Drosophila.}, journal = {Frontiers in microbiology}, volume = {15}, number = {}, pages = {1364009}, pmid = {38591028}, issn = {1664-302X}, abstract = {INTRODUCTION: Endosymbiotic Wolbachia bacteria are widespread in nature, present in half of all insect species. The success of Wolbachia is supported by a commensal lifestyle. Unlike bacterial pathogens that overreplicate and harm host cells, Wolbachia infections have a relatively innocuous intracellular lifestyle. This raises important questions about how Wolbachia infection is regulated. Little is known about how Wolbachia abundance is controlled at an organismal scale.

METHODS: This study demonstrates methodology for rigorous identification of cellular processes that affect whole-body Wolbachia abundance, as indicated by absolute counts of the Wolbachia surface protein (wsp) gene.

RESULTS: Candidate pathways, associated with well-described infection scenarios, were identified. Wolbachia-infected fruit flies were exposed to small molecule inhibitors known for targeting those same pathways. Sequential tests in D. melanogaster and D. simulans yielded a subset of chemical inhibitors that significantly affected whole-body Wolbachia abundance, including the Wnt pathway disruptor, IWR-1 and the mTOR pathway inhibitor, Rapamycin. The implicated pathways were genetically retested for effects in D. melanogaster, using inducible RNAi expression driven by constitutive as well as chemically-induced somatic GAL4 expression. Genetic disruptions of armadillo, tor, and ATG6 significantly affected whole-body Wolbachia abundance.

DISCUSSION: As such, the data corroborate reagent targeting and pathway relevance to whole-body Wolbachia infection. The results also implicate Wnt and mTOR regulation of autophagy as important for regulation of Wolbachia titer.}, } @article {pmid38588812, year = {2024}, author = {Nascimento da Silva, J and Conceição, CC and Ramos de Brito, GC and Renato de Oliveira Daumas Filho, C and Walter Nuno, AB and Talyuli, OAC and Arcanjo, A and de Oliveira, PL and Moreira, LA and Vaz, IDS and Logullo, C}, title = {Immunometabolic crosstalk in Aedes fluviatilis and Wolbachia pipientis symbiosis.}, journal = {The Journal of biological chemistry}, volume = {300}, number = {6}, pages = {107272}, pmid = {38588812}, issn = {1083-351X}, mesh = {*Wolbachia/physiology/metabolism ; *Symbiosis ; *Aedes/microbiology/immunology/metabolism ; Animals ; Glycogen Synthase Kinase 3 beta/metabolism/genetics ; Insect Proteins/metabolism/genetics ; Glycogen/metabolism ; }, abstract = {Wolbachia pipientis is a maternally transmitted symbiotic bacterium that mainly colonizes arthropods, potentially affecting different aspects of the host's physiology, e.g., reproduction, immunity, and metabolism. It has been shown that Wolbachia modulates glycogen metabolism in mosquito Aedes fluviatilis (Ae. fluviatilis). Glycogen synthesis is controlled by the enzyme GSK3, which is also involved in immune responses in both vertebrate and invertebrate organisms. Here we investigated the mechanisms behind immune changes mediated by glycogen synthase kinase β (GSK3β) in the symbiosis between Ae. fluviatilis and W. pipientis using a GSK3β inhibitor or RNAi-mediated gene silencing. GSK3β inhibition or knockdown increased glycogen content and Wolbachia population, together with a reduction in Relish2 and gambicin transcripts. Furthermore, knockdown of Relish2 or Caspar revealed that the immunodeficiency pathway acts to control Wolbachia numbers in the host. In conclusion, we describe for the first time the involvement of GSK3β in Ae. fluviatilis immune response, acting to control the Wolbachia endosymbiotic population.}, } @article {pmid38585949, year = {2024}, author = {Mirchandani, C and Wang, P and Jacobs, J and Genetti, M and Pepper-Tunick, E and Sullivan, WT and Corbett-Detig, R and Russell, SL}, title = {Mixed Wolbachia infections resolve rapidly during in vitro evolution.}, journal = {bioRxiv : the preprint server for biology}, volume = {}, number = {}, pages = {}, pmid = {38585949}, issn = {2692-8205}, support = {R00 GM135583/GM/NIGMS NIH HHS/United States ; R35 GM128932/GM/NIGMS NIH HHS/United States ; R35 GM139595/GM/NIGMS NIH HHS/United States ; T32 HG012344/HG/NHGRI NIH HHS/United States ; }, abstract = {The intracellular symbiont Wolbachia pipientis evolved after the divergence of arthropods and nematodes, but it reached high prevalence in many of these taxa through its abilities to infect new hosts and their germlines. Some strains exhibit long-term patterns of co-evolution with their hosts, while other strains are capable of switching hosts. This makes strain selection an important factor in symbiont-based biological control. However, little is known about the ecological and evolutionary interactions that occur when a promiscuous strain colonizes an infected host. Here, we study what occurs when two strains come into contact in host cells following horizontal transmission and infection. We focus on the faithful wMel strain from Drosophila melanogaster and the promiscuous wRi strain from Drosophila simulans using an in vitro cell culture system with multiple host cell types and combinatorial infection states. Mixing D. melanogaster cell lines stably infected with wMel and wRi revealed that wMel outcompetes wRi quickly and reproducibly. Furthermore, wMel was able to competitively exclude wRi even from minuscule starting quantities, indicating that this is a nearly deterministic outcome, independent of the starting infection frequency. This competitive advantage was not exclusive to wMel's native D. melanogaster cell background, as wMel also outgrew wRi in D. simulans cells. Overall, wRi is less adept at in vitro growth and survival than wMel and its in vivo state, revealing differences between cellular and humoral regulation. These attributes may underlie the observed low rate of mixed infections in nature and the relatively rare rate of host-switching in most strains. Our in vitro experimental framework for estimating cellular growth dynamics of Wolbachia strains in different host species, tissues, and cell types provides the first strategy for parameterizing endosymbiont and host cell biology at high resolution. This toolset will be crucial to our application of these bacteria as biological control agents in novel hosts and ecosystems.}, } @article {pmid38571791, year = {2024}, author = {Fisher, AM and Knell, RJ and Price, TAR and Bonsall, MB}, title = {Sex ratio distorting microbes exacerbate arthropod extinction risk in variable environments.}, journal = {Ecology and evolution}, volume = {14}, number = {4}, pages = {e11216}, pmid = {38571791}, issn = {2045-7758}, abstract = {Maternally-inherited sex ratio distorting microbes (SRDMs) are common among arthropod species. Typically, these microbes cause female-biased sex ratios in host broods, either by; killing male offspring, feminising male offspring, or inducing parthenogenesis. As a result, infected populations can experience drastic ecological and evolutionary change. The mechanism by which SRDMs operate is likely to alter their impact on host evolutionary ecology; despite this, the current literature is heavily biased towards a single mechanism of sex ratio distortion, male-killing. Furthermore, amidst the growing concerns surrounding the loss of arthropod diversity, research into the impact of SRDMs on the viability of arthropod populations is generally lacking. In this study, using a theoretical approach, we model the epidemiology of an understudied mechanism of microbially-induced sex ratio distortion-feminisation-to ask an understudied question-how do SRDMs impact extinction risk in a changing environment? We constructed an individual-based model and measured host population extinction risk under various environmental and epidemiological scenarios. We also used our model to identify the precise mechanism modulating extinction. We find that the presence of feminisers increases host population extinction risk, an effect that is exacerbated in highly variable environments. We also identified transmission rate as the dominant epidemiological trait responsible for driving extinction. Finally, our model shows that sex ratio skew is the mechanism driving extinction. We highlight feminisers and, more broadly, SRDMs as important determinants of the resilience of arthropod populations to environmental change.}, } @article {pmid38570719, year = {2024}, author = {Rawle, DJ and Hugo, LE and Cox, AL and Devine, GJ and Suhrbier, A}, title = {Generating prophylactic immunity against arboviruses in vertebrates and invertebrates.}, journal = {Nature reviews. Immunology}, volume = {24}, number = {9}, pages = {621-636}, pmid = {38570719}, issn = {1474-1741}, mesh = {Animals ; Humans ; *Arboviruses/immunology ; *Arbovirus Infections/immunology/prevention & control ; Vertebrates/immunology ; Viral Vaccines/immunology ; Invertebrates/immunology ; Mosquito Vectors/immunology/virology ; }, abstract = {The World Health Organization recently declared a global initiative to control arboviral diseases. These are mainly caused by pathogenic flaviviruses (such as dengue, yellow fever and Zika viruses) and alphaviruses (such as chikungunya and Venezuelan equine encephalitis viruses). Vaccines represent key interventions for these viruses, with licensed human and/or veterinary vaccines being available for several members of both genera. However, a hurdle for the licensing of new vaccines is the epidemic nature of many arboviruses, which presents logistical challenges for phase III efficacy trials. Furthermore, our ability to predict or measure the post-vaccination immune responses that are sufficient for subclinical outcomes post-infection is limited. Given that arboviruses are also subject to control by the immune system of their insect vectors, several approaches are now emerging that aim to augment antiviral immunity in mosquitoes, including Wolbachia infection, transgenic mosquitoes, insect-specific viruses and paratransgenesis. In this Review, we discuss recent advances, current challenges and future prospects in exploiting both vertebrate and invertebrate immune systems for the control of flaviviral and alphaviral diseases.}, } @article {pmid38569988, year = {2024}, author = {Amala, M and Nagarajan, H and Ahila, M and Nachiappan, M and Veerapandiyan, M and Vetrivel, U and Jeyakanthan, J}, title = {Unveiling the intricacies of allosteric regulation in aspartate kinase from the Wolbachia endosymbiont of Brugia Malayi: Mechanistic and therapeutic insights.}, journal = {International journal of biological macromolecules}, volume = {267}, number = {Pt 1}, pages = {131326}, doi = {10.1016/j.ijbiomac.2024.131326}, pmid = {38569988}, issn = {1879-0003}, mesh = {*Brugia malayi/enzymology/microbiology ; Allosteric Regulation ; *Wolbachia ; Animals ; *Molecular Dynamics Simulation ; *Aspartate Kinase/metabolism/genetics/chemistry ; Symbiosis ; Adenosine Triphosphate/metabolism ; Lysine/chemistry/metabolism ; }, abstract = {Aspartate kinase (AK), an enzyme from the Wolbachia endosymbiont of Brugia malayi (WBm), plays a pivotal role in the bacterial cell wall and amino acid biosynthesis, rendering it an attractive candidate for therapeutic intervention. Allosteric inhibition of aspartate kinase is a prevalent mode of regulation across microorganisms and plants, often modulated by end products such as lysine, threonine, methionine, or meso-diaminopimelate. The intricate and diverse nature of microbial allosteric regulation underscores the need for rigorous investigation. This study employs a combined experimental and computational approach to decipher the allosteric regulation of WBmAK. Molecular Dynamics (MD) simulations elucidate that ATP (cofactor) and ASP (substrate) binding induce a closed conformation, promoting enzymatic activity. In contrast, the binding of lysine (allosteric inhibitor) leads to enzyme inactivation and an open conformation. The enzymatic assay demonstrates the optimal activity of WBmAK at 28 °C and a pH of 8.0. Notably, the allosteric inhibition study highlights lysine as a more potent inhibitor compared to threonine. Importantly, this investigation sheds light on the allosteric mechanism governing WBmAK and imparts novel insights into structure-based drug discovery, paving the way for the development of effective inhibitors against filarial pathogens.}, } @article {pmid38562961, year = {2024}, author = {Reyes, JIL and Suzuki, T and Suzuki, Y and Watanabe, K}, title = {Detection and quantification of natural Wolbachia in Aedes aegypti in Metropolitan Manila, Philippines using locally designed primers.}, journal = {Frontiers in cellular and infection microbiology}, volume = {14}, number = {}, pages = {1360438}, pmid = {38562961}, issn = {2235-2988}, mesh = {Animals ; Humans ; *Aedes/microbiology ; *Wolbachia/genetics ; Philippines ; RNA, Ribosomal, 16S/genetics ; Mosquito Vectors ; Phylogeny ; }, abstract = {BACKGROUND: The Philippines bears health and economic burden caused by high dengue cases annually. Presently, the Philippines still lack an effective and sustainable vector management. The use of Wolbachia, a maternally transmitted bacterium, that mitigate arbovirus transmission has been recommended. Cytoplasmic incompatibility and viral blocking, two characteristics that make Wolbachia suitable for vector control, depend on infection prevalence and density. There are no current Wolbachia release programs in the Philippines, and studies regarding the safety of this intervention. Here, we screened for Wolbachia in Aedes aegypti collected from Metropolitan Manila, Philippines. We designed location-specific primers for qPCR to test whether this improved Wolbachia detection in Ae. aegypti. We explored if host sex and Wolbachia strain could be potential factors affecting Wolbachia density.

METHODS: Ae. aegypti mosquitoes (n=429) were screened for natural Wolbachia by taqman qPCR using location-specific Wolbachia surface protein primers (wspAAML) and known 16S rRNA primers. Samples positive for wspAAML (n=267) were processed for Sanger sequencing. We constructed a phylogenetic tree using IQ-TREE 2 to further characterize Wolbachia present in the Philippine Ae. aegypti. We then compared Wolbachia densities between Wolbachia groups and host sex. Statistical analyses were done using GraphPad Prism 9.0.

RESULTS: Wolbachia prevalence for 16S rRNA (40%) and wspAAML (62%) markers were high. Wolbachia relative densities for 16S rRNA ranged from -3.84 to 2.71 and wspAAML from -4.02 to 1.81. Densities were higher in male than female mosquitoes. Wolbachia strains detected in Ae. aegypti clustered into supergroup B. Some 54% (123/226) of these sequences clustered under a group referred to here as "wAegML," that belongs to the supergroup B, which had a significantly lower density than wAegB/wAlbB, and wAlbA strains.

CONCLUSION: Location-specific primers improved detection of natural Wolbachia in Ae. aegypti and allowed for relative quantification. Wolbachia density is relatively low, and differed between host sexes and Wolbachia strains. An economical way of confirming sporadic or transient Wolbachia in Ae. aegypti is necessary while considering host sex and bacterial strain.}, } @article {pmid38558489, year = {2024}, author = {Ferguson, LF and Ross, PA and van Heerwaarden, B}, title = {Wolbachia infection negatively impacts Drosophila simulans heat tolerance in a strain- and trait-specific manner.}, journal = {Environmental microbiology}, volume = {26}, number = {4}, pages = {e16609}, doi = {10.1111/1462-2920.16609}, pmid = {38558489}, issn = {1462-2920}, support = {DE230100067//Australian Research Council/ ; FT200100025//Australian Research Council/ ; }, mesh = {Animals ; Male ; Drosophila/physiology ; Drosophila simulans/genetics ; *Wolbachia/genetics ; *Thermotolerance ; Fertility ; }, abstract = {The susceptibility of insects to rising temperatures has largely been measured by their ability to survive thermal extremes. However, the capacity for maternally inherited endosymbionts to influence insect heat tolerance has been overlooked. Further, while some studies have addressed the impact of heat on traits like fertility, which can decline at temperatures below lethal thermal limits, none have considered the impact of endosymbionts. Here, we assess the impact of three Wolbachia strains (wRi, wAu and wNo) on the survival and fertility of Drosophila simulans exposed to heat stress during development or as adults. The effect of Wolbachia infection on heat tolerance was generally small and trait/strain specific. Only the wNo infection significantly reduced the survival of adult males after a heat shock. When exposed to fluctuating heat stress during development, the wRi and wAu strains reduced egg-to-adult survival but only the wNo infection reduced male fertility. Wolbachia densities of all three strains decreased under developmental heat stress, but reductions occurred at temperatures above those that reduced host fertility. These findings emphasize the necessity to account for endosymbionts and their effect on both survival and fertility when investigating insect responses to heat stress.}, } @article {pmid38553514, year = {2024}, author = {Konecka, E and Szymkowiak, P}, title = {Wolbachia supergroup A in Enoplognatha latimana (Araneae: Theridiidae) in Poland as an example of possible horizontal transfer of bacteria.}, journal = {Scientific reports}, volume = {14}, number = {1}, pages = {7486}, pmid = {38553514}, issn = {2045-2322}, mesh = {Animals ; Bacterial Proteins/genetics ; *Wolbachia/genetics ; RNA, Ribosomal, 16S/genetics ; Poland ; *Spiders/genetics ; Phylogeny ; }, abstract = {Wolbachia (phylum Pseudomonadota, class Alfaproteobacteria, order Rickettsiales, family Ehrlichiaceae) is a maternally inherited bacterial symbiont infecting more than half of arthropod species worldwide and constituting an important force in the evolution, biology, and ecology of invertebrate hosts. Our study contributes to the limited knowledge regarding the presence of intracellular symbiotic bacteria in spiders. Specifically, we investigated the occurrence of Wolbachia infection in the spider species Enoplognatha latimana Hippa and Oksala, 1982 (Araneae: Theridiidae) using a sample collected in north-western Poland. To the best of our knowledge, this is the first report of Wolbachia infection in E. latimana. A phylogeny based on the sequence analysis of multiple genes, including 16S rRNA, coxA, fbpA, ftsZ, gatB, gltA, groEL, hcpA, and wsp revealed that Wolbachia from the spider represented supergroup A and was related to bacterial endosymbionts discovered in other spider hosts, as well as insects of the orders Diptera and Hymenoptera. A sequence unique for Wolbachia supergroup A was detected for the ftsZ gene. The sequences of Wolbachia housekeeping genes have been deposited in publicly available databases and are an important source of molecular data for comparative studies. The etiology of Wolbachia infection in E. latimana is discussed.}, } @article {pmid38547310, year = {2024}, author = {Martin, M and López-Madrigal, S and Newton, ILG}, title = {The Wolbachia WalE1 effector alters Drosophila endocytosis.}, journal = {PLoS pathogens}, volume = {20}, number = {3}, pages = {e1011245}, pmid = {38547310}, issn = {1553-7374}, support = {P40 OD018537/OD/NIH HHS/United States ; R01 AI144430/AI/NIAID NIH HHS/United States ; }, mesh = {Male ; Animals ; *Drosophila/microbiology ; *Wolbachia/metabolism ; Seeds ; Reproduction ; Endocytosis ; Drosophila melanogaster ; Symbiosis/genetics ; }, abstract = {The most common intracellular bacterial infection is Wolbachia pipientis, a microbe that manipulates host reproduction and is used in control of insect vectors. Phenotypes induced by Wolbachia have been studied for decades and range from sperm-egg incompatibility to male killing. How Wolbachia alters host biology is less well understood. Previously, we characterized the first Wolbachia effector-WalE1, which encodes an alpha-synuclein domain at the N terminus. Purified WalE1 sediments with and bundles actin and when heterologously expressed in flies, increases Wolbachia titer in the developing oocyte. In this work, we first identify the native expression of WalE1 by Wolbachia infecting both fly cells and whole animals. WalE1 appears as aggregates in the host cell cytosol. We next show that WalE1 co-immunoprecipitates with the host protein Past1, although might not directly interact with it, and that WalE1 manipulates host endocytosis. Yeast expressing WalE1 show deficiency in uptake of FM4-64 dye, and flies harboring mutations in Past1 or overexpressing WalE1 are sensitive to AgNO3, a hallmark of endocytosis defects. We also show that flies expressing WalE1 suffer from endocytosis defects in larval nephrocytes. Finally, we also show that Past1 null flies harbor more Wolbachia overall and in late egg chambers. Our results identify interactions between Wolbachia and a host protein involved in endocytosis and point to yet another important host cell process impinged upon by Wolbachia's WalE1 effector.}, } @article {pmid38547237, year = {2024}, author = {Kaur, R and Meier, CJ and McGraw, EA and Hillyer, JF and Bordenstein, SR}, title = {The mechanism of cytoplasmic incompatibility is conserved in Wolbachia-infected Aedes aegypti mosquitoes deployed for arbovirus control.}, journal = {PLoS biology}, volume = {22}, number = {3}, pages = {e3002573}, pmid = {38547237}, issn = {1545-7885}, support = {R01 AI132581/AI/NIAID NIH HHS/United States ; R01 AI143725/AI/NIAID NIH HHS/United States ; R56 AI155573/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Female ; Male ; *Aedes ; Drosophila melanogaster/genetics ; *Wolbachia ; Histones/genetics ; *Arboviruses ; Mosquito Vectors ; Semen ; Drosophila/genetics ; Chromatin ; Protamines/genetics ; }, abstract = {The rising interest and success in deploying inherited microorganisms and cytoplasmic incompatibility (CI) for vector control strategies necessitate an explanation of the CI mechanism. Wolbachia-induced CI manifests in the form of embryonic lethality when sperm from Wolbachia-bearing testes fertilize eggs from uninfected females. Embryos from infected females however survive to sustain the maternally inherited symbiont. Previously in Drosophila melanogaster flies, we demonstrated that CI modifies chromatin integrity in developing sperm to bestow the embryonic lethality. Here, we validate these findings using wMel-transinfected Aedes aegypti mosquitoes released to control vector-borne diseases. Once again, the prophage WO CI proteins, CifA and CifB, target male gametic nuclei to modify chromatin integrity via an aberrant histone-to-protamine transition. Cifs are not detected in the embryo, and thus elicit CI via the nucleoprotein modifications established pre-fertilization. The rescue protein CifA in oogenesis localizes to stem cell, nurse cell, and oocyte nuclei, as well as embryonic DNA during embryogenesis. Discovery of the nuclear targeting Cifs and altered histone-to-protamine transition in both Aedes aegypti mosquitoes and D. melanogaster flies affirm the Host Modification Model of CI is conserved across these host species. The study also newly uncovers the cell biology of Cif proteins in the ovaries, CifA localization in the embryos, and an impaired histone-to-protamine transition during spermiogenesis of any mosquito species. Overall, these sperm modification findings may enable future optimization of CI efficacy in vectors or pests that are refractory to Wolbachia transinfections.}, } @article {pmid38543596, year = {2024}, author = {Ilbeigi Khamseh Nejad, M and Cappelli, A and Damiani, C and Falcinelli, M and Catapano, PL and Nanfack-Minkeu, F and Mayi, MPA and Currà, C and Ricci, I and Favia, G}, title = {Wolbachia and Asaia Distribution among Different Mosquito Vectors Is Affected by Tissue Localization and Host Species.}, journal = {Microorganisms}, volume = {12}, number = {3}, pages = {}, pmid = {38543596}, issn = {2076-2607}, support = {PRIN 2022 PNRR MUR P20225TJWB//European Union - NextGenerationEU/ ; }, abstract = {Microbial communities play an important role in the fitness of mosquito hosts. However, the factors shaping microbial communities in wild populations, with regard to interactions among microbial species, are still largely unknown. Previous research has demonstrated that two of the most studied mosquito symbionts, the bacteria Wolbachia and Asaia, seem to compete or not compete, depending on the genetic background of the reference mosquito host. The large diversity of Wolbachia-Asaia strain combinations that infect natural populations of mosquitoes may offer a relevant opportunity to select suitable phenotypes for the suppression of pathogen transmission and for the manipulation of host reproduction. We surveyed Wolbachia and Asaia in 44 mosquito populations belonging to 11 different species of the genera Anopheles, Aedes, and Culex using qualitative PCR. Through quantitative PCR, the amounts of both bacteria were assessed in different mosquito organs, and through metagenomics, we determined the microbiota compositions in some selected mosquito populations. We show that variation in microbial community structure is likely associated with the species/strain of mosquito, its geographical position, and tissue localization. Together, our results shed light on the interactions among different bacterial species in the microbial communities of mosquito vectors, and this can aid the development and/or improvement of methods for symbiotic control of insect vectors.}, } @article {pmid38535401, year = {2024}, author = {Cholvi, M and Trelis, M and Bueno-Marí, R and Khoubbane, M and Gil, R and Marcilla, A and Moretti, R}, title = {Wolbachia Infection through Hybridization to Enhance an Incompatible Insect Technique-Based Suppression of Aedes albopictus in Eastern Spain.}, journal = {Insects}, volume = {15}, number = {3}, pages = {}, pmid = {38535401}, issn = {2075-4450}, abstract = {The emergence of insecticide resistance in arbovirus vectors is putting the focus on the development of new strategies for control. In this regard, the exploitation of Wolbachia endosymbionts is receiving increasing attention due to its demonstrated effectiveness in reducing the vectorial capacity of Aedes mosquitoes. Here, we describe the establishment of a naïve Wolbachia infection in a wild Aedes albopictus population of eastern Spain through a hybridization approach to obtain males capable of sterilizing wild females. The obtained lines were compared with the Wolbachia donor, Ae. albopictus ARwP, previously artificially infected with Wolbachia wPip, regarding immature and adult survival, female fecundity, egg fertility, and level of induced sterility. Our results did not show significant differences between lines in any of the biological parameters analyzed, indicating the full suitability of the hybrids to be used as a control tool against Ae. albopictus. In particular, hybrid males induced 99.9% sterility in the eggs of wild females without the need for any preliminary treatment. Being harmless to non-target organisms and the environment, the use of this bacterium for the control of Ae. albopictus deserves further exploration. This is especially relevant in areas such as eastern Spain, where this mosquito species has recently spread and may represent a serious threat due to its competence as a vector for dengue, chikungunya, and Zika viruses.}, } @article {pmid38534451, year = {2024}, author = {Miranda, LS and Rudd, SR and Mena, O and Hudspeth, PE and Barboza-Corona, JE and Park, HW and Bideshi, DK}, title = {The Perpetual Vector Mosquito Threat and Its Eco-Friendly Nemeses.}, journal = {Biology}, volume = {13}, number = {3}, pages = {}, pmid = {38534451}, issn = {2079-7737}, support = {na//an Internal Research Grant through the Office of the Provost and the Faculty Development Committee at California Baptist University./ ; }, abstract = {Mosquitoes are the most notorious arthropod vectors of viral and parasitic diseases for which approximately half the world's population, ~4,000,000,000, is at risk. Integrated pest management programs (IPMPs) have achieved some success in mitigating the regional transmission and persistence of these diseases. However, as many vector-borne diseases remain pervasive, it is obvious that IPMP successes have not been absolute in eradicating the threat imposed by mosquitoes. Moreover, the expanding mosquito geographic ranges caused by factors related to climate change and globalization (travel, trade, and migration), and the evolution of resistance to synthetic pesticides, present ongoing challenges to reducing or eliminating the local and global burden of these diseases, especially in economically and medically disadvantaged societies. Abatement strategies include the control of vector populations with synthetic pesticides and eco-friendly technologies. These "green" technologies include SIT, IIT, RIDL, CRISPR/Cas9 gene drive, and biological control that specifically targets the aquatic larval stages of mosquitoes. Regarding the latter, the most effective continues to be the widespread use of Lysinibacillus sphaericus (Ls) and Bacillus thuringiensis subsp. israelensis (Bti). Here, we present a review of the health issues elicited by vector mosquitoes, control strategies, and lastly, focus on the biology of Ls and Bti, with an emphasis on the latter, to which no resistance has been observed in the field.}, } @article {pmid38534421, year = {2024}, author = {Hyder, M and Lodhi, AM and Wang, Z and Bukero, A and Gao, J and Mao, R}, title = {Wolbachia Interactions with Diverse Insect Hosts: From Reproductive Modulations to Sustainable Pest Management Strategies.}, journal = {Biology}, volume = {13}, number = {3}, pages = {}, pmid = {38534421}, issn = {2079-7737}, support = {32202276//National Science Foundation of China/ ; 2022GDASZH-2022010106, 2022GDASZH-2022030501-08//GDAS Special Project of Science and Technology Development/ ; KTP20210352//Guangdong Province Rural Science and Technology Commissioner Project/ ; 2023SDZG06//Top Ten Critical Priorities of Agricultural Science and Technology Innovation for the 14th Five-Year Plan of Guangdong Province/ ; }, abstract = {Effective in a variety of insect orders, including dipteran, lepidopteran, and hemipteran, Wolbachia-based control tactics are investigated, noting the importance of sterile and incompatible insect techniques. Encouraging approaches for controlling Aedes mosquitoes are necessary, as demonstrated by the evaluation of a new SIT/IIT combination and the incorporation of SIT into Drosophila suzukii management. For example, Wolbachia may protect plants from rice pests, demonstrating its potential for agricultural biological vector management. Maternal transmission and cytoplasmic incompatibility dynamics are explored, while Wolbachia phenotypic impacts on mosquito and rice pest management are examined. The importance of host evolutionary distance is emphasised in recent scale insect research that addresses host-shifting. Using greater information, a suggested method for comprehending Wolbachia host variations in various contexts emphasises ecological connectivity. Endosymbionts passed on maternally in nematodes and arthropods, Wolbachia are widely distributed around the world and have evolved both mutualistic and parasitic traits. Wolbachia is positioned as a paradigm for microbial symbiosis due to advancements in multiomics, gene functional assays, and its effect on human health. The challenges and opportunities facing Wolbachia research include scale issues, ecological implications, ethical conundrums, and the possibility of customising strains through genetic engineering. It is thought that cooperative efforts are required to include Wolbachia-based therapies into pest management techniques while ensuring responsible and sustainable ways.}, } @article {pmid38530785, year = {2024}, author = {Fricke, LC and Lindsey, ARI}, title = {Identification of Parthenogenesis-Inducing Effector Proteins in Wolbachia.}, journal = {Genome biology and evolution}, volume = {16}, number = {4}, pages = {}, pmid = {38530785}, issn = {1759-6653}, support = {R35 GM150991/GM/NIGMS NIH HHS/United States ; R35GM150991/NH/NIH HHS/United States ; }, mesh = {Male ; Animals ; Female ; *Wolbachia/genetics ; Parthenogenesis/genetics ; *Wasps/genetics ; Bacterial Proteins/genetics ; Genomics ; Symbiosis ; }, abstract = {Bacteria in the genus Wolbachia have evolved numerous strategies to manipulate arthropod sex, including the conversion of would-be male offspring to asexually reproducing females. This so-called "parthenogenesis induction" phenotype can be found in a number of Wolbachia strains that infect arthropods with haplodiploid sex determination systems, including parasitoid wasps. Despite the discovery of microbe-mediated parthenogenesis more than 30 yr ago, the underlying genetic mechanisms have remained elusive. We used a suite of genomic, computational, and molecular tools to identify and characterize two proteins that are uniquely found in parthenogenesis-inducing Wolbachia and have strong signatures of host-associated bacterial effector proteins. These putative parthenogenesis-inducing proteins have structural homology to eukaryotic protein domains including nucleoporins, the key insect sex determining factor Transformer, and a eukaryotic-like serine-threonine kinase with leucine-rich repeats. Furthermore, these proteins significantly impact eukaryotic cell biology in the model Saccharomyces cerevisiae. We suggest that these proteins are parthenogenesis-inducing factors and our results indicate that this would be made possible by a novel mechanism of bacterial-host interaction.}, } @article {pmid38528862, year = {2024}, author = {Soo, WF and Gunasekaran, K and Ng, DX and Kwek, K and Tan, NC}, title = {Literacy and attitude of Asian youths on dengue and its prevention in an endemic developed community.}, journal = {Frontiers in public health}, volume = {12}, number = {}, pages = {1361717}, pmid = {38528862}, issn = {2296-2565}, mesh = {Young Adult ; Humans ; Adolescent ; Female ; Male ; *Literacy ; Cross-Sectional Studies ; *Dengue/epidemiology/prevention & control ; Surveys and Questionnaires ; Regression Analysis ; }, abstract = {BACKGROUND: Over the past few decades, the incidence of dengue fever has considerably increased. Effective vector control strategies and specific protection using dengue vaccine are thought to be the key elements to combat dengue. The dengue incidence among the Singapore youths (15-24 years) was second only to that of adults (25-44 years). This study evaluated the knowledge and attitude of Singapore youths on dengue and its preventive measures.

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

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

CONCLUSION: The overall knowledge of the youths on dengue and its prevention was satisfactory. Future health promotion campaigns targeting the youths should focus on transforming the knowledge into practice.}, } @article {pmid38525276, year = {2024}, author = {Guse, K and Pietri, JE}, title = {Endosymbiont and gut bacterial communities of the brown-banded cockroach, Supella longipalpa.}, journal = {PeerJ}, volume = {12}, number = {}, pages = {e17095}, pmid = {38525276}, issn = {2167-8359}, support = {R01 AI171014/AI/NIAID NIH HHS/United States ; }, mesh = {Male ; Animals ; Female ; Adult ; Humans ; *Blattellidae/genetics ; *Gastrointestinal Microbiome ; RNA, Ribosomal, 16S/genetics ; *Flavobacteriaceae/genetics ; Symbiosis/genetics ; }, abstract = {The brown-banded cockroach (Supella longipalpa) is a widespread nuisance and public health pest. Like the German cockroach (Blattella germanica), this species is adapted to the indoor biome and completes the entirety of its life cycle in human-built structures. Recently, understanding the contributions of commensal and symbiotic microbes to the biology of cockroach pests, as well as the applications of targeting these microbes for pest control, have garnered significant scientific interest. However, relative to B. germanica, the biology of S. longipalpa, including its microbial associations, is understudied. Therefore, the goal of the present study was to quantitatively examine and characterize both the endosymbiont and gut bacterial communities of S. longipalpa for the first time. To do so, bacterial 16S rRNA gene amplicon sequencing was conducted on DNA extracts from whole adult females and males, early instar nymphs, and late instar nymphs. The results demonstrate that the gut microbiome is dominated by two genera of bacteria known to have beneficial probiotic effects in other organisms, namely Lactobacillus and Akkermansia. Furthermore, our data show a significant effect of nymphal development on diversity and variation in the gut microbiome. Lastly, we reveal significant negative correlations between the two intracellular endosymbionts, Blattabacterium and Wolbachia, as well as between Blattabacterium and the gut microbiome, suggesting that Blattabacterium endosymbionts could directly or indirectly influence the composition of other bacterial populations. These findings have implications for understanding the adaptation of S. longipalpa to the indoor biome, its divergence from other indoor cockroach pest species such as B. germanica, the development of novel control approaches that target the microbiome, and fundamental insect-microbe interactions more broadly.}, } @article {pmid38507220, year = {2024}, author = {Hollingsworth, BD and Cho, C and Vella, M and Roh, H and Sass, J and Lloyd, AL and Brown, ZS}, title = {Economic optimization of Wolbachia-infected Aedes aegypti release to prevent dengue.}, journal = {Pest management science}, volume = {80}, number = {8}, pages = {3829-3838}, doi = {10.1002/ps.8086}, pmid = {38507220}, issn = {1526-4998}, mesh = {*Aedes/microbiology/virology ; *Wolbachia/physiology ; Animals ; *Dengue/prevention & control/transmission ; *Mosquito Control/methods/economics ; *Mosquito Vectors/microbiology ; }, abstract = {BACKGROUND: Dengue virus, primarily transmitted by the Aedes aegypti mosquito, is a major public health concern affecting ≈3.83 billion people worldwide. Recent releases of Wolbachia-transinfected Ae. aegypti in several cities worldwide have shown that it can reduce dengue transmission. However, these releases are costly, and, to date, no framework has been proposed for determining economically optimal release strategies that account for both costs associated with disease risk and releases.

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

CONCLUSIONS: These results reinforce that Wolbachia-transinfected mosquitoes can reduce the cost of dengue epidemics. Furthermore, they emphasize the importance of determining the size of the target population and fitness costs associated with Wolbachia before releases occur. © 2024 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.}, } @article {pmid38504126, year = {2024}, author = {Du Toit, A}, title = {Drosophila sperm sabotage by Wolbachia prophage.}, journal = {Nature reviews. Microbiology}, volume = {22}, number = {5}, pages = {257}, pmid = {38504126}, issn = {1740-1534}, mesh = {Animals ; Male ; *Drosophila ; *Wolbachia ; Prophages ; Semen ; Spermatozoa ; Drosophila melanogaster ; }, } @article {pmid38502456, year = {2024}, author = {Singh, AS and Pathak, D and Devi, MS and Anifowoshe, AT and Nongthomba, U}, title = {Antibiotic alters host's gut microbiota, fertility, and antimicrobial peptide gene expression vis-à-vis ampicillin treatment on model organism Drosophila melanogaster.}, journal = {International microbiology : the official journal of the Spanish Society for Microbiology}, volume = {}, number = {}, pages = {}, pmid = {38502456}, issn = {1618-1905}, support = {DBT-RA/2022/January NE/994//Department of Biotechnology, Government of India./ ; DBT-RA/2022/January NE/994//Department of Biotechnology, Government of India./ ; }, abstract = {Antibiotics are commonly used to treat infectious diseases; however, persistence is often expressed by the pathogenic bacteria and their long-term relative effect on the host have been neglected. The present study investigated the impact of antibiotics in gut microbiota (GM) and metabolism of host. The effect of ampicillin antibiotics on GM of Drosophila melanogaster was analyzed through deep sequencing of 16S rRNA amplicon gene. The dominant phyla consisted of Proteobacteria, Bacteroidetes, Firmicutes, Actinobacteria, Planctomycetes, Chloroflexi, Euryarchaeota, Acedobacteria, Verrucomicrobia, and Cyanobacteria. It was found that the composition of GM was significantly altered on administration of antibiotics. On antibiotic treatments, there were decline in relative abundance of Proteobacteria and Firmicutes, while there were increase in relative abundance of Chlorophyta and Bacteroidota. High abundance of 14 genera, viz., Wolbachia, Lactobacillus, Bacillus, Pseudomonas, Thiolamprovum, Pseudoalteromonas, Vibrio, Romboutsia, Staphylococcus, Alteromonas, Clostridium, Lysinibacillus, Litoricola, and Cellulophaga were significant (p ≤ 0.05) upon antibiotic treatment. Particularly, the abundance of Acetobacter was significantly (p ≤ 0.05) declined but increased for Wolbachia. Further, a significant (p ≤ 0.05) increase in Wolbachia endosymbiont of D. melanogaster, Wolbachia endosymbiont of Curculio okumai, and Wolbachia pipientis and a decrease in the Acinetobacter sp. were observed. We observed an increase in functional capacity for biosynthesis of certain nucleotides and the enzyme activities. Further, the decrease in antimicrobial peptide production in the treated group and potential effects on the host's defense mechanisms were observed. This study helps shed light on an often-overlooked dimension, namely the persistence of antibiotics' effects on the host.}, } @article {pmid38496649, year = {2024}, author = {Hague, MTJ and Wheeler, TB and Cooper, BS}, title = {Comparative analysis of Wolbachia maternal transmission and localization in host ovaries.}, journal = {bioRxiv : the preprint server for biology}, volume = {}, number = {}, pages = {}, pmid = {38496649}, issn = {2692-8205}, support = {R35 GM124701/GM/NIGMS NIH HHS/United States ; }, abstract = {Many insects and other animals carry microbial endosymbionts that influence their reproduction and fitness. These relationships only persist if endosymbionts are reliably transmitted from one host generation to the next. Wolbachia are maternally transmitted endosymbionts found in most insect species, but transmission rates can vary across environments. Maternal transmission of wMel Wolbachia depends on temperature in natural Drosophila melanogaster hosts and in transinfected Aedes aegypti, where wMel is used to block pathogens that cause human disease. In D. melanogaster, wMel transmission declines in the cold as Wolbachia become less abundant in host ovaries and at the posterior pole plasm (the site of germline formation) in mature oocytes. Here, we assess how temperature affects maternal transmission and underlying patterns of Wolbachia localization across 10 Wolbachia strains diverged up to 50 million years-including strains closely related to wMel-and their natural Drosophila hosts. Many Wolbachia maintain high transmission rates across temperatures, despite highly variable (and sometimes low) levels of Wolbachia in the ovaries and at the developing germline in late-stage oocytes. Identifying strains like closely related wMel-like Wolbachia with stable transmission across variable environmental conditions may improve the efficacy of Wolbachia-based biocontrol efforts as they expand into globally diverse environments.}, } @article {pmid38493661, year = {2024}, author = {González, MA and Ruiz-Arrondo, I and Bravo-Barriga, D and Cervera-Acedo, C and Santibáñez, P and Oteo, JA and Miranda, MÁ and Barceló, C}, title = {Surveillance and screening of Stomoxyinae flies from Mallorca Island (Spain) reveal the absence of selected pathogens but confirm the presence of the endosymbiotic bacterium Wolbachia pipientis.}, journal = {Research in veterinary science}, volume = {171}, number = {}, pages = {105206}, doi = {10.1016/j.rvsc.2024.105206}, pmid = {38493661}, issn = {1532-2661}, mesh = {Animals ; *Wolbachia/genetics ; Spain ; RNA, Ribosomal, 16S/genetics ; *Muscidae/genetics/microbiology/parasitology ; Bacteria/genetics ; }, abstract = {Adult brachycera biting flies can significantly impact livestock through both direct effects (reduction of food intake, disturbance, painful bites, and blood loss) and indirect effects (pathogen transmission), leading to substantial economic losses and production damage. This study aimed to assess the presence of blood-sucking flies in six mixed-animal farm environments on the island of Mallorca (Balearic Islands, Spain) by employing multiple trapping methods. Additionally, distribution maps of brachycera biting fly species recorded in Spain were created, based on data extracted thorough review of scientific literature and citizen digital databases. Investigation of several pathogens, including equine infectious anemia virus (EIAV), Anaplasmataceae bacteria, and piroplasm protozoa, was carried out using different PCR targets (18S rRNA, 16S rRNA, groESL, and tat genes). Citizen science databases and literature review corroborated the consistent distribution trend for two Stomoxyinae species, underscoring the importance of citizen collaboration as a complement to traditional entomological surveillance. Our study confirmed the presence of two biting Stomoxyinae species: the prevalent stable fly Stomoxys calcitrans across all sampled farms, and the horn fly Haematobia irritans, which turned out to be less abundant. DNA barcoding techniques validated the identification of the two species. Neither EIAV nor bacterial/protozoan pathogens were detected using the selected PCR targets in either fly species. However, Wolbachia pipientis (clustered in the supergroup A together with the only sequence of W. pipientis from the USA) was identified through PCR targeting 16S rRNA, groESL and wsp genes in all pools of H. irritans (n = 13) collected from two of the examined farms. This study represents the first attempt to investigate pathogens in Stomoxyinae biting flies in Spain. The discovery of the endosymbiotic Wolbachia organism in H. irritans represents the first record in Spain and the second from Europe. This finding holds significant implications for future research on the applications of this bacterium in biocontrol programs.}, } @article {pmid38489098, year = {2024}, author = {Calleros-González, P and Ibarra-Juarez, A and Lamelas, A and Suárez-Moo, P}, title = {How host species and body part determine the microbial communities of five ambrosia beetle species.}, journal = {International microbiology : the official journal of the Spanish Society for Microbiology}, volume = {}, number = {}, pages = {}, pmid = {38489098}, issn = {1618-1905}, abstract = {The ambrosia beetles are farming insects that feed mainly on their cultivated fungi, which in some occasions are pathogens from forest and fruit trees. We used a culture-independent approach based on 16S and 18S rRNA gene metabarcoding analysis to investigate the diversity and composition of the bacterial and fungal communities associated with five ambrosia beetle species: four species native to America (Monarthrum dimidiatum, Dryocoetoides capucinus, Euwallacea discretus, Corthylus consimilis) and an introduced species (Xylosandrus morigerus). For the bacterial community, the beetle species hosted a broad diversity with 1,579 amplicon sequence variants (ASVs) and 66 genera, while for the fungal community they hosted 288 ASVs and 39 genera. Some microbial groups dominated the community within a host species or a body part (Wolbachia in the head-thorax of E. discretus; Ambrosiella in the head-thorax and abdomen of X. morigerus). The taxonomic composition and structure of the microbial communities appeared to differ between beetle species; this was supported by beta-diversity analysis, which indicated that bacterial and fungal communities were clustered mainly by host species. This study characterizes for the first time the microbial communities associated with unexplored ambrosia beetle species, as well as the factors that affect the composition and taxonomic diversity per se, contributing to the knowledge of the ambrosia beetle system.}, } @article {pmid38484041, year = {2024}, author = {Simmons, CP and Donald, W and Tagavi, L and Tarivonda, L and Quai, T and Tavoa, R and Noran, T and Manikaoti, E and Kareaua, L and Abwai, TT and Chand, D and Rama, V and Deo, V and Deo, KK and Tavuii, A and Valentine, W and Prasad, R and Seru, E and Naituku, L and Ratu, A and Hesketh, M and Kenny, N and Beebe, SC and Goundar, AA and McCaw, A and Buntine, M and Green, B and Frossard, T and Gilles, JRL and Joubert, DA and Wilson, G and Duong, LQ and Bouvier, JB and Stanford, D and Forder, C and Duyvestyn, JM and Pacidônio, EC and Flores, HA and Wittmeier, N and Retzki, K and Ryan, PA and Denton, JA and Smithyman, R and Tanamas, SK and Kyrylos, P and Dong, Y and Khalid, A and Hodgson, L and Anders, KL and O'Neill, SL}, title = {Successful introgression of wMel Wolbachia into Aedes aegypti populations in Fiji, Vanuatu and Kiribati.}, journal = {PLoS neglected tropical diseases}, volume = {18}, number = {3}, pages = {e0012022}, pmid = {38484041}, issn = {1935-2735}, mesh = {Animals ; Humans ; *Dengue Virus ; *Aedes/genetics/microbiology ; Mosquito Vectors/genetics/microbiology ; *Wolbachia/genetics ; Fiji/epidemiology ; Vanuatu ; *Zika Virus Infection ; *Zika Virus ; *Dengue ; }, abstract = {Pacific Island countries have experienced periodic dengue, chikungunya and Zika outbreaks for decades. The prevention and control of these mosquito-borne diseases rely heavily on control of Aedes aegypti mosquitoes, which in most settings are the primary vector. Introgression of the intracellular bacterium Wolbachia pipientis (wMel strain) into Ae. aegypti populations reduces their vector competence and consequently lowers dengue incidence in the human population. Here we describe successful area-wide deployments of wMel-infected Ae. aegypti in Suva, Lautoka, Nadi (Fiji), Port Vila (Vanuatu) and South Tarawa (Kiribati). With community support, weekly releases of wMel-infected Ae. aegypti mosquitoes for between 2 to 5 months resulted in wMel introgression in nearly all locations. Long term monitoring confirmed a high, self-sustaining prevalence of wMel infecting mosquitoes in almost all deployment areas. Measurement of public health outcomes were disrupted by the Covid19 pandemic but are expected to emerge in the coming years.}, } @article {pmid38483475, year = {2024}, author = {Maciel-de-Freitas, R and Sauer, FG and Kliemke, K and Garcia, GA and Pavan, MG and David, MR and Schmidt-Chanasit, J and Hoffmann, A and Lühken, R}, title = {Wolbachia strains wMel and wAlbB differentially affect Aedes aegypti traits related to fecundity.}, journal = {Microbiology spectrum}, volume = {12}, number = {4}, pages = {e0012824}, pmid = {38483475}, issn = {2165-0497}, support = {MA 9541/1-1//Deutsche Forschungsgemeinschaft (DFG)/ ; NEED 01Kl2022//Bundesministerium für Bildung und Forschung (BMBF)/ ; E-14/2019//Fundação Carlos Chagas Filho de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ)/ ; }, mesh = {Animals ; Female ; *Aedes ; *Wolbachia ; *Zika Virus ; *Zika Virus Infection ; Fertility ; *Dengue/prevention & control ; }, abstract = {Two Wolbachia strains, wMel and wAlbB, have been transinfected into Aedes aegypti mosquitoes for population replacement with the aim of reducing dengue transmission. Epidemiological data from various endemic sites suggest a pronounced decrease in dengue transmission after implementing this strategy. In this study, we investigated the impact of the Wolbachia strains wMel and wAlbB on Ae. aegypti fitness in a common genetic background. We found that Ae. aegypti females infected with the wMel strain exhibited several significant differences compared with those infected with the wAlbB strain. Specifically, wMel-infected females laid significantly fewer eggs, ingested a lower amount of blood, had a reduced egg production rate, and exhibited a decreased Wolbachia density at a later age compared with mosquitoes infected with the wAlbB strain. Conversely, the wAlbB strain showed only mild negative effects when compared with Wolbachia-uninfected specimens. These differential effects on Ae. aegypti fitness following infection with either wMel or wAlbB may have important implications for the success of population replacement strategies in invading native Ae. aegypti populations in endemic settings. Further research is needed to better understand the underlying mechanisms responsible for these differences in fitness effects and their potential impact on the long-term efficacy of Wolbachia-based dengue control programs.IMPORTANCEThe transmission of arboviruses such as dengue, Zika, and chikungunya is on the rise globally. Among the most promising strategies to reduce arbovirus burden is the release of one out of two strains of Wolbachia-infected Aedes aegypti: wMel and wAlbB. One critical aspect of whether this approach will succeed involves the fitness cost of either Wolbachia strains on mosquito life history traits. For instance, we found that wMel-infected Ae. aegypti females laid significantly fewer eggs, ingested a lower amount of blood, had a reduced egg production rate, and exhibited a decreased Wolbachia density at a later age compared with mosquitoes infected with the wAlbB strain. Conversely, the wAlbB strain showed only mild negative effects when compared with Wolbachia-uninfected specimens. These differential effects on mosquito fitness following infection with either wMel or wAlbB may have important implications for the success of population replacement strategies in invading native Ae. aegypti populations.}, } @article {pmid38470882, year = {2024}, author = {Auguste, A and Ris, N and Belgaidi, Z and Kremmer, L and Mouton, L and Fauvergue, X}, title = {Insect population dynamics under Wolbachia-induced cytoplasmic incompatibility: Puzzle more than buzz in Drosophila suzukii.}, journal = {PloS one}, volume = {19}, number = {3}, pages = {e0300248}, pmid = {38470882}, issn = {1932-6203}, mesh = {Humans ; Animals ; *Drosophila ; *Wolbachia ; Cytoplasm ; Population Dynamics ; Insecta ; }, abstract = {In theory, the introduction of individuals infected with an incompatible strain of Wolbachia pipientis into a recipient host population should result in the symbiont invasion and reproductive failures caused by cytoplasmic incompatibility (CI). Modelling studies combining Wolbachia invasion and host population dynamics show that these two processes could interact to cause a transient population decline and, in some conditions, extinction. However, these effects could be sensitive to density dependence, with the Allee effect increasing the probability of extinction, and competition reducing the demographic impact of CI. We tested these predictions with laboratory experiments in the fruit fly Drosophila suzukii and the transinfected Wolbachia strain wTei. Surprisingly, the introduction of wTei into D. suzukii populations at carrying capacity did not result in the expected wTei invasion and transient population decline. In parallel, we found no Allee effect but strong negative density dependence. From these results, we propose that competition interacts in an antagonistic way with Wolbachia-induced cytoplasmic incompatibility on insect population dynamics. If future models and data support this hypothesis, pest management strategies using Wolbachia-induced CI should target populations with negligible competition but a potential Allee effect, for instance at the beginning of the reproductive season.}, } @article {pmid38469495, year = {2023}, author = {Field, EN and Smith, RC}, title = {Seasonality influences key physiological components contributing to Culex pipiens vector competence.}, journal = {Frontiers in insect science}, volume = {3}, number = {}, pages = {1144072}, pmid = {38469495}, issn = {2673-8600}, abstract = {Mosquitoes are the most important animal vector of disease on the planet, transmitting a variety of pathogens of both medical and veterinary importance. Mosquito-borne diseases display distinct seasonal patterns driven by both environmental and biological variables. However, an important, yet unexplored component of these patterns is the potential for seasonal influences on mosquito physiology that may ultimately influence vector competence. To address this question, we selected Culex pipiens, a primary vector of the West Nile virus (WNV) in the temperate United States, to examine the seasonal impacts on mosquito physiology by examining known immune and bacterial components implicated in mosquito arbovirus infection. Semi-field experiments were performed under spring, summer, and late-summer conditions, corresponding to historically low-, medium-, and high-intensity periods of WNV transmission, respectively. Through these experiments, we observed differences in the expression of immune genes and RNA interference (RNAi) pathway components, as well as changes in the distribution and abundance of Wolbachia in the mosquitoes across seasonal cohorts. Together, these findings support the conclusion that seasonal changes significantly influence mosquito physiology and components of the mosquito microbiome, suggesting that seasonality may impact mosquito susceptibility to pathogen infection, which could account for the temporal patterns in mosquito-borne disease transmission.}, } @article {pmid38468292, year = {2024}, author = {Ahouandjinou, MJ and Sovi, A and Sidick, A and Sewadé, W and Koukpo, CZ and Chitou, S and Towakinou, L and Adjottin, B and Hougbe, S and Tokponnon, F and Padonou, GG and Akogbéto, M and Messenger, LA and Ossè, RA}, title = {First report of natural infection of Anopheles gambiae s.s. and Anopheles coluzzii by Wolbachia and Microsporidia in Benin: a cross-sectional study.}, journal = {Malaria journal}, volume = {23}, number = {1}, pages = {72}, pmid = {38468292}, issn = {1475-2875}, mesh = {Animals ; Humans ; Benin/epidemiology ; *Anopheles ; *Wolbachia ; Cross-Sectional Studies ; Mosquito Vectors ; *Malaria, Falciparum/epidemiology ; *Pyrethrins ; Sporozoites ; }, abstract = {BACKGROUND: Recently, bacterial endosymbiont, including Wolbachia and Microsporidia were found to limit the infection of Anopheles mosquitoes with Plasmodium falciparum. This study aimed to investigate the natural presence of key transmission-blocking endosymbionts in Anopheles gambiae and Anopheles coluzzii in Southern Benin.

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

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

CONCLUSIONS: The present study is the first report of natural infections of field-collected An. gambiae s.l. populations from Benin with Wolbachia and Microsporidia. Sustained efforts should be made to widen the spectrum of bacteria identified in mosquitoes, with the potential to develop endosymbiont-based control tools; such interventions could be the game-changer in the control of malaria and arboviral disease transmission.}, } @article {pmid38461924, year = {2024}, author = {Ateutchia-Ngouanet, S and Nanfack-Minkeu, F and Mavridis, K and Wanji, S and Demanou, M and Vontas, J and Djouaka, R}, title = {Monitoring Aedes populations for arboviruses, Wolbachia, insecticide resistance and its mechanisms in various agroecosystems in Benin.}, journal = {Acta tropica}, volume = {253}, number = {}, pages = {107178}, doi = {10.1016/j.actatropica.2024.107178}, pmid = {38461924}, issn = {1873-6254}, support = {001/WHO_/World Health Organization/International ; }, mesh = {Animals ; Humans ; *Insecticides/pharmacology ; Insecticide Resistance/genetics ; *Pyrethrins/pharmacology ; *Aedes ; *Arboviruses/genetics ; *Wolbachia/genetics ; Permethrin/pharmacology ; Benin ; *Zika Virus Infection ; Mosquito Vectors ; *Zika Virus ; Mutation ; }, abstract = {Aedes mosquitoes are the main vectors of arboviruses in Benin. Cases of dengue have been reported in Benin with all four serotypes of the virus actively circulating in this region. Some agricultural settings are known to harbor Aedes vectors responsible for the transmission of arboviruses. The massive use of certain insecticides in agricultural settings has probably contributed to insecticide resistance in these vectors. In Benin, the susceptibility of arbovirus vectors to insecticides is poorly studied. In addition, the distribution of Wolbachia spp., which is used against some arboviruses is unknown. Moreover, there is limited information regarding the vectors responsible for the transmission of arboviruses in Benin. This present study monitored the species composition, arboviruses, and Wolbachia symbiont status, as well as the phenotypic and molecular insecticide resistance profile of Aedes populations from three agroecosystems in Benin. Aedes species identification was performed morphologically and confirmed using qPCR. (RT)-qPCR assay was applied for monitoring the presence of DENV, CHIKV, ZIKV, and WNV pathogens as well as for naturally occurring Wolbachia symbionts. Insecticide resistance was assessed phenotypically, by permethrin (0.75%) exposure of Adults (F0) using World Health Organization (WHO) bioassay protocols, and at the molecular level, using TaqMan (RT)-qPCR assays for assessing knock-down resistance (kdr) mutations (F1534C, V1016G/I, and S989P) and the expression levels of eight detoxification genes (P450s from the CYP9 and CYP6 families, carboxylesterases and glutathione-S-transferases). Aedes aegypti (Ae. aegypti) mosquitoes were the most abundant (93.9%) in the three agroecosystems studied, followed by Aedes albopictus (Ae. albopictus) mosquitoes (6.1%). No arboviruses were detected in the study's mosquito populations. Naturally occurring Wolbachia symbionts were present in 7 pools out of 15 pools tested. This could influence the effectiveness of vector control strategies based on exogenously introduced Wolbachia, all present in the three agroecosystems. Full susceptibility to permethrin was observed in all tested populations of Ae. albopictus. On the contrary, Ae. aegypti were found to be resistant in all three agroecosystem sites except for banana plantation sites, where full susceptibility was observed. Molecular analysis revealed that individual target site resistance kdr mutations F1534C and V1016G/I were detected in most Ae. aegypti populations. Additionally, double mutant (F1534C + V1016G/I) mosquitoes were found in some populations, and in one case, triple mutant (F1534C + V1016G/I + S989P) mosquitoes were detected. Metabolic resistance, as reflected by overexpression of three P450 genes (CYP6BB2, CYP9J26, and CYP9J32), was also detected in Ae. aegypti mosquitoes. Our study provides information that could be used to strategize future vector control strategies and highlights the importance of continuing vector surveillance. Future studies should assess the effect of piperonyl butoxide (PBO) on metabolic resistance and identify the different strains of Wolbachia spp., to choose the best vector control strategies in Benin.}, } @article {pmid38460359, year = {2024}, author = {Zurita, A and Trujillo, I and Cutillas, C}, title = {New records of pathogenic bacteria in different species of fleas collected from domestic and peridomestic animals in Spain. A potential zoonotic threat?.}, journal = {Comparative immunology, microbiology and infectious diseases}, volume = {107}, number = {}, pages = {102153}, doi = {10.1016/j.cimid.2024.102153}, pmid = {38460359}, issn = {1878-1667}, mesh = {Animals ; Dogs ; *Siphonaptera/microbiology ; Spain/epidemiology ; *Ctenocephalides/genetics ; *Rickettsia felis/genetics ; *Flea Infestations/epidemiology/veterinary/microbiology ; *Bartonella/genetics ; *Rickettsia ; *Felis ; }, abstract = {Climate change is causing many vectors of infectious diseases to expand their geographic distribution as well as the pathogens they transmit are also conditioned by temperature for their multiplication. Within this context, it is worth highlighting the significant role that fleas can play as vectors of important pathogenic bacteria. For this purpose, our efforts focused on detecting and identifying a total of 9 bacterial genera (Rickettsia sp.; Bartonella sp.; Yersinia sp.; Wolbachia sp., Mycobacterium sp., Leishmania sp., Borrelia sp., Francisella sp. and Coxiella sp.) within fleas isolated from domestic and peridomestic animals in the southwestern region of Spain (Andalusia). Over a 19-months period, we obtained flea samples from dogs, cats and hedgehogs. A total of 812 fleas was collected for this study. Five different species were morphologically identified, including C. felis, C. canis, S. cuniculi, P. irritans, and A. erinacei. Wolbachia sp. was detected in all five species identified in our study which a total prevalence of 86%. Within Rickettsia genus, two different species, R. felis and R. asembonensis were mainly identified in C. felis and A. erinacei, respectively. On the other hand, our results revealed a total of 131 fleas testing positive for the presence of Bartonella sp., representing a prevalence rate of 16% for this genus identifying two species B. henselae and B. clarridgeiae. Lastly, both Y. pestis and L. infantum were detected in DNA of P. irritans and C. felis, respectively isolated from dogs. With these data we update the list of bacterial zoonotic agents found in fleas in Spain, emphasizing the need to continue conducting future experimental studies to assess and confirm the potential vectorial role of certain synanthropic fleas.}, } @article {pmid38458883, year = {2024}, author = {Moore, CO and André, MR and Šlapeta, J and Breitschwerdt, EB}, title = {Vector biology of the cat flea Ctenocephalides felis.}, journal = {Trends in parasitology}, volume = {40}, number = {4}, pages = {324-337}, pmid = {38458883}, issn = {1471-5007}, support = {T32 GM133366/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Cats ; *Ctenocephalides/microbiology ; *Flea Infestations/veterinary/epidemiology/microbiology ; *Rickettsia ; *Bartonella ; Biology ; *Felis ; *Rickettsia felis/genetics ; *Cat Diseases ; *Siphonaptera/microbiology ; }, abstract = {Ctenocephalides felis, the cat flea, is among the most prevalent and widely dispersed vectors worldwide. Unfortunately, research on C. felis and associated pathogens (Bartonella and Rickettsia spp.) lags behind that of other vectors and vector-borne pathogens. Therefore, we aimed to review fundamental aspects of C. felis as a vector (behavior, epidemiology, phylogenetics, immunology, and microbiome composition) with an emphasis on key techniques and research avenues employed in other vector species. Future laboratory C. felis experimental infections with Bartonella, Rickettsia, and Wolbachia species/strains should examine the vector-pathogen interface utilizing contemporary visualization, transcriptomic, and gene-editing techniques. Further environmental sampling will inform the range and prevalence of C. felis and associated pathogens, improving the accuracy of vector and pathogen modeling to improve infection/infestation risk assessment and diagnostic recommendations.}, } @article {pmid38454541, year = {2024}, author = {Carlassara, M and Khorramnejad, A and Oker, H and Bahrami, R and Lozada-Chávez, AN and Mancini, MV and Quaranta, S and Body, MJA and Lahondère, C and Bonizzoni, M}, title = {Population-specific responses to developmental temperature in the arboviral vector Aedes albopictus: Implications for climate change.}, journal = {Global change biology}, volume = {30}, number = {3}, pages = {e17226}, doi = {10.1111/gcb.17226}, pmid = {38454541}, issn = {1365-2486}, support = {//2022J45MLL/ ; PE00000007//NextGeneration EU-MUR PNRR Extended Partnership/ ; }, mesh = {Animals ; *Climate Change ; Temperature ; *Aedes/physiology ; Global Warming ; Larva/physiology ; }, abstract = {The increase of environmental temperature due to current global warming is not only favouring the expansion of the distribution range of many insect species, but it is also changing their phenology. Insect phenology is tightly linked to developmental timing, which is regulated by environmental temperatures. However, the degree to which the effects of developmental temperatures extend across developmental stages and their inter-stage relationships have not been thoroughly quantified in mosquitoes. Here, we used the mosquito Aedes albopictus, which is an aggressive invasive species and an arboviral vector, to study how developmental temperature influences fitness across developmental stages, thermal traits, energy reserves, transcriptome and Wolbachia prevalence in laboratory-reared populations originally collected from either temperate or tropical regions. We show that hatchability, larval and pupal viability and developmental speed are strongly influenced by temperature, and these effects extend to wing length, body mass, longevity and content of water, protein and lipids in adults in a population-specific manner. On the contrary, neither adult thermal preference nor heat resistance significantly change with temperature. Wolbachia density was generally lower in adult mosquitoes reared at 18°C than at other tested temperatures, and transcriptome analysis showed enrichment for functions linked to stress responses (i.e. cuticle proteins and chitin, cytochrome p450 and heat shock proteins) in mosquitoes reared at both 18 and 32°C. Our data showed an overall reduced vector fitness performance when mosquitoes were reared at 32°C, and the absence of isomorphy in the relationship between developmental stages and temperature in the laboratory population deriving from larvae collected in northern Italy. Altogether, these results have important implications for reliable model projections of the invasion potentials of Ae. albopictus and its epidemiological impact.}, } @article {pmid38452081, year = {2024}, author = {Kaur, R and McGarry, A and Shropshire, JD and Leigh, BA and Bordenstein, SR}, title = {Prophage proteins alter long noncoding RNA and DNA of developing sperm to induce a paternal-effect lethality.}, journal = {Science (New York, N.Y.)}, volume = {383}, number = {6687}, pages = {1111-1117}, pmid = {38452081}, issn = {1095-9203}, support = {F32 AI140694/AI/NIAID NIH HHS/United States ; R01 AI132581/AI/NIAID NIH HHS/United States ; R01 AI143725/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Male ; Cytoplasm/metabolism ; DNA/metabolism ; *Prophages/genetics/metabolism ; *RNA, Long Noncoding/metabolism ; *Spermatozoa/growth & development/metabolism ; *Wolbachia/metabolism/virology ; *Paternal Inheritance ; *Viral Proteins/metabolism ; *Drosophila melanogaster/genetics/microbiology ; *Bacterial Proteins/metabolism ; *Deoxyribonucleases/metabolism ; }, abstract = {The extent to which prophage proteins interact with eukaryotic macromolecules is largely unknown. In this work, we show that cytoplasmic incompatibility factor A (CifA) and B (CifB) proteins, encoded by prophage WO of the endosymbiont Wolbachia, alter long noncoding RNA (lncRNA) and DNA during Drosophila sperm development to establish a paternal-effect embryonic lethality known as cytoplasmic incompatibility (CI). CifA is a ribonuclease (RNase) that depletes a spermatocyte lncRNA important for the histone-to-protamine transition of spermiogenesis. Both CifA and CifB are deoxyribonucleases (DNases) that elevate DNA damage in late spermiogenesis. lncRNA knockdown enhances CI, and mutagenesis links lncRNA depletion and subsequent sperm chromatin integrity changes to embryonic DNA damage and CI. Hence, prophage proteins interact with eukaryotic macromolecules during gametogenesis to create a symbiosis that is fundamental to insect evolution and vector control.}, } @article {pmid38450861, year = {2024}, author = {Urakova, N and Joseph, RE and Huntsinger, A and Macias, VM and Jones, MJ and Sigle, LT and Li, M and Akbari, OS and Xi, Z and Lymperopoulos, K and Sayre, RT and McGraw, EA and Rasgon, JL}, title = {Alpha-mannosidase-2 modulates arbovirus infection in a pathogen- and Wolbachia-specific manner in Aedes aegypti mosquitoes.}, journal = {Insect molecular biology}, volume = {33}, number = {4}, pages = {362-371}, pmid = {38450861}, issn = {1365-2583}, support = {1645331//National Science Foundation/ ; R01 AI151004/AI/NIAID NIH HHS/United States ; HR0011-17-2-0047//Defense Advanced Research Projects Agency/ ; R01AI150251/NH/NIH HHS/United States ; //Pebble Labs/ ; //Dorothy Foehr Huck and J. Lloyd Huck Endowment/ ; R01 AI143758/AI/NIAID NIH HHS/United States ; R01AI116636/NH/NIH HHS/United States ; R21 AI156078/AI/NIAID NIH HHS/United States ; R01 AI150251/AI/NIAID NIH HHS/United States ; R01 AI116636/AI/NIAID NIH HHS/United States ; R01AI143758/NH/NIH HHS/United States ; R21AI156078/NH/NIH HHS/United States ; R01AI151004/NH/NIH HHS/United States ; AID-OAA-F-16-00082//United States Agency for International Development/ ; DP2 AI152071/AI/NIAID NIH HHS/United States ; DP2AI152071/NH/NIH HHS/United States ; }, mesh = {Animals ; *Aedes/microbiology/virology/genetics ; *Wolbachia/physiology ; *alpha-Mannosidase/metabolism/genetics ; Dengue Virus/physiology ; Arboviruses/physiology ; Mosquito Vectors/microbiology/virology/genetics ; Female ; Arbovirus Infections/transmission ; Insect Proteins/metabolism/genetics ; CRISPR-Cas Systems ; }, abstract = {Multiple Wolbachia strains can block pathogen infection, replication and/or transmission in Aedes aegypti mosquitoes under both laboratory and field conditions. However, Wolbachia effects on pathogens can be highly variable across systems and the factors governing this variability are not well understood. It is increasingly clear that the mosquito host is not a passive player in which Wolbachia governs pathogen transmission phenotypes; rather, the genetics of the host can significantly modulate Wolbachia-mediated pathogen blocking. Specifically, previous work linked variation in Wolbachia pathogen blocking to polymorphisms in the mosquito alpha-mannosidase-2 (αMan2) gene. Here we use CRISPR-Cas9 mutagenesis to functionally test this association. We developed αMan2 knockouts and examined effects on both Wolbachia and virus levels, using dengue virus (DENV; Flaviviridae) and Mayaro virus (MAYV; Togaviridae). Wolbachia titres were significantly elevated in αMan2 knockout (KO) mosquitoes, but there were complex interactions with virus infection and replication. In Wolbachia-uninfected mosquitoes, the αMan2 KO mutation was associated with decreased DENV titres, but in a Wolbachia-infected background, the αMan2 KO mutation significantly increased virus titres. In contrast, the αMan2 KO mutation significantly increased MAYV replication in Wolbachia-uninfected mosquitoes and did not affect Wolbachia-mediated virus blocking. These results demonstrate that αMan2 modulates arbovirus infection in A. aegypti mosquitoes in a pathogen- and Wolbachia-specific manner, and that Wolbachia-mediated pathogen blocking is a complex phenotype dependent on the mosquito host genotype and the pathogen. These results have a significant impact for the design and use of Wolbachia-based strategies to control vector-borne pathogens.}, } @article {pmid38450576, year = {2024}, author = {Mancini, MV and Murdochy, SM and Bilgo, E and Ant, TH and Gingell, D and Gnambani, EJ and Failloux, AB and Diabate, A and Sinkins, SP}, title = {Wolbachia strain wAlbB shows favourable characteristics for dengue control use in Aedes aegypti from Burkina Faso.}, journal = {Environmental microbiology}, volume = {26}, number = {3}, pages = {e16588}, doi = {10.1111/1462-2920.16588}, pmid = {38450576}, issn = {1462-2920}, support = {AV/PP0025/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; 108508/WT_/Wellcome Trust/United Kingdom ; 226166/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Animals ; Burkina Faso/epidemiology ; *Aedes ; *Wolbachia/genetics ; Asia ; *Dengue/prevention & control ; }, abstract = {Dengue represents an increasing public health burden worldwide. In Africa, underreporting and misdiagnosis often mask its true epidemiology, and dengue is likely to be both more widespread than reported data suggest and increasing in incidence and distribution. Wolbachia-based dengue control is underway in Asia and the Americas but has not to date been deployed in Africa. Due to the genetic heterogeneity of African Aedes aegypti populations and the complexity of the host-symbiont interactions, characterization of key parameters of Wolbachia-carrying mosquitoes is paramount for determining the potential of the system as a control tool for dengue in Africa. The wAlbB Wolbachia strain was stably introduced into an African Ae. aegypti population by introgression, and showed high intracellular density in whole bodies and different mosquito tissues; high intracellular density was also maintained following larval rearing at high temperatures. No effect on the adult lifespan induced by Wolbachia presence was detected. Moreover, the ability of this strain to strongly inhibit DENV-2 dissemination and transmission in the host was also demonstrated in the African background. Our findings suggest the potential of harnessing Wolbachia for dengue control for African populations of Ae. aegypti.}, } @article {pmid38446807, year = {2024}, author = {Pérez-Guerra, CL and Rosado-Santiago, C and Ramos, SA and Marrero-Santos, KM and González-Zeno, G and Partridge, SK and Rivera-Amill, V and Paz-Bailey, G and Sánchez-González, L and Hayden, MH}, title = {Acceptability of emergent Aedes aegypti vector control methods in Ponce, Puerto Rico: A qualitative assessment.}, journal = {PLOS global public health}, volume = {4}, number = {3}, pages = {e0002744}, pmid = {38446807}, issn = {2767-3375}, support = {U01 CK000580/CK/NCEZID CDC HHS/United States ; U01CK000437/ACL/ACL HHS/United States ; }, abstract = {Aedes aegypti control has been fraught with challenges in Puerto Rico. The government has implemented commonly used vector control methods, but arboviral epidemics still occur. It is necessary to explore new Ae. aegypti control methods. This study aimed to understand the perceptions of community members in Ponce, Puerto Rico about emergent and traditional Ae. aegypti vector control methods and determine their acceptability and support for these methods. We identified the type of information needed to increase support for emergent vector control methods, and the preferred strategies to disseminate this information. Four group discussions were conducted with a total of 32 participants representing eight of the 14 clusters participating in the Communities Organized for the Prevention of Arboviruses (COPA), a project designed to mobilize communities in Ponce, Puerto Rico to prevent diseases transmitted by mosquitoes. Group discussions began with an overview of different methods used for controlling Ae. aegypti mosquitoes. These overviews facilitated participant understanding of the mosquito control methods presented. Use of source reduction, autocidal gravid ovitraps (AGO), and manual application of larvicide for arboviral mosquito control received support from almost all participants. Vector control methods that use more familiar techniques in Puerto Rico such as truck-mounted larvicide spraying (TMLS) and insecticide residual spraying received support from most participants. More than half of participants supported the use of emergent mosquito control methods including Wolbachia suppression, Wolbachia replacement, or genetically modified mosquitoes (GMM). Participants preferred to receive vector control information through house-to-house visits with the distribution of written materials, followed by dissemination of information through traditional (i.e., radio, television) and social media. The detailed information resulting from this study was used to develop messages for a communications campaign to garner future community support. Community acceptance and support are critical for the success of vector control programs using emergent mosquito control methods.}, } @article {pmid38437189, year = {2024}, author = {Dye, D and Cain, JW}, title = {Efficacy of Wolbachia-based mosquito control: Predictions of a spatially discrete mathematical model.}, journal = {PloS one}, volume = {19}, number = {3}, pages = {e0297964}, pmid = {38437189}, issn = {1932-6203}, mesh = {Female ; Humans ; Animals ; Male ; *Wolbachia ; *Aedes ; *Charadriiformes ; Cytoplasm ; Cytosol ; }, abstract = {Wolbachia is an endosymbiont bacterium present in many insect species. When Wolbachia-carrying male Aedes aegypti mosquitoes mate with non-carrier females, their embryos are not viable due to cytoplasmic incompatibility. This phenomenon has been exploited successfully for the purpose of controlling mosquito populations and the spread of mosquito-borne illnesses: Wolbachia carriers are bred and released into the environment. Because Wolbachia is not harmful to humans, this method of mosquito control is regarded as a safer alternative to pesticide spraying. In this article, we introduce a mathematical framework for exploring (i) whether a one-time release of Wolbachia carriers can elicit a sustained presence of carriers near the release site, and (ii) the extent to which spatial propagation of carriers may allow them to establish fixation in other territories. While some prior studies have formulated mosquito dispersal models using advection-reaction-diffusion PDEs, the predictive power of such models requires careful ecological mapping: advection and diffusion coefficients exhibit significant spatial dependence due to heterogeneity of resources and topography. Here, we adopt a courser-grained view, regarding the environment as a network of discrete, diffusively-coupled "habitats"-distinct zones of high mosquito density such as stagnant ponds. We extend two previously published single-habitat mosquito models to multiple habitats, and calculate rates of migration between pairs of habitats using dispersal kernels. Our primary results are quantitative estimates regarding how the success of carrier fixation in one or more habitats is determined by: the number of carriers released, sizes of habitats, distances between habitats, and the rate of migration between habitats. Besides yielding sensible and potentially useful predictions regarding the success of Wolbachia-based control, our framework applies to other approaches (e.g., gene drives) and contexts beyond the realm of insect pest control.}, } @article {pmid38431055, year = {2024}, author = {Walt, HK and King, JG and Sheele, JM and Meyer, F and Pietri, JE and Hoffmann, FG}, title = {Do bed bugs transmit human viruses, or do humans spread bed bugs and their viruses? A worldwide survey of the bed bug RNA virosphere.}, journal = {Virus research}, volume = {343}, number = {}, pages = {199349}, pmid = {38431055}, issn = {1872-7492}, mesh = {Animals ; Humans ; *Bedbugs ; Feeding Behavior ; Disease Vectors ; *Arthropods ; *Viruses ; }, abstract = {BED BUGS: (Hemiptera: Cimicidae) are a globally distributed hematophagous pest that routinely feed on humans. Unlike many blood-sucking arthropods, they have never been linked to pathogen transmission in a natural setting, and despite increasing interest in their role as disease vectors, little is known about the viruses that bed bugs naturally harbor. Here, we present a global-scale survey of the bed bug RNA virosphere. We sequenced the metatranscriptomes of 22 individual bed bugs (Cimex lectularius and Cimex hemipterus) from 8 locations around the world. We detected sequences from two known bed bug viruses (Shuangao bedbug virus 1 and Shuangao bedbug virus 2) which extends their geographical range. We identified three novel bed bug virus sequences from a tenui-like virus (Bunyavirales), a toti-like virus (Ghabrivirales), and a luteo-like virus (Tolivirales). Interestingly, some of the bed bug viruses branch near to insect-transmitted plant-infecting viruses, opening questions regarding the evolution of plant virus infection. When we analyzed the viral sequences by their host's collection location, we found unexpected patterns of geographical diversity that may reflect humans' role in bed bug dispersal. Additionally, we investigated the effect that Wolbachia, the primary bed bug endosymbiont, may have on viral abundance and found that Wolbachia infection neither promotes nor inhibits viral infection. Finally, our results provide no evidence that bed bugs transmit any known human pathogenic viruses.}, } @article {pmid38427588, year = {2024}, author = {Aldridge, RL and Gibson, S and Linthicum, KJ}, title = {Aedes aegypti Controls AE. Aegypti: SIT and IIT-An Overview.}, journal = {Journal of the American Mosquito Control Association}, volume = {40}, number = {1}, pages = {32-49}, doi = {10.2987/23-7154}, pmid = {38427588}, issn = {1943-6270}, mesh = {Animals ; Humans ; *Aedes ; Mosquito Control/methods ; *Wolbachia ; Mosquito Vectors ; Insecta ; }, abstract = {The sterile insect technique (SIT) and the incompatible insect technique (IIT) are emerging and potentially revolutionary tools for controlling Aedes aegypti (L.), a prominent worldwide mosquito vector threat to humans that is notoriously difficult to reduce or eliminate in intervention areas using traditional integrated vector management (IVM) approaches. Here we provide an overview of the discovery, development, and application of SIT and IIT to Ae. aegypti control, and innovations and advances in technology, including transgenics, that could elevate these techniques to a worldwide sustainable solution to Ae. aegypti when combined with other IVM practices.}, } @article {pmid38426058, year = {2024}, author = {Cantin, LJ and Dunning Hotopp, JC and Foster, JM}, title = {Improved metagenome assemblies through selective enrichment of bacterial genomic DNA from eukaryotic host genomic DNA using ATAC-seq.}, journal = {Frontiers in microbiology}, volume = {15}, number = {}, pages = {1352378}, pmid = {38426058}, issn = {1664-302X}, abstract = {Genomics can be used to study the complex relationships between hosts and their microbiota. Many bacteria cannot be cultured in the laboratory, making it difficult to obtain adequate amounts of bacterial DNA and to limit host DNA contamination for the construction of metagenome-assembled genomes (MAGs). For example, Wolbachia is a genus of exclusively obligate intracellular bacteria that live in a wide range of arthropods and some nematodes. While Wolbachia endosymbionts are frequently described as facultative reproductive parasites in arthropods, the bacteria are obligate mutualistic endosymbionts of filarial worms. Here, we achieve 50-fold enrichment of bacterial sequences using ATAC-seq (Assay for Transposase-Accessible Chromatin using sequencing) with Brugia malayi nematodes, containing Wolbachia (wBm). ATAC-seq uses the Tn5 transposase to cut and attach Illumina sequencing adapters to accessible DNA lacking histones, typically thought to be open chromatin. Bacterial and mitochondrial DNA in the lysates are also cut preferentially since they lack histones, leading to the enrichment of these sequences. The benefits of this include minimal tissue input (<1 mg of tissue), a quick protocol (<4 h), low sequencing costs, less bias, correct assembly of lateral gene transfers and no prior sequence knowledge required. We assembled the wBm genome with as few as 1 million Illumina short paired-end reads with >97% coverage of the published genome, compared to only 12% coverage with the standard gDNA libraries. We found significant bacterial sequence enrichment that facilitated genome assembly in previously published ATAC-seq data sets from human cells infected with Mycobacterium tuberculosis and C. elegans contaminated with their food source, the OP50 strain of E. coli. These results demonstrate the feasibility and benefits of using ATAC-seq to easily obtain bacterial genomes to aid in symbiosis, infectious disease, and microbiome research.}, } @article {pmid38412904, year = {2024}, author = {Liu, K and Fang, S and Li, Q and Lou, Y}, title = {Effectiveness evaluation of mosquito suppression strategies on dengue transmission under changing temperature and precipitation.}, journal = {Acta tropica}, volume = {253}, number = {}, pages = {107159}, doi = {10.1016/j.actatropica.2024.107159}, pmid = {38412904}, issn = {1873-6254}, mesh = {Animals ; Male ; *Dengue/epidemiology ; Temperature ; *Aedes ; Mosquito Control ; Models, Theoretical ; Mosquito Vectors ; *Wolbachia ; }, abstract = {Widespread resurgence of dengue outbreaks has seriously threatened the global health. Due to lack of treatments and vaccines, one key strategy in dengue control is to reduce the vector population size. As an environment-friendly mosquito control approach, releasing male mosquitoes transinfected with specific Wolbachia strain into the field to suppress the wild mosquito population size has become wildly accepted. The current study evaluates the effectiveness of this suppression strategy on dengue control under changing temperature and precipitation profiles. We formulate a mathematical model which includes larval intra-specific competition, the maturation period for mosquitoes, the extrinsic incubation period (EIP) and intrinsic incubation period (IIP). The persistence of mosquitoes and disease is discussed in terms of two basic reproduction numbers (RM and R0) and the release ratio pw. Further numerical simulations are carried out to not only validate theoretical results, but also provide interesting quantitative observations. Sensitivity analysis on the reproduction numbers, peak size, peak time and the final epidemic size is performed with respect to model parameters, which highlights effective control measures against dengue transmission. Moreover, by assuming temperature and precipitation dependent mosquito-related parameters, the model can be used to project the effectiveness of releasing Wolbachia-carrying males under climatic variations. It is shown that the effectiveness of various control strategies is highly dependent on the changing temperature and precipitation profiles. In particular, the model projects that it is most challenging to control the disease at the favorable temperature (around 27∼30[∘]C) and precipitation (5∼8mm/day) range, during which the basic reproduction number R0 is very high and more Wolbachia-infected males should be released.}, } @article {pmid38404602, year = {2024}, author = {Yan, J and Green, K and Noel, K and Kim, CH and Stone, CM}, title = {Effects of seasonality and developed land cover on Culex mosquito abundance and microbiome diversity.}, journal = {Frontiers in microbiology}, volume = {15}, number = {}, pages = {1332970}, pmid = {38404602}, issn = {1664-302X}, abstract = {The vectorial capacity of mosquitoes, which influences the dynamics of vector-borne disease transmission, is intricately linked to mosquito abundance and the composition and diversity of their associated microbiomes. However, the influence of environmental factors on mosquito populations and microbiome diversity remains underexplored. Here we examined the effects of seasonality and developed land cover on Culex mosquito abundance and bacterial diversity. Biweekly field sampling of female Culex mosquitoes was conducted using gravid and CDC light traps, spanning summer to autumn across varying developed land cover levels in two urban areas in Central Illinois. Mosquito abundance was assessed by the number of mosquitoes captured per trap night and compared across seasons and developed levels. The mean mosquito abundance for gravid and light traps was 12.96 ± 2.15 and 7.67 ± 1.44, respectively. Notably, higher levels of developed land cover exhibited higher Culex abundance than the low level for light traps, but no significant difference was found between summer and early autumn. In gravid traps, no significant differences were detected across seasons or developed levels. Microbial analysis of the mosquito microbiome revealed that Proteobacteria and Wolbachia, with a mean relative abundance of 80.77 and 52.66% respectively, were identified as the most dominant bacterial phylum and genus. Their relative abundance remained consistent across seasons and developed land cover levels, with negligible variations. Alpha diversity, as measured by observed species, Chao1, Shannon, and Simpson, showed slightly higher values in early-autumn compared to late-summer. A notable pattern of bacterial diversity, as indicated by all four diversity indexes, is evident across varying levels of land development. Significantly, high or intermediate developed levels consistently showed reduced alpha diversity when compared to the lower level. This underscores the pronounced impact of anthropogenic ecological disturbances in shaping mosquito microbiomes. Beta diversity analysis revealed no significant dissimilarities in bacterial community composition across seasons and developed levels, although some separation was noted among different levels of developed land cover. These findings highlight the significant role of environmental factors in shaping mosquito abundance and their associated microbiomes, with potential implications for the vectorial capacity in the transmission of vector-borne diseases.}, } @article {pmid38399990, year = {2024}, author = {Corrêa-Antônio, J and David, MR and Couto-Lima, D and Garcia, GA and Keirsebelik, MSG and Maciel-de-Freitas, R and Pavan, MG}, title = {DENV-1 Titer Impacts Viral Blocking in wMel Aedes aegypti with Brazilian Genetic Background.}, journal = {Viruses}, volume = {16}, number = {2}, pages = {}, pmid = {38399990}, issn = {1999-4915}, support = {E26/2001.844/2017//Fundação Carlos Chagas Filho de Amparo à Pesquisa do Estado do Rio de Janeiro/ ; E26/210.335/2022//Fundação Carlos Chagas Filho de Amparo à Pesquisa do Estado do Rio de Janeiro/ ; E26/201.370/2022//Fundação Carlos Chagas Filho de Amparo à Pesquisa do Estado do Rio de Janeiro/ ; E26/211.159/2019//Fundação Carlos Chagas Filho de Amparo à Pesquisa do Estado do Rio de Janeiro/ ; finance code 001//Coordenação de Aperfeicoamento de Pessoal de Nível Superior/ ; }, mesh = {Animals ; Humans ; *Aedes ; *Dengue Virus/genetics ; Brazil ; Mosquito Vectors ; *Zika Virus ; *Zika Virus Infection ; *Wolbachia/genetics ; *Dengue ; }, abstract = {Several countries have been using Wolbachia deployments to replace highly competent native Aedes aegypti populations with Wolbachia-carrying mosquitoes with lower susceptibility to arboviruses such as dengue, Zika, and chikungunya. In Rio de Janeiro, Wolbachia deployments started in 2015 and still present a moderate introgression with a modest reduction in dengue cases in humans (38%). Here, we evaluated the vector competence of wild-type and wMel-infected Ae. aegypti with a Brazilian genetic background to investigate whether virus leakage could contribute to the observed outcomes in Brazil. We collected the specimens in three areas of Rio de Janeiro with distinct frequencies of mosquitoes with wMel strain and two areas with wild Ae. aegypti. The mosquitoes were orally exposed to two titers of DENV-1 and the saliva of DENV-1-infected Ae. aegypti was microinjected into wMel-free mosquitoes to check their infectivity. When infected with the high DENV-1 titer, the presence of wMel did not avoid viral infection in mosquitoes' bodies and saliva but DENV-1-infected wMel mosquitoes produced lower viral loads than wMel-free mosquitoes. On the other hand, wMel mosquitoes infected with the low DENV-1 titer were less susceptible to virus infection than wMel-free mosquitoes, although once infected, wMel and wMel-free mosquitoes exhibited similar viral loads in the body and the saliva. Our results showed viral leakage in 60% of the saliva of wMel mosquitoes with Brazilian background; thus, sustained surveillance is imperative to monitor the presence of other circulating DENV-1 strains capable of overcoming the Wolbachia blocking phenotype, enabling timely implementation of action plans.}, } @article {pmid38399310, year = {2024}, author = {Voronin, D and Tricoche, N and Peguero, R and Kaminska, AM and Ghedin, E and Sakanari, JA and Lustigman, S}, title = {Repurposed Drugs That Activate Autophagy in Filarial Worms Act as Effective Macrofilaricides.}, journal = {Pharmaceutics}, volume = {16}, number = {2}, pages = {}, pmid = {38399310}, issn = {1999-4923}, support = {R21AI153649//National Institute of Allergy and Infectious Diseases/ ; ZIA-AI001322//Division of Intramural Research of the NIAID/NIH/ ; }, abstract = {Onchocerciasis and lymphatic filariasis are two neglected tropical diseases caused by filarial nematodes that utilize insect vectors for transmission to their human hosts. Current control strategies are based on annual or biannual mass drug administration (MDA) of the drugs Ivermectin or Ivermectin plus Albendazole, respectively. These drug regimens kill the first-stage larvae of filarial worms (i.e., microfilariae) and interrupt the transmission of infections. MDA programs for these microfilaricidal drugs must be given over the lifetime of the filarial adult worms, which can reach 15 years in the case of Onchocerca volvulus. This is problematic because of suboptimal responses to ivermectin in various endemic regions and inefficient reduction of transmission even after decades of MDA. There is an urgent need for the development of novel alternative treatments to support the 2030 elimination goals of onchocerciasis and lymphatic filariasis. One successful approach has been to target Wolbachia, obligatory endosymbiotic bacteria on which filarial worms are dependent for their survival and reproduction within the human host. A 4-6-week antibiotic therapy with doxycycline, for example, resulted in the loss of Wolbachia that subsequently led to extensive apoptosis of somatic cells, germline, embryos, and microfilariae, as well as inhibition of fourth-stage larval development. However, this long-course regimen has limited use in MDA programs. As an alternative approach to the use of bacteriostatic antibiotics, in this study, we focused on autophagy-inducing compounds, which we hypothesized could disturb various pathways involved in the interdependency between Wolbachia and filarial worms. We demonstrated that several such compounds, including Niclosamide, an FDA-approved drug, Niclosamide ethanolamine (NEN), and Rottlerin, a natural product derived from Kamala trees, significantly reduced the levels of Wolbachia in vitro. Moreover, when these compounds were used in vivo to treat Brugia pahangi-infected gerbils, Niclosamide and NEN significantly decreased adult worm survival, reduced the release of microfilariae, and decreased embryonic development depending on the regimen and dose used. All three drugs given orally significantly reduced Wolbachia loads and induced an increase in levels of lysosome-associated membrane protein in worms from treated animals, suggesting that Niclosamide, NEN, and Rottlerin were effective in causing drug-induced autophagy in these filarial worms. These repurposed drugs provide a new avenue for the clearance of adult worms in filarial infections.}, } @article {pmid38394822, year = {2024}, author = {Zhang, X and Ferree, PM}, title = {PSRs: Selfish chromosomes that manipulate reproductive development.}, journal = {Seminars in cell & developmental biology}, volume = {159-160}, number = {}, pages = {66-73}, doi = {10.1016/j.semcdb.2024.01.008}, pmid = {38394822}, issn = {1096-3634}, mesh = {Humans ; Animals ; Male ; Female ; *Wasps/genetics ; Semen ; Chromosomes/genetics ; Genome ; Base Sequence ; }, abstract = {B chromosomes are intriguing "selfish" genetic elements, many of which exhibit higher-than-Mendelian transmission. This perspective highlights a group of B chromosomes known as Paternal Sex Ratio chromosomes (PSRs), which are found in several insects with haplo-diploid reproduction. PSRs harshly alter the organism's reproduction to facilitate their own inheritance. A manifestation of this effect is the conversion of female destined individuals into males. Key to this conversion is the mysterious ability of PSRs to cause elimination of the sperm-inherited half of the genome during zygote formation. Here we discuss how PSRs were discovered, what is known about how they alter paternal chromatin dynamics to cause sex conversion, and how PSR-induced genome elimination is different from other forms of programmed genome elimination in different insects. PSRs also stand out because their DNA sequence compositions differ in remarkable ways from their insect's essential chromosomes, a characteristic suggestive of interspecies origins. Broadly, we also highlight poorly understood aspects of PSR dynamics that need to be investigated.}, } @article {pmid38392845, year = {2024}, author = {Lamichhane, B and Brockway, C and Evasco, K and Nicholson, J and Neville, PJ and Levy, A and Smith, D and Imrie, A}, title = {Metatranscriptomic Sequencing of Medically Important Mosquitoes Reveals Extensive Diversity of RNA Viruses and Other Microbial Communities in Western Australia.}, journal = {Pathogens (Basel, Switzerland)}, volume = {13}, number = {2}, pages = {}, pmid = {38392845}, issn = {2076-0817}, abstract = {Mosquitoes harbor a wide diversity of microorganisms, including viruses that are human pathogens, or that are insect specific. We used metatranscriptomics, an unbiased high-throughput molecular approach, to describe the composition of viral and other microbial communities in six medically important mosquito species from across Western Australia: Aedes vigilax, Culex annulirostris, Cx. australicus, Cx. globocoxitus, Cx. pipiens biotype molestus, and Cx. quinquefasciatus. We identified 42 viral species, including 13 novel viruses, from 19 families. Culex mosquitoes exhibited a significantly higher diversity of viruses than Aedes mosquitoes, and no virus was shared between the two genera. Comparison of mosquito populations revealed a heterogenous distribution of viruses between geographical regions and between closely related species, suggesting that geography and host species may play a role in shaping virome composition. We also detected bacterial and parasitic microorganisms, among which Wolbachia bacteria were detected in three members of the Cx. pipiens complex, Cx. australicus, Cx. pipiens biotype molestus, and Cx. quinquefasciatus. In summary, our unbiased metatranscriptomics approach provides important insights into viral and other microbial diversity in Western Australian mosquitoes that vector medically important viruses.}, } @article {pmid38392551, year = {2024}, author = {Zhu, X and Li, J and He, A and Gurr, GM and You, M and You, S}, title = {Developmental Shifts in the Microbiome of a Cosmopolitan Pest: Unraveling the Role of Wolbachia and Dominant Bacteria.}, journal = {Insects}, volume = {15}, number = {2}, pages = {}, pmid = {38392551}, issn = {2075-4450}, support = {2022J06013//The Fujian Natural Science Fund for Distinguished Young Scholars/ ; 2022L3087//Central Guided Local Science and Technology Development Project/ ; }, abstract = {Wolbachia bacteria (phylum Proteobacteria) are ubiquitous intracellular parasites of diverse invertebrates. In insects, coevolution has forged mutualistic associations with Wolbachia species, influencing reproduction, immunity, development, pathogen resistance, and overall fitness. However, the impact of Wolbachia on other microbial associates within the insect microbiome, which are crucial for host fitness, remains less explored. The diamondback moth (Plutella xylostella), a major pest of cruciferous vegetables worldwide, harbors the dominant Wolbachia strain plutWB1, known to distort its sex ratio. This study investigated the bacterial community diversity and dynamics across different developmental life stages and Wolbachia infection states in P. xylostella using high-throughput 16S rDNA amplicon sequencing. Proteobacteria and Firmicutes dominated the P. xylostella microbiome regardless of life stage or Wolbachia infection. However, the relative abundance of dominant genera, including an unclassified genus of Enterobacteriaceae, Wolbachia, Carnobacterium, and Delftia tsuruhatensis, displayed significant stage-specific variations. While significant differences in bacterial diversity and composition were observed across life stages, Wolbachia infection had no substantial impact on overall diversity. Nonetheless, relative abundances of specific genera differed between infection states. Notably, Wolbachia exhibited a stable, high relative abundance across all stages and negatively correlated with an unclassified genus of Enterobacteriaceae, Delftia tsuruhatensis, and Carnobacterium. Our findings provide a foundational understanding of the complex interplay between the host, Wolbachia, and the associated microbiome in P. xylostella, paving the way for a deeper understanding of their complex interactions and potential implications for pest control strategies.}, } @article {pmid38392506, year = {2024}, author = {Budrys, E and Orlovskytė, S and Budrienė, A}, title = {Ecological Speciation without Morphological Differentiation? A New Cryptic Species of Diodontus Curtis (Hymenoptera, Pemphredonidae) from the Centre of Europe.}, journal = {Insects}, volume = {15}, number = {2}, pages = {}, pmid = {38392506}, issn = {2075-4450}, support = {Contract No S-MIP-20-23//Lietuvos Mokslo Taryba/ ; }, abstract = {Upon exploring the mitotype diversity of the aphid-hunting wasp, Diodontus tristis, we revealed specimens with highly divergent mitotypes from two localities in Lithuania and nesting in clayey substrate, while the specimens with typical mitotypes were found nesting in sandy sites. The comparison of inter- and intra-specific distances and application of delimitation algorithms supported the species status of the clay-nesting populations. Using a set of DNA markers that included complete or partial sequences of six mitochondrial genes, three markers of ribosomal operon, two homeobox genes, and four other nuclear genes, we clarified the phylogenetic relationships of the new cryptic species. The endosymbiotic bacteria infestation was checked, considering the option that the divergent populations may represent clades isolated by Wolbachia infection; however, it did not demonstrate any specificity. We found only subtle morphological differences in the new clay-nesting species, D. argillicola sp. nov.; the discriminant analysis of morphometric measurements did not reliably segregate it as well. Thus, we provide the molecular characters of the cryptic species, which allow confident identification, its phylogenetic position within the genus, and an updated identification key for the D. tristis species group.}, } @article {pmid38392339, year = {2024}, author = {Reese, C and Graber, LC and Ramalho, MO and Moreau, CS}, title = {The Diversity of Wolbachia across the Turtle Ants (Formicidae: Cephalotes spp.).}, journal = {Biology}, volume = {13}, number = {2}, pages = {}, pmid = {38392339}, issn = {2079-7737}, support = {DGE-1650441; NSF DEB 1900357//National Science Foundation/ ; }, abstract = {Wolbachia is a widespread and well-known bacterium that can induce a wide range of changes within its host. Ants specifically harbor a great deal of Wolbachia diversity and are useful systems to study endosymbiosis. The turtle ants (Cephalotes) are a widespread group of tropical ants that rely on gut microbes to support their herbivorous diet for their survival, yet little is known of the extent of this diversity. Therefore, studying their endosymbionts and categorizing the diversity of bacteria within Cephalotes hosts could help to delimit species and identify new strains and can help lead to a further understanding of how the microbiome leads to survival and speciation in the wild. In our study, 116 individual samples were initially tested for positive infection with the wsp gene. Of the initial 116 samples, 9 samples were infected with only one strain of Wolbachia, and 7 were able to be used successfully for multilocus sequence typing (MLST). We used the new MLST data to infer a phylogeny with other Formicidae samples from the MLST online database to identify new Wolbachia strains and related genes, of which only one came back as an exact match. The 18 Wolbachia-positive samples ranged across 15 different species and 7 different countries, which we further test for species identity and geographic correlation. This study is the first comprehensive look into the diversity of Wolbachia in the turtle ants, providing insight into how endosymbionts are oriented in widespread species and providing a strong foundation for further research in host-microbe interactions.}, } @article {pmid38388801, year = {2024}, author = {Ye, QT and Gong, X and Liu, HH and Wu, BX and Peng, CW and Hong, XY and Bing, XL}, title = {The symbiont Wolbachia alleviates pesticide susceptibility in the two-spotted spider mite Tetranychus urticae through enhanced host detoxification pathways.}, journal = {Insect science}, volume = {}, number = {}, pages = {}, doi = {10.1111/1744-7917.13341}, pmid = {38388801}, issn = {1744-7917}, support = {32020103011//National Natural Science Foundation of China/ ; 32372528//National Natural Science Foundation of China/ ; TJ-2023-038//Young Elite Scientists Sponsorship Program by Jiangsu Association for Science and Technology/ ; BK20211213//Natural Science Foundation of Jiangsu Province/ ; }, abstract = {The two-spotted spider mite (Tetranychus urticae) is one of the most well-known pesticide-resistant agricultural pests, with resistance often attributed to changes such as target-site mutations and detoxification activation. Recent studies show that pesticide resistance can also be influenced by symbionts, but their involvement in this process in spider mites remains uncertain. Here, we found that infection with Wolbachia, a well-known bacterial reproductive manipulator, significantly increased mite survival after exposure to the insecticides abamectin, cyflumetofen, and pyridaben. Wolbachia-infected (WI) mites showed higher expression of detoxification genes such as P450, glutathione-S-transferase (GST), ABC transporters, and carboxyl/cholinesterases. RNA interference experiments confirmed the role of the two above-mentioned detoxification genes, TuCYP392D2 and TuGSTd05, in pesticide resistance. Increased GST activities were also observed in abamectin-treated WI mites. In addition, when wild populations were treated with abamectin, WI mites generally showed better survival than uninfected mites. However, genetically homogeneous mites with different Wolbachia strains showed similar survival. Finally, abamectin treatment increased Wolbachia abundance without altering the mite's bacterial community. This finding highlights the role of Wolbachia in orchestrating pesticide resistance by modulating host detoxification. By unraveling the intricate interplay between symbionts and pesticide resistance, our study lays the groundwork for pioneering strategies to combat agricultural pests.}, } @article {pmid38381797, year = {2024}, author = {Mendoza-Roldan, JA and Perles, L and Filippi, E and Szafranski, N and Montinaro, G and Carbonara, M and Scalera, R and de Abreu Teles, PP and Walochnik, J and Otranto, D}, title = {Parasites and microorganisms associated with the snakes collected for the "festa Dei serpari" in Cocullo, Italy.}, journal = {PLoS neglected tropical diseases}, volume = {18}, number = {2}, pages = {e0011973}, pmid = {38381797}, issn = {1935-2735}, mesh = {Animals ; Humans ; *Parasites ; *Cryptosporidiosis ; *Tick-Borne Diseases/microbiology ; *Cryptosporidium ; *Rickettsia ; Italy/epidemiology ; }, abstract = {While in much of the Western world snakes are feared, in the small, rural, mountainous town of Cocullo, in the middle of central Italy, snakes are annually collected and celebrated in a sacro-profane ritual. Every 1st of May, Serpari (snake catchers) capture and showcase dozens of non-venomous snakes to celebrate the ritual of San Domenico. In order to detect potential zoonotic pathogens within this unique epidemiological context, parasites and microorganisms of snakes harvested for the "festa dei serpari" ritual were investigated. Snakes (n = 112) were examined and ectoparasites collected, as well as blood and feces sampled. Ectoparasites were identified morpho-molecularly, and coprological examination conducted through direct smear and flotation. Molecular screenings were performed to identify parasites and microorganisms in collected samples (i.e., Mesostigmata mites, Anaplasma/Ehrlichia spp., Rickettsia spp., Borrelia burgdorferi sensu lato, Coxiella burnetii, Babesia/Theileria spp., Cryptosporidium spp., Giardia spp., Leishmania spp. and helminths). Overall, 28.5% (32/112) of snakes were molecularly positive for at least one parasite and/or microorganism. Endosymbiont Wolbachia bacteria were identified from Macronyssidae mites and zoonotic vector-borne pathogens (e.g., Rickettsia, Leishmania), as well as orally transmitted pathogens (i.e., Cryptosporidium, Giardia, Proteus vulgaris, Pseudomonas), were detected from blood and feces. Thus, given the central role of the snakes in the tradition of Cocullo, surveys of their parasitic fauna and associated zoonotic pathogens may aid to generate conservation policies to benefit the human-snake interactions, whilst preserving the cultural patrimony of this event.}, } @article {pmid38380092, year = {2024}, author = {Pramono, AK and Hidayanti, AK and Tagami, Y and Ando, H}, title = {Bacterial community and genome analysis of cytoplasmic incompatibility-inducing Wolbachia in American serpentine leafminer, Liriomyza trifolii.}, journal = {Frontiers in microbiology}, volume = {15}, number = {}, pages = {1304401}, pmid = {38380092}, issn = {1664-302X}, abstract = {Liriomyza trifolii, an agricultural pest, is occasionally infected by Wolbachia. A Wolbachia strain present in Liriomyza trifolii is associated with cytoplasmic incompatibility (CI) effects, leading to the death of embryos resulting from incompatible crosses between antibiotic-treated or naturally Wolbachia-free strain females and Wolbachia-infected males. In this study, high-throughput sequencing of hypervariable rRNA genes was employed to characterize the bacterial community in Wolbachia-infected L. trifolii without antibiotic treatment. The analysis revealed that Wolbachia dominates the bacterial community in L. trifolii, with minor presence of Acinetobacter, Pseudomonas, and Limnobacter. To elucidate the genetic basis of the CI phenotype, metagenomic sequencing was also conducted to assemble the genome of the Wolbachia strain. The draft-genome of the Wolbachia strain wLtri was 1.35 Mbp with 34% GC content and contained 1,487 predicted genes. Notably, within the wLtri genome, there are three distinct types of cytoplasmic incompatibility factor (cif) genes: Type I, Type III, and Type V cifA;B. These genes are likely responsible for inducing the strong cytoplasmic incompatibility observed in L. trifolii.}, } @article {pmid38374896, year = {2024}, author = {Mazel, F and Pitteloud, C and Guisan, A and Pellissier, L}, title = {Contrasted host specificity of gut and endosymbiont bacterial communities in alpine grasshoppers and crickets.}, journal = {ISME communications}, volume = {4}, number = {1}, pages = {ycad013}, pmid = {38374896}, issn = {2730-6151}, abstract = {Bacteria colonize the body of macroorganisms to form associations ranging from parasitic to mutualistic. Endosymbiont and gut symbiont communities are distinct microbiomes whose compositions are influenced by host ecology and evolution. Although the composition of horizontally acquired symbiont communities can correlate to host species identity (i.e. harbor host specificity) and host phylogeny (i.e. harbor phylosymbiosis), we hypothesize that the microbiota structure of vertically inherited symbionts (e.g. endosymbionts like Wolbachia) is more strongly associated with the host species identity and phylogeny than horizontally acquired symbionts (e.g. most gut symbionts). Here, using 16S metabarcoding on 336 guts from 24 orthopteran species (grasshoppers and crickets) in the Alps, we observed that microbiota correlated to host species identity, i.e. hosts from the same species had more similar microbiota than hosts from different species. This effect was ~5 times stronger for endosymbionts than for putative gut symbionts. Although elevation correlated with microbiome composition, we did not detect phylosymbiosis for endosymbionts and putative gut symbionts: closely related host species did not harbor more similar microbiota than distantly related species. Our findings indicate that gut microbiota of studied orthopteran species is more correlated to host identity and habitat than to the host phylogeny. The higher host specificity in endosymbionts corroborates the idea that-everything else being equal-vertically transmitted microbes harbor stronger host specificity signal, but the absence of phylosymbiosis suggests that host specificity changes quickly on evolutionary time scales.}, } @article {pmid38362501, year = {2024}, author = {Hegde, S and Marriott, AE and Pionnier, N and Steven, A and Bulman, C and Gunderson, E and Vogel, I and Koschel, M and Ehrens, A and Lustigman, S and Voronin, D and Tricoche, N and Hoerauf, A and Hübner, MP and Sakanari, J and Aljayyoussi, G and Gusovsky, F and Dagley, J and Hong, DW and O'Neill, P and Ward, SA and Taylor, MJ and Turner, JD}, title = {Combinations of the azaquinazoline anti-Wolbachia agent, AWZ1066S, with benzimidazole anthelmintics synergise to mediate sub-seven-day sterilising and curative efficacies in experimental models of filariasis.}, journal = {Frontiers in microbiology}, volume = {15}, number = {}, pages = {1346068}, pmid = {38362501}, issn = {1664-302X}, support = {MC_PC_18055/MRC_/Medical Research Council/United Kingdom ; }, abstract = {Lymphatic filariasis and onchocerciasis are two major neglected tropical diseases that are responsible for causing severe disability in 50 million people worldwide, whilst veterinary filariasis (heartworm) is a potentially lethal parasitic infection of companion animals. There is an urgent need for safe, short-course curative (macrofilaricidal) drugs to eliminate these debilitating parasite infections. We investigated combination treatments of the novel anti-Wolbachia azaquinazoline small molecule, AWZ1066S, with benzimidazole drugs (albendazole or oxfendazole) in up to four different rodent filariasis infection models: Brugia malayi-CB.17 SCID mice, B. malayi-Mongolian gerbils, B. pahangi-Mongolian gerbils, and Litomosoides sigmodontis-Mongolian gerbils. Combination treatments synergised to elicit threshold (>90%) Wolbachia depletion from female worms in 5 days of treatment, using 2-fold lower dose-exposures of AWZ1066S than monotherapy. Short-course lowered dose AWZ1066S-albendazole combination treatments also delivered partial adulticidal activities and/or long-lasting inhibition of embryogenesis, resulting in complete transmission blockade in B. pahangi and L. sigmodontis gerbil models. We determined that short-course AWZ1066S-albendazole co-treatment significantly augmented the depletion of Wolbachia populations within both germline and hypodermal tissues of B. malayi female worms and in hypodermal tissues in male worms, indicating that anti-Wolbachia synergy is not limited to targeting female embryonic tissues. Our data provides pre-clinical proof-of-concept that sub-seven-day combinations of rapid-acting novel anti-Wolbachia agents with benzimidazole anthelmintics are a promising curative and transmission-blocking drug treatment strategy for filarial diseases of medical and veterinary importance.}, } @article {pmid38348307, year = {2023}, author = {Yang, K and Zhang, HY and Wang, P and Jin, GX and Chu, D}, title = {Both symbionts and environmental factors contribute to shape the microbiota in a pest insect, Sogatella furcifera.}, journal = {Frontiers in microbiology}, volume = {14}, number = {}, pages = {1336345}, pmid = {38348307}, issn = {1664-302X}, abstract = {INTRODUCTION: Bacterial symbionts are prevalent in arthropods globally and play a vital role in the fitness and resistance of hosts. While several symbiont infections have been identified in the white-backed planthopper Sogatella furcifera, the impact of environmental factors on the microbiota within S. furcifera remains elusive.

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

RESULTS AND DISCUSSION: Based on the results, in S. furcifera, the presence of symbionts Wolbachia and Cardinium negatively influenced the abundance of other bacteria, including Enterobacter, Acinetobacter, and Lysinibacillus, while Wolbachia infection significantly decreased the diversity of the microbial community. Moreover, several environmental factors, including longitude, latitude, temperature, and precipitation, affected the abundance of symbionts and microbiota diversity in S. furcifera. These results collectively highlight the vital role of Wolbachia in S. furcifera microbiota, as well as the intricate effects of environmental factors on the bacterial communities of S. furcifera.}, } @article {pmid38342109, year = {2024}, author = {Lim, JT and Bansal, S and Chong, CS and Dickens, B and Ng, Y and Deng, L and Lee, C and Tan, LY and Chain, G and Ma, P and Sim, S and Tan, CH and Cook, AR and Ng, LC}, title = {Efficacy of Wolbachia-mediated sterility to reduce the incidence of dengue: a synthetic control study in Singapore.}, journal = {The Lancet. Microbe}, volume = {5}, number = {5}, pages = {e422-e432}, doi = {10.1016/S2666-5247(23)00397-X}, pmid = {38342109}, issn = {2666-5247}, mesh = {*Wolbachia/physiology ; *Dengue/prevention & control/epidemiology/transmission ; Singapore/epidemiology ; Animals ; *Aedes/microbiology/virology ; Incidence ; Female ; Male ; *Mosquito Control/methods ; *Mosquito Vectors/microbiology/virology ; Humans ; Dengue Virus ; Pest Control, Biological/methods ; }, abstract = {BACKGROUND: Due to the absence of available therapeutics and good vaccines, vector control solutions are needed to mitigate the spread of dengue. Matings between male Aedes aegypti mosquitoes infected with the wAlbB strain of Wolbachia and wildtype females yield non-viable eggs. We evaluated the efficacy of releasing wAlbB-infected A aegypti male mosquitoes to suppress dengue incidence.

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

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

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

FUNDING: Singapore Ministry of Finance, Ministry of Sustainability, and the National Environment Agency, and the Singapore National Robotics Program.}, } @article {pmid38327789, year = {2024}, author = {Hoffmann, AA and Ahmad, NW and Keong, WM and Ling, CY and Ahmad, NA and Golding, N and Tierney, N and Jelip, J and Putit, PW and Mokhtar, N and Sandhu, SS and Ming, LS and Khairuddin, K and Denim, K and Rosli, NM and Shahar, H and Omar, T and Ridhuan Ghazali, MK and Aqmar Mohd Zabari, NZ and Abdul Karim, MA and Saidin, MI and Mohd Nasir, MN and Aris, T and Sinkins, SP}, title = {Introduction of Aedes aegypti mosquitoes carrying wAlbB Wolbachia sharply decreases dengue incidence in disease hotspots.}, journal = {iScience}, volume = {27}, number = {2}, pages = {108942}, pmid = {38327789}, issn = {2589-0042}, support = {MC_UU_00034/4/MRC_/Medical Research Council/United Kingdom ; 108508/A/15/Z/WT_/Wellcome Trust/United Kingdom ; 202888/Z/16/Z/WT_/Wellcome Trust/United Kingdom ; MC_UU_12014/8/MRC_/Medical Research Council/United Kingdom ; /WT_/Wellcome Trust/United Kingdom ; }, abstract = {Partial replacement of resident Aedes aegypti mosquitoes with introduced mosquitoes carrying certain strains of inherited Wolbachia symbionts can result in transmission blocking of dengue and other viruses of public health importance. Wolbachia strain wAlbB is an effective transmission blocker and stable at high temperatures, making it particularly suitable for hot tropical climates. Following trial field releases in Malaysia, releases using wAlbB Ae. aegypti have become operationalized by the Malaysian health authorities. We report here on an average reduction in dengue fever of 62.4% (confidence intervals 50-71%) in 20 releases sites when compared to 76 control sites in high-rise residential areas. Importantly the level of dengue reduction increased with Wolbachia frequency, with 75.8% reduction (61-87%) estimated at 100% Wolbachia frequency. These findings indicate large impacts of wAlbB Wolbachia invasions on dengue fever incidence in an operational setting, with incidence expected to further decrease as wider areas are invaded.}, } @article {pmid38327358, year = {2023}, author = {Liu, Y and Zhao, L and Qiu, Z and Yuan, H}, title = {The gut microbiota diversity of five Orthoptera (Insecta, Polyneoptera) insects determined by DNA metabarcoding.}, journal = {Biodiversity data journal}, volume = {11}, number = {}, pages = {e98162}, pmid = {38327358}, issn = {1314-2828}, abstract = {Most orthopteran insects are phytophagous and some are important pests in agriculture and forests. Many intestinal microflora of Orthoptera insects have been reported, primarily from Acridoidea and there have been few reports of other taxa. In this study, we collected 15 individuals representing five species (Ruspolialineosa, Tetrixjaponica, Erianthusversicolor, Gryllotalpaorientalis and Teleogryllusemma) belonging to five orthopteran superfamilies (Tettigonioidea, Tetrigoidea, Eumastacoidea, Gryllotalpoidea and Grylloidea) to characterise and compare the gut microbiota with greater taxonomic width by performing sequencing analysis of the 16S rRNA V4 region in gut material. A total of 606,053 high-quality sequences and 3,105 OTUs were acquired from 15 gut samples representing 24 phyla, 48 classes, 69 orders, 133 families and 219 genera. Firmicutes and bacteria were the most abundant phyla, followed by Bacteroidetes, Cyanobacteria, Actinobacteria and Acidobacteria. At the genus level, Serratia, Citrobacter, Wolbachia, Lactobacillus and Parabacteroides were the most predominant genera in R.lineosa, T.japonica, E.versicolor, G.orientalis and T.emma, respectively. Both Principal Coordinates Analysis (PCoA) and heatmap results revealed significant differences in bacterial community composition across species. Additionally, alpha diversity analysis indicated the bacterial richness was significantly different amongst the five species.}, } @article {pmid38315724, year = {2024}, author = {Namias, A and Ngaku, A and Makoundou, P and Unal, S and Sicard, M and Weill, M}, title = {Intra-lineage microevolution of Wolbachia leads to the emergence of new cytoplasmic incompatibility patterns.}, journal = {PLoS biology}, volume = {22}, number = {2}, pages = {e3002493}, pmid = {38315724}, issn = {1545-7885}, mesh = {Animals ; Female ; Male ; *Wolbachia/genetics ; Antidotes/metabolism ; Mosquito Vectors/genetics ; Cytoplasm ; *Culex ; }, abstract = {Mosquitoes of the Culex pipiens complex are worldwide vectors of arbovirus, filarial nematodes, and avian malaria agents. In these hosts, the endosymbiotic bacteria Wolbachia induce cytoplasmic incompatibility (CI), i.e., reduced embryo viability in so-called incompatible crosses. Wolbachia infecting Culex pipiens (wPip) cause CI patterns of unparalleled complexity, associated with the amplification and diversification of cidA and cidB genes, with up to 6 different gene copies described in a single wPip genome. In wPip, CI is thought to function as a toxin-antidote (TA) system where compatibility relies on having the right antidotes (CidA) in the female to bind and neutralize the male's toxins (CidB). By repeating crosses between Culex isofemale lines over a 17 years period, we documented the emergence of a new compatibility type in real time and linked it to a change in cid genes genotype. We showed that loss of specific cidA gene copies in some wPip genomes results in a loss of compatibility. More precisely, we found that this lost antidote had an original sequence at its binding interface, corresponding to the original sequence at the toxin's binding interface. We showed that these original cid variants are recombinant, supporting a role for recombination rather than point mutations in rapid CI evolution. These results strongly support the TA model in natura, adding to all previous data acquired with transgenes expression.}, } @article {pmid38296097, year = {2024}, author = {Li, JH and Liu, XH and Liang, GR and Gao, HT and Guo, SH and Zhou, XY and Xing, D and Zhao, T and Li, CX}, title = {Microplastics affect mosquito from aquatic to terrestrial lifestyles and are transferred to mammals through mosquito bites.}, journal = {The Science of the total environment}, volume = {917}, number = {}, pages = {170547}, doi = {10.1016/j.scitotenv.2024.170547}, pmid = {38296097}, issn = {1879-1026}, mesh = {Animals ; Mice ; Microplastics ; Plastics ; Ecosystem ; *Insect Bites and Stings ; *Insecticides/toxicity ; Larva ; Mammals ; Mosquito Control ; *Culex ; *Nitriles ; *Pyrethrins ; }, abstract = {Microplastics (MPs) transfer from the environment to living organisms is a nonignorable global problem. As a complete metamorphosis insect, the larvae and adult Culex quinquefasciatus mosquito live in aquatic and terrestrial environments, respectively, where they easily access MPs. However, little is known about mosquitoes' potential role in MPs accumulation throughout ecosystems. Therefore, we conducted a study with different MPs particle sizes (0.1/1/10 μm) and concentrations (0.5/5/50 μg/mL) on Cx. quinquefasciatus to address this issue. Once exposed at the young larval stage, MPs could accompany the mosquitoes their entire life. The fluorescence signals of MPs in the larvae were mainly located in the intestines. Its intensity increased (from 3.72 × 10[6] AU to 5.45 × 10[7] AU) as the concentrations of MPs increases. The fluorescence signals of MPs were also detected in the blood and skin tissues of mice bitten by adult mosquitoes with MPs containing in their bodies. Mosquitos exposed to MPs showed longer larval pupation and eclosion time as well as lower adult body weight. In addition, MPs significantly reduced the lethal effect of pyrethroid insecticides (97.77 % vs. 48.88 %, p < 0.05) with 15.1 % removal of the deltamethrin concentration. After MPs exposure, the relative abundance of the Cx. quinquefasciatus gut microbiome, such as Wolbachia spp., Elizabethkingia spp., and Asaia spp., changed as the MPs size and concentration changes. Mosquitoes provide a new pathway for MPs accumulation and transfer to higher-level living organisms. Moreover, MPs significantly reduce the control effect of deltamethrin, providing new guidelines for mosquito insecticide application in MPs contamination circumstances.}, } @article {pmid38287029, year = {2024}, author = {McNamara, CJ and Ant, TH and Harvey-Samuel, T and White-Cooper, H and Martinez, J and Alphey, L and Sinkins, SP}, title = {Transgenic expression of cif genes from Wolbachia strain wAlbB recapitulates cytoplasmic incompatibility in Aedes aegypti.}, journal = {Nature communications}, volume = {15}, number = {1}, pages = {869}, pmid = {38287029}, issn = {2041-1723}, support = {/WT_/Wellcome Trust/United Kingdom ; MC_UU_00034/4/MRC_/Medical Research Council/United Kingdom ; MC_UU_12014/8/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Animals ; Male ; Female ; *Aedes ; *Wolbachia ; Mosquito Vectors/genetics ; Animals, Genetically Modified ; *Infertility ; }, abstract = {The endosymbiotic bacteria Wolbachia can invade insect populations by modifying host reproduction through cytoplasmic incompatibility (CI), an effect that results in embryonic lethality when Wolbachia-carrying males mate with Wolbachia-free females. Here we describe a transgenic system for recreating CI in the major arbovirus vector Aedes aegypti using CI factor (cif) genes from wAlbB, a Wolbachia strain currently being deployed to reduce dengue transmission. CI-like sterility is induced when cifA and cifB are co-expressed in testes; this sterility is rescued by maternal cifA expression, thereby reproducing the pattern of Wolbachia-induced CI. Expression of cifB alone is associated with extensive DNA damage and disrupted spermatogenesis. The strength of rescue by maternal cifA expression is dependent on the comparative levels of cifA/cifB expression in males. These findings are consistent with CifB acting as a toxin and CifA as an antitoxin, with CifA attenuating CifB toxicity in both the male germline and in developing embryos. These findings provide important insights into the interactions between cif genes and their mechanism of activity and provide a foundation for the building of a cif gene-based drive system in Ae. aegypti.}, } @article {pmid38278818, year = {2024}, author = {Piccinno, R and Tatti, A and Avosani, S and Galla, G and Lazazzara, V and Pedrazzoli, F and Zadra, N and Rodeghiero, M and Seljak, G and Özgen, İ and Hauffe, HC and Verrastro, V and Stacconi, MVR and Mazzoni, V and Rota-Stabelli, O}, title = {A multidisciplinary approach to tackling invasive species: barcoding, morphology, and metataxonomy of the leafhopper Arboridia adanae.}, journal = {Scientific reports}, volume = {14}, number = {1}, pages = {2229}, pmid = {38278818}, issn = {2045-2322}, mesh = {Humans ; Animals ; *Introduced Species ; *Hemiptera ; RNA, Ribosomal, 16S/genetics ; Phylogeny ; Greece ; }, abstract = {The leafhopper genus Arboridia includes several species that feed on Vitis vinifera and cause leaf chlorosis. We report the first alien Arboridia infestation in Italy in 2021 in an Apulian vineyard. To confirm the taxonomic status of the species responsible for crop damage, and reconstruct its demographic history, we barcoded individuals from Apulia together with Arboridia spp. from Crete (Greece), A. adanae from Central Turkey and other specimens of the presumed sister species, A. dalmatina from Dalmatia (Croatia). Molecular phylogenies and barcoding gap analysis identified clades not associated with sampling locations. This result is incongruent with classical specimen assignment and is further supported by morphological analyses, which did not reveal significant differences among the populations. Therefore, we propose A. dalmatina as a junior synonym of A. adanae, which would become the only grapevine-related Arboridia species in the eastern Mediterranean. To further characterise A. adanae evolution, we performed a molecular clock analysis that suggested a radiation during the Pleistocene glaciations. Finally, to assess whether the Apulian individuals carried microorganisms of agricultural relevance, we sequenced their bacterial microbiota using 16S rRNA amplicon sequencing identifying three phytopathogens not generally associated with Arboridia activities as well as Wolbachia in one Apulian haplogroup. We discuss the agricultural implications of this infestation.}, } @article {pmid38257815, year = {2024}, author = {Rainey, SM and Lefteri, DA and Darby, C and Kohl, A and Merits, A and Sinkins, SP}, title = {Evidence of Differences in Cellular Regulation of Wolbachia-Mediated Viral Inhibition between Alphaviruses and Flaviviruses.}, journal = {Viruses}, volume = {16}, number = {1}, pages = {}, pmid = {38257815}, issn = {1999-4915}, support = {108508/A/15/Z/WT_/Wellcome Trust/United Kingdom ; WT094664MA/WT_/Wellcome Trust/United Kingdom ; 108508/WT_/Wellcome Trust/United Kingdom ; /WT_/Wellcome Trust/United Kingdom ; 202888/WT_/Wellcome Trust/United Kingdom ; MC_UU_00034/4/MRC_/Medical Research Council/United Kingdom ; MC_UU_12014/8/MRC_/Medical Research Council/United Kingdom ; 202888/Z/16/Z/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Animals ; *Alphavirus ; *Flavivirus ; *Wolbachia ; *Aedes ; *Zika Virus ; Cholesterol ; Antiviral Agents/pharmacology ; *Zika Virus Infection ; }, abstract = {The intracellular bacterium Wolbachia is increasingly being utilised in control programs to limit the spread of arboviruses by Aedes mosquitoes. Achieving a better understanding of how Wolbachia strains can reduce viral replication/spread could be important for the long-term success of such programs. Previous studies have indicated that for some strains of Wolbachia, perturbations in lipid metabolism and cholesterol storage are vital in Wolbachia-mediated antiviral activity against the flaviviruses dengue and Zika; however, it has not yet been examined whether arboviruses in the alphavirus group are affected in the same way. Here, using the reporters for the alphavirus Semliki Forest virus (SFV) in Aedes albopictus cells, we found that Wolbachia strains wMel, wAu and wAlbB blocked viral replication/translation early in infection and that storage of cholesterol in lipid droplets is not key to this inhibition. Another alphavirus, o'nyong nyong virus (ONNV), was tested in both Aedes albopictus cells and in vivo in stable, transinfected Aedes aegypti mosquito lines. The strains wMel, wAu and wAlbB show strong antiviral activity against ONNV both in vitro and in vivo. Again, 2-hydroxypropyl-β-cyclodextrin (2HPCD) was not able to rescue ONNV replication in cell lines, suggesting that the release of stored cholesterol caused by wMel is not able to rescue blockage of ONNV. Taken together, this study shows that alphaviruses appear to be inhibited early in replication/translation and that there may be differences in how alphaviruses are inhibited by Wolbachia in comparison to flaviviruses.}, } @article {pmid38254452, year = {2024}, author = {Cheng, Z and Liu, Q and Huang, X}, title = {Partial Correspondence between Host Plant-Related Differentiation and Symbiotic Bacterial Community in a Polyphagous Insect.}, journal = {Animals : an open access journal from MDPI}, volume = {14}, number = {2}, pages = {}, pmid = {38254452}, issn = {2076-2615}, support = {31970446//National Natural Science Foundation of China/ ; }, abstract = {Host plants play a vital role in insect population differentiation, while symbiotic associations between bacteria and insects are ubiquitous in nature. However, existing studies have given limited attention to the connection between host-related differentiation and symbiotic bacterial communities in phytophagous insects. In this study, we collected 58 samples of Aphis odinae from different host plants in southern China and constructed phylogenetic trees to investigate their differentiation in relation to host plants. We also selected aphid samples from the five most preferred host plants and analyzed their symbiotic bacterial composition using Illumina sequencing of the V3-V4 hypervariable region of the 16S rRNA gene. The phylogeny and symbiotic bacterial community structure of A. odinae populations on different host plants showed that samples from Triadica sebifera (Euphorbiaceae) had a consistent presence of Wolbachia as the predominant secondary symbiont and suggested the possibility of undergoing differentiation. Conversely, although differentiation was observed in samples from Rhus chinensis (Anacardiaceae), no consistent presence of predominant secondary symbionts was found. Additionally, the samples from Heptapleurum heptaphyllum (Araliaceae) consistently carried Serratia, but no host differentiation was evident. In summary, this study reveals a partial correspondence between symbiotic bacterial communities and host-related differentiation in A. odinae. The findings contribute to our understanding of the microevolutionary influencing the macroevolutionary relationships between bacterial symbionts and phytophagous insects. The identification of specific symbionts associated with host-related differentiation provides valuable insights into the intricate dynamics of insect-bacteria interactions.}, } @article {pmid38243558, year = {2024}, author = {Sohail, A and Anders, KL and McGuinness, SL and Leder, K}, title = {The epidemiology of imported and locally acquired dengue in Australia, 2012-2022.}, journal = {Journal of travel medicine}, volume = {31}, number = {2}, pages = {}, pmid = {38243558}, issn = {1708-8305}, support = {/WT_/Wellcome Trust/United Kingdom ; 224459/Z/21/Z/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Animals ; Humans ; Australia/epidemiology ; *Arbovirus Infections/epidemiology ; Queensland/epidemiology ; *Culicidae ; *Dengue/epidemiology ; }, abstract = {BACKGROUND: Dengue is the most important arboviral disease globally and poses ongoing challenges for control including in non-endemic countries with competent mosquito vectors at risk of local transmission through imported cases. We examined recent epidemiological trends in imported and locally acquired dengue in Australia, where the Wolbachia mosquito population replacement method was implemented throughout dengue-prone areas of northern Queensland between 2011 and 2019.

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

RESULTS: Between 2012 and 2022, 13 343 dengue cases were reported in Australia (median 1466 annual cases); 12 568 cases (94.2%) were imported, 584 (4.4%) were locally acquired and 191 (1.4%) had no origin recorded. Locally acquired cases decreased from a peak in 2013 (n = 236) to zero in 2021-22. Annual incidence of imported dengue ranged from 8.29/100 000 (n = 917 cases) to 22.10/100 000 (n = 2203) annual traveller movements between 2012 and 2019, decreased in 2020 (6.74/100 000 traveller movements; n = 191) and 2021 (3.32/100 000 traveller movements; n = 10) during COVID-19-related border closures, then rose to 34.79/100 000 traveller movements (n = 504) in 2022. Imported cases were primarily acquired in Southeast Asia (n = 9323; 74%), Southern and Central Asia (n = 1555; 12%) and Oceania (n = 1341; 11%). Indonesia (n = 5778; 46%) and Thailand (n = 1483; 12%) were top acquisition countries. DENV-2 (n = 2147; 42%) and DENV-1 (n = 1526; 30%) were predominant serotypes.

CONCLUSION: Our analysis highlights Australia's successful control of locally acquired dengue with Wolbachia. Imported dengue trends reflect both Australian travel destinations and patterns and local epidemiology in endemic countries.}, } @article {pmid38235434, year = {2023}, author = {Mantilla-Granados, JS and Castellanos, JE and Velandia-Romero, ML}, title = {A tangled threesome: understanding arbovirus infection in Aedes spp. and the effect of the mosquito microbiota.}, journal = {Frontiers in microbiology}, volume = {14}, number = {}, pages = {1287519}, pmid = {38235434}, issn = {1664-302X}, abstract = {Arboviral infections transmitted by Aedes spp. mosquitoes are a major threat to human health, particularly in tropical regions but are expanding to temperate regions. The ability of Aedes aegypti and Aedes albopictus to transmit multiple arboviruses involves a complex relationship between mosquitoes and the virus, with recent discoveries shedding light on it. Furthermore, this relationship is not solely between mosquitoes and arboviruses, but also involves the mosquito microbiome. Here, we aimed to construct a comprehensive review of the latest information about the arbovirus infection process in A. aegypti and A. albopictus, the source of mosquito microbiota, and its interaction with the arbovirus infection process, in terms of its implications for vectorial competence. First, we summarized studies showing a new mechanism for arbovirus infection at the cellular level, recently described innate immunological pathways, and the mechanism of adaptive response in mosquitoes. Second, we addressed the general sources of the Aedes mosquito microbiota (bacteria, fungi, and viruses) during their life cycle, and the geographical reports of the most common microbiota in adults mosquitoes. How the microbiota interacts directly or indirectly with arbovirus transmission, thereby modifying vectorial competence. We highlight the complexity of this tripartite relationship, influenced by intrinsic and extrinsic conditions at different geographical scales, with many gaps to fill and promising directions for developing strategies to control arbovirus transmission and to gain a better understanding of vectorial competence. The interactions between mosquitoes, arboviruses and their associated microbiota are yet to be investigated in depth.}, } @article {pmid38230418, year = {2024}, author = {Nindrea, RD}, title = {Wolbachia: New Hopes for the Prevention of Dengue Hemorrhagic Fever in Indonesia.}, journal = {Asia-Pacific journal of public health}, volume = {36}, number = {2-3}, pages = {280}, doi = {10.1177/10105395231226296}, pmid = {38230418}, issn = {1941-2479}, mesh = {Humans ; Animals ; *Severe Dengue ; *Wolbachia ; Indonesia/epidemiology ; *Dengue/prevention & control ; *Dengue Virus ; *Aedes ; }, } @article {pmid38204789, year = {2024}, author = {Arai, H and Watada, M and Kageyama, D}, title = {Two male-killing Wolbachia from Drosophila birauraia that are closely related but distinct in genome structure.}, journal = {Royal Society open science}, volume = {11}, number = {1}, pages = {231502}, pmid = {38204789}, issn = {2054-5703}, abstract = {Insects harbour diverse maternally inherited bacteria and viruses, some of which have evolved to kill the male progeny of their hosts (male killing: MK). The fly species Drosophila biauraria carries a maternally transmitted MK-inducing partiti-like virus, but it was unknown if it carries other MK-inducing endosymbionts. Here, we identified two male-killing Wolbachia strains (wBiau1 and wBiau2) from D. biauraria and compared their genomes to elucidate their evolutionary processes. The two strains were genetically closely related but had exceptionally different genome structures with considerable rearrangements compared with combinations of other Wolbachia strains. Despite substantial changes in the genome structure, the two Wolbachia strains did not experience gene losses that would disrupt the male-killing expression or persistence in the host population. The two Wolbachia-infected matrilines carried distinct mitochondrial haplotypes, suggesting that wBiau1 and wBiau2 have invaded D. biauraria independently and undergone considerable genome changes owing to unknown selective pressures in evolutionary history. This study demonstrated the presence of three male-killers from two distinct origins in one fly species and highlighted the diverse and rapid genome evolution of MK Wolbachia in the host.}, } @article {pmid38196945, year = {2023}, author = {Côrtes, N and Lira, A and Prates-Syed, W and Dinis Silva, J and Vuitika, L and Cabral-Miranda, W and Durães-Carvalho, R and Balan, A and Cabral-Marques, O and Cabral-Miranda, G}, title = {Integrated control strategies for dengue, Zika, and Chikungunya virus infections.}, journal = {Frontiers in immunology}, volume = {14}, number = {}, pages = {1281667}, pmid = {38196945}, issn = {1664-3224}, mesh = {Animals ; Humans ; *Zika Virus ; *Chikungunya Fever/epidemiology/prevention & control ; Mosquito Vectors ; *Zika Virus Infection/epidemiology/prevention & control ; Antiviral Agents ; *Dengue/epidemiology/prevention & control ; }, abstract = {Arboviruses are a major threat to public health in tropical regions, encompassing over 534 distinct species, with 134 capable of causing diseases in humans. These viruses are transmitted through arthropod vectors that cause symptoms such as fever, headache, joint pains, and rash, in addition to more serious cases that can lead to death. Among the arboviruses, dengue virus stands out as the most prevalent, annually affecting approximately 16.2 million individuals solely in the Americas. Furthermore, the re-emergence of the Zika virus and the recurrent outbreaks of chikungunya in Africa, Asia, Europe, and the Americas, with one million cases reported annually, underscore the urgency of addressing this public health challenge. In this manuscript we discuss the epidemiology, viral structure, pathogenicity and integrated control strategies to combat arboviruses, and the most used tools, such as vaccines, monoclonal antibodies, treatment, etc., in addition to presenting future perspectives for the control of arboviruses. Currently, specific medications for treating arbovirus infections are lacking, and symptom management remains the primary approach. However, promising advancements have been made in certain treatments, such as Chloroquine, Niclosamide, and Isatin derivatives, which have demonstrated notable antiviral properties against these arboviruses in vitro and in vivo experiments. Additionally, various strategies within vector control approaches have shown significant promise in reducing arbovirus transmission rates. These encompass public education initiatives, targeted insecticide applications, and innovative approaches like manipulating mosquito bacterial symbionts, such as Wolbachia. In conclusion, combatting the global threat of arbovirus diseases needs a comprehensive approach integrating antiviral research, vaccination, and vector control. The continued efforts of research communities, alongside collaborative partnerships with public health authorities, are imperative to effectively address and mitigate the impact of these arboviral infections on public health worldwide.}, } @article {pmid38193019, year = {2024}, author = {Mat Udin, AS and Uni, S and Rodrigues, J and Martin, C and Junker, K and Agatsuma, T and Low, VL and Saijuntha, W and Omar, H and Zainuri, NA and Fukuda, M and Matsubayashi, M and Kimura, D and Takaoka, H and Ramli, R}, title = {Redescription, molecular characterisation and Wolbachia endosymbionts of Mansonella (Tupainema) dunni (Mullin & Orihel, 1972) (Spirurida: Onchocercidae) from the common treeshrew Tupaia glis Diard & Duvaucel (Mammalia: Scandentia) in Peninsular Malaysia.}, journal = {Current research in parasitology & vector-borne diseases}, volume = {5}, number = {}, pages = {100154}, pmid = {38193019}, issn = {2667-114X}, abstract = {The genus Mansonella Faust, 1929 includes 29 species, mainly parasites of platyrrhine monkeys in South America and anthropoid apes in Africa. In Malaysia, Mansonella (Tupainema) dunni (Mullin & Orihel, 1972) was described from the common treeshrew Tupaia glis Diard & Duvaucel (Scandentia). In a recent classification of the genus Mansonella, seven subgenera were proposed, with M. (Tup.) dunni as a monotypic species in the subgenus Tupainema. In this study, we collected new material of M. (Tup.) dunni from common treeshrews in Peninsular Malaysia and redescribed the morphological features of this species. We found that M. (Tup.) dunni differs from M. (Cutifilaria) perforata Uni et al., 2004 from sika deer Cervus nippon (Cetartiodactyla) in Japan, with regards to morphological features and predilection sites in their respective hosts. Based on multi-locus sequence analyses, we examined the molecular phylogeny of M. (Tup.) dunni and its Wolbachia genotype. Species of the genus Mansonella grouped monophyletically in clade ONC5 and M. (Tup.) dunni was placed in the most derived position within this genus. Mansonella (Tup.) dunni was closely related to M. (M.) ozzardi (Manson, 1897) from humans in Central and South America, and most distant from M. (C.) perforata. The calculated p-distances between the cox1 gene sequences for M. (Tup.) dunni and its congeners were 13.09% for M. (M.) ozzardi and 15.6-16.15% for M. (C.) perforata. The molecular phylogeny of Mansonella spp. thus corroborates their morphological differences. We determined that M. (Tup.) dunni harbours Wolbachia endosymbionts of the supergroup F genotype, in keeping with all other Mansonella species screened to date.}, } @article {pmid38192184, year = {2024}, author = {Strunov, A and Schönherr, C and Kapun, M}, title = {Wolbachia effects on thermal preference of natural Drosophila melanogaster are influenced by host genetic background, Wolbachia type, and bacterial titer.}, journal = {Environmental microbiology}, volume = {26}, number = {2}, pages = {e16579}, doi = {10.1111/1462-2920.16579}, pmid = {38192184}, issn = {1462-2920}, support = {FWF P32275//Austrian Science Fund/ ; }, mesh = {Animals ; *Drosophila melanogaster/genetics ; *Wolbachia/genetics ; Drosophila/genetics/microbiology ; Reproduction ; Biological Evolution ; Symbiosis ; }, abstract = {Temperature plays a fundamental role in the fitness of all organisms. In particular, it strongly affects metabolism and reproduction in ectotherms that have limited physiological capabilities to regulate their body temperature. The influence of temperature variation on the physiology and behaviour of ectotherms is well studied but we still know little about the influence of symbiotic interactions on thermal preference (Tp) of the host. A growing number of studies focusing on the Wolbachia-Drosophila host-symbiont system found that Wolbachia can influence Tp in Drosophila laboratory strains. Here, we investigated the effect of Wolbachia on Tp in wild-type D. melanogaster flies recently collected from nature. Consistent with previous data, we found reduced Tp compared to an uninfected control in one of two fly strains infected with the wMelCS Wolbachia type. Additionally, we, for the first time, found that Wolbachia titer variation influences the thermal preference of the host fly. These data indicate that the interaction of Wolbachia and Drosophila resulting in behavioural variation is strongly influenced by the genetic background of the host and symbiont. More studies are needed to better understand the evolutionary significance of Tp variation influenced by Wolbachia in natural Drosophila populations.}, } @article {pmid38172384, year = {2024}, author = {Volpe, HXL and Carmo-Sousa, M and Luvizotto, RAG and de Freitas, R and Esperança, V and Darolt, JC and Pegoraro, AAL and Magalhães, DM and Favaris, AP and Wulff, NA and Miranda, MP and Bento, JMS and Leal, WS}, title = {The greening-causing agent alters the behavioral and electrophysiological responses of the Asian citrus psyllid to a putative sex pheromone.}, journal = {Scientific reports}, volume = {14}, number = {1}, pages = {455}, pmid = {38172384}, issn = {2045-2322}, support = {201600147//National Institute of Science and Technology of Semiochemicals in Agriculture (INCT)/ ; 2014/50871-0//Fundação de Amparo à Pesquisa do Estado de São Paulo/ ; 201600147//FUNDECITRUS/ University of California-Davis Research Agreement/ ; }, mesh = {Female ; Male ; Animals ; *Hemiptera/physiology ; *Sex Attractants/pharmacology ; Brazil ; *Citrus/microbiology ; *Citrus sinensis ; *Wolbachia ; *Rhizobiaceae/physiology ; Acetates ; Plant Diseases/microbiology ; }, abstract = {The Asian Citrus Psyllid (ACP), Diaphorina citri, is a vector of the pathological bacterium Candidatus Liberibacter asiaticus (CLas), which causes the most devastating disease to the citrus industry worldwide, known as greening or huanglongbing (HLB). Earlier field tests with an acetic acid-based lure in greening-free, 'Valencia' citrus orange groves in California showed promising results. The same type of lures tested in São Paulo, Brazil, showed unsettling results. During the unsuccessful trials, we noticed a relatively large proportion of females in the field, ultimately leading us to test field-collected males and females for Wolbachia and CLas. The results showed high rates of Wolbachia and CLas infection in field populations. We then compared the olfactory responses of laboratory-raised, CLas-free, and CLas-infected males to acetic acid. As previously reported, CLas-uninfected males responded to acetic acid at 1 µg. Surprisingly, CLas-infected males required 50 × higher doses of the putative sex pheromone, thus explaining the failure to capture CLas-infected males in the field. CLas infection was also manifested in electrophysiological responses. Electroantennogram responses from CLas-infected ACP males were significantly higher than those obtained with uninfected males. To the best of our knowledge, this is the first report of a pathogen infection affecting a vector's response to a sex attractant.}, } @article {pmid38162582, year = {2023}, author = {Muharromah, AF and Reyes, JIL and Kagia, N and Watanabe, K}, title = {Genome-wide detection of Wolbachia in natural Aedes aegypti populations using ddRAD-Seq.}, journal = {Frontiers in cellular and infection microbiology}, volume = {13}, number = {}, pages = {1252656}, pmid = {38162582}, issn = {2235-2988}, mesh = {Humans ; Animals ; Male ; Female ; *Aedes/microbiology ; *Wolbachia/genetics ; RNA, Ribosomal, 16S/genetics ; Philippines ; Ribosomal Proteins ; }, abstract = {BACKGROUND: Wolbachia, an endosymbiotic bacterium, is globally used to control arboviruses because of its ability to block arboviral replication and manipulate the reproduction of Wolbachia host, Aedes aegypti. Polymerase chain reaction (PCR)-based Wolbachia detection has been recently reported from natural Ae. aegypti populations. However, due to the technical limitations of PCR, such as primer incompatibility, PCR-based assays are not sufficiently reliable or accurate. In this study, we examined double digestion restriction site-associated DNA sequencing (ddRAD-Seq) efficiency and limitations in Wolbachia detection and quantification in field-collected Ae. aegypti natural populations in Metro Manila, the Philippines, compared with PCR-based assays.

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

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

CONCLUSIONS: Taken together, our results demonstrate the feasibility of ddRAD-Seq-based Wolbachia detection from field-collected Ae. aegypti mosquitoes.}, } @article {pmid38157329, year = {2023}, author = {, }, title = {Expression of Concern: A Potential Role for the Interaction of Wolbachia Surface Proteins with the Brugia malayi Glycolytic Enzymes and Cytoskeleton in Maintenance of Endosymbiosis.}, journal = {PLoS neglected tropical diseases}, volume = {17}, number = {12}, pages = {e0011860}, pmid = {38157329}, issn = {1935-2735}, } @article {pmid38143905, year = {2023}, author = {Mouillaud, T and Berger, A and Buysse, M and Rahola, N and Daron, J and Agbor, JP and Sango, SN and Neafsey, DE and Duron, O and Ayala, D}, title = {Limited association between Wolbachia and Plasmodium falciparum infections in natural populations of the major malaria mosquito Anopheles moucheti.}, journal = {Evolutionary applications}, volume = {16}, number = {12}, pages = {1999-2006}, pmid = {38143905}, issn = {1752-4571}, support = {U19 AI110818/AI/NIAID NIH HHS/United States ; }, abstract = {Since the discovery of natural malaria vector populations infected by the endosymbiont bacterium Wolbachia, a renewed interest has arisen for using this bacterium as an alternative for malaria control. Among naturally infected mosquitoes, Anopheles moucheti, a major malaria mosquito in Central Africa, exhibits one of the highest prevalences of Wolbachia infection. To better understand whether this maternally inherited bacterium could be used for malaria control, we investigated Wolbachia influence in An. moucheti populations naturally infected by the malaria parasite Plasmodium falciparum. To this end, we collected mosquitoes in a village from Cameroon, Central Africa, where this mosquito is the main malaria vector. We found that the prevalence of Wolbachia bacterium was almost fixed in the studied mosquito population, and was higher than previously recorded. We also quantified Wolbachia in whole mosquitoes and dissected abdomens, confirming that the bacterium is also elsewhere than in the abdomen, but at lower density. Finally, we analyzed the association of Wolbachia presence and density on P. falciparum infection. Wolbachia density was slightly higher in mosquitoes infected with the malaria parasite than in uninfected mosquitoes. However, we observed no correlation between the P. falciparum and Wolbachia densities. In conclusion, our study indicates that naturally occurring Wolbachia infection is not associated to P. falciparum development within An. moucheti mosquitoes.}, } @article {pmid38139239, year = {2023}, author = {Gruntenko, NE and Deryuzhenko, MA and Andreenkova, OV and Shishkina, OD and Bobrovskikh, MA and Shatskaya, NV and Vasiliev, GV}, title = {Drosophila melanogaster Transcriptome Response to Different Wolbachia Strains.}, journal = {International journal of molecular sciences}, volume = {24}, number = {24}, pages = {}, pmid = {38139239}, issn = {1422-0067}, support = {21-14-00090//Russian Science Foundation/ ; }, mesh = {Animals ; Female ; *Drosophila melanogaster/physiology ; *Wolbachia/genetics ; Transcriptome ; Genotype ; }, abstract = {Wolbachia is a maternally inherited, intercellular bacterial symbiont of insects and some other invertebrates. Here, we investigated the effect of two different Wolbachia strains, differing in a large chromosomal inversion, on the differential expression of genes in D. melanogaster females. We revealed significant changes in the transcriptome of the infected flies compared to the uninfected ones, as well as in the transcriptome of flies infected with the Wolbachia strain, wMelPlus, compared to flies infected with the wMelCS[112] strain. We linked differentially expressed genes (DEGs) from two pairwise comparisons, "uninfected-wMelPlus-infected" and "uninfected-wMelCS[112]-infected", into two gene networks, in which the following functional groups were designated: "Proteolysis", "Carbohydrate transport and metabolism", "Oxidation-reduction process", "Embryogenesis", "Transmembrane transport", "Response to stress" and "Alkaline phosphatases". Our data emphasized similarities and differences between infections by different strains under study: a wMelPlus infection results in more than double the number of upregulated DEGs and half the number of downregulated DEGs compared to a wMelCS[112] infection. Thus, we demonstrated that Wolbachia made a significant contribution to differential expression of host genes and that the bacterial genotype plays a vital role in establishing the character of this contribution.}, } @article {pmid38132636, year = {2024}, author = {Loterio, RK and Monson, EA and Templin, R and de Bruyne, JT and Flores, HA and Mackenzie, JM and Ramm, G and Helbig, KJ and Simmons, CP and Fraser, JE}, title = {Antiviral Wolbachia strains associate with Aedes aegypti endoplasmic reticulum membranes and induce lipid droplet formation to restrict dengue virus replication.}, journal = {mBio}, volume = {15}, number = {2}, pages = {e0249523}, pmid = {38132636}, issn = {2150-7511}, support = {DP220102997//Department of Education and Training | Australian Research Council (ARC)/ ; APP1182432//DHAC | National Health and Medical Research Council (NHMRC)/ ; }, mesh = {Animals ; Humans ; *Dengue Virus/physiology ; *Aedes ; *Wolbachia/physiology ; Lipid Droplets ; Virus Replication ; *Dengue ; Endoplasmic Reticulum ; Antiviral Agents ; Lipids ; }, abstract = {Wolbachia are a genus of insect endosymbiotic bacteria which includes strains wMel and wAlbB that are being utilized as a biocontrol tool to reduce the incidence of Aedes aegypti-transmitted viral diseases like dengue. However, the precise mechanisms underpinning the antiviral activity of these Wolbachia strains are not well defined. Here, we generated a panel of Ae. aegypti-derived cell lines infected with antiviral strains wMel and wAlbB or the non-antiviral Wolbachia strain wPip to understand host cell morphological changes specifically induced by antiviral strains. Antiviral strains were frequently found to be entirely wrapped by the host endoplasmic reticulum (ER) membrane, while wPip bacteria clustered separately in the host cell cytoplasm. ER-derived lipid droplets (LDs) increased in volume in wMel- and wAlbB-infected cell lines and mosquito tissues compared to cells infected with wPip or Wolbachia-free controls. Inhibition of fatty acid synthase (required for triacylglycerol biosynthesis) reduced LD formation and significantly restored ER-associated dengue virus replication in cells occupied by wMel. Together, this suggests that antiviral Wolbachia strains may specifically alter the lipid composition of the ER to preclude the establishment of dengue virus (DENV) replication complexes. Defining Wolbachia's antiviral mechanisms will support the application and longevity of this effective biocontrol tool that is already being used at scale.IMPORTANCEAedes aegypti transmits a range of important human pathogenic viruses like dengue. However, infection of Ae. aegypti with the insect endosymbiotic bacterium, Wolbachia, reduces the risk of mosquito to human viral transmission. Wolbachia is being utilized at field sites across more than 13 countries to reduce the incidence of viruses like dengue, but it is not well understood how Wolbachia induces its antiviral effects. To examine this at the subcellular level, we compared how different strains of Wolbachia with varying antiviral strengths associate with and modify host cell structures. Strongly antiviral strains were found to specifically associate with the host endoplasmic reticulum and induce striking impacts on host cell lipid droplets. Inhibiting Wolbachia-induced lipid redistribution partially restored dengue virus replication demonstrating this is a contributing role for Wolbachia's antiviral activity. These findings provide new insights into how antiviral Wolbachia strains associate with and modify Ae. aegypti host cells.}, } @article {pmid38132616, year = {2023}, author = {Shapoval, NA and Kir'yanov, AV and Krupitsky, AV and Yakovlev, RV and Romanovich, AE and Zhang, J and Cong, Q and Grishin, NV and Kovalenko, MG and Shapoval, GN}, title = {Phylogeography of Two Enigmatic Sulphur Butterflies, Colias mongola Alphéraky, 1897 and Colias tamerlana Staudinger, 1897 (Lepidoptera, Pieridae), with Relations to Wolbachia Infection.}, journal = {Insects}, volume = {14}, number = {12}, pages = {}, pmid = {38132616}, issn = {2075-4450}, support = {22-24-01086//Russian Science Foundation/ ; }, abstract = {The genus Colias Fabricius, 1807 includes numerous taxa and forms with uncertain status and taxonomic position. Among such taxa are Colias mongola Alphéraky, 1897 and Colias tamerlana Staudinger, 1897, interpreted in the literature either as conspecific forms, as subspecies of different but morphologically somewhat similar Colias species or as distinct species-level taxa. Based on mitochondrial and nuclear DNA markers, we reconstructed a phylogeographic pattern of the taxa in question. We recover and include in our analysis DNA barcodes of the century-old type specimens, the lectotype of C. tamerlana deposited in the Natural History Museum (Museum für Naturkunde), Berlin, Germany (ZMHU) and the paralectotype of C. tamerlana and the lectotype of C. mongola deposited in the Zoological Institute, Russian Academy of Sciences, St. Petersburg, Russia (ZISP). Our analysis grouped all specimens within four (HP_I-HP_IV) deeply divergent but geographically poorly structured clades which did not support nonconspecifity of C. mongola-C. tamerlana. We also show that all studied females of the widely distributed haplogroup HP_II were infected with a single Wolbachia strain belonging to the supergroup B, while the males of this haplogroup, as well as all other investigated specimens of both sexes, were not infected. Our data highlight the relevance of large-scale sampling dataset analysis and the need for testing for Wolbachia infection to avoid erroneous phylogenetic reconstructions and species misidentification.}, } @article {pmid38132601, year = {2023}, author = {Adonyeva, NV and Efimov, VM and Gruntenko, NE}, title = {The Effect of Genotype Combinations of Wolbachia and Its Drosophila melanogaster Host on Fertility, Developmental Rate and Heat Stress Resistance of Flies.}, journal = {Insects}, volume = {14}, number = {12}, pages = {}, pmid = {38132601}, issn = {2075-4450}, support = {21-14-00090//Russian Science Foundation/ ; }, abstract = {The best-known effect of the intracellular bacterium Wolbachia is its mostly negative influence on the reproduction of the host. However, there is evidence of a positive influence of Wolbachia on the host's resistance to stress, pathogens, and viruses. Here, we analyzed the effects of two Wolbachia strains belonging to wMel and wMelCS genotypes on D. melanogaster traits, such as fertility, survival under acute heat stress, and developmental rate. We found that D. melanogaster lines under study differ significantly in the above-mentioned characteristics, both when the natural infection was preserved, and when it was eliminated. One of Wolbachia strains, wMel, did not affect any of the studied traits. Another strain, wMelPlus, had a significant effect on the development time. Moreover, this effect is observed not only in the line in which it was discovered but also in the one it was transferred to. When transferred to a new line, wMelPlus also caused changes in survival under heat stress. Thus, it could be concluded that Wolbachia-Drosophila interaction depends on the genotypes of both the host and the symbiont, but some Wolbachia effects could depend not on the genotypes, but on the fact of recent transfer of the symbiont.}, } @article {pmid38132578, year = {2023}, author = {Zadra, N and Tatti, A and Silverj, A and Piccinno, R and Devilliers, J and Lewis, C and Arnoldi, D and Montarsi, F and Escuer, P and Fusco, G and De Sanctis, V and Feuda, R and Sánchez-Gracia, A and Rizzoli, A and Rota-Stabelli, O}, title = {Shallow Whole-Genome Sequencing of Aedes japonicus and Aedes koreicus from Italy and an Updated Picture of Their Evolution Based on Mitogenomics and Barcoding.}, journal = {Insects}, volume = {14}, number = {12}, pages = {}, pmid = {38132578}, issn = {2075-4450}, support = {PRIN 2022 Prot. n 2022YNL8ZH//Partially funded by the European Union under NextGenerationEU/ ; }, abstract = {Aedes japonicus and Aedes koreicus are two invasive mosquitoes native to East Asia that are quickly establishing in temperate regions of Europe. Both species are vectors of arboviruses, but we currently lack a clear understanding of their evolution. Here, we present new short-read, shallow genome sequencing of A. japonicus and A. koreicus individuals from northern Italy, which we used for downstream phylogenetic and barcode analyses. We explored associated microbial DNA and found high occurrences of Delftia bacteria in both samples, but neither Asaia nor Wolbachia. We then assembled complete mitogenomes and used these data to infer divergence times estimating the split of A. japonicus from A. koreicus in the Oligocene, which was more recent than that previously reported using mitochondrial markers. We recover a younger age for most other nodes within Aedini and other Culicidae. COI barcoding and phylogenetic analyses indicate that A. japonicus yaeyamensis, A. japonicus amamiensis, and the two A. koreicus sampled from Europe should be considered as separate species within a monophyletic species complex. Our studies further clarify the evolution of A. japonicus and A. koreicus, and indicate the need to obtain whole-genome data from putative species in order to disentangle their complex patterns of evolution.}, } @article {pmid38132308, year = {2023}, author = {Lin, YJ and Yeh, CH and Wu, CZ and Wu, LH}, title = {Climate and Wolbachia Impacts on Anoplolepis gracilipes (Hymenoptera: Formicidae).}, journal = {Biology}, volume = {12}, number = {12}, pages = {}, pmid = {38132308}, issn = {2079-7737}, support = {111-2313-B-020-003-MY3//National Science and Technology Council, Taiwan/ ; 112AS-5.5.3-BQ-B1//Animal and Plant Health Inspection Agency (APHIA), Ministry of Agriculture (MOA), Taiwan/ ; }, abstract = {The yellow crazy ant (Anoplolepis gracilipes (Smith, 1857)) is a prominent invasive species exhibiting variable population dynamics. Through collecting long-term climate data and validating field surveys with MaxEnt SDM projections, our results indicated that winter temperature and precipitation accumulation strongly influence the population dynamics. An aggression analysis showed that A. gracilipes nests with higher aggression levels (over 2.5 scores) experienced a higher mean maximum temperature (31.84 ± 0.43 °C) and lower prevalence of wAgra (84.8 ± 4.70%) in A. gracilipes from June to October. The nest manipulation and aggression experiments confirmed that temperature increases aggression (1.3 to 2.8 scores) among A. gracilipes workers due to the reduced prevalence of wAgra. To the best of our knowledge, this is the first case of a notable reduction in the prevalence of Wolbachia (100 to 66%) within a colony of A. gracilipes while maintaining stable nests for further experiments.}, } @article {pmid38125201, year = {2024}, author = {Ochieng, FO}, title = {SEIRS model for malaria transmission dynamics incorporating seasonality and awareness campaign.}, journal = {Infectious Disease Modelling}, volume = {9}, number = {1}, pages = {84-102}, pmid = {38125201}, issn = {2468-0427}, abstract = {Malaria, a devastating disease caused by the Plasmodium parasite and transmitted through the bites of female Anopheles mosquitoes, remains a significant public health concern, claiming over 600,000 lives annually, predominantly among children. Novel tools, including the application of Wolbachia, are being developed to combat malaria-transmitting mosquitoes. This study presents a modified susceptible-exposed-infectious-recovered-susceptible (SEIRS) compartmental mathematical model to evaluate the impact of awareness-based control measures on malaria transmission dynamics, incorporating mosquito interactions and seasonality. Employing the next-generation matrix approach, we calculated a basic reproduction number (R0) of 2.4537, indicating that without robust control measures, the disease will persist in the human population. The model equations were solved numerically using fourth and fifth-order Runge-Kutta methods. The model was fitted to malaria incidence data from Kenya spanning 2000 to 2021 using least squares curve fitting. The fitting algorithm yielded a mean absolute error (MAE) of 2.6463 when comparing the actual data points to the simulated values of infectious human population (Ih). This finding indicates that the proposed mathematical model closely aligns with the recorded malaria incidence data. The optimal values of the model parameters were estimated from the fitting algorithm, and future malaria dynamics were projected for the next decade. The research findings suggest that social media-based awareness campaigns, coupled with specific optimization control measures and effective management methods, offer the most cost-effective approach to managing malaria.}, } @article {pmid38105949, year = {2024}, author = {Shropshire, JD and Conner, WR and Vanderpool, D and Hoffmann, AA and Turelli, M and Cooper, BS}, title = {Rapid host switching of Wolbachia and even more rapid turnover of their phages and incompatibility-causing loci.}, journal = {bioRxiv : the preprint server for biology}, volume = {}, number = {}, pages = {}, pmid = {38105949}, issn = {2692-8205}, support = {R35 GM124701/GM/NIGMS NIH HHS/United States ; }, abstract = {About half of all insect species carry maternally inherited Wolbachia alphaproteobacteria, making Wolbachia the most common endosymbionts known in nature. Often Wolbachia spread to high frequencies within populations due to cytoplasmic incompatibility (CI), a Wolbachia-induced sperm modification caused by prophage-associated genes (cifs) that kill embryos without Wolbachia. Several Wolbachia variants also block viruses, including wMel from Drosophila melanogaster when transinfected into the mosquito Aedes aegypti. CI enables the establishment and stable maintenance of pathogen-blocking wMel in natural Ae. aegypti populations. These transinfections are reducing dengue disease incidence on multiple continents. While it has long been known that closely related Wolbachia occupy distantly related hosts, the timing of Wolbachia host switching and molecular evolution has not been widely quantified. We provide a new, conservative calibration for Wolbachia chronograms based on examples of co-divergence of Wolbachia and their insect hosts. Synthesizing publicly available and new genomic data, we use our calibration to demonstrate that wMel-like variants separated by only about 370,000 years have naturally colonized holometabolous dipteran and hymenopteran insects that diverged approximately 350 million years ago. Data from Wolbachia variants closely related to those currently dominant in D. melanogaster and D. simulans illustrate that cifs are rapidly acquired and lost among Wolbachia genomes, on a time scale of 10[4]-10[5] years. This turnover occurs with and without the Wovirus prophages that contain them, with closely related cifs found in distantly related phages and distantly related cifs found in closely related phages. We present evidence for purifying selection on CI rescue function and on particular Cif protein domains. Our results quantify the tempo and mode of rapid host switching and horizontal gene transfer that underlie the spread and diversity of Wolbachia sampled from diverse host species. The wMel variants we highlight from hosts in different climates may offer new options for broadening Wolbachia-based biocontrol of diseases and pests.}, } @article {pmid38105374, year = {2023}, author = {Cain, JL and Norris, JK and Swan, MP and Nielsen, MK}, title = {A diverse microbial community and common core microbiota associated with the gonad of female Parascaris spp.}, journal = {Parasitology research}, volume = {123}, number = {1}, pages = {56}, pmid = {38105374}, issn = {1432-1955}, mesh = {Animals ; Horses ; Female ; *Ascaridoidea/genetics ; *Horse Diseases/parasitology ; *Ascaridida Infections/veterinary/parasitology ; Drug Resistance ; Feces/parasitology ; *Parasites ; *Helminths ; Gonads ; }, abstract = {The microbiome plays an important role in health, where changes in microbiota composition can have significant downstream effects within the host, and host-microbiota relationships can be exploited to affect health outcomes. Parasitic helminths affect animals globally, but an exploration of their microbiota has been limited, despite the development of anti-Wolbachia drugs to help control infections with some filarial nematodes. The equine ascarids, Parascaris spp., are considered the most pathogenic nematodes affecting juvenile horses and are also the only ascarid parasite to have developed widespread anthelmintic resistance. The aim of this study was to characterize the microbiota of this helminth, focusing on the female gonad, determine a core microbiota for this organ, identify bacterial species, and show bacterial localization to the female gonad via in situ hybridization (ISH). A total of 22 gonads were isolated from female Parascaris spp. collected from three foals, and 9 female parasites were formalin-fixed and paraffin-embedded for ISH. Next-generation sequencing was performed using V3-V4 primers as well as the Swift Amplicon™ 16S+ ITS Panel. Overall, ten genera were identified as members of the Parascaris spp. female gonad and twelve bacterial species were identified. The most prevalent genus was Mycoplasma, followed by Reyranella, and there were no differences in alpha diversity between parasites from different horses. Specific eubacteria staining was identified in both the intestine and within the gonad using ISH. Overall, this study provided in-depth information regarding the female Parascaris spp. microbiota and was the first to identify the core microbiota within a specific parasite organ.}, } @article {pmid38104431, year = {2024}, author = {Lau, DC and Power, RI and Šlapeta, J}, title = {Exploring multiplex qPCR as a diagnostic tool for detecting microfilarial DNA in dogs infected with Dirofilaria immitis: A comparative analysis with the modified Knott's test.}, journal = {Veterinary parasitology}, volume = {325}, number = {}, pages = {110097}, doi = {10.1016/j.vetpar.2023.110097}, pmid = {38104431}, issn = {1873-2550}, mesh = {Animals ; Dogs ; *Dirofilaria immitis/genetics ; Microfilariae/genetics ; Australia ; *Dog Diseases/diagnosis ; *Dirofilariasis/diagnosis ; DNA ; Lactones ; }, abstract = {Current recommendations to diagnose cardiopulmonary dirofilariosis in dogs caused by Dirofilaria immitis involves tandem antigen and circulating microfilariae tests. The modified Knott's test is an important tool in heartworm diagnosis, allowing identification of circulating microfilariae. However, the subjective nature of the modified Knott's test affects its accuracy and diagnostic laboratories usually do not provide a quantitative outcome. Quantitative enumeration of microfilariae enables clinicians to track treatment progress and acts as a proxy for detecting emerging macrocyclic lactone resistance. There is a need for better diagnostic tools suitable for routine use to efficiently and accurately quantify the presence of D. immitis microfilaremia. The aim of this study was to determine whether the quantitative modified Knott's test can be substituted by multiplex quantitative polymerase chain reaction (qPCR) targeting D. immitis and associated Wolbachia endosymbiont DNA in canine blood samples. To do this, genomic DNA samples (n = 161) from Australian dogs, collected as part of a previous 2021 study, were assessed in a TaqMan qPCR targeting DNA of D. immitis, Wolbachia sp. and Canis lupus familiaris. Of the 161 genomic DNA samples, eight were considered positive for D. immitis microfilariae. The qPCR assay demonstrated good efficiency (E = 90 to 110%, R[2] > 0.94). Considering the performance and efficient use of bench time, this TaqMan qPCR assay is a suitable alternative to the modified Knott's test for quantitative enumeration of microfilariae (Cohen's kappa coefficient [κ]: κ = 1 using D. immitis qPCR marker, κ = 0.93 using Wolbachia qPCR marker). The qPCR data demonstrated a comparable result to that of the quantitative modified Knott's test in a 2022 survey of D. immitis in Australian dogs (n = 23) before and after macrocyclic lactone (ML) administration. Improving the detection and diagnosis of canine heartworm infections will assist veterinarians in better managing and controlling disease outcomes and will be valuable for tracking the spread of ML resistance in Australia.}, } @article {pmid38100392, year = {2023}, author = {Malavige, GN and Sjö, P and Singh, K and Piedagnel, JM and Mowbray, C and Estani, S and Lim, SCL and Siquierra, AM and Ogg, GS and Fraisse, L and Ribeiro, I}, title = {Facing the escalating burden of dengue: Challenges and perspectives.}, journal = {PLOS global public health}, volume = {3}, number = {12}, pages = {e0002598}, pmid = {38100392}, issn = {2767-3375}, support = {MC_UU_00008/5/MRC_/Medical Research Council/United Kingdom ; MC_UU_00036/2/MRC_/Medical Research Council/United Kingdom ; }, abstract = {Dengue is the most rapidly emerging mosquito-borne infection and, due to climate change and unplanned urbanization, it is predicted that the global burden of dengue will rise further as the infection spreads to new geographical locations. Dengue-endemic countries are often unable to cope with such increases, with health care facilities becoming overwhelmed during each dengue season. Furthermore, although dengue has been predominantly a childhood illness in the past, it currently mostly affects adults in many countries, with higher incidence of severe disease and mortality rates in pregnant women and in those with comorbidities. As there is currently no specific treatment for dengue and no early biomarker to identify those who will progress to develop vascular leakage, all individuals with dengue are closely monitored in case they need fluid management. Furthermore, diagnosing patients with acute dengue is challenging due to the similarity of clinical symptoms during early illness and poor sensitivity and specificity of point-of-care diagnostic tests. Novel vector control methods, such as the release of Wolbachia-infected mosquitoes, have shown promising results by reducing vector density and dengue incidence in clinical trial settings. A new dengue vaccine, TAK-003, had an efficacy of 61.2% against virologically confirmed dengue, 84.1% efficacy against hospitalizations and a 70% efficacy against development of dengue haemorrhagic fever (DHF) at 54 months. While vaccines and mosquito control methods are welcome, they alone are unlikely to fully reduce the burden of dengue, and a treatment for dengue is therefore essential. Several novel antiviral drugs are currently being evaluated along with drugs that inhibit host mediators, such as mast cell products. Although viral proteins such as NS1 contribute to the vascular leak observed in severe dengue, the host immune response to the viral infection also plays a significant role in progression to severe disease. There is an urgent need to discover safe and effective treatments for dengue to prevent disease progression.}, } @article {pmid38076953, year = {2023}, author = {Fricke, LC and Lindsey, AR}, title = {Identification of parthenogenesis-inducing effector proteins in Wolbachia.}, journal = {bioRxiv : the preprint server for biology}, volume = {}, number = {}, pages = {}, pmid = {38076953}, issn = {2692-8205}, support = {R35 GM150991/GM/NIGMS NIH HHS/United States ; }, abstract = {Bacteria in the genus Wolbachia have evolved numerous strategies to manipulate arthropod sex, including the conversion of would-be male offspring to asexually reproducing females. This so-called "parthenogenesis-induction" phenotype can be found in a number of Wolbachia strains that infect arthropods with haplodiploid sex determination systems, including parasitoid wasps. Despite the discovery of microbe-mediated parthenogenesis more than 30 years ago, the underlying genetic mechanisms have remained elusive. We used a suite of genomic, computational, and molecular tools to identify and characterize two proteins that are uniquely found in parthenogenesis-inducing Wolbachia and have strong signatures of host-associated bacterial effector proteins. These putative parthenogenesis-inducing proteins have structural homology to eukaryotic protein domains including nucleoporins, the key insect sex-determining factor Transformer, and a eukaryotic-like serine-threonine kinase with leucine rich repeats. Furthermore, these proteins significantly impact eukaryotic cell biology in the model, Saccharomyces cerevisiae. We suggest these proteins are parthenogenesis-inducing factors and our results indicate this would be made possible by a novel mechanism of bacterial-host interaction.}, } @article {pmid38071646, year = {2024}, author = {Urairi, C and Fujito, S}, title = {Interbiotype hybridization between biotypes A and B of Liriomyza chinensis (Diptera: Agromyzidae).}, journal = {Journal of economic entomology}, volume = {117}, number = {1}, pages = {240-250}, doi = {10.1093/jee/toad223}, pmid = {38071646}, issn = {1938-291X}, mesh = {Female ; Male ; Animals ; *Diptera/genetics ; *Hemiptera ; Reproduction ; Hybridization, Genetic ; Fertility ; }, abstract = {Liriomyza chinensis (Kato) is a formidable pest of Allium species, especially the Japanese bunching onion Allium fistulosum L. Recently, a novel biotype of L. chinensis (biotype B) has emerged, which causes more severe damage than the native biotype A. It has been reported that biotype B has frequently displaced biotype A in the Japanese bunching onion fields in Japan. As interbiotype hybridization is a possible factor that influences such displacement, interbiotype hybridization was conducted between L. chinensis biotypes A and B. Eggs were not laid under one-by-one crossing conditions; however, adult hybrid progeny of both sexes emerged from no-choice mating combinations-when multiple males and females were present. The fertility of F1 hybrid adults was also investigated, and backcrossed adults emerged from F1 females in both mating combinations. F1 males might have exhibited reproductive abnormalities because only a small number of backcross progeny emerged from the mating combinations using F1 males. Additionally, 3 representative endosymbionts (Wolbachia, Spiroplasma, and Cardinium) were investigated, and both biotypes were found to be infected by the same strain of Wolbachia. In addition, the courtship signals (tapping) of male adults differed between biotypes A and B as well as between F1 hybrids; the F1 males exhibited tapping behavior that was intermediate between biotypes A and B. Therefore, mating sounds serve as a form of premating reproductive isolation between biotypes A and B.}, } @article {pmid38070178, year = {2024}, author = {Arai, H and Ueda, M and Hirano, T and Akizuki, N and Lin, SR and Hanh, DK and Widada, J and Rohman, MS and Nakai, M and Kunimi, Y and Vang, LV and Wijonarko, A and Inoue, MN}, title = {Conserved infections and reproductive phenotypes of Wolbachia symbionts in Asian tortrix moths.}, journal = {Environmental microbiology reports}, volume = {16}, number = {1}, pages = {e13219}, pmid = {38070178}, issn = {1758-2229}, support = {19J13123//Japan Society for the Promotion of Science/ ; 21J00895//Japan Society for the Promotion of Science/ ; 21KK0105//Japan Society for the Promotion of Science/ ; 1539057329//JSPS Overseas Challenge Program for Young Researchers (2019) with RISTEK Foreign Research Permit/ ; }, mesh = {Animals ; Female ; Male ; *Moths/genetics/microbiology ; *Wolbachia/genetics ; Reproduction ; Phenotype ; Bacteria ; Symbiosis ; }, abstract = {Wolbachia is a ubiquitous endosymbiotic bacterium that manipulates insect reproduction. A notable feature of Wolbachia is male killing (MK), whereby sons of infected females are killed during development; however, the evolutionary processes by which Wolbachia acquired the MK ability remain unclear. The tea tortrix moth Homona magnanima (Tortricidae) harbours three non-MK Wolbachia strains (wHm-a, wHm-b and wHm-c) and an MK strain wHm-t. Although wHm-t and wHm-c are closely related, only wHm-t has an MK-associated prophage region. To understand the evolutionary processes underlying the emergence of MK wHm-t, we examined Wolbachia infections and phenotypes in 62 tortricid species collected from 39 localities across Japan, Taiwan, Vietnam and Indonesia. PCR assays detected wHm-c relatives in 51 species and triple infection of wHm-a, wHm-b and wHm-c in 31 species. Apart from Taiwanese H. magnanima, no species exhibited the MK phenotype and were positive for the wHm-t-specific prophage. While wHm-t infection was dominant in Taiwanese H. magnanima, wHm-a, wHm-b and wHm-c were dominant in Japanese H. magnanima populations. These results suggest that wHm-a, wHm-b and wHm-c strains descended from a common ancestor with repeated infection loss and that wHm-t evolved from the wHm-c acquiring MK ability in allopatric populations of H. magnanima.}, } @article {pmid38060903, year = {2023}, author = {Valdez-Delgado, KM and Ríos-Delgado, JC and Nettel-Cruz, JA and Angulo-Kladt, R and Villarreal-Treviño, C}, title = {Aerial release of Aedes aegypti male mosquitoes using an unmanned aerial vehicle: a novel control strategy.}, journal = {Salud publica de Mexico}, volume = {65}, number = {4, jul-ago}, pages = {387-393}, doi = {10.21149/14466}, pmid = {38060903}, issn = {1606-7916}, mesh = {Humans ; Animals ; Female ; Male ; *Aedes ; Unmanned Aerial Devices ; Temperature ; Sexual Behavior ; *Wolbachia ; Mosquito Control/methods ; }, abstract = {OBJECTIVE: To development of a methodology for the chilling, handling, transport, and release of male Aedes aegypti mosquitoes, reared in insectary conditions to release in the field with unmanned vehicles to compete sexually with wild males in the field.

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

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

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

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

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

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

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

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

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

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

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

CONCLUSION: These results, from the largest contiguous releases of wMel Wolbachia mosquitoes to date, highlight the operational feasibility of implementing the method in large urban settings. Based on results from previous studies, we expect that Wolbachia establishment will be sustained long term. Ongoing monitoring will confirm Wolbachia persistence in local mosquito populations and track its establishment in the remaining areas.}, } @article {pmid38021190, year = {2023}, author = {Bawm, S and Khaing, Y and Chel, HM and Hmoon, MM and Win, SY and Bo, M and Naing, T and Htun, LL}, title = {Molecular detection of Dirofilaria immitis and its Wolbachia endosymbionts in dogs from Myanmar.}, journal = {Current research in parasitology & vector-borne diseases}, volume = {4}, number = {}, pages = {100148}, pmid = {38021190}, issn = {2667-114X}, abstract = {Heartworm disease in dogs and cats caused by Dirofilaria immitis continues to be a major clinical issue globally. This study focused on dogs suspicious of having tick-borne diseases (TBD) brought to a clinic and a veterinary teaching hospital in Myanmar. Blood samples were collected and initially screened using SNAP® 4Dx® Plus test kit. All dog blood samples were subjected to conventional PCR to detect both Dirofilaria spp. (cox1 gene) and Wolbachia spp. (16S rDNA) infections. Infection with D. immitis was detected in 14 (28.0%) of 50 examined samples, while the detection rate of TBD causative agents, including Anaplasma phagocytophilum and Ehrlichia canis, was 26.0% (13/50) and 26.0% (13/50), respectively, as determined by ELISA rapid test. In this study, D. immitis infection was moderately but significantly correlated with TBD infections (Pearson's r = 0.397, P = 0.008). Comparative sequence and phylogenetic analyses provided molecular identification of D. immitis in Myanmar and confirmed the identity of its Wolbachia endosymbiont with Wolbachia endosymbionts isolated from D. immitis, Rhipicephalus sanguineus and Aedes aegypti. The present study contributes to our understanding of the coexistence of D. immitis and Wolbachia endosymbiosis in dogs, and the findings may benefit the future prevention and control of dirofilariasis in dogs.}, } @article {pmid38020682, year = {2023}, author = {McPherson, AE and Abram, PK and Curtis, CI and Wannop, ER and Dudzic, JP and Perlman, SJ}, title = {Dynamic changes in Wolbachia infection over a single generation of Drosophila suzukii, across a wide range of resource availability.}, journal = {Ecology and evolution}, volume = {13}, number = {11}, pages = {e10722}, pmid = {38020682}, issn = {2045-7758}, abstract = {Wolbachia bacteria are maternally inherited symbionts that commonly infect terrestrial arthropods. Many Wolbachia reach high frequencies in their hosts by manipulating their reproduction, for example by causing reproductive incompatibilities between infected male and uninfected female hosts. However, not all strains manipulate reproduction, and a key unresolved question is how these non-manipulative Wolbachia persist in their hosts, often at intermediate to high frequencies. One such strain, wSuz, infects the invasive fruit pest Drosophila suzukii, spotted-wing drosophila. Here, we tested the hypothesis that wSuz infection provides a competitive benefit when resources are limited. Over the course of one season, we established population cages with varying amounts of food in a semi-field setting and seeded them with a 50:50 mixture of flies with and without Wolbachia. We predicted that Wolbachia-infected individuals should have higher survival and faster development than their uninfected counterparts when there was little available food. We found that while food availability strongly impacted fly fitness, there was no difference in development times or survival between Wolbachia-infected and uninfected flies. Interestingly, however, Wolbachia infection frequencies changed dramatically, with infections either increasing or decreasing by as much as 30% in a single generation, suggesting the possibility of unidentified factors shaping Wolbachia infection over the course of the season.}, } @article {pmid38018822, year = {2023}, author = {Dai, X and Quan, Q and Jiao, J}, title = {Modelling and analysis of periodic impulsive releases of the Nilaparvata lugens infected with wStri-Wolbachia.}, journal = {Journal of biological dynamics}, volume = {17}, number = {1}, pages = {2287077}, doi = {10.1080/17513758.2023.2287077}, pmid = {38018822}, issn = {1751-3766}, mesh = {Animals ; *Wolbachia ; Models, Biological ; Population Dynamics ; *Hemiptera ; }, abstract = {In this paper, we formulate a population suppression model and a population replacement model with periodic impulsive releases of Nilaparvata lugens infected with wStri. The conditions for the stability of wild-N.lugens-eradication periodic solution of two systems are obtained by applying the Floquet theorem and comparison theorem. And the sufficient conditions for the persistence in the mean of wild N.lugens are also given. In addition, the sufficient conditions for the extinction and persistence of the wild N.lugens in the subsystem without wLug are also obtained. Finally, we give numerical analysis which shows that increasing the release amount or decreasing the release period are beneficial for controlling the wild N.lugens, and the efficiency of population replacement strategy in controlling wild populations is higher than that of population suppression strategy under the same release conditions.}, } @article {pmid38016137, year = {2024}, author = {Azarm, A and Koosha, M and Dalimi, A and Zahraie-Ramazani, A and Akhavan, AA and Saeidi, Z and Mohebali, M and Azam, K and Vatandoost, H and Oshaghi, MA}, title = {Association Between Wolbachia Infection and Susceptibility to Deltamethrin Insecticide in Phlebotomus papatasi (Diptera: Psychodidae), the Main Vector of Zoonotic Cutaneous Leishmaniasis.}, journal = {Vector borne and zoonotic diseases (Larchmont, N.Y.)}, volume = {24}, number = {3}, pages = {159-165}, doi = {10.1089/vbz.2023.0085}, pmid = {38016137}, issn = {1557-7759}, mesh = {Animals ; Humans ; *Phlebotomus/microbiology ; *Psychodidae ; *Insecticides/pharmacology ; *Wolbachia/genetics ; *Leishmaniasis, Cutaneous/veterinary ; *Nitriles ; *Pyrethrins ; }, abstract = {Background: Phlebotomus papatasi (Diptera: Psychodidae) is the main vector of zoonotic cutaneous leishmaniasis. Wolbachia is a symbiotic alphaproteobacteria of arthropods that can be involved in susceptibility or resistance. This study aimed to investigate the relationship between Wolbachia and Deltamethrin susceptibility/resistance in Ph. papatasi. Deltamethrin filter papers (0.00002%) were used to test sand fly field collected from southern Iran. After the test, PCR amplification of the Wolbachia surface protein gene (wsp) was used to measure Wolbachia infection rate in the killed, surviving, and control groups. Result: The rates of infection by Wolbachia strain (wPap, super group A) differed between killed (susceptible) and surviving (resistant) Ph. papatasi specimens. The rate of Wolbachia infection in susceptible individuals was more than twice (2.3) (39% vs. 17%) in resistant individuals with the same genetic background. This difference was highly significant (p < 0.001), indicating a positive association between Wolbachia infection and susceptibility to Deltamethrin. In addition, the results showed that Deltamethrin can act as a PCR inhibitor during detection of Wolbachia in Ph. papatasi. Conclusion: Results of this study show that Wolbachia is associated with Deltamethrin susceptibility level in Ph. papatasi. Also, as Deltamethrin has been identified as a PCR inhibitor, great care must be taken in interpreting Wolbachia infection status in infected populations. The results of this study may provide information for a better understanding of the host-symbiont relationship, as well as application of host symbiosis in pest management.}, } @article {pmid38012718, year = {2023}, author = {Zhang, H and Trueman, E and Hou, X and Chew, X and Deng, L and Liew, J and Chia, T and Xi, Z and Tan, CH and Cai, Y}, title = {Different mechanisms of X-ray irradiation-induced male and female sterility in Aedes aegypti.}, journal = {BMC biology}, volume = {21}, number = {1}, pages = {274}, pmid = {38012718}, issn = {1741-7007}, mesh = {Humans ; Animals ; Male ; Female ; *Infertility, Female ; X-Rays ; *Aedes ; Mosquito Vectors ; Mosquito Control/methods ; *Wolbachia ; Insecta ; *Zika Virus ; *Zika Virus Infection ; }, abstract = {BACKGROUND: Aedes aegypti (Ae. aegypti) is the major vector that transmits many diseases including dengue, Zika, and filariasis in tropical and subtropical regions. Due to the growing resistance to chemical-based insecticides, biological control methods have become an emerging direction to control mosquito populations. The sterile insect technique (SIT) deploys high doses of ionizing radiation to sterilize male mosquitoes before the release. The Wolbachia-based population suppression method of the incompatible insect technique (IIT) involves the release of Wolbachia-infected males to sterilize uninfected field females. Due to the lack of perfect sex separation tools, a low percentage of female contamination is detected in the male population. To prevent the unintentional release of these Wolbachia-infected females which might result in population replacement, a low dose of X-ray irradiation is deployed to sterilize any female escapees. However, it remains unclear whether these irradiation-induced male and female sterilizations share common mechanisms.

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

CONCLUSIONS: Our findings thus reveal the distinct mechanisms of ionizing X-ray irradiation-induced male or female sterility in Ae. aegypti mosquitoes, which may help the design of X-ray irradiation-based vector control methods.}, } @article {pmid38011160, year = {2023}, author = {Oliveira, TMP and Rojas, MVR and Amorim, JA and Alonso, DP and de Carvalho, DP and Ribeiro, KAN and Sallum, MAM}, title = {Bacterial diversity on larval and female Mansonia spp. from different localities of Porto Velho, Rondonia, Brazil.}, journal = {PloS one}, volume = {18}, number = {11}, pages = {e0293946}, pmid = {38011160}, issn = {1932-6203}, mesh = {Female ; Animals ; Larva/microbiology ; Brazil ; RNA, Ribosomal, 16S/genetics ; *Culicidae/genetics ; Bacteria/genetics ; }, abstract = {Studies based on the bacterial diversity present in Mansonia spp. are limited; therefore, the aim of this study was to investigate the bacterial diversity in females and larvae of Mansonia spp., describe the differences between the groups identified, and compare the microbiota of larvae from different collection sites. Sequences of the 16S rRNA region from the larvae and females of Mansonia spp. were analyzed. Diversity analyzes were performed to verify the possible bacterial differences between the groups and the collection sites. The results showed Pseudomonas was the most abundant genus in both females and larvae, followed by Wolbachia in females and Rikenellaceae and Desulfovibrio in larvae. Desulfovibrio and Sulfurospirillum, sulfate- and sulfur-reducing bacteria, respectively, were abundant on the larvae. Aminomonas, an amino acid-degrading bacterium, was found only in larvae, whereas Rickettsia was identified in females. Bacterial diversity was observed between females and larvae of Mansonia and between larvae from different collection sites. In addition, the results suggest that the environment influenced bacterial diversity.}, } @article {pmid38006562, year = {2024}, author = {Serbus, LR}, title = {A Light in the Dark: Uncovering Wolbachia-Host Interactions Using Fluorescence Imaging.}, journal = {Methods in molecular biology (Clifton, N.J.)}, volume = {2739}, number = {}, pages = {349-373}, pmid = {38006562}, issn = {1940-6029}, mesh = {Animals ; *Wolbachia/genetics ; *Arthropods/microbiology ; *Nematoda ; Symbiosis ; Optical Imaging ; }, abstract = {The success of microbial endosymbionts, which reside naturally within a eukaryotic "host" organism, requires effective microbial interaction with, and manipulation of, the host cells. Fluorescence microscopy has played a key role in elucidating the molecular mechanisms of endosymbiosis. For 30 years, fluorescence analyses have been a cornerstone in studies of endosymbiotic Wolbachia bacteria, focused on host colonization, maternal transmission, reproductive parasitism, horizontal gene transfer, viral suppression, and metabolic interactions in arthropods and nematodes. Fluorescence-based studies stand to continue informing Wolbachia-host interactions in increasingly detailed and innovative ways.}, } @article {pmid38006561, year = {2024}, author = {Bordenstein, SR}, title = {Isolation of Phage WO Particles from Wolbachia-Infected Arthropods.}, journal = {Methods in molecular biology (Clifton, N.J.)}, volume = {2739}, number = {}, pages = {337-348}, pmid = {38006561}, issn = {1940-6029}, mesh = {Animals ; *Arthropods ; *Wolbachia/genetics ; *Bacteriophages/genetics ; Prophages ; Myoviridae ; }, abstract = {Nearly all arthropod-associated Wolbachia contain intact and/or genomic remnants of phage WO, temperate bacteriophages that facilitate horizontal gene transfer, genomic rearrangement of the bacterial chromosome, and symbiotic interactions between Wolbachia and their arthropod hosts. Integrated prophage WO genomes produce active, lytic particles; but the lack of a cell-free culturing system for Wolbachia render them difficult to purify and study. This chapter describes polyethylene glycol (PEG) precipitation of phage particles from Wolbachia-infected arthropods, followed by confirmation of phage WO isolation and purification using electron microscopy and PCR.}, } @article {pmid38006560, year = {2024}, author = {Murphy, RO and Beckmann, JF}, title = {Using Baker's Yeast to Determine Functions of Novel Wolbachia (and Other Prokaryotic) Effectors.}, journal = {Methods in molecular biology (Clifton, N.J.)}, volume = {2739}, number = {}, pages = {321-336}, pmid = {38006560}, issn = {1940-6029}, mesh = {Animals ; Humans ; *Saccharomyces cerevisiae/genetics/metabolism ; *Wolbachia/genetics ; Genome ; Protein Interaction Mapping ; Insecta/genetics ; Symbiosis ; }, abstract = {Yeasts are single-celled eukaryotic organisms classified as fungi, mostly in the phylum Ascomycota. Of about 1500 named species, Saccharomyces cerevisiae, also known as baker's yeast, domesticated by humans in the context of cooking and brewing, is a profound genetic tool for exploring functions of novel effector proteins from Wolbachia and prokaryotes in general. Wolbachia is a Gram-negative alpha-proteobacterium that infects up to ~75% of all insects as an obligate intracellular microbe (Jeyaprakash A, Hoy MA, Insect Mol Biol 9:393-405, 2000). Wolbachia's lifestyle presents unique challenges for researchers. Wolbachia cannot be axenically cultured and has never been genetically manipulated. Furthermore, many Wolbachia genes have no known function or well-annotated orthologs in other genomes. Yet given the effects of Wolbachia on host phenotypes, which have considerable practical applications for pest control, they undoubtedly involve secreted effector proteins that interact with host gene products. Studying these effectors is challenging with Wolbachia's current genetic limitations. However, some of the constraints to working with Wolbachia can be overcome by expressing candidate proteins in S. cerevisiae. This approach capitalizes on yeast's small genome (~6500 genes), typical eukaryotic cellular organization, and the sophisticated suite of genetic tools available for its manipulation in culture. Thus, yeast can serve as a powerful mock eukaryotic host background to study Wolbachia effector function. Specifically, yeast is used for recombinant protein expression, drug discovery, protein localization studies, protein interaction mapping (yeast two-hybrid system), modeling chromosomal evolution, and examining interactions between proteins responsible for complex phenotypes in less tractable prokaryotic systems. As an example, the paired genes responsible for Wolbachia-mediated cytoplasmic incompatibility (CI) encode novel proteins with limited homology to other known proteins, and no obvious function. This article details how S. cerevisiae was used as an initial staging ground to explore the molecular basis of one of Wolbachia's trademark phenotypes (CI).}, } @article {pmid38006559, year = {2024}, author = {Cortez, CT and Murphy, RO and Owens, IM and Beckmann, JF}, title = {Use of Drosophila Transgenics to Identify Functions for Symbiont Effectors.}, journal = {Methods in molecular biology (Clifton, N.J.)}, volume = {2739}, number = {}, pages = {301-320}, doi = {10.1007/978-1-0716-3553-7_18}, pmid = {38006559}, issn = {1940-6029}, mesh = {Animals ; *Drosophila ; Drosophila melanogaster/genetics ; Animals, Genetically Modified ; Biological Evolution ; Insecta ; *Wolbachia/genetics ; Symbiosis/genetics ; }, abstract = {Wolbachia, one of the most successful and studied insect symbionts, and Drosophila, one of the most understood model insects, can be exploited as complementary tools to unravel mechanisms of insect symbiosis. Although Wolbachia itself cannot be grown axenically as clonal isolates or genetically manipulated by standard methods, its reproductive phenotypes, including cytoplasmic incompatibility (CI), have been elucidated using well-developed molecular tools and precise transgenic manipulations available for Drosophila melanogaster. Current research only scratches the surface of how Drosophila can provide a tool for understanding Wolbachia's evolutionary success and the molecular roles of its genetic elements. Here, we briefly outline basic methodologies inherent to transgenic Drosophila systems that have already contributed significant advances in understanding CI, but may be unfamiliar to those who lack experience in Drosophila genetics. In the future, these approaches will continue providing significant insights into Wolbachia that undoubtedly will be extended to other insect symbionts and their biological capabilities.}, } @article {pmid38006558, year = {2024}, author = {Valerio, F and Twort, VG and Duplouy, A}, title = {A Worked Example of Screening Genomic Material for the Presence of Wolbachia Infection.}, journal = {Methods in molecular biology (Clifton, N.J.)}, volume = {2739}, number = {}, pages = {275-299}, doi = {10.1007/978-1-0716-3553-7_17}, pmid = {38006558}, issn = {1940-6029}, mesh = {Genome, Bacterial ; Genomics ; Phylogeny ; Symbiosis/genetics ; *Wolbachia/genetics ; }, abstract = {This chapter gives a brief overview of how to screen existing host genomic data for the presence of endosymbionts, such as Wolbachia. The various programs used provide test examples, and the corresponding manuals and discussion boards provide invaluable information. Please do consult these resources.}, } @article {pmid38006557, year = {2024}, author = {Valerio, F and Twort, VG and Duplouy, A}, title = {Screening Host Genomic Data for Wolbachia Infections.}, journal = {Methods in molecular biology (Clifton, N.J.)}, volume = {2739}, number = {}, pages = {251-274}, pmid = {38006557}, issn = {1940-6029}, mesh = {*Wolbachia/genetics ; Genome, Bacterial ; Genomics ; Base Sequence ; DNA, Bacterial/genetics ; Symbiosis/genetics ; }, abstract = {Less than a decade ago, the production of Wolbachia genomic assemblies was tedious, time-consuming, and expensive. The production of Wolbachia genomic DNA free of contamination from host DNA, as required for Wolbachia-targeted sequencing, was then only possible after the amplification and extraction of a large amount of clonal Wolbachia DNA. However, as an endosymbiotic bacterium, Wolbachia does not grow outside the host cell environment, and large-scale recovery of the bacteria required mass rearing of their host, preferably clones of a single individual to avoid strain genetic diversity, or amplification of cell cultures infected with a single Wolbachia strain. Bacterial DNA could be separated from host DNA based on genomic size. Nowadays, the production of full Wolbachia genomes does not require the physical isolation of the bacterial strains from their respective hosts, and the bacterium is often sequenced as a by-catch of host genomic projects. Here, we provide a step-by-step protocol to (1) identify whether host genome projects contain reads from associated Wolbachia and (2) isolate/retrieve the Wolbachia reads from the rest of the sequenced material. We hope this simple protocol will support many projects aiming at studying diverse Wolbachia genome assemblies.}, } @article {pmid38006555, year = {2024}, author = {Chrostek, E}, title = {Procedures for the Detection of Wolbachia-Conferred Antiviral Protection in Drosophila melanogaster.}, journal = {Methods in molecular biology (Clifton, N.J.)}, volume = {2739}, number = {}, pages = {219-237}, pmid = {38006555}, issn = {1940-6029}, mesh = {Animals ; Drosophila melanogaster/genetics ; *Wolbachia/genetics ; Host-Pathogen Interactions/genetics ; *Virus Diseases ; Antiviral Agents ; Symbiosis ; }, abstract = {Spread of Wolbachia infections in host populations may be enhanced by Wolbachia-conferred protection from viral pathogens. Wolbachia-infected Drosophila melanogaster survive the pathogenic effects of positive-sense single-stranded RNA virus infections at a higher rate than the flies without Wolbachia. The protection can occur with or without detectable reduction in virus titer. For the comparisons to be meaningful, Wolbachia-harboring and Wolbachia-free insects need to be genetically matched, and original populations of gut microbiota need to be restored after the removal of Wolbachia using antibiotics. Here, I describe the procedures needed to detect Wolbachia-conferred antiviral protection against Drosophila C virus measured as the difference in survival and viral titer between flies with and without Wolbachia.}, } @article {pmid38006554, year = {2024}, author = {Walker, T}, title = {Detection of Natural Wolbachia Strains in Anopheles Mosquitoes.}, journal = {Methods in molecular biology (Clifton, N.J.)}, volume = {2739}, number = {}, pages = {205-218}, pmid = {38006554}, issn = {1940-6029}, mesh = {Animals ; Humans ; *Anopheles/genetics ; *Wolbachia/genetics ; In Situ Hybridization, Fluorescence ; Mosquito Vectors ; *Malaria ; }, abstract = {Wolbachia is an endosymbiotic bacterium that naturally infects many insect species, including mosquitoes that transmit human diseases. Wolbachia strains have been shown to inhibit the transmission of both arboviruses and malaria Plasmodium parasites. The existence of natural strains in wild Anopheles (An.) mosquitoes, the vectors of malaria parasites, in an endosymbiotic relationship is still to be fully determined. Although Wolbachia has been reported to be present in wild populations of the An. gambiae complex, the primary vectors of malaria in Sub-Saharan Africa, Wolbachia DNA sequence density and infection frequencies are low. As most studies have used highly sensitive nested PCR as the only detection method, more robust evidence is required to determine whether Wolbachia strains are established as endosymbionts in Anopheles species. Techniques such as fluorescent in situ hybridization, microbiome sequencing, and Wolbachia whole genome sequencing have provided concrete evidence for genuine Wolbachia strains in two mosquito species: An. moucheti and An. demeilloni. In this chapter, the current methodology used to determine if resident strains exist in Anopheles mosquitoes will be reviewed, including both PCR- and non-PCR-based protocols.}, } @article {pmid38006553, year = {2024}, author = {Ross, PA}, title = {Measuring Host Fitness Effects and Transmission of Wolbachia Strains in Aedes aegypti Mosquitoes.}, journal = {Methods in molecular biology (Clifton, N.J.)}, volume = {2739}, number = {}, pages = {189-203}, pmid = {38006553}, issn = {1940-6029}, mesh = {Animals ; Male ; *Aedes ; *Wolbachia ; Mosquito Vectors ; *Arboviruses ; }, abstract = {Lines of Aedes aegypti mosquitoes infected with heritable Wolbachia bacteria are being developed and released for arbovirus control. Coordinated releases of lab-reared Wolbachia mosquitoes have reduced local disease incidence by spreading virus-blocking Wolbachia strains and by crashing mosquito populations through incompatible male releases. The phenotypic effects of Wolbachia are diverse and depend on both genetics and the environment. Accurate assessments of Wolbachia effects in mosquitoes are essential, as such effects can make the difference between success and failure of a Wolbachia release program. This chapter provides guidelines for testing key Wolbachia host effects and transmission in Aedes aegypti: the most important arbovirus vector and the most common target of Wolbachia release programs. The protocols should be useful for evaluating mosquito strains prior to field release.}, } @article {pmid38006552, year = {2024}, author = {Zhang, M and Xi, Z}, title = {Wolbachia Transinfection Via Embryonic Microinjection.}, journal = {Methods in molecular biology (Clifton, N.J.)}, volume = {2739}, number = {}, pages = {175-188}, pmid = {38006552}, issn = {1940-6029}, mesh = {Animals ; Mosquito Vectors ; *Wolbachia ; Microinjections ; Cytoplasm ; Cytosol ; *Aedes ; Insecta ; }, abstract = {The process of transferring Wolbachia from one species to another to establish a stable, maternally inherited infection in the target species is known as transinfection. The success of transinfection is primarily achieved through embryonic microinjection, which is the most direct and efficient means of delivering Wolbachia into the germline of the target species and establishing stable maternal transmission. For the fundamental studies, transinfection is often used to characterize Wolbachia-host interactions, including Wolbachia host range, the role of host or bacterial factors in symbiosis, and evolution of Wolbachia-host associations. For the applied studies, use of transinfection to generate a novel infection in the target species is the first step to build the weapon for both population replacement and population suppression for controlling insect pests or their transmitted diseases. For the primary dengue vector Aedes aegypti and Anopheles vectors of malaria, which either do not naturally carry Wolbachia or are infected with strains that lack necessary features for implementation, transinfection can be established by introducing a novel strain capable of inducing both cytoplasmic incompatibility (CI) and pathogen blocking. For A. albopictus and Culex mosquito species, which naturally harbor CI-inducing Wolbachia, transinfection can be achieved by either introducing a novel strain to generate superinfection or replacing the native infection with a different Wolbachia strain in a symbiont-free line, which is derived from antibiotic treatment of the wild type. Here, we use A. aegypti as an example to describe the Wolbachia transinfection method, which can be adapted to other insect species, such as planthoppers, according to their specific developmental requirements.}, } @article {pmid38006551, year = {2024}, author = {Fallon, AM}, title = {Preparation of Infectious Wolbachia from a Mosquito Cell Line.}, journal = {Methods in molecular biology (Clifton, N.J.)}, volume = {2739}, number = {}, pages = {157-171}, pmid = {38006551}, issn = {1940-6029}, mesh = {Animals ; *Wolbachia/genetics ; Mitomycin ; Cell Line ; *Aedes ; }, abstract = {Eventual genetic engineering of Wolbachia will require maximizing recovery of infectious bacteria, maintaining Wolbachia in a viable state for efficient manipulation, and reinfection of host cells for propagation and expansion of recombinant progeny. Challenges to manipulating Wolbachia arise from its obligate intracellular lifestyle and inability to divide outside a host cell, requiring modifications of standard bacteriological methods. The Aedes albopictus C7-10 cell line has proven to be a good recipient for the Wolbachia supergroup B strain, wStri, from the planthopper Laodelphax striatellus; the persistently infected C/wStri1 population provides a source of wStri inoculum that can be used systematically to explore conditions that increase yields of infectious material from input Wolbachia and identify conditions conducive to Wolbachia replication. After reintroduction into naive, uninfected C7-10 cells, wStri recovery, relative to the input inoculum, is influenced by diverse conditions, such as the cell cycle arrest that follows treatment of infected host cells with the insect steroid hormone, 20-hydroxyecdysone. Pretreatment of recipient cells with mitomycin C, which cross-links DNA and inhibits host cell replication, can improve recovery from low levels of input Wolbachia. This protocol describes preparation of infectious inoculum from Aedes albopictus C/wStri1 cells and amplification of Wolbachia in mitomycin C-treated, uninfected C7-10 cells, followed by a brief description of conditions used for various small-scale manipulations of Wolbachia in infected cells.}, } @article {pmid38006550, year = {2024}, author = {Bishop, C and Asgari, S}, title = {Use of Antibiotics to Eliminate Wolbachia from Mosquitoes and Cell Culture.}, journal = {Methods in molecular biology (Clifton, N.J.)}, volume = {2739}, number = {}, pages = {147-156}, pmid = {38006550}, issn = {1940-6029}, mesh = {Animals ; Anti-Bacterial Agents/pharmacology ; *Wolbachia/genetics ; *Aedes ; Cell Line ; Cell Culture Techniques ; }, abstract = {Removal of Wolbachia from infected insects is required in most experimental settings when the effects of Wolbachia on biological traits, pathogen blocking, reproduction, and fitness are assessed. This is to ensure that the genetic backgrounds of Wolbachia-infected and uninfected insects are the same. Here, we describe methodologies used for the elimination of Wolbachia from insects and insect cell lines with antibiotics.}, } @article {pmid38006549, year = {2024}, author = {Madhav, M and James, P}, title = {Developing Wolbachia-Infected Cell Lines from Insects.}, journal = {Methods in molecular biology (Clifton, N.J.)}, volume = {2739}, number = {}, pages = {137-145}, pmid = {38006549}, issn = {1940-6029}, mesh = {Animals ; *Wolbachia ; Insecta ; Cell Line ; }, abstract = {Wolbachia, an intracellular bacterium of arthropods, is an ideal candidate for use in the biological control of insect pests. The inability of Wolbachia to grow in the extracellular environment requires maintenance in live insects or insect cell cultures. Growing and adapting high-density Wolbachia in the targeted host cells improves the possibility of cross-species transinfection. Here, we describe a method for the establishment of a primary cell culture from insect embryos and its transinfection with Wolbachia.}, } @article {pmid38006548, year = {2024}, author = {Kaur, A and Brown, AMV}, title = {Detection and Analysis of Wolbachia in Plant-Parasitic Nematodes and Insights into Wolbachia Evolution.}, journal = {Methods in molecular biology (Clifton, N.J.)}, volume = {2739}, number = {}, pages = {115-134}, pmid = {38006548}, issn = {1940-6029}, mesh = {Animals ; *Wolbachia/genetics ; *Nematoda/genetics ; Plants/parasitology ; Genomics ; Metagenomics ; }, abstract = {Since the discovery of Wolbachia in plant-parasitic nematodes (PPNs), there has been increased interest in this earliest branching clade that may hold important clues to early transitions in Wolbachia function in the Ecdysozoa. However, due to the specialized skills and equipment of nematology and the difficulty in culturing most PPNs, these PPN-type Wolbachia remain undersampled and poorly understood. To date, there are few established laboratory methods for working with PPN-type Wolbachia strains, and most research has relied on chance discovery and comparative genomics. Here, we address this challenge by providing detailed methods to assist researchers with more efficiently collecting PPNs and screen these communities, populations, or single nematodes with a newly developed PPN-type Wolbachia-specific PCR assay. We provide an overview of the typical yields and outcomes of these methods, to facilitate further targeted cultivation or experimental methods, and finally we provide a short introduction to some of the specific challenges and solutions in following through with comparative or population genomics on PPN-type Wolbachia strains.}, } @article {pmid38006547, year = {2024}, author = {Kakumanu, ML and Hickin, ML and Schal, C}, title = {Detection, Quantification, and Elimination of Wolbachia in Bed Bugs.}, journal = {Methods in molecular biology (Clifton, N.J.)}, volume = {2739}, number = {}, pages = {97-114}, pmid = {38006547}, issn = {1940-6029}, support = {P30 ES025128/ES/NIEHS NIH HHS/United States ; }, mesh = {Animals ; *Bedbugs/genetics ; *Wolbachia/genetics ; Anti-Bacterial Agents ; }, abstract = {Wolbachia is an obligatory nutritional symbiont of the common bed bug, Cimex lectularius, providing B-vitamins to its host. The biological significance of Wolbachia to bed bugs is investigated primarily by eliminating the symbiont with antibiotics, which is followed by confirmation with molecular assays. In this chapter, we describe a protocol for eliminating Wolbachia in bed bugs using the ansamycin antibiotic rifampicin (also known as rifampin) and three molecular methods to accurately detect and quantify the Wolbachia gene copies in bed bug samples. We describe the digital droplet PCR (ddPCR), a highly sensitive technique for absolute quantification of low abundance target genes, which has proven to be a valuable technique for confirmation of the elimination of Wolbachia.}, } @article {pmid38006546, year = {2024}, author = {Bertaux, J and Raimond, M and Grève, P}, title = {Pillbug: A Toolbox for Feminisation.}, journal = {Methods in molecular biology (Clifton, N.J.)}, volume = {2739}, number = {}, pages = {69-95}, pmid = {38006546}, issn = {1940-6029}, mesh = {Animals ; Male ; Humans ; Female ; *Isopoda/genetics/metabolism ; Feminization ; *Butterflies ; Sex Differentiation ; Androgens/metabolism ; *Wolbachia ; }, abstract = {Wolbachia-induced feminisation is an extended phenotype that has been observed in only three models: isopods, butterflies and leafhoppers. Even among these, the process of feminisation differs markedly, especially since sexual differentiation is coordinated body-wide by a hormone in isopods, whereas in insects it is a chromosomally determined process and enforced cell by cell. For isopods, Armadillidium vulgare has contributed substantially to an understanding of Wolbachia-mediated feminisation, because a wide array of know-how and methods has been tailored to this model. Here, we describe establishment and maintenance of Wolbachia-infected laboratory lineages from animals collected in the field, the grafting of an androgenic gland to demonstrate the mode of action of Wolbachia (i.e. the disruption of the Androgenic Hormone pathway), and transinfection of naïve adults with Wolbachia. Finally, we describe selection of a ZZ genetic background in lineages created from transinfected WZ females, which is necessary to benefit from a property intrinsic to the naturally infected lineages: In a ZZ background, the vertical transmission rate of Wolbachia directly equates to the female ratio in the progeny. These protocols provide the essential framework for investigating molecular aspects of Wolbachia-induced feminisation.}, } @article {pmid38006545, year = {2024}, author = {Fricke, LC and Lindsey, ARI}, title = {Examining Wolbachia-Induced Parthenogenesis in Hymenoptera.}, journal = {Methods in molecular biology (Clifton, N.J.)}, volume = {2739}, number = {}, pages = {55-68}, pmid = {38006545}, issn = {1940-6029}, support = {R35 GM150991/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Male ; Female ; *Wolbachia/genetics ; Parthenogenesis/genetics ; *Wasps/genetics ; Reproduction ; }, abstract = {The maternally transmitted reproductive manipulator Wolbachia can impact sex ratios of its arthropod host by different mechanisms, ultimately promoting the spread of infection across a population. One of these reproductive phenotypes, parthenogenesis induction (PI), is characterized by the asexual production of female offspring, which in many cases results in an entirely female population. Cases of Wolbachia-mediated PI are most common in the order Hymenoptera, specifically in parasitoid wasps. The complex sex determination pathways of hymenopterans, their diverse life histories, the multiple cytogenetic mechanisms of PI, and the lack of males make functional studies of parthenogenesis induction challenging. Here, we describe the mechanisms of PI, outline methods to recognize and cure PI-Wolbachia infection, and note possible complications when working with PI-Wolbachia strains and their parthenogenetic hosts.}, } @article {pmid38006544, year = {2024}, author = {Schedl, ME and Nguyen, NH and Unckless, RL and Perlmutter, JI}, title = {Maintenance and Evaluation of Wolbachia Male-Killers of Dipterans.}, journal = {Methods in molecular biology (Clifton, N.J.)}, volume = {2739}, number = {}, pages = {29-53}, pmid = {38006544}, issn = {1940-6029}, mesh = {Male ; Animals ; *Wolbachia ; Drosophila ; Sex Ratio ; Embryonic Development ; }, abstract = {As research on Wolbachia male-killers of dipterans increases over time, several standard methods have emerged for the validation and characterization of these Wolbachia strains. Here, we describe typical steps for maintenance and evaluation of male-killers in the laboratory. Drosophila that host Wolbachia male-killers include both mushroom-feeders and flies reared on standard media, which require different maintenance protocols. In addition, male killing in flies typically occurs during embryonic development, and as such, many experiments require embryo tissue samples. We provide step-by-step instructions for standard methods of isolation, rearing, confirmation of Wolbachia male-killing as the basis of a sex ratio bias, and collection of embryos from these species. Thus, anyone intending to study a male-killer in the lab will have the complete set of protocols necessary to collect and rear them, validate them, and collect embryonic tissue for further study.}, } @article {pmid38006543, year = {2024}, author = {Namias, A and Atyame, C and Pasteur, N and Sicard, M and Weill, M}, title = {Investigation of Cytoplasmic Incompatibility Patterns in Culex pipiens: From Field Samples to Laboratory Isofemale Lines.}, journal = {Methods in molecular biology (Clifton, N.J.)}, volume = {2739}, number = {}, pages = {17-27}, pmid = {38006543}, issn = {1940-6029}, mesh = {Animals ; *Culex/genetics ; Cytoplasm ; Cytosol ; }, abstract = {This protocol describes approaches to qualify Wolbachia-induced cytoplasmic incompatibility (CI) patterns (compatible, uni or bidirectional) in crosses between two or more Culex pipiens isofemale lines, hosting different Wolbachia (wPip) strains. Here, we describe how to (1) collect the larvae in the field and grow them to the adult stage in the insectary, (2) set up isofemale lines in the insectary, (3) genetically characterize the wPip group of these lines, and (4) perform reciprocal crosses to qualify CI.}, } @article {pmid38006542, year = {2024}, author = {Fallon, AM}, title = {Wolbachia: Advancing into a Second Century.}, journal = {Methods in molecular biology (Clifton, N.J.)}, volume = {2739}, number = {}, pages = {1-13}, pmid = {38006542}, issn = {1940-6029}, mesh = {Animals ; *Wolbachia/genetics ; *Filarioidea ; *Arthropods ; Symbiosis ; }, abstract = {Wolbachia pipientis had its scientific debut nearly a century ago and has recently emerged as a target for therapeutic treatment of filarial infections and an attractive tool for control of arthropod pests. Wolbachia was known as a biological entity before DNA was recognized as the molecule that carries the genetic information on which life depends, and before arthropods and nematodes were grouped in the Ecdysozoa. Today, some investigators consider Wolbachia the most abundant endosymbiont on earth, given the numbers of its hosts and its diverse mutualistic, commensal, and parasitic roles in their life histories. Recent advances in molecular technologies have revolutionized our understanding of Wolbachia and its associated reproductive phenotypes. New models have emerged for its investigation, and substantial progress has been made towards Wolbachia-based interventions in medicine and agriculture. Here I introduce Wolbachia, with a focus on aspects of its biology that are covered in greater detail in subsequent chapters.}, } @article {pmid38003752, year = {2023}, author = {Garrigós, M and Garrido, M and Panisse, G and Veiga, J and Martínez-de la Puente, J}, title = {Interactions between West Nile Virus and the Microbiota of Culex pipiens Vectors: A Literature Review.}, journal = {Pathogens (Basel, Switzerland)}, volume = {12}, number = {11}, pages = {}, pmid = {38003752}, issn = {2076-0817}, support = {PID2020-118205GB-I00//Ministerio Español de Ciencia e Innovación/ ; PRE2021-098544//Ministerio Español de Ciencia e Innovación/ ; FJC2021-048057-I//Ministerio Español de Ciencia e Innovación/ ; María Zambrano//Ministerio Español de Universidades/ ; Margarita Salas//Ministerio Español de Universidades/ ; P21_00049//Junta de Andalucía, Consejería de Universidad, Investigación e Innovación/ ; }, abstract = {The flavivirus West Nile virus (WNV) naturally circulates between mosquitoes and birds, potentially affecting humans and horses. Different species of mosquitoes play a role as vectors of WNV, with those of the Culex pipiens complex being particularly crucial for its circulation. Different biotic and abiotic factors determine the capacity of mosquitoes for pathogen transmission, with the mosquito gut microbiota being recognized as an important one. Here, we review the published studies on the interactions between the microbiota of the Culex pipiens complex and WNV infections in mosquitoes. Most articles published so far studied the interactions between bacteria of the genus Wolbachia and WNV infections, obtaining variable results regarding the directionality of this relationship. In contrast, only a few studies investigate the role of the whole microbiome or other bacterial taxa in WNV infections. These studies suggest that bacteria of the genera Serratia and Enterobacter may enhance WNV development. Thus, due to the relevance of WNV in human and animal health and the important role of mosquitoes of the Cx. pipiens complex in its transmission, more research is needed to unravel the role of mosquito microbiota and those factors affecting this microbiota on pathogen epidemiology. In this respect, we finally propose future lines of research lines on this topic.}, } @article {pmid37999039, year = {2023}, author = {El Yamlahi, Y and Bel Mokhtar, N and Maurady, A and Britel, MR and Batargias, C and Mutembei, DE and Nyingilili, HS and Malulu, DJ and Malele, II and Asimakis, E and Stathopoulou, P and Tsiamis, G}, title = {Characterization of the Bacterial Profile from Natural and Laboratory Glossina Populations.}, journal = {Insects}, volume = {14}, number = {11}, pages = {}, pmid = {37999039}, issn = {2075-4450}, support = {URT-22658//FAO/IAEA Agriculture and Biotechnology Laboratories/ ; }, abstract = {Tsetse flies (Glossina spp.; Diptera: Glossinidae) are viviparous flies that feed on blood and are found exclusively in sub-Saharan Africa. They are the only cyclic vectors of African trypanosomes, responsible for human African trypanosomiasis (HAT) and animal African trypanosomiasis (AAT). In this study, we employed high throughput sequencing of the 16S rRNA gene to unravel the diversity of symbiotic bacteria in five wild and three laboratory populations of tsetse species (Glossina pallidipes, G. morsitans, G. swynnertoni, and G. austeni). The aim was to assess the dynamics of bacterial diversity both within each laboratory and wild population in relation to the developmental stage, insect age, gender, and location. Our results indicated that the bacterial communities associated with the four studied Glossina species were significantly influenced by their region of origin, with wild samples being more diverse compared to the laboratory samples. We also observed that the larval microbiota was significantly different than the adults. Furthermore, the sex and the species did not significantly influence the formation of the bacterial profile of the laboratory colonies once these populations were kept under the same rearing conditions. In addition, Wigglesworthia, Acinetobacter, and Sodalis were the most abundant bacterial genera in all the samples, while Wolbachia was significantly abundant in G. morsitans compared to the other studied species. The operational taxonomic unit (OTU) co-occurrence network for each location (VVBD insectary, Doma, Makao, and Msubugwe) indicated a high variability between G. pallidipes and the other species in terms of the number of mutual exclusion and copresence interactions. In particular, some bacterial genera, like Wigglesworthia and Sodalis, with high relative abundance, were also characterized by a high degree of interactions.}, } @article {pmid37995370, year = {2023}, author = {Strunov, A and Kirchner, S and Schindelar, J and Kruckenhauser, L and Haring, E and Kapun, M}, title = {Historic Museum Samples Provide Evidence for a Recent Replacement of Wolbachia Types in European Drosophila melanogaster.}, journal = {Molecular biology and evolution}, volume = {40}, number = {12}, pages = {}, pmid = {37995370}, issn = {1537-1719}, support = {P 32275/FWF_/Austrian Science Fund FWF/Austria ; }, mesh = {Animals ; *Drosophila melanogaster/genetics ; *Wolbachia/genetics ; Museums ; Biological Evolution ; Reproduction ; Symbiosis ; }, abstract = {Wolbachia is one of the most common bacterial endosymbionts, which is frequently found in numerous arthropods and nematode taxa. Wolbachia infections can have a strong influence on the evolutionary dynamics of their hosts since these bacteria are reproductive manipulators that affect the fitness and life history of their host species for their own benefit. Host-symbiont interactions with Wolbachia are perhaps best studied in the model organism Drosophila melanogaster, which is naturally infected with at least 5 different variants among which wMel and wMelCS are the most frequent ones. Comparisons of infection types between natural flies and long-term lab stocks have previously indicated that wMelCS represents the ancestral type, which was only very recently replaced by the nowadays dominant wMel in most natural populations. In this study, we took advantage of recently sequenced museum specimens of D. melanogaster that have been collected 90 to 200 yr ago in Northern Europe to test this hypothesis. Our comparison to contemporary Wolbachia samples provides compelling support for the replacement hypothesis. Our analyses show that sequencing data from historic museum specimens and their bycatch are an emerging and unprecedented resource to address fundamental questions about evolutionary dynamics in host-symbiont interactions. However, we also identified contamination with DNA from crickets that resulted in co-contamination with cricket-specific Wolbachia in several samples. These results underpin the need for rigorous quality assessments of museomic data sets to account for contamination as a source of error that may strongly influence biological interpretations if it remains undetected.}, } @article {pmid37994906, year = {2023}, author = {Meyer, DF and Moumène, A and Rodrigues, V}, title = {Microbe Profile: Ehrlichia ruminantium - stealthy as it goes.}, journal = {Microbiology (Reading, England)}, volume = {169}, number = {11}, pages = {}, pmid = {37994906}, issn = {1465-2080}, mesh = {*Ehrlichia ruminantium ; }, abstract = {Ehrlichia ruminantium is an obligate intracellular pathogenic bacterium that causes heartwater, a fatal disease of ruminants in tropical areas. Some human cases have also been reported. This globally important pathogen is primarily transmitted by ticks of the Amblyomma genus and threatens American mainland. E. ruminantium replicates within eukaryotic mammal or tick cell is a membrane-bound vacuole, where it undergoes a biphasic developmental growth cycle and differentiates from noninfectious replicative form into infectious elementary bodies. The ability of E. ruminantium to hijack host cellular processes and avoid innate immunity is a fundamental, but not yet fully understood, virulence trait of this stealth pathogen in the genomic era.}, } @article {pmid37989350, year = {2023}, author = {Bannister-Tyrrell, M and Hillman, A and Indriani, C and Ahmad, RA and Utarini, A and Simmons, CP and Anders, KL and Sergeant, E}, title = {Utility of surveillance data for planning for dengue elimination in Yogyakarta, Indonesia: a scenario-tree modelling approach.}, journal = {BMJ global health}, volume = {8}, number = {11}, pages = {}, pmid = {37989350}, issn = {2059-7908}, support = {/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Animals ; Humans ; *Dengue Virus ; Indonesia/epidemiology ; *Aedes/microbiology ; Incidence ; *Dengue/epidemiology/prevention & control ; }, abstract = {INTRODUCTION: Field trials and modelling studies suggest that elimination of dengue transmission may be possible through widespread release of Aedes aegypti mosquitoes infected with the insect bacterium Wolbachia pipientis (wMel strain), in conjunction with routine dengue control activities. This study aimed to develop a modelling framework to guide planning for the potential elimination of locally acquired dengue in Yogyakarta, a city of almost 400 000 people in Java, Indonesia.

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

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

CONCLUSION: This study suggests that with a combination of hospital-based surveillance and enhanced clinic-based surveillance for dengue, an acceptable level of confidence (80% probability) in the elimination of locally acquired dengue can be reached within 2 years. Increasing the surveillance system sensitivity could shorten the time to first ascertainment of elimination of dengue and increase the level of confidence in elimination.}, } @article {pmid37985236, year = {2024}, author = {Fukui, T and Kiuchi, T and Tomihara, K and Muro, T and Matsuda-Imai, N and Katsuma, S}, title = {Expression of the Wolbachia male-killing factor Oscar impairs dosage compensation in lepidopteran embryos.}, journal = {FEBS letters}, volume = {598}, number = {3}, pages = {331-337}, doi = {10.1002/1873-3468.14777}, pmid = {37985236}, issn = {1873-3468}, support = {2023//G-7 Scholarship Foundation/ ; 17H06431//Japan Society for the Promotion of Science/ ; 22H00366//Japan Society for the Promotion of Science/ ; JP21J12325//Japan Society for the Promotion of Science/ ; }, mesh = {Animals ; Male ; *Wolbachia/genetics/metabolism ; *Moths/genetics/metabolism ; *Bombyx/genetics/metabolism ; Insect Proteins/genetics/metabolism ; Gene Expression Profiling ; }, abstract = {Wolbachia are intracellular bacteria in insects that can manipulate the sexual development and reproduction by male killing or other methods. We have recently identified a Wolbachia protein named Oscar that acts as a male-killing factor for lepidopteran insects. Oscar interacts with the Masculinizer (Masc) protein, which is required for both masculinization and dosage compensation (DC) in lepidopteran insects. Embryonic expression of Oscar inhibits masculinization and causes male killing in two lepidopteran species, Ostrinia furnacalis and Bombyx mori. However, it remains unknown whether Oscar-induced male killing is caused by a failure of DC. Here, we performed a transcriptome analysis of Oscar complementary RNA-injected O. furnacalis and B. mori embryos, and found that Oscar primarily targets the Masc protein, resulting in male killing by interfering with DC in lepidopteran insects.}, } @article {pmid37978413, year = {2023}, author = {Pascar, J and Middleton, H and Dorus, S}, title = {Aedes aegypti microbiome composition covaries with the density of Wolbachia infection.}, journal = {Microbiome}, volume = {11}, number = {1}, pages = {255}, pmid = {37978413}, issn = {2049-2618}, support = {R21 HD088910/HD/NICHD NIH HHS/United States ; }, mesh = {Humans ; Animals ; Female ; *Dengue Virus ; *Aedes ; *Wolbachia/genetics ; Mosquito Vectors/microbiology ; Drosophila melanogaster/microbiology ; *Microbiota ; }, abstract = {BACKGROUND: Wolbachia is a widespread bacterial endosymbiont that can inhibit vector competency when stably transinfected into the mosquito, Aedes aegypti, a primary vector of the dengue virus (DENV) and other arboviruses. Although a complete mechanistic understanding of pathogen blocking is lacking, it is likely to involve host immunity induction and resource competition between Wolbachia and DENV, both of which may be impacted by microbiome composition. The potential impact of Wolbachia transinfection on host fitness is also of importance given the widespread release of mosquitos infected with the Drosophila melanogaster strain of Wolbachia (wMel) in wild populations. Here, population-level genomic data from Ae. aegypti was surveyed to establish the relationship between the density of wMel infection and the composition of the host microbiome.

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

CONCLUSION: Our bioinformatic survey provided a species-level characterization of Ae. aegypti microbiome composition and variation. wMel load varied substantially across populations and individuals and, importantly, wMel was a major hub of a negative interactions across the microbiome. These interactions may be an inherent consequence of heightened pathogen blocking in densely infected individuals or, alternatively, may result from antagonistic Wolbachia-incompatible bacteria that could impede the efficacy of wMel as a biological control agent in future applications. The relationship between wMel infection variation and the microbiome warrants further investigation in the context of developing wMel as a multivalent control agent against other arboviruses. Video Abstract.}, } @article {pmid37968789, year = {2023}, author = {Valdivia, C and Newton, JA and von Beeren, C and O'Donnell, S and Kronauer, DJC and Russell, JA and Łukasik, P}, title = {Microbial symbionts are shared between ants and their associated beetles.}, journal = {Environmental microbiology}, volume = {25}, number = {12}, pages = {3466-3483}, doi = {10.1111/1462-2920.16544}, pmid = {37968789}, issn = {1462-2920}, support = {//"Excellence Initiative - Research University" at the Faculty of Biology of the Jagiellonian University in Kraków, Poland/ ; PPN/PPO/2018/1/00015//Narodowa Agencja Wymiany Akademickiej/ ; 2018/31/B/NZ8/01158//Narodowe Centrum Nauki/ ; 1050360//National Science Foundation/ ; }, mesh = {Animals ; *Ants/genetics/microbiology ; Phylogeny ; *Coleoptera ; RNA, Ribosomal, 16S/genetics ; Larva ; *Microbiota ; Bacteria/genetics ; Symbiosis ; }, abstract = {The transmission of microbial symbionts across animal species could strongly affect their biology and evolution, but our understanding of transmission patterns and dynamics is limited. Army ants (Formicidae: Dorylinae) and their hundreds of closely associated insect guest species (myrmecophiles) can provide unique insights into interspecific microbial symbiont sharing. Here, we compared the microbiota of workers and larvae of the army ant Eciton burchellii with those of 13 myrmecophile beetle species using 16S rRNA amplicon sequencing. We found that the previously characterized specialized bacterial symbionts of army ant workers were largely absent from ant larvae and myrmecophiles, whose microbial communities were usually dominated by Rickettsia, Wolbachia, Rickettsiella and/or Weissella. Strikingly, different species of myrmecophiles and ant larvae often shared identical 16S rRNA genotypes of these common bacteria. Protein-coding gene sequences confirmed the close relationship of Weissella strains colonizing army ant larvae, some workers and several myrmecophile species. Unexpectedly, these strains were also similar to strains infecting dissimilar animals inhabiting very different habitats: trout and whales. Together, our data show that closely interacting species can share much of their microbiota, and some versatile microbial species can inhabit and possibly transmit across a diverse range of hosts and environments.}, } @article {pmid37957741, year = {2023}, author = {Moldovan, OT and Carrell, AA and Bulzu, PA and Levei, E and Bucur, R and Sitar, C and Faur, L and Mirea, IC and Șenilă, M and Cadar, O and Podar, M}, title = {The gut microbiome mediates adaptation to scarce food in Coleoptera.}, journal = {Environmental microbiome}, volume = {18}, number = {1}, pages = {80}, pmid = {37957741}, issn = {2524-6372}, support = {R01 DE024463/DE/NIDCR NIH HHS/United States ; R01DE024463/NH/NIH HHS/United States ; }, abstract = {Beetles are ubiquitous cave invertebrates worldwide that adapted to scarce subterranean resources when they colonized caves. Here, we investigated the potential role of gut microbiota in the adaptation of beetles to caves from different climatic regions of the Carpathians. The beetles' microbiota was host-specific, reflecting phylogenetic and nutritional adaptation. The microbial community structure further resolved conspecific beetles by caves suggesting microbiota-host coevolution and influences by local environmental factors. The detritivore species hosted a variety of bacteria known to decompose and ferment organic matter, suggesting turnover and host cooperative digestion of the sedimentary microbiota and allochthonous-derived nutrients. The cave Carabidae, with strong mandibula, adapted to predation and scavenging of animal and plant remains, had distinct microbiota dominated by symbiotic lineages Spiroplasma or Wolbachia. All beetles had relatively high levels of fermentative Carnobacterium and Vagococcus involved in lipid accumulation and a reduction of metabolic activity, and both features characterize adaptation to caves.}, } @article {pmid37952351, year = {2024}, author = {El Hamss, H and Maruthi, MN and Omongo, CA and Wang, HL and van Brunschot, S and Colvin, J and Delatte, H}, title = {Microbiome diversity and composition in Bemisia tabaci SSA1-SG1 whitefly are influenced by their host's life stage.}, journal = {Microbiological research}, volume = {278}, number = {}, pages = {127538}, doi = {10.1016/j.micres.2023.127538}, pmid = {37952351}, issn = {1618-0623}, mesh = {Animals ; *Hemiptera/microbiology ; *Microbiota ; Life Cycle Stages ; }, abstract = {Within the Bemisia tabaci group of cryptic whitefly species, many are damaging agricultural pests and plant-virus vectors, conferring upon this group the status of one of the world's top 100 most invasive and destructive species, affecting farmers' income and threatening their livelihoods. Studies on the microbiome of whitefly life stages are scarce, although their composition and diversity greatly influence whitefly fitness and development. We used high-throughput sequencing to understand microbiome diversity in different developmental stages of the B. tabaci sub-Saharan Africa 1 (SSA1-SG1) species of the whitefly from Uganda. Endosymbionts (Portiera, Arsenophonus, Wolbachia, and Hemipteriphilus were detected but excluded from further statistical analysis as they were not influenced by life stage using Permutational Multivariate Analysis of Variance Using Distance Matrices (ADONIS, p = 0.925 and Bray, p = 0.903). Our results showed significant differences in the meta microbiome composition in different life stages of SSA1-SG1. The diversity was significantly higher in eggs (Shannon, p = 0.024; Simpson, p = 0.047) than that in nymphs and pupae, while the number of microbial species observed by the amplicon sequence variant (ASV) was not significant (n(ASV), p = 0.094). At the phylum and genus levels, the dominant constituents in the microbiome changed significantly during various developmental stages, with Halomonas being present in eggs, whereas Bacillus and Caldalkalibacillus were consistently found across all life stages. These findings provide the first description of differing meta microbiome diversity in the life stage of whiteflies, suggesting their putative role in whitefly development.}, } @article {pmid37949964, year = {2024}, author = {Bickerstaff, JRM and Jordal, BH and Riegler, M}, title = {Two sympatric lineages of Australian Cnestus solidus share Ambrosiella symbionts but not Wolbachia.}, journal = {Heredity}, volume = {132}, number = {1}, pages = {43-53}, pmid = {37949964}, issn = {1365-2540}, mesh = {Animals ; *Weevils/microbiology ; Phylogeny ; *Wolbachia/genetics ; Australia ; *Ascomycota/genetics ; }, abstract = {Sympatric lineages of inbreeding species provide an excellent opportunity to investigate species divergence patterns and processes. Many ambrosia beetle lineages (Curculionidae: Scolytinae) reproduce by predominant inbreeding through sib mating in nests excavated in woody plant parts wherein they cultivate symbiotic ambrosia fungi as their sole source of nutrition. The Xyleborini ambrosia beetle species Cnestus solidus and Cnestus pseudosolidus are sympatrically distributed across eastern Australia and have overlapping morphological variation. Using multilocus sequencing analysis of individuals collected from 19 sites spanning their sympatric distribution, we assessed their phylogenetic relationships, taxonomic status and microbial symbionts. We found no genetic differentiation between individuals morphologically identified as C. solidus and C. pseudosolidus confirming previous suggestions that C. pseudosolidus is synonymous to C. solidus. However, within C. solidus we unexpectedly discovered the sympatric coexistence of two morphologically indistinguishable but genetically distinct lineages with small nuclear yet large mitochondrial divergence. At all sites except one, individuals of both lineages carried the same primary fungal symbiont, a new Ambrosiella species, indicating that fungal symbiont differentiation may not be involved in lineage divergence. One strain of the maternally inherited bacterial endosymbiont Wolbachia was found at high prevalence in individuals of the more common lineage but not in the other, suggesting that it may influence host fitness. Our data suggest that the two Australian Cnestus lineages diverged allopatrically, and one lineage then acquired Wolbachia. Predominant inbreeding and Wolbachia infection may have reinforced reproductive barriers between these two lineages after their secondary contact contributing to their current sympatric distribution.}, } @article {pmid37948354, year = {2024}, author = {Hussain, M and Zhong, Y and Tao, T and Xiu, B and Ye, F and Gao, J and Mao, R}, title = {Effect of tree height and spraying methods on Diaphorina citri kuwayama endosymbionts in the context of Huanglongbing disease management in citrus orchards.}, journal = {Pest management science}, volume = {80}, number = {3}, pages = {1484-1500}, doi = {10.1002/ps.7880}, pmid = {37948354}, issn = {1526-4998}, support = {2021GDASYL-20210103051//The GDAS Special Project of Science and Technology Development/ ; 2022GDASZH-2022030501//The GDAS Special Project of Science and Technology Development/ ; QN2022030005//Foreign Youth Talent Programs/ ; 2023SDZG06//the open competition program of top ten critical priorities of Agricultural Science and Technology Innovation for the 14th Five-year plan of Guangdong province/ ; 2022SDZG06//the open competition program of top ten critical priorities of Agricultural Science and Technology Innovation for the 14th Five-year plan of Guangdong province/ ; 20220610043//the Research Programs of Guangzhou/ ; 202103000065//the Research Programs of Guangzhou/ ; KTP20210352//Science and Technology Planning Project of Guangdong Province/ ; }, mesh = {Animals ; Trees ; *Citrus ; *Hemiptera ; Disease Management ; Plant Diseases ; *Rhizobiaceae ; *Liberibacter ; }, abstract = {BACKGROUND: Huanglongbing (HLB) (caused by Candidatus Liberibacter asiaticus) is the most damaging disease of citrus around the world. This study investigated the effects of citrus tree height on Diaphorina citri Kuwayama mortality, endosymbiont responses, and HLB distribution.

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

CONCLUSION: This research provides valuable insights into the complex interaction between citrus tree age, psyllid endosymbionts responses, and HLB distribution. These results emphasize effective HLB management strategies, especially in orchards with diverse tree age populations, ultimately contributing to the long-term sustainability of citrus cultivation. © 2023 Society of Chemical Industry.}, } @article {pmid37936139, year = {2023}, author = {Hakobyan, A and Velte, S and Sickel, W and Quandt, D and Stoll, A and Knief, C}, title = {Tillandsia landbeckii phyllosphere and laimosphere as refugia for bacterial life in a hyperarid desert environment.}, journal = {Microbiome}, volume = {11}, number = {1}, pages = {246}, pmid = {37936139}, issn = {2049-2618}, mesh = {Humans ; Soil Microbiology ; *Tillandsia ; Refugium ; *Microbiota ; Bacteria/genetics ; Plants/microbiology ; Soil ; Desert Climate ; }, abstract = {BACKGROUND: The lack of water is a major constraint for microbial life in hyperarid deserts. Consequently, the abundance and diversity of microorganisms in common habitats such as soil are strongly reduced, and colonization occurs primarily by specifically adapted microorganisms that thrive in particular refugia to escape the harsh conditions that prevail in these deserts. We suggest that plants provide another refugium for microbial life in hyperarid deserts. We studied the bacterial colonization of Tillandsia landbeckii (Bromeliaceae) plants, which occur in the hyperarid regions of the Atacama Desert in Chile, one of the driest and oldest deserts on Earth.

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

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

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

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

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

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

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

CONCLUSIONS: Wolbachia infection is commonly prevalent in Ae. albopictus and Cx. pipiens pallens from Henan Province, and Wolbachia strains wAlbB and wAlbA are predominant in Ae. albopictus, while wPip strain is predominant in Cx. pipiens pallens. This is the first report to present Wolbachia wMors strain infection in Cx. pipiens pallens in Henan Province.}, } @article {pmid37926385, year = {2024}, author = {Rodríguez-Rojas, JJ and Lozano-Sardaneta, YN and Fernández-Salas, I and Sánchez-Casas, RM and Becker, I}, title = {Species diversity, barcode, detection of pathogens and blood meal pattern in Phlebotominae (Diptera: Psychodidae) from northeastern Mexico.}, journal = {Acta tropica}, volume = {249}, number = {}, pages = {107064}, doi = {10.1016/j.actatropica.2023.107064}, pmid = {37926385}, issn = {1873-6254}, mesh = {Animals ; Male ; Female ; Humans ; *Psychodidae/parasitology ; Mexico ; Insect Vectors/parasitology ; *Leishmania/genetics ; *Leishmaniasis ; Feeding Behavior ; }, abstract = {More than 90 species of phlebotomines are vectors of parasites, bacteria, and viruses, which cause disease in animals and humans. Therefore, their study is necessary to establish prevention and control strategies. Mexico is an endemic country for leishmaniasis, mostly in the center and southern regions of the country, yet only few studies have been conducted in the northern part of the country. The present study aims to: (a) assess the alpha diversity of Phlebotominae in an annual cycle, (b) to correlate climatic variables with abundance, (c) to generate barcodes of these insects as part of the integrative taxonomy, and (d) to detect Leishmania, Wolbachia and blood sources in an area close to where a case of autochthonous leishmaniasis has been detected in Nuevo Leon, Mexico. A systematic sampling was conducted during three consecutive nights from 17:00 to 22:00 h., placing Shannon traps, CDC traps with incandescent light, and BG Sentinel 2 + BG Lure traps. A total catch effort of 660 nights/traps/hours was achieved, in which a total number of 707 phlebotomines (58% female and 42% male) of six species were collected and identified. The most abundant species were Psathyromyia cratifer (57%) and Psathyromyia shannoni sensu stricto (26%). The highest abundance (72%; 507/707) was collected during March, April and May 2021. Barcodes were generated for four species of phlebotomines, which represent new records for Mexico. For the molecular detection of microorganisms, 302 specimens were analyzed, although no specimens were positive for Leishmania spp. Wolbachia strains were detected in phlebotomines with an infection rate of 1.32% (4/302) and found in Pa. cratifer and Lu. cruciata. Likewise, human DNA was identified in female Lu. cruciata and Pa. cratifer phlebotomines. These findings indicate the presence of potential vector species of the parasite Leishmania spp. This result shows the need for further entomological surveillance to elucidate the transmission mechanisms in these northern areas of the country.}, } @article {pmid37926187, year = {2024}, author = {Badger, JH and Giordano, R and Zimin, A and Wappel, R and Eskipehlivan, SM and Muller, S and Donthu, R and Soto-Adames, F and Vieira, P and Zasada, I and Goodwin, S}, title = {Direct sequencing of insect symbionts via nanopore adaptive sampling.}, journal = {Current opinion in insect science}, volume = {61}, number = {}, pages = {101135}, doi = {10.1016/j.cois.2023.101135}, pmid = {37926187}, issn = {2214-5753}, support = {P30 CA045508/CA/NCI NIH HHS/United States ; }, mesh = {Animals ; *Nanopores ; *Buchnera/genetics ; DNA Transposable Elements ; Insecta/genetics ; }, abstract = {Insect symbionts can alter their host phenotype and their effects can range from beneficial to pathogenic. Moreover, many insects exhibit co-infections, making their study more challenging. Less than 1% of insect species have high-quality referenced genomes available and fewer still also have their symbionts sequenced. Two methods are commonly used to sequence symbionts: whole-genome sequencing to concomitantly capture the host and bacterial genomes, or isolation of the symbiont's genome before sequencing. These methods are limited when dealing with rare or poorly characterized symbionts. Long-read technology is an important tool to generate high-quality genomes as they can overcome high levels of heterozygosity, repeat content, and transposable elements that confound short-read methods. Oxford Nanopore (ONT) adaptive sampling allows a sequencing instrument to select or reject sequences in real time. We describe a method based on ONT adaptive sampling (subtractive) approach that readily permitted the sequencing of the complete genomes of mitochondria, Buchnera and its plasmids (pLeu, pTrp), and Wolbachia genomes in two aphid species, Aphis glycines and Pentalonia nigronervosa. Adaptive sampling is able to retrieve organelles such as mitochondria and symbionts that have high representation in their hosts such as Buchnera and Wolbachia, but is less successful at retrieving symbionts in low concentrations.}, } @article {pmid37923779, year = {2023}, author = {Flores, GAM and Lopez, RP and Cerrudo, CS and Perotti, MA and Consolo, VF and Berón, CM}, title = {Wolbachia dominance influences the Culex quinquefasciatus microbiota.}, journal = {Scientific reports}, volume = {13}, number = {1}, pages = {18980}, pmid = {37923779}, issn = {2045-2322}, mesh = {Animals ; *Aedes/genetics ; Bacteria/genetics ; *Culex/genetics ; *Microbiota ; Mosquito Vectors/microbiology ; RNA, Ribosomal, 16S/genetics ; *Wolbachia/genetics ; }, abstract = {Microorganisms present in mosquitoes and their interactions are key factors affecting insect development. Among them, Wolbachia is closely associated with the host and affects several fitness parameters. In this study, the bacterial and fungal microbiota from two laboratory Culex quinquefasciatus isolines (wild type and tetracycline-cured) were characterized by metagenome amplicon sequencing of the ITS2 and 16S rRNA genes at different developmental stages and feeding conditions. We identified 572 bacterial and 61 fungal OTUs. Both isolines presented variable bacterial communities and different trends in the distribution of diversity among the groups. The lowest bacterial richness was detected in sugar-fed adults of the cured isoline, whereas fungal richness was highly reduced in blood-fed mosquitoes. Beta diversity analysis indicated that isolines are an important factor in the differentiation of mosquito bacterial communities. Considering composition, Penicillium was the dominant fungal genus, whereas Wolbachia dominance was inversely related to that of Enterobacteria (mainly Thorsellia and Serratia). This study provides a more complete overview of the mosquito microbiome, emphasizing specific highly abundant components that should be considered in microorganism manipulation approaches to control vector-borne diseases.}, } @article {pmid37921460, year = {2023}, author = {Zhang, Y and Liu, S and Huang, X-y and Zi, H-b and Gao, T and Ji, R-j and Sheng, J and Zhi, D and Zhang, Y-l and Gong, C-m and Yang, Y-q}, title = {Altitude as a key environmental factor shaping microbial communities of tea green leafhoppers (Matsumurasca onukii).}, journal = {Microbiology spectrum}, volume = {11}, number = {6}, pages = {e0100923}, pmid = {37921460}, issn = {2165-0497}, support = {No.2021YFD1601105//MOST | National Key Research and Development Program of China (NKPs)/ ; No.32172635//MOST | National Natural Science Foundation of China (NSFC)/ ; }, mesh = {Animals ; *Hemiptera ; Altitude ; Tea ; }, abstract = {Host-associated microbial communities play an important role in the fitness of insect hosts. However, the factors shaping microbial communities in wild populations, including environmental factors and interactions among microbial species, remain largely unknown. The tea green leafhopper has a wide geographical distribution and is highly adaptable, providing a suitable model for studying the effect of ecological drivers on microbiomes. This is the first large-scale culture-independent study investigating the microbial communities of M. onukii sampled from different locations. Altitude as a key environmental factor may have shaped microbial communities of M. onukii by affecting the relative abundance of endosymbionts, especially Wolbachia. The results of this study, therefore, offer not only an in-depth view of the microbial diversity of this species but also an insight into the influence of environmental factors.}, } @article {pmid37914998, year = {2023}, author = {Liu, W and Xia, X and Hoffmann, AA and Ding, Y and Fang, JC and Yu, H}, title = {Evolution of Wolbachia reproductive and nutritional mutualism: insights from the genomes of two novel strains that double infect the pollinator of dioecious Ficus hirta.}, journal = {BMC genomics}, volume = {24}, number = {1}, pages = {657}, pmid = {37914998}, issn = {1471-2164}, support = {2021A1515110981//Guangdong Basic and Applied Basic Research Foundation/ ; 2022ZB773//Jiangsu Funding Program for Excellent Postdoctoral Talent/ ; 2022VBA0002//The Chinese Academy of Sciences PIFI Fellowship for Visiting Scientists/ ; 2023YFE0100540//National Key R & D Program of China/ ; 202206010058//Guangzhou Collaborative Innovation Center on Science-tech of Ecology and Landscape/ ; }, mesh = {*Ficus/genetics ; *Wolbachia/genetics ; Biotin/genetics ; Symbiosis/genetics ; Phylogeny ; DNA Transposable Elements/genetics ; Multilocus Sequence Typing ; Prophages/genetics ; Reproduction ; }, abstract = {Wolbachia is a genus of maternally inherited endosymbionts that can affect reproduction of their hosts and influence metabolic processes. The pollinator, Valisia javana, is common in the male syconium of the dioecious fig Ficus hirta. Based on a high-quality chromosome-level V. javana genome with PacBio long-read and Illumina short-read sequencing, we discovered a sizeable proportion of Wolbachia sequences and used these to assemble two novel Wolbachia strains belonging to supergroup A. We explored its phylogenetic relationship with described Wolbachia strains based on MLST sequences and the possibility of induction of CI (cytoplasmic incompatibility) in this strain by examining the presence of cif genes known to be responsible for CI in other insects. We also identified mobile genetic elements including prophages and insertion sequences, genes related to biotin synthesis and metabolism. A total of two prophages and 256 insertion sequences were found. The prophage WOjav1 is cryptic (structure incomplete) and WOjav2 is relatively intact. IS5 is the dominant transposon family. At least three pairs of type I cif genes with three copies were found which may cause strong CI although this needs experimental verification; we also considered possible nutritional effects of the Wolbachia by identifying genes related to biotin production, absorption and metabolism. This study provides a resource for further studies of Wolbachia-pollinator-host plant interactions.}, } @article {pmid37907954, year = {2023}, author = {Pfarr, KM and Krome, AK and Al-Obaidi, I and Batchelor, H and Vaillant, M and Hoerauf, A and Opoku, NO and Kuesel, AC}, title = {The pipeline for drugs for control and elimination of neglected tropical diseases: 2. Oral anti-infective drugs and drug combinations for off-label use.}, journal = {Parasites & vectors}, volume = {16}, number = {1}, pages = {394}, pmid = {37907954}, issn = {1756-3305}, support = {001/WHO_/World Health Organization/International ; }, mesh = {Humans ; *Ivermectin/therapeutic use ; Rifampin ; Doxycycline ; Fluconazole ; Off-Label Use ; *Anti-Infective Agents/therapeutic use ; Drug Combinations ; Neglected Diseases/drug therapy/prevention & control ; }, abstract = {In its 'Road map for neglected tropical diseases 2021-2030', the World Health Organization outlined its targets for control and elimination of neglected tropical diseases (NTDs) and research needed to achieve them. For many NTDs, this includes research for new treatment options for case management and/or preventive chemotherapy. Our review of small-molecule anti-infective drugs recently approved by a stringent regulatory authority (SRA) or in at least Phase 2 clinical development for regulatory approval showed that this pipeline cannot deliver all new treatments needed. WHO guidelines and country policies show that drugs may be recommended for control and elimination for NTDs for which they are not SRA approved (i.e. for 'off-label' use) if efficacy and safety data for the relevant NTD are considered sufficient by WHO and country authorities. Here, we are providing an overview of clinical research in the past 10 years evaluating the anti-infective efficacy of oral small-molecule drugs for NTD(s) for which they are neither SRA approved, nor included in current WHO strategies nor, considering the research sponsors, likely to be registered with a SRA for that NTD, if found to be effective and safe. No such research has been done for yaws, guinea worm, Trypanosoma brucei gambiense human African trypanosomiasis (HAT), rabies, trachoma, visceral leishmaniasis, mycetoma, T. b. rhodesiense HAT, echinococcosis, taeniasis/cysticercosis or scabies. Oral drugs evaluated include sparfloxacin and acedapsone for leprosy; rifampicin, rifapentin and moxifloxacin for onchocerciasis; imatinib and levamisole for loiasis; itraconazole, fluconazole, ketoconazole, posaconazole, ravuconazole and disulfiram for Chagas disease, doxycycline and rifampicin for lymphatic filariasis; arterolane, piperaquine, artesunate, artemether, lumefantrine and mefloquine for schistosomiasis; ivermectin, tribendimidine, pyrantel, oxantel and nitazoxanide for soil-transmitted helminths including strongyloidiasis; chloroquine, ivermectin, balapiravir, ribavirin, celgosivir, UV-4B, ivermectin and doxycycline for dengue; streptomycin, amoxicillin, clavulanate for Buruli ulcer; fluconazole and isavuconazonium for mycoses; clarithromycin and dapsone for cutaneous leishmaniasis; and tribendimidine, albendazole, mebendazole and nitazoxanide for foodborne trematodiasis. Additional paths to identification of new treatment options are needed. One promising path is exploitation of the worldwide experience with 'off-label' treatment of diseases with insufficient treatment options as pursued by the 'CURE ID' initiative.}, } @article {pmid37901801, year = {2023}, author = {Minwuyelet, A and Petronio, GP and Yewhalaw, D and Sciarretta, A and Magnifico, I and Nicolosi, D and Di Marco, R and Atenafu, G}, title = {Symbiotic Wolbachia in mosquitoes and its role in reducing the transmission of mosquito-borne diseases: updates and prospects.}, journal = {Frontiers in microbiology}, volume = {14}, number = {}, pages = {1267832}, pmid = {37901801}, issn = {1664-302X}, abstract = {Mosquito-borne diseases such as malaria, dengue fever, West Nile virus, chikungunya, Zika fever, and filariasis have the greatest health and economic impact. These mosquito-borne diseases are a major cause of morbidity and mortality in tropical and sub-tropical areas. Due to the lack of effective vector containment strategies, the prevalence and severity of these diseases are increasing in endemic regions. Nowadays, mosquito infection by the endosymbiotic Wolbachia represents a promising new bio-control strategy. Wild-infected mosquitoes had been developing cytoplasmic incompatibility (CI), phenotypic alterations, and nutrition competition with pathogens. These reduce adult vector lifespan, interfere with reproduction, inhibit other pathogen growth in the vector, and increase insecticide susceptibility of the vector. Wild, uninfected mosquitoes can also establish stable infections through trans-infection and have the advantage of adaptability through pathogen defense, thereby selectively infecting uninfected mosquitoes and spreading to the entire population. This review aimed to evaluate the role of the Wolbachia symbiont with the mosquitoes (Aedes, Anopheles, and Culex) in reducing mosquito-borne diseases. Global databases such as PubMed, Web of Sciences, Scopus, and pro-Quest were accessed to search for potentially relevant articles. We used keywords: Wolbachia, Anopheles, Aedes, Culex, and mosquito were used alone or in combination during the literature search. Data were extracted from 56 articles' texts, figures, and tables of the included article.}, } @article {pmid37891252, year = {2023}, author = {Lenharo, M}, title = {Dengue rates drop after release of modified mosquitoes in Colombia.}, journal = {Nature}, volume = {623}, number = {7986}, pages = {235-236}, pmid = {37891252}, issn = {1476-4687}, mesh = {Animals ; Humans ; *Aedes/microbiology/virology ; Colombia/epidemiology ; *Dengue/epidemiology/prevention & control/transmission/virology ; *Dengue Virus ; *Wolbachia ; *Biological Control Agents ; }, } @article {pmid37887819, year = {2023}, author = {Xoconostle-Cázares, B and Ramírez-Pool, JA and Núñez-Muñoz, LA and Calderón-Pérez, B and Vargas-Hernández, BY and Bujanos-Muñiz, R and Ruiz-Medrano, R}, title = {The Characterization of Melanaphis sacchari Microbiota and Antibiotic Treatment Effect on Insects.}, journal = {Insects}, volume = {14}, number = {10}, pages = {}, pmid = {37887819}, issn = {2075-4450}, support = {grant CF-2023-G-731//Consejo Nacional de Humanidades Ciencias y Tecnologias/ ; }, abstract = {Insects are under constant selective pressure, which has resulted in adaptations to novel niches such as crops. This is the case of the pest Melanaphis sacchari, the sugarcane aphid, native to Africa and currently spreading worldwide. The aphid undergoes successful parthenogenesis, causing important damage to a variety of crops and leading to important economic losses for farmers. A natural M. sacchari population grown in sorghum was studied to identify its microbiome through the sequencing of its 16S rDNA metagenome. A high proportion of Proteobacteria, followed by Firmicutes, Bacteroidetes, and Actinobacteria, was observed. We also detected Wolbachia, which correlates with the asexual reproduction of its host. M. sacchari was challenged in a bioassay with the antibiotics oxytetracycline and streptomycin, resulting in a dose-dependent decay of its survival rate. The possibility of controlling this pest by altering its microbiota is proposed.}, } @article {pmid37887804, year = {2023}, author = {Liu, Y and Liu, J and Zhang, X and Yun, Y}, title = {Diversity of Bacteria Associated with Guts and Gonads in Three Spider Species and Potential Transmission Pathways of Microbes within the Same Spider Host.}, journal = {Insects}, volume = {14}, number = {10}, pages = {}, pmid = {37887804}, issn = {2075-4450}, support = {2018YFA0901101//The National Key Technology R&D Program of China/ ; 2016AHB003//Key Scientific and Technological Projects of Hubei/ ; }, abstract = {Microbial symbiosis plays a crucial role in the ecological and evolutionary processes of animals. It is well known that spiders, with their unique and diverse predatory adaptations, assume an indispensable role in maintaining ecological balance and the food chain. However, our current understanding of spider microbiomes remains relatively limited. The gut microbiota and gonad microbiota of spiders can both potentially influence their physiology, ecology, and behavior, including aspects such as digestion, immunity, reproductive health, and reproductive behavior. In the current study, based on high-throughput sequencing of the 16S rRNA V3 and V4 regions, we detected the gut and gonad microbiota communities of three spider species captured from the same habitat, namely, Eriovixia cavaleriei, Larinioides cornutus, and Pardosa pseudoannulata. In these three species, we observed that, at the phylum level classification, the gut and gonad of E. cavaleriei are primarily composed of Proteobacteria, while those of L. cornutus and P. pseudoannulata are primarily composed of Firmicutes. At the genus level of classification, we identified 372 and 360 genera from the gut and gonad bacterial communities. It is noteworthy that the gut and gonad bacterial flora of E. cavaleriei and L. cornutus were dominated by Wolbachia and Spiroplasma. Results show that there were no differences in microbial communities between females and males of the same spider species. Furthermore, there is similarity between the gut and ovary microbial communities of female spiders, implying a potential avenue for microbial transmission between the gut and gonad within female spiders. By comprehensively studying these two microbial communities, we can establish the theoretical foundation for exploring the relationship between gut and gonad microbiota and their host, as well as the mechanisms through which microbes exert their effects.}, } @article {pmid37882523, year = {2023}, author = {Głowska, E and Gerth, M}, title = {Draft genome sequence of a Wolbachia endosymbiont from Syringophilopsis turdi (Fritsch, 1958) (Acari, Syringophilidae).}, journal = {Microbiology resource announcements}, volume = {12}, number = {11}, pages = {e0060523}, pmid = {37882523}, issn = {2576-098X}, support = {UMO-2015/19/D/NZ8/00191//Narodowe Centrum Nauki (NCN)/ ; GE 2824/1-1//Deutsche Forschungsgemeinschaft (DFG)/ ; }, abstract = {We present the draft genome of a Wolbachia endosymbiont from quill mites. This is the first representative of a recently discovered distinct Wolbachia lineage (supergroup P). We hope the genome will be a useful resource for comparative evolutionary and genomic studies across the globally distributed symbiont Wolbachia.}, } @article {pmid37874788, year = {2023}, author = {Russell, SL and Castillo, JR and Sullivan, WT}, title = {Wolbachia endosymbionts manipulate the self-renewal and differentiation of germline stem cells to reinforce fertility of their fruit fly host.}, journal = {PLoS biology}, volume = {21}, number = {10}, pages = {e3002335}, pmid = {37874788}, issn = {1545-7885}, support = {K99 GM135583/GM/NIGMS NIH HHS/United States ; R00 GM135583/GM/NIGMS NIH HHS/United States ; R35 GM139595/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; *Wolbachia ; Drosophila/metabolism ; Fertility ; Cell Differentiation ; Germ Cells/metabolism ; Stem Cells/metabolism ; RNA, Messenger/metabolism ; Drosophila melanogaster/genetics ; RNA-Binding Proteins/genetics ; *Drosophila Proteins/metabolism ; }, abstract = {The alphaproteobacterium Wolbachia pipientis infects arthropod and nematode species worldwide, making it a key target for host biological control. Wolbachia-driven host reproductive manipulations, such as cytoplasmic incompatibility (CI), are credited for catapulting these intracellular bacteria to high frequencies in host populations. Positive, perhaps mutualistic, reproductive manipulations also increase infection frequencies, but are not well understood. Here, we identify molecular and cellular mechanisms by which Wolbachia influences the molecularly distinct processes of germline stem cell (GSC) self-renewal and differentiation. We demonstrate that wMel infection rescues the fertility of flies lacking the translational regulator mei-P26 and is sufficient to sustain infertile homozygous mei-P26-knockdown stocks indefinitely. Cytology revealed that wMel mitigates the impact of mei-P26 loss through restoring proper pMad, Bam, Sxl, and Orb expression. In Oregon R files with wild-type fertility, wMel infection elevates lifetime egg hatch rates. Exploring these phenotypes through dual-RNAseq quantification of eukaryotic and bacterial transcripts revealed that wMel infection rescues and offsets many gene expression changes induced by mei-P26 loss at the mRNA level. Overall, we show that wMel infection beneficially reinforces host fertility at mRNA, protein, and phenotypic levels, and these mechanisms may promote the emergence of mutualism and the breakdown of host reproductive manipulations.}, } @article {pmid37873081, year = {2023}, author = {Perlmutter, JI and Atadurdyyeva, A and Schedl, ME and Unckless, RL}, title = {Wolbachia enhances the survival of Drosophila infected with fungal pathogens.}, journal = {bioRxiv : the preprint server for biology}, volume = {}, number = {}, pages = {}, pmid = {37873081}, issn = {2692-8205}, support = {P20 GM103418/GM/NIGMS NIH HHS/United States ; R01 AI139154/AI/NIAID NIH HHS/United States ; }, abstract = {Wolbachia bacteria of arthropods are at the forefront of basic and translational research on multipartite host-symbiont-pathogen interactions. These microbes are vertically inherited from mother to offspring via the cytoplasm. They are the most widespread endosymbionts on the planet due to their infamous ability to manipulate the reproduction of their hosts to spread themselves in a population, and to provide a variety of fitness benefits to their hosts. Importantly, some strains of Wolbachia can inhibit viral pathogenesis within and between arthropod hosts. Mosquitoes carrying the wMel Wolbachia strain of Drosophila melanogaster have a greatly reduced capacity to spread viruses like dengue and Zika to humans. Therefore, Wolbachia are the basis of several global vector control initiatives. While significant research efforts have focused on viruses, relatively little attention has been given to Wolbachia-fungal interactions despite the ubiquity of fungal entomopathogens in nature. Here, we demonstrate that Wolbachia increase the longevity of their Drosophila melanogaster hosts when challenged with a spectrum of yeast and filamentous fungal pathogens. We find that this pattern can vary based on host genotype, sex, and fungal species. Further, Wolbachia correlates with higher fertility and reduced pathogen titers during initial fungal infection, indicating a significant fitness benefit. This study demonstrates Wolbachia's role in diverse fungal pathogen interactions and determines that the phenotype is broad, but with several variables that influence both the presence and strength of the phenotype. These results enhance our knowledge of the strategies Wolbachia uses that likely contribute to such a high global symbiont prevalence.}, } @article {pmid37871129, year = {2023}, author = {Wenzel, M and Aquadro, CF}, title = {Wolbachia infection at least partially rescues the fertility and ovary defects of several new Drosophila melanogaster bag of marbles protein-coding mutants.}, journal = {PLoS genetics}, volume = {19}, number = {10}, pages = {e1011009}, pmid = {37871129}, issn = {1553-7404}, support = {R01 GM095793/GM/NIGMS NIH HHS/United States ; S10 OD018516/OD/NIH HHS/United States ; }, mesh = {Animals ; Female ; Male ; *Drosophila melanogaster/genetics/microbiology ; *Drosophila Proteins/genetics/metabolism ; Fertility/genetics ; Ovary/metabolism ; *Wolbachia/genetics/metabolism ; }, abstract = {The D. melanogaster protein coding gene bag of marbles (bam) plays a key role in early male and female reproduction by forming complexes with partner proteins to promote differentiation in gametogenesis. Like another germline gene, Sex lethal, bam genetically interacts with the endosymbiont Wolbachia, as Wolbachia rescues the reduced fertility of a bam hypomorphic mutant. Here, we explored the specificity of the bam-Wolbachia interaction by generating 22 new bam mutants, with ten mutants displaying fertility defects. Nine of these mutants trend towards rescue by the wMel Wolbachia variant, with eight statistically significant at the fertility and/or cytological level. In some cases, fertility was increased a striking 20-fold. There is no specificity between the rescue and the known binding regions of bam, suggesting wMel does not interact with one singular bam partner to rescue the reproductive phenotype. We further tested if wMel interacts with bam in a non-specific way, by increasing bam transcript levels or acting upstream in germline stem cells. A fertility assessment of a bam RNAi knockdown mutant reveals that wMel rescue is specific to functionally mutant bam alleles and we find no obvious evidence of wMel interaction with germline stem cells in bam mutants.}, } @article {pmid37860089, year = {2023}, author = {Rushidi, MNA and Azhari, MLH and Yaakop, S and Hazmi, IR}, title = {Detection and Characterisation of Endosymbiont Wolbachia (Rickettsiales: Anaplasmataceae) in Elaeidobius kamerunicus (Coleoptera: Curculionoidea), Pollinating Agent of Oil Palm, and Its Relationships between Populations.}, journal = {Tropical life sciences research}, volume = {34}, number = {3}, pages = {95-111}, pmid = {37860089}, issn = {1985-3718}, abstract = {Elaeidobius kamerunicus is the most efficient pollinator of oil palm. Wolbachia is an endosymbiotic bacteria associated with E. kamerunicus that has a potential to affect the fecundity and fitness of the E. kamerunicus. Despite their importance, no studies have been conducted to investigate its prevalence in E. kamerunicus. The objectives of this study were to detect and characterise Wolbachia in E. kamerunicus and determine the phylogenetic relationship of Wolbachia strains that infect E. kamerunicus by using three genetic markers namely Filamenting temperature-sensitive mutant Z (ftsZ), Chaperonin folding protein (groEL), and Citrate Synthase Coding Gene (gltA). DNA was extracted from 210 individuals of E. kamerunicus and the Wolbachia infections were detected using the wsp marker. The infected samples (n = 25, 11.9%) were then sequenced using ftsZ, gltA and groEL markers for strain characterization. In this study, a combination of four markers was used to construct the phylogeny of Wolbachia. Similar topologies were shown in all trees; Neighbour-Joining (NJ), Maximum Parsimony (MP), and Bayesian Inference (BI), which showed the mixing of individuals that harbor Wolbachia between populations. Interestingly, Wolbachia on E. kamerunicus was claded together with the species Drosophila simulans under supergroup B. This is the first report of Wolbachia infecting E. kamerunicus which is very valuable and significant as one of the parameters to evaluate the quality of the E. kamerunicus population for sustaining its function as a great pollinator for oil palm.}, } @article {pmid37859959, year = {2023}, author = {Ridha, MR and Marlinae, L and Zubaidah, T and Fadillah, NA and Widjaja, J and Rosadi, D and Rahayu, N and Ningsih, M and Desimal, I and Sofyandi, A}, title = {Control methods for invasive mosquitoes of Aedes aegypti and Aedes albopictus (Diptera: Culicidae) in Indonesia.}, journal = {Veterinary world}, volume = {16}, number = {9}, pages = {1952-1963}, pmid = {37859959}, issn = {0972-8988}, abstract = {The two invasive mosquito species in Indonesia are Aedes aegypti and Ae. albopictus. These mosquitoes are a serious nuisance to humans and are also the primary vectors of several foreign pathogens, such as dengue, Zika, and chikungunya viruses. Efforts must be made to reduce the possibility of mosquito bites and the potential for disease transmission. Given the invasion of these two Aedes species, this approach should be considered as part of an integrated strategy to manage them. This review discusses existing and developing control techniques for invasive Ae. aegypti and Ae. albopictus, with an emphasis on those that have been and are being used in Indonesia. Environmental, mechanical, biological (e.g., Bacillus thuringiensis and Wolbachia), and chemical (e.g., insect growth regulators and pyrethroids) approaches are discussed in this review, considering their effectiveness, sustainability, and control methods.}, } @article {pmid37838705, year = {2023}, author = {Sounart, H and Voronin, D and Masarapu, Y and Chung, M and Saarenpää, S and Ghedin, E and Giacomello, S}, title = {Miniature spatial transcriptomics for studying parasite-endosymbiont relationships at the micro scale.}, journal = {Nature communications}, volume = {14}, number = {1}, pages = {6500}, pmid = {37838705}, issn = {2041-1723}, mesh = {Animals ; Female ; Humans ; *Parasites/genetics ; Transcriptome ; Anti-Bacterial Agents/metabolism ; Gene Expression Profiling ; *Communicable Diseases ; *Wolbachia/genetics/metabolism ; Symbiosis/genetics ; }, abstract = {Several important human infectious diseases are caused by microscale-sized parasitic nematodes like filarial worms. Filarial worms have their own spatial tissue organization; to uncover this tissue structure, we need methods that can spatially resolve these miniature specimens. Most filarial worms evolved a mutualistic association with endosymbiotic bacteria Wolbachia. However, the mechanisms underlying the dependency of filarial worms on the fitness of these bacteria remain unknown. As Wolbachia is essential for the development, reproduction, and survival of filarial worms, we spatially explored how Wolbachia interacts with the worm's reproductive system by performing a spatial characterization using Spatial Transcriptomics (ST) across a posterior region containing reproductive tissue and developing embryos of adult female Brugia malayi worms. We provide a proof-of-concept for miniature-ST to explore spatial gene expression patterns in small sample types, demonstrating the method's ability to uncover nuanced tissue region expression patterns, observe the spatial localization of key B. malayi - Wolbachia pathway genes, and co-localize the B. malayi spatial transcriptome in Wolbachia tissue regions, also under antibiotic treatment. We envision our approach will open up new avenues for the study of infectious diseases caused by micro-scale parasitic worms.}, } @article {pmid37830551, year = {2023}, author = {Riparbelli, MG and Pratelli, A and Callaini, G}, title = {Wolbachia Induces Structural Defects Harmful to Drosophila simulans Riverside Spermiogenesis.}, journal = {Cells}, volume = {12}, number = {19}, pages = {}, pmid = {37830551}, issn = {2073-4409}, mesh = {Animals ; Male ; *Drosophila simulans ; Drosophila ; *Wolbachia ; Semen ; Spermatogenesis ; }, abstract = {The relationship between cytoplasmic incompatibility and the obligate intracellular alphaproteobacteria Wolbachia has for a long time been reported. Although the molecular mechanisms responsible for this reproductive alteration are beginning to be understood, the effects of Wolbachia on germ cell structure and dynamics have not yet been fully investigated. We report here that the presence of Wolbachia in infected cysts of elongating spermatids is associated with major structural defects that become more evident in mature sperm. We find mitochondrial defects, an improper axoneme structure, reduced sperm numbers, and individualization failures. The large heterogeneous variety of the ultrastructural defects found in elongating spermatids and mature sperm provide the first cytological evidence for the reduced fertility associated with Wolbachia infection in Drosophila simulans males. The observed abnormalities could be the result of the mechanical stress induced by the high bacteria numbers during the process of spermatid elongation, rather than the result of the released factors affecting the proper morphogenesis of the germ cells. Moreover, high Wolbachia densities in male germ cells may not be appropriate for causing cytoplasmic incompatibility as the bacteria are harmful for spermatid differentiation, leading to abnormal sperm that is unlikely to be functional.}, } @article {pmid37822704, year = {2023}, author = {Pavan, MG and Garcia, GA and David, MR and Maciel-de-Freitas, R}, title = {The double-edged sword effect of expanding Wolbachia deployment in dengue endemic settings.}, journal = {Lancet regional health. Americas}, volume = {27}, number = {}, pages = {100610}, pmid = {37822704}, issn = {2667-193X}, } @article {pmid37819592, year = {2023}, author = {Haghshenas-Gorgabi, N and Poorjavd, N and Khajehali, J and Wybouw, N}, title = {Cardinium symbionts are pervasive in Iranian populations of the spider mite Panonychus ulmi despite inducing an infection cost and no demonstrable reproductive phenotypes when Wolbachia is a symbiotic partner.}, journal = {Experimental & applied acarology}, volume = {91}, number = {3}, pages = {369-380}, pmid = {37819592}, issn = {1572-9702}, mesh = {Female ; Male ; Animals ; *Wolbachia ; *Tetranychidae/genetics ; Iran ; Seeds ; Reproduction ; Symbiosis ; Bacteroidetes ; }, abstract = {Maternally transmitted symbionts such as Cardinium and Wolbachia are widespread in arthropods. Both Cardinium and Wolbachia can cause cytoplasmic incompatibility, a reproductive phenotype that interferes with the development of uninfected eggs that are fertilized by infected sperm. In haplodiploid hosts, these symbionts can also distort sex allocation to facilitate their spread through host populations. Without other fitness effects, symbionts that induce strong reproductive phenotypes tend to spread to high and stable infection frequencies, whereas variants that induce weak reproductive phenotypes are typically associated with intermediate and variable frequencies. To study the spread of Cardinium in a haplodiploid host, we sampled Iranian populations of the economically important spider mite Panonychus ulmi in apple orchards. Within several field populations, we also studied the Wolbachia infection frequencies. All P. ulmi field populations carried a Cardinium infection and exhibited high infection frequencies. In contrast, Wolbachia frequency ranged between ca. 10% and ca. 70% and was only found in co-infected mites. To test whether Cardinium induce reproductive phenotypes in P. ulmi, a Cardinium-cured derived line was generated by antibiotic treatment from a co-infected field population. Genetic crosses indicated that Cardinium do not induce demonstrable levels of cytoplasmic incompatibility and sex allocation distortion in co-infected P. ulmi. However, Cardinium infection was associated with a longer developmental time and reduced total fecundity for co-infected females. We hypothesize that Cardinium spread through P. ulmi populations via uncharacterized fitness effects and that co-infection with Wolbachia might impact these drive mechanisms.}, } @article {pmid37811984, year = {2023}, author = {Edwards, B and Ghedin, E and Voronin, D}, title = {Wolbachia interferes with Zika virus replication by hijacking cholesterol metabolism in mosquito cells.}, journal = {Microbiology spectrum}, volume = {11}, number = {6}, pages = {e0218023}, pmid = {37811984}, issn = {2165-0497}, mesh = {Animals ; Humans ; *Zika Virus ; *Wolbachia ; *Aedes/microbiology ; Mosquito Vectors/microbiology ; *Zika Virus Infection ; *Arboviruses ; Virus Replication ; Cholesterol ; }, abstract = {Arthropod-borne viruses are emerging pathogens that are spread widely by mosquitos. Zika virus is an arbovirus that can infect humans and be transmitted from an infected mother to the fetus, potentially leading to microcephaly in infants. One promising strategy to prevent disease caused by arboviruses is to target the insect vector population. Recent field studies have shown that mosquito populations infected with Wolbachia bacteria suppress arbovirus replication and transmission. Here, we describe how intracellular bacteria redirect resources within their host cells and suppress Zika virus replication at the cellular level. Understanding the mechanism behind Wolbachia-induced interference of arbovirus replication could help advance strategies to control arbovirus pathogens in insect vectors and human populations.}, } @article {pmid37808371, year = {2023}, author = {Sharma, AK and Som, A}, title = {Assigning new supergroups V and W to the Wolbachia diversity.}, journal = {Bioinformation}, volume = {19}, number = {3}, pages = {336-340}, pmid = {37808371}, issn = {0973-2063}, abstract = {Wolbachia are endosymbiotic and alphaproteobacteria that belong to the order Rickettsiales. They are known to infect half of the insect population and cause host manipulation, and have been categorized into 19 monophyletic lineages called supergroups. Recently, two strains, wCfeJ and wCfeT were isolated from cat fleas (Ctenocephalides felis), but their supergroup relationships were not assigned. In this article, we have attempted to classify these two novel strains and establish their evolutionary lineage (i.e., supergroup designation). For this we performed 16S rRNA similarity analysis and reconstructed 16S rRNA phylogeny of 52 Wolbachia strains (including two novel strains) belong to 19 supergroups. We also performed average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) studies to measure genomic similarity between the two novel genomes. The results revealed that 16S rRNA similarity between the two novel strains is 97.94%, which is below the threshold value of 98.6% and phylogeny shows that they are placed at the two different positions (i.e., showing distinct evolutionary lineages). Further, genomic similarity analysis revealed that the novel genomes have ANI and dDDH values 79% and 22.4% respectively, which were below the threshold value of ANI (95%) and dDDH (70%). These results suggested that the novel strains neither shared a species boundary between them nor with any other previously identified supergroups, which designate them as two new supergroups, namely supergroup V (strain wCfeJ) and supergroup W (strain wCfeT).}, } @article {pmid37808301, year = {2023}, author = {Cui, X and Liu, Y and Zhang, J and Hu, P and Zheng, Z and Deng, X and Xu, M}, title = {Variation of endosymbiont and citrus tristeza virus (CTV) titers in the Huanglongbing insect vector, Diaphorina citri, on CTV-infected plants.}, journal = {Frontiers in microbiology}, volume = {14}, number = {}, pages = {1236731}, pmid = {37808301}, issn = {1664-302X}, abstract = {"Candidatus Liberibacter asiaticus" (CLas) is a notorious agent that causes Citrus Huanglongbing (HLB), which is transmitted by Diaphorina citri (D. citri). We recently found that the acquisition and transmission of CLas by D. citri was facilitated by Citrus tristeza virus (CTV), a widely distributed virus in the field. In this study, we further studied whether different CTV strains manipulate the host preference of D. citri, and whether endosymbionts variation is related to CTV strains in D. citri. The results showed that the non-viruliferous D. citri preferred to select the shoots infected with CTV, without strain differences was observed in the selection. However, the viruliferous D. citri prefered to select the mixed strain that is similar to the field's. Furthermore, D. citri effectively acquired the CTV within 2-12 h depending on the strains of the virus. The persistence period of CTV in D. citri was longer than 24 days, without reduction of the CTV titers being observed. These results provide a foundation for understanding the transmission mode of D. citri on CTV. During the process of CTV acquisition and persistence, the titers of main endosymbionts in D. citri showed similar variation trend, but their relative titers were different at different time points. The titers of the "Candidatus Profftella armatura" and CTV tended to be positively correlated, and the titers of Wolbachia and "Candidatus Carsonella ruddii" were mostly negatively related with titers of CT31. These results showed the relationship among D. citri, endosymbionts, and CTV and provided useful information for further research on the interactions between D. citri and CLas, which may benefit the development of approaches for the prevention of CLas transmission and control of citrus HLB.}, } @article {pmid37808105, year = {2023}, author = {Pikula, J and Piacek, V and Bandouchova, H and Bartlova, M and Bednarikova, S and Burianova, R and Danek, O and Jedlicka, P and Masova, S and Nemcova, M and Seidlova, V and Zukalova, K and Zukal, J}, title = {Case report: Filarial infection of a parti-coloured bat: Litomosa sp. adult worms in abdominal cavity and microfilariae in bat semen.}, journal = {Frontiers in veterinary science}, volume = {10}, number = {}, pages = {1284025}, pmid = {37808105}, issn = {2297-1769}, abstract = {BACKGROUND: Filarial infections have been understudied in bats. Likewise, little is known about pathogens associated with the reproductive system in chiropterans. While semen quality is critical for reproductive success, semen-borne pathogens may contribute to reproductive failure.

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

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

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

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

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

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

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

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

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

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

CONCLUSIONS/SIGNIFICANCE: This is the first report that Belgian Culex mosquitoes can transmit both WNV and USUV, forewarning the risk of human transmission. More research is needed to understand the potential influence of Wolbachia on arbovirus transmission in Culex modestus mosquitoes.}, } @article {pmid37720159, year = {2023}, author = {Giordano, R and Weber, EP and Mitacek, R and Flores, A and Ledesma, A and De, AK and Herman, TK and Soto-Adames, FN and Nguyen, MQ and Hill, CB and Hartman, GL}, title = {Patterns of asexual reproduction of the soybean aphid, Aphis glycines (Matsumura), with and without the secondary symbionts Wolbachia and Arsenophonus, on susceptible and resistant soybean genotypes.}, journal = {Frontiers in microbiology}, volume = {14}, number = {}, pages = {1209595}, pmid = {37720159}, issn = {1664-302X}, abstract = {Plant breeding is used to develop crops with host resistance to aphids, however, virulent biotypes often develop that overcome host resistance genes. We tested whether the symbionts, Arsenophonus (A) and Wolbachia (W), affect virulence and fecundity in soybean aphid biotypes Bt1 and Bt3 cultured on whole plants and detached leaves of three resistant, Rag1, Rag2 and Rag1 + 2, and one susceptible, W82, soybean genotypes. Whole plants and individual aphid experiments of A. glycines with and without Arsenophonus and Wolbachia did not show differences in overall fecundity. Differences were observed in peak fecundity, first day of deposition, and day of maximum nymph deposition of individual aphids on detached leaves. Bt3 had higher fecundity than Bt1 on detached leaves of all plant genotypes regardless of bacterial profile. Symbionts did not affect peak fecundity of Bt1 but increased it in Bt3 (A+W+) and all Bt3 strains began to deposit nymphs earlier than the Bt1 (A+W-). Arsenophonus in Bt1 delayed the first day of nymph deposition in comparison to aposymbiotic Bt1 except when reared on Rag1 + 2. For the Bt1 and Bt3 strains, symbionts did not result in a significant difference in the day they deposited the maximum number of nymphs nor was there a difference in survival or variability in number of nymphs deposited. Variability of number of aphids deposited was higher in aphids feeding on resistant plant genotypes. The impact of Arsenophonus on soybean aphid patterns of fecundity was dependent on the aphid biotype and plant genotype. Wolbachia alone had no detectable impact but may have contributed to the increased fecundity of Bt3 (A+W+). An individual based model, using data from the detached leaves experiment and with intraspecific competition removed, found patterns similar to those observed in the greenhouse and growth chamber experiments including a significant interaction between soybean genotype and aphid strain. Combining individual data with the individual based model of population growth isolated the impact of fecundity and host resistance from intraspecific competition and host health. Changes to patterns of fecundity, influenced by the composition and concentration of symbionts, may contribute to competitive interactions among aphid genotypes and influence selection on virulent aphid populations.}, } @article {pmid37716961, year = {2023}, author = {Mfopit, YM and Engel, JS and Chechet, GD and Ibrahim, MAM and Signaboubo, D and Achukwi, DM and Mamman, M and Balogun, EO and Shuaibu, MN and Kabir, J and Kelm, S}, title = {Molecular detection of Sodalis glossinidius, Spiroplasma species and Wolbachia endosymbionts in wild population of tsetse flies collected in Cameroon, Chad and Nigeria.}, journal = {BMC microbiology}, volume = {23}, number = {1}, pages = {260}, pmid = {37716961}, issn = {1471-2180}, support = {K43 TW012015/TW/FIC NIH HHS/United States ; }, mesh = {Animals ; *Tsetse Flies ; *Wolbachia/genetics ; Cameroon ; Chad ; Nigeria ; *Glossinidae ; *Trypanosomiasis, African ; *Spiroplasma/genetics ; }, abstract = {BACKGROUND: Tsetse flies are cyclical vectors of African trypanosomiasis (AT). The flies have established symbiotic associations with different bacteria that influence certain aspects of their physiology. Vector competence of tsetse flies for different trypanosome species is highly variable and is suggested to be affected by bacterial endosymbionts amongst other factors. Symbiotic interactions may provide an avenue for AT control. The current study provided prevalence of three tsetse symbionts in Glossina species from Cameroon, Chad and Nigeria.

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

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

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

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

CONCLUSIONS: The results reinforce the view that VT is a rare event in wild-type mosquitoes and that infection with wMel is effective in reducing VT.}, } @article {pmid37652566, year = {2023}, author = {Cavany, S and Huber, JH and Wieler, A and Tran, QM and Alkuzweny, M and Elliott, M and España, G and Moore, SM and Perkins, TA}, title = {Does ignoring transmission dynamics lead to underestimation of the impact of interventions against mosquito-borne disease?.}, journal = {BMJ global health}, volume = {8}, number = {8}, pages = {}, pmid = {37652566}, issn = {2059-7908}, support = {R35 GM143029/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Humans ; *Vector Borne Diseases/prevention & control/transmission ; Culicidae ; Bias ; Models, Biological ; }, abstract = {New vector-control technologies to fight mosquito-borne diseases are urgently needed, the adoption of which depends on efficacy estimates from large-scale cluster-randomised trials (CRTs). The release of Wolbachia-infected mosquitoes is one promising strategy to curb dengue virus (DENV) transmission, and a recent CRT reported impressive reductions in dengue incidence following the release of these mosquitoes. Such trials can be affected by multiple sources of bias, however. We used mathematical models of DENV transmission during a CRT of Wolbachia-infected mosquitoes to explore three such biases: human movement, mosquito movement and coupled transmission dynamics between trial arms. We show that failure to account for each of these biases would lead to underestimated efficacy, and that the majority of this underestimation is due to a heretofore unrecognised bias caused by transmission coupling. Taken together, our findings suggest that Wolbachia-infected mosquitoes could be even more promising than the recent CRT suggested. By emphasising the importance of accounting for transmission coupling between arms, which requires a mathematical model, we highlight the key role that models can play in interpreting and extrapolating the results from trials of vector control interventions.}, } @article {pmid37649415, year = {2023}, author = {Martínez-Burgos, M and Lozano-Sardaneta, YN and Rodríguez-Rojas, JJ and Gómez-Rivera, ÁS and Canto-Mis, KL and Flores-Escobar, E and Mis-Ávila, PC and Correa-Morales, F and Becker, I}, title = {Species diversity and detection of pathogens in phlebotomine sand flies collected from forest management areas of Quintana Roo, Mexico.}, journal = {Medical and veterinary entomology}, volume = {37}, number = {4}, pages = {845-858}, doi = {10.1111/mve.12691}, pmid = {37649415}, issn = {1365-2915}, mesh = {Animals ; *Psychodidae ; Mexico ; Insect Vectors ; *Phlebotomus ; Forests ; }, abstract = {Sand flies have expanded their areas of distribution, thereby increasing the risk of pathogen transmission in non-endemic areas. To establish efficient prevention and control strategies for the transmission of vector-borne pathogens, it is important to understand seasonal dynamics of their vectors. In Mexico, there are several areas where the contact between sand flies, hosts and reservoirs favours the transmission of the pathogen. We compared sand fly communities in a forest management area and a conserved area in Noh-Bec, Quintana Roo, Mexico. The analysis included species diversity, activity peaks and molecular detection of pathogens. Sand flies were collected from November to December 2021 and April to May 2022, during 84 night-traps. The conserved area showed higher numbers and greater species heterogeneity of sand flies as compared with the other sites. The β-diversity analysis revealed that sites disturbed by logging (S1, S2, S3) had greater similarity (90%) in their sand fly species composition than a conserved area (S4) (similarity = 36%). Although none of the specimens were infected with Leishmania, we detected Wolbachia (19.4%) in all four sites, as well as Bartonella (3.25%) only in the disturbed sites. Further studies on the dynamics of sand fly populations and their association with pathogens are necessary.}, } @article {pmid37647311, year = {2023}, author = {Wei, TL and Zheng, YP and Wang, ZH and Shang, YX and Pei, MS and Liu, HN and Yu, YH and Shi, QF and Jiang, DM and Guo, DL}, title = {Comparative microbiome analysis reveals the variation in microbial communities between 'Kyoho' grape and its bud mutant variety.}, journal = {PloS one}, volume = {18}, number = {8}, pages = {e0290853}, pmid = {37647311}, issn = {1932-6203}, mesh = {*Chlorobi ; *Vitis ; *Microbiota/genetics ; *Anaplasmataceae ; Enterobacteriaceae ; }, abstract = {Microbes are an important part of the vineyard ecosystem, which significantly influence the quality of grapes. Previously, we identified a bud mutant variety (named 'Fengzao') from 'Kyoho' grapes. The variation of microbial communities in grape and its bud mutant variety has not been studied yet. So, in this study, with the samples of both 'Fengzao' and 'Kyoho', we conducted high-throughput microbiome sequencing and investigated their microbial communities in different tissues. Obvious differences were observed in the microbial communities between 'Fengzao' and 'Kyoho'. The fruit and the stem are the tissues with relatively higher abundance of microbes, while the leaves contained less microbes. The fruit and the stem of 'Kyoho' and the stem of 'Fengzao' had relatively higher species diversity based on the alpha diversity analysis. Proteobacteria, Enterobacteriaceae and Rhodobacteraceae had significantly high abundance in 'Fengzao'. Firmicutes and Pseudomonas were highly abundant in the stems of 'Kyoho', and family of Spirochaetaceae, Anaplasmataceae, Chlorobiaceae, and Sphingomonadaceae, and genera of Spirochaeta, Sphingomonas, Chlorobaculum and Wolbachia were abundant in the fruits of 'Kyoho'. These identified microbes are main components of the microbial communities, and could be important regulators of grapevine growth and development. This study revealed the differences in the microbial compositions between 'Kyoho' and its bud mutant, and these identified microbes will be significant resources for the future researches on the quality regulation and disease control of grapevines.}, } @article {pmid37645949, year = {2023}, author = {Wenzel, M and Aquadro, CF}, title = {Wolbachia infection at least partially rescues the fertility and ovary defects of several new Drosophila melanogaster bag of marbles protein-coding mutants.}, journal = {bioRxiv : the preprint server for biology}, volume = {}, number = {}, pages = {}, doi = {10.1101/2023.03.20.532813}, pmid = {37645949}, issn = {2692-8205}, abstract = {UNLABELLED: The D. melanogaster protein coding gene bag of marbles (bam) plays a key role in early male and female reproduction by forming complexes with partner proteins to promote differentiation in gametogenesis. Like another germline gene, Sex lethal , bam genetically interacts with the endosymbiont Wolbachia , as Wolbachia rescues the reduced fertility of a bam hypomorphic mutant. Here, we explored the specificity of the bam-Wolbachia interaction by generating 22 new bam mutants, with ten mutants displaying fertility defects. Nine of these mutants trend towards rescue by the w Mel Wolbachia variant, with eight statistically significant at the fertility and/or cytological level. In some cases, fertility was increased a striking 20-fold. There is no specificity between the rescue and the known binding regions of bam , suggesting w Mel does not interact with one singular bam partner to rescue the reproductive phenotype. We further tested if w Mel interacts with bam in a non-specific way, by increasing bam transcript levels or acting upstream in germline stem cells. A fertility assessment of a bam RNAi knockdown mutant reveals that w Mel rescue is specific to functionally mutant bam alleles and we find no obvious evidence of w Mel interaction with germline stem cells in bam mutants.

AUTHOR SUMMARY: Reproduction in the Drosophila melanogaster fruit fly is dependent on the bag of marbles (bam) gene, which acts early in the process of generating eggs and sperm. Mutations to this gene negatively impact the fertility of the fly, causing it to be sterile or have fewer progeny. Interestingly, we find that the bacteria Wolbachia , which resides within reproductive cells across a wide range of insects, partially restores the fertility and ovary phenotype of several bam mutants of which the resultant Bam protein is altered from wildtype. The protein function of Bam is further suggested to be important by the lack of rescue for a fly that has a fertility defect due to low expression of a non-mutated bam gene. Previous work makes similar conclusions about Wolbachia with another reproductive gene, Sex lethal (Sxl), highlighting the potential for rescue of fertility mutants to occur in a similar way across different genes. An understanding of the ways in which Wolbachia can affect host reproduction provides us with context with which to frame Wolbachia 's impact on host genes, such as bam and Sxl, and consider the evolutionary implications of Wolbachia 's infection in D. melanogaster fruit flies.}, } @article {pmid37630471, year = {2023}, author = {Chao, LL and Shih, CM}, title = {First Detection and Genetic Identification of Wolbachia Endosymbiont in Field-Caught Aedes aegypti (Diptera: Culicidae) Mosquitoes Collected from Southern Taiwan.}, journal = {Microorganisms}, volume = {11}, number = {8}, pages = {}, pmid = {37630471}, issn = {2076-2607}, support = {MOST 111-2314-B-037-031//Ministry of Science and Technology, Taiwan/ ; }, abstract = {The prevalence and genetic character of Wolbachia endosymbionts in field-collected Aedes aegypti mosquitoes were examined for the first time in Taiwan. A total of 665 Ae. aegypti were screened for Wolbachia infection using a PCR assay targeting the Wolbachia surface protein (wsp) gene. In general, the prevalence of Wolbachia infection was detected in 3.3% Ae. aegypti specimens (2.0% female and 5.2% male). Group-specific Wolbachia infection was detected with an infection rate of 1.8%, 0.8% and 0.8% in groups A, B and A&B, respectively. Genetic analysis demonstrated that all Wolbachia strains from Taiwan were phylogenetically affiliated with Wolbachia belonging to the supergroups A and B, with high sequence similarities of 99.4-100% and 99.2-100%, respectively. Phylogenetic relationships can be easily distinguished by maximum likelihood (ML) analysis and were congruent with the unweighted pair group with the arithmetic mean (UPGMA) method. The intra- and inter-group analysis of genetic distance (GD) values revealed a lower level within the Taiwan strains (GD < 0.006 for group A and GD < 0.008 for group B) and a higher level (GD > 0.498 for group A and GD > 0.286 for group B) as compared with other Wolbachia strains. Our results describe the first detection and molecular identification of Wolbachia endosymbiont in field-caught Ae. aegypti mosquitoes collected from Taiwan, and showed a low Wolbachia infection rate belonging to supergroups A and B in Ae. aegypti mosquitoes.}, } @article {pmid37628597, year = {2023}, author = {Corpuz, RL and Bellinger, MR and Veillet, A and Magnacca, KN and Price, DK}, title = {The Transmission Patterns of the Endosymbiont Wolbachia within the Hawaiian Drosophilidae Adaptive Radiation.}, journal = {Genes}, volume = {14}, number = {8}, pages = {}, pmid = {37628597}, issn = {2073-4425}, mesh = {Female ; Animals ; *Ecosystem ; Hawaii ; Phylogeny ; *Wolbachia/genetics ; Drosophila/genetics ; }, abstract = {The evolution of endosymbionts and their hosts can lead to highly dynamic interactions with varying fitness effects for both the endosymbiont and host species. Wolbachia, a ubiquitous endosymbiont of arthropods and nematodes, can have both beneficial and detrimental effects on host fitness. We documented the occurrence and patterns of transmission of Wolbachia within the Hawaiian Drosophilidae and examined the potential contributions of Wolbachia to the rapid diversification of their hosts. Screens for Wolbachia infections across a minimum of 140 species of Hawaiian Drosophila and Scaptomyza revealed species-level infections of 20.0%, and across all 399 samples, a general infection rate of 10.3%. Among the 44 Wolbachia strains we identified using a modified Wolbachia multi-locus strain typing scheme, 30 (68.18%) belonged to supergroup B, five (11.36%) belonged to supergroup A, and nine (20.45%) had alleles with conflicting supergroup assignments. Co-phylogenetic reconciliation analysis indicated that Wolbachia strain diversity within their endemic Hawaiian Drosophilidae hosts can be explained by vertical (e.g., co-speciation) and horizontal (e.g., host switch) modes of transmission. Results from stochastic character trait mapping suggest that horizontal transmission is associated with the preferred oviposition substrate of the host, but not the host's plant family or island of occurrence. For Hawaiian Drosophilid species of conservation concern, with 13 species listed as endangered and 1 listed as threatened, knowledge of Wolbachia strain types, infection status, and potential for superinfection could assist with conservation breeding programs designed to bolster population sizes, especially when wild populations are supplemented with laboratory-reared, translocated individuals. Future research aimed at improving the understanding of the mechanisms of Wolbachia transmission in nature, their impact on the host, and their role in host species formation may shed light on the influence of Wolbachia as an evolutionary driver, especially in Hawaiian ecosystems.}, } @article {pmid37622600, year = {2023}, author = {Ciocchetta, S and Frentiu, FD and Montarsi, F and Capelli, G and Devine, GJ}, title = {Investigation on key aspects of mating biology in the mosquito Aedes koreicus.}, journal = {Medical and veterinary entomology}, volume = {37}, number = {4}, pages = {826-833}, doi = {10.1111/mve.12687}, pmid = {37622600}, issn = {1365-2915}, mesh = {Female ; Male ; Animals ; *Aedes ; Reproduction ; Insemination ; Italy ; Biology ; Introduced Species ; Mosquito Vectors ; }, abstract = {Aedes koreicus Edwards, 1917 (Hulecoetomyia koreica) is a mosquito (Diptera: Culicidae) from Northeast Asia with a rapidly expanding presence outside its original native range. Over the years, the species has been discovered in several new countries, either spreading after first introduction or remaining localised to limited areas. Notably, recent studies have demonstrated the ability of the species to transmit zoonotic parasites and viruses both in the field and in laboratory settings. Combined with its invasive potential, the possible role of Ae. koreicus in pathogen transmission highlights the public health risks resulting from its invasion. In this study, we used a recently established population from Italy to investigate aspects of biology that influence reproductive success in Ae. koreicus: autogeny, mating behaviour, mating disruption by the sympatric invasive species Aedes albopictus Skuse, 1894, and the presence of the endosymbiont Wolbachia pipientis Hertig, 1936. Our laboratory population did not exhibit autogenic behaviour and required a bloodmeal to complete its ovarian cycle. When we exposed Ae. koreicus females to males of Ae. albopictus, we observed repeated attempts at insemination and an aggressive, disruptive mating behaviour initiated by male Ae. albopictus. Despite this, no sperm was identified in Ae. koreicus spermathecae. Wolbachia, an endosymbiotic bacterium capable of influencing mosquito reproductive behaviour, was not detected in this Ae. koreicus population and, therefore, had no effect on Ae. koreicus reproduction.}, } @article {pmid37612420, year = {2023}, author = {Strunov, A and Schoenherr, C and Kapun, M}, title = {Wolbachia has subtle effects on thermal preference in highly inbred Drosophila melanogaster which vary with life stage and environmental conditions.}, journal = {Scientific reports}, volume = {13}, number = {1}, pages = {13792}, pmid = {37612420}, issn = {2045-2322}, support = {P 32275/FWF_/Austrian Science Fund FWF/Austria ; }, mesh = {Animals ; Drosophila melanogaster ; *Wolbachia ; Drosophila ; *Arthropods ; Body Temperature ; }, abstract = {Temperature fluctuations are challenging for ectotherms which are not able to regulate body temperature by physiological means and thus have to adjust their thermal environment via behavior. However, little is yet known about whether microbial symbionts influence thermal preference (Tp) in ectotherms by modulating their physiology. Several recent studies have demonstrated substantial effects of Wolbachia infections on host Tp in different Drosophila species. These data indicate that the direction and strength of thermal preference variation is strongly dependent on host and symbiont genotypes and highly variable among studies. By employing highly controlled experiments, we investigated the impact of several environmental factors including humidity, food quality, light exposure, and experimental setup that may influence Tp measurements in adult Drosophila melanogaster flies. Additionally, we assessed the effects of Wolbachia infection on Tp of Drosophila at different developmental stages, which has not been done before. We find only subtle effects of Wolbachia on host Tp which are strongly affected by experimental variation in adult, but not during juvenile life stages. Our in-depth analyses show that environmental variation has a substantial influence on Tp which demonstrates the necessity of careful experimental design and cautious interpretations of Tp measurements together with a thorough description of the methods and equipment used to conduct behavioral studies.}, } @article {pmid37612118, year = {2023}, author = {Maruyama, J and Inoue, H and Hirose, Y and Nakabachi, A}, title = {16S rRNA Gene Sequencing of Six Psyllid Species of the Family Carsidaridae Identified Various Bacteria Including Symbiopectobacterium.}, journal = {Microbes and environments}, volume = {38}, number = {3}, pages = {}, pmid = {37612118}, issn = {1347-4405}, mesh = {Animals ; *Hemiptera ; RNA, Ribosomal, 16S/genetics ; Genes, rRNA ; Bacteria/genetics ; Enterobacteriaceae/genetics ; *Gammaproteobacteria/genetics ; }, abstract = {Psyllids (Hemiptera: Sternorrhyncha: Psylloidea) are plant sap-sucking insects that are closely associated with various microbes. To obtain a more detailed understanding of the ecological and evolutionary behaviors of microbes in Psylloidea, the bacterial populations of six psyllid species, belonging to the family Carsidaridae, were analyzed using high-throughput amplicon sequencing of the 16S rRNA gene. The majority of the secondary symbionts identified in the present study were gammaproteobacteria, particularly those of the order Enterobacterales, including Arsenophonus and Sodalis, which are lineages found in a wide variety of insect hosts. Additionally, Symbiopectobacterium, another Enterobacterales lineage, which has recently been recognized and increasingly shown to be vertically transmitted and mutualistic in various invertebrates, was identified for the first time in Psylloidea. This lineage is closely related to Pectobacterium spp., which are plant pathogens, but forms a distinct clade exhibiting no pathogenicity to plants. Non-Enterobacterales gammaproteobacteria found in the present study were Acinetobacter, Pseudomonas (both Pseudomonadales), Delftia, Comamonas (both Burkholderiales), and Xanthomonas (Xanthomonadales), a putative plant pathogen. Regarding alphaproteobacteria, three Wolbachia (Rickettsiales) lineages belonging to supergroup B, the major group in insect lineages, were detected in four psyllid species. In addition, a Wolbachia lineage of supergroup O, a minor group recently found for the first time in Psylloidea, was detected in one psyllid species. These results suggest the pervasive transfer of bacterial symbionts among animals and plants, providing deeper insights into the evolution of the interactions among these organisms.}, } @article {pmid37604924, year = {2023}, author = {Osorio, J and Villa-Arias, S and Camargo, C and Ramírez-Sánchez, LF and Barrientos, LM and Bedoya, C and Rúa-Uribe, G and Dorus, S and Alfonso-Parra, C and Avila, FW}, title = {wMel Wolbachia alters female post-mating behaviors and physiology in the dengue vector mosquito Aedes aegypti.}, journal = {Communications biology}, volume = {6}, number = {1}, pages = {865}, pmid = {37604924}, issn = {2399-3642}, mesh = {Animals ; Male ; Female ; Humans ; *Aedes ; Proteomics ; *Wolbachia ; Semen ; Mosquito Vectors ; *Dengue/prevention & control ; }, abstract = {Globally invasive Aedes aegypti disseminate numerous arboviruses that impact human health. One promising method to control Ae. aegypti populations is transinfection with Wolbachia pipientis, which naturally infects ~40-52% of insects but not Ae. aegypti. Transinfection of Ae. aegypti with the wMel Wolbachia strain induces cytoplasmic incompatibility (CI), allows infected individuals to invade native populations, and inhibits transmission of medically relevant arboviruses by females. Female insects undergo post-mating physiological and behavioral changes-referred to as the female post-mating response (PMR)-required for optimal fertility. PMRs are typically elicited by male seminal fluid proteins (SFPs) transferred with sperm during mating but can be modified by other factors, including microbiome composition. Wolbachia has modest effects on Ae. aegypti fertility, but its influence on other PMRs is unknown. Here, we show that Wolbachia influences female fecundity, fertility, and re-mating incidence and significantly extends the longevity of virgin females. Using proteomic methods to examine the seminal proteome of infected males, we found that Wolbachia moderately affects SFP composition. However, we identified 125 paternally transferred Wolbachia proteins, but the CI factor proteins (Cifs) were not among them. Our findings indicate that Wolbachia infection of Ae. aegypti alters female PMRs, potentially influencing control programs that utilize Wolbachia-infected individuals.}, } @article {pmid37603352, year = {2023}, author = {Zhu, Z and Hui, Y and Hu, L}, title = {The impact of predators of mosquito larvae on Wolbachia spreading dynamics.}, journal = {Journal of biological dynamics}, volume = {17}, number = {1}, pages = {2249024}, doi = {10.1080/17513758.2023.2249024}, pmid = {37603352}, issn = {1751-3766}, mesh = {Animals ; *Wolbachia ; Models, Biological ; Mosquito Vectors ; *Culicidae ; Larva ; }, abstract = {Dengue fever creates more than 390 million cases worldwide yearly. The most effective way to deal with this mosquito-borne disease is to control the vectors. In this work we consider two weapons, the endosymbiotic bacteria Wolbachia and predators of mosquito larvae, for combating the disease. As Wolbachia-infected mosquitoes are less able to transmit dengue virus, releasing infected mosquitoes to invade wild mosquito populations helps to reduce dengue transmission. Besides this measure, the introduction of predators of mosquito larvae can control mosquito population. To evaluate the impact of the predators on Wolbachia spreading dynamics, we develop a stage-structured five-dimensional model, which links the predator-prey dynamics with the Wolbachia spreading. By comparatively analysing the dynamics of the models without and with predators, we observe that the introduction of the predators augments the number of coexistence equilibria and impedes Wolbachia spreading. Some numerical simulations are presented to support and expand our theoretical results.}, } @article {pmid37602844, year = {2023}, author = {Cull, B and Wang, XR}, title = {Methods for the Study of Ticks, Mosquitoes, and their Transmitted Pathogens: Toward a Greater Understanding of Vector Biology and Arthropod-Microbe Interactions.}, journal = {Journal of visualized experiments : JoVE}, volume = {}, number = {193}, pages = {}, doi = {10.3791/64986}, pmid = {37602844}, issn = {1940-087X}, support = {R01 AI049424/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; *Arthropods ; Mosquito Vectors ; *Ixodes ; *Aedes ; Biology ; }, abstract = {Chen, L., Xiao, Q., Shi, M., Cheng, J., Wu, J. Detecting Wolbachia strain wAlbB in Aedes albopictus cell lines. Journal of Visualized Experiments. (184), e63662 (2022). Haziqah-Rashid, A. et al. Determining temperature preference of mosquitoes and other ectotherms. Journal of Visualized Experiments. (187), e64356 (2022). Huang, D. et al. Mosquito-associated virus isolation from field-collected mosquitoes. Journal of Visualized Experiments. (186), e63852 (2022). Khoo, B., Cull, B., Oliver, J. D. Tick artificial membrane feeding for Ixodes scapularis. Journal of Visualized Experiments. 64553 (2022). Leal-Galvan, B., Harvey, C., Thomas, D., Saelao, P., Oliva Chavez, A. S. Isolation of microRNAs from tick ex vivo salivary gland cultures and extracellular vesicles. Journal of Visualized Experiments. (182), e63618 (2022). Liang, Q. et al. Control of Aedes albopictus mosquito larvae with Carpesium abrotanoides L. Journal of Visualized Experiments. (186), e63976 (2022). Wang, F. et al. Experimental viral infection in adult mosquitoes by oral feeding and microinjection. Journal of Visualized Experiments. (185), e63830 (2022). Wang, X. R., Burkhardt, N. Y., Price, L. D., Munderloh, U. G. An electroporation method to transform Rickettsia spp. with a fluorescent protein-expressing shuttle vector in tick cell lines. Journal of Visualized Experiments. (188), e64562 (2022).}, } @article {pmid37601442, year = {2023}, author = {Archer, J and Hurst, GDD and Hornett, EA}, title = {Male-killer symbiont screening reveals novel associations in Adalia ladybirds.}, journal = {Access microbiology}, volume = {5}, number = {7}, pages = {}, pmid = {37601442}, issn = {2516-8290}, abstract = {While male-killing bacteria are known to infect across arthropods, ladybird beetles represent a hotspot for these symbioses. In some host species, there are multiple different symbionts that vary in presence and frequency between populations. To further our understanding of spatial and frequency variation, we tested for the presence of three male-killing bacteria: Wolbachia , Rickettsia and Spiroplasma , in two Adalia ladybird species from a previously unexplored UK population. The two-spot ladybird, A. bipunctata, is known to harbour all three male-killers, and we identified Spiroplasma infection in the Merseyside population for the first time. However, in contrast to previous studies on two-spot ladybirds from continental Europe, evidence from egg-hatch rates indicates the Spiroplasma strain present in the Merseyside population does not cause embryonic male-killing. In the related ten-spot ladybird, A. decempunctata, there is only one previous record of a male-killing symbiont, a Rickettsia , which we did not detect in the Merseyside sample. However, PCR assays indicated the presence of a Spiroplasma in a single A. decempunctata specimen. Marker sequence indicated that this Spiroplasma was divergent from that found in sympatric A. bipunctata. Genome sequencing of the Spiroplasma -infected A. decempunctata additionally revealed the presence of cobionts in the form of a Centistes parasitoid wasp and the parasitic fungi Beauveria. Further study of A. decempunctata from this population is needed to resolve whether it is the ladybird or wasp cobiont that harbours Spiroplasma , and to establish the phenotype of this strain. These data indicate first that microbial symbiont phenotype should not be assumed from past studies conducted in different locations, and second that cobiont presence may confound screening studies aimed to detect the frequency of a symbiont in field collected material from a focal host species.}, } @article {pmid37584011, year = {2023}, author = {Perles, L and Otranto, D and Barreto, WTG and de Macedo, GC and Lia, RP and Mendoza-Roldan, JA and Herrera, HM and de Oliveira, CE and Machado, RZ and André, MR}, title = {Mansonella sp. and associated Wolbachia endosymbionts in ring-tailed coatis (Nasua nasua) in periurban areas from Midwestern Brazil.}, journal = {International journal for parasitology. Parasites and wildlife}, volume = {22}, number = {}, pages = {14-19}, pmid = {37584011}, issn = {2213-2244}, abstract = {Coatis (Nasua nasua) are wild carnivorous well adapted to anthropized environments especially important because they act as reservoirs hosts for many arthropod-borne zoonotic pathogens. Information about filarioids from coatis and associated Wolbachia spp. in Brazil is scant. To investigate the diversity of filarial nematodes, blood samples (n = 100 animals) were obtained from two urban areas in midwestern Brazil and analyzed using blood smears and buffy coats and cPCR assays based on the cox1, 12S rRNA, 18S rRNA, hsp70 and myoHC genes for nematodes and 16S rRNA for Wolbachia. When analyzing coati blood smears and buffy coats, 30% and 80% of the samples presented at least one microfilaria, respectively. Twenty-five cox1 sequences were obtained showing 89% nucleotide identity with Mansonella ozzardi. Phylogenetic analyses clustered cox1 sequences herein obtained within the Mansonella spp. clade. Sequences of both myoHC and two hsp70 genes showed 99.8% nucleotide identity with Mansonella sp. and clustered into a clade within Mansonella sp., previously detected in coatis from Brazil. Two blood samples were positive for Wolbachia, with a 99% nucleotide identity with Wolbachia previously found in Mansonella perstans, Mansonella ozzardi and Mansonella atelensis and in ectoparasites of the genus Pseudolynchia, Melophagus and Cimex. The study showed a high prevalence of Mansonella sp. in the coati population examined, suggesting that this animal species play a role as reservoirs of a novel, yet to be described, species within the Onchocercidae family.}, } @article {pmid37577425, year = {2023}, author = {Garrido, M and Veiga, J and Garrigós, M and Martínez-de la Puente, J}, title = {The interplay between vector microbial community and pathogen transmission on the invasive Asian tiger mosquito, Aedes albopictus: current knowledge and future directions.}, journal = {Frontiers in microbiology}, volume = {14}, number = {}, pages = {1208633}, pmid = {37577425}, issn = {1664-302X}, abstract = {The invasive Asian tiger mosquito Aedes albopictus is nowadays broadly distributed with established populations in all continents except Antarctica. In the invaded areas, this species represents an important nuisance for humans and, more relevant, it is involved in the local transmission of pathogens relevant under a public health perspective. Aedes albopictus is a competent vector of parasites such as Dirofilaria and viruses including dengue virus, Zika virus, and chikungunya virus, among others. The mosquito microbiota has been identified as one of the major drivers of vector competence, acting upon relevant vector functions as development or immunity. Here, we review the available literature on the interaction between Ae. albopictus microbiota and pathogen transmission and identify the knowledge gaps on the topic. Most studies are strictly focused on the interplay between pathogens and Wolbachia endosymbiont while studies screening whole microbiota are still scarce but increasing in recent years, supported on Next-generation sequencing tools. Most experimental trials use lab-reared mosquitoes or cell lines, exploring the molecular mechanisms of the microbiota-pathogen interaction. Yet, correlational studies on wild populations are underrepresented. Consequently, we still lack sufficient evidence to reveal whether the microbiota of introduced populations of Ae. albopictus differ from those of native populations, or how microbiota is shaped by different environmental and anthropic factors, but especially, how these changes affect the ability of Ae. albopictus to transmit pathogens and favor the occurrence of outbreaks in the colonized areas. Finally, we propose future research directions on this research topic.}, } @article {pmid37567493, year = {2023}, author = {Kamkong, P and Jitsamai, W and Thongmeesee, K and Ratthawongjirakul, P and Taweethavonsawat, P}, title = {Genetic diversity and characterization of Wolbachia endosymbiont in canine filariasis.}, journal = {Acta tropica}, volume = {246}, number = {}, pages = {107000}, doi = {10.1016/j.actatropica.2023.107000}, pmid = {37567493}, issn = {1873-6254}, mesh = {Animals ; Dogs ; *Wolbachia/genetics ; RNA, Ribosomal, 16S/genetics ; Phylogeny ; *Dirofilariasis ; *Dirofilaria immitis/genetics ; Dirofilaria ; *Filarioidea/genetics ; *Heart Diseases ; Genetic Variation ; *Dog Diseases ; }, abstract = {Canine filariasis is caused by nematodes from the family Onchocercidae, which is transmitted by arthropod vectors. The disease is commonly found in Southeast Asia and exists worldwide. Some filarial nematodes are associated with intracellular bacteria of the genus Wolbachia, which plays an important role in embryogenesis, molting, and the long-term survival of adult worms. This study aims to characterize Wolbachia sp. and determine the association between Wolbachia and canine filarial nematode species in Thailand. A total of 46 dog blood samples that were naturally infected with filarial nematodes were obtained to identify filarial nematode species by Giemsa stained under a light microscope and confirmed using the molecular technique. In order to characterize Wolbachia sp., the nested PCR assay targeting the 16S rRNA gene showed that all samples of Dirofilaria immitis and fifteen samples of Candidatus Dirofilaria hongkongensis were grouped into Wolbachia supergroup C. In addition, all samples of Brugia spp. and five samples of Candidatus Dirofilaria hongkongensis were classified into Wolbachia supergroup D. The genetic diversity analysis conducted using the 16S rRNA gene revealed a similar result when analyzed through phylogenetic tree analysis. This is the first genetic diversity study of Wolbachia of Candidatus Dirofilaria hongkongensis in infected dogs in Thailand.}, } @article {pmid37546688, year = {2023}, author = {Vásquez, VN and Kueppers, LM and Rašić, G and Marshall, JM}, title = {wMel replacement of dengue-competent mosquitoes is robust to near-term change.}, journal = {Nature climate change}, volume = {13}, number = {8}, pages = {848-855}, pmid = {37546688}, issn = {1758-678X}, support = {R01 AI143698/AI/NIAID NIH HHS/United States ; }, abstract = {Rising temperatures are impacting the range and prevalence of mosquito-borne diseases. A promising biocontrol technology replaces wild mosquitoes with those carrying the virus-blocking Wolbachia bacterium. Because the most widely used strain, wMel, is adversely affected by heat stress, we examined how global warming may influence wMel-based replacement. We simulated interventions in two locations with successful field trials using Coupled Model Intercomparison Project Phase 5 climate projections and historical temperature records, integrating empirical data on wMel's thermal sensitivity into a model of Aedes aegypti population dynamics to evaluate introgression and persistence over one year. We show that in Cairns, Australia, climatic futures necessitate operational adaptations for heatwaves exceeding two weeks. In Nha Trang, Vietnam, projected heatwaves of three weeks and longer eliminate wMel under the most stringent assumptions of that symbiont's thermal limits. We conclude that this technology is generally robust to near-term (2030s) climate change. Accelerated warming may challenge this in the 2050s and beyond.}, } @article {pmid37533804, year = {2023}, author = {Namias, A and Sahlin, K and Makoundou, P and Bonnici, I and Sicard, M and Belkhir, K and Weill, M}, title = {Nanopore sequencing of PCR products enables multicopy gene family reconstruction.}, journal = {Computational and structural biotechnology journal}, volume = {21}, number = {}, pages = {3656-3664}, pmid = {37533804}, issn = {2001-0370}, abstract = {The importance of gene amplifications in evolution is more and more recognized. Yet, tools to study multi-copy gene families are still scarce, and many such families are overlooked using common sequencing methods. Haplotype reconstruction is even harder for polymorphic multi-copy gene families. Here, we show that all variants (or haplotypes) of a multi-copy gene family present in a single genome, can be obtained using Oxford Nanopore Technologies sequencing of PCR products, followed by steps of mapping, SNP calling and haplotyping. As a proof of concept, we acquired the sequences of highly similar variants of the cidA and cidB genes present in the genome of the Wolbachia wPip, a bacterium infecting Culex pipiens mosquitoes. Our method relies on a wide database of cid genes, previously acquired by cloning and Sanger sequencing. We addressed problems commonly faced when using mapping approaches for multi-copy gene families with highly similar variants. In addition, we confirmed that PCR amplification causes frequent chimeras which have to be carefully considered when working on families of recombinant genes. We tested the robustness of the method using a combination of bioinformatics (read simulations) and molecular biology approaches (sequence acquisitions through cloning and Sanger sequencing, specific PCRs and digital droplet PCR). When different haplotypes present within a single genome cannot be reconstructed from short reads sequencing, this pipeline confers a high throughput acquisition, gives reliable results as well as insights of the relative copy numbers of the different variants.}, } @article {pmid37532686, year = {2023}, author = {Hasnaoui, B and Bérenger, JM and Delaunay, P and Diarra, AZ and Ndiaye, EHI and M'madi, SA and Masotti, N and Sevestre, J and Parola, P}, title = {Survey of bed bug infestations in homeless shelters in southern France.}, journal = {Scientific reports}, volume = {13}, number = {1}, pages = {12557}, pmid = {37532686}, issn = {2045-2322}, mesh = {Animals ; Humans ; *Bedbugs ; Surveys and Questionnaires ; Public Health ; *Ectoparasitic Infestations ; France ; *Ill-Housed Persons ; }, abstract = {Bed bug has become a major public health pest worldwide. Infestation may result in numerous negative health effects. Homeless shelters are one of the most habitats that can be infested with bed bugs, a few studies have focused on bed bug infestations in these settings. We conducted a survey of infestations of bed bugs in a homeless shelter in southern France, using an innovative seven-level scale (0-6) to assess the degree of infestation, MALDI TOF-MS to identify bed bugs, and a biomolecular tool to detect bacteria. Bed bug infestations were documented in 13% (9/68) of investigated rooms. A total of 184 bed bugs were collected and morphologically identified as Cimex lectularius. MALDI TOF-MS analysis allowed us to obtain high-quality MS spectra for all 184 specimens, to correctly identify all specimens, and included 178/184 (97%) Log Score Values higher than 1.8. Among the bacteria tested, Wolbachia sp. DNA was found in 149/184 (81%) of the bed bugs, and one sample was positive for Coxiella burnetii, the agent of Q fever. Our study is the first of its kind that offers new perspectives for increasing public awareness of the conditions in homeless shelters.}, } @article {pmid37525959, year = {2023}, author = {Kolasa, M and Kajtoch, Ł and Michalik, A and Maryańska-Nadachowska, A and Łukasik, P}, title = {Till evolution do us part: The diversity of symbiotic associations across populations of Philaenus spittlebugs.}, journal = {Environmental microbiology}, volume = {25}, number = {11}, pages = {2431-2446}, doi = {10.1111/1462-2920.16473}, pmid = {37525959}, issn = {1462-2920}, mesh = {Humans ; Animals ; RNA, Ribosomal, 16S/genetics ; Phylogeny ; Enterobacteriaceae/genetics ; Bacteria/genetics ; *Hemiptera/microbiology ; Symbiosis/genetics ; *Wolbachia/genetics ; }, abstract = {Symbiotic bacteria have played crucial roles in the evolution of sap-feeding insects and can strongly affect host function. However, their diversity and distribution within species are not well understood; we do not know to what extent environmental factors or associations with other species may affect microbial community profiles. We addressed this question in Philaenus spittlebugs by surveying both insect and bacterial marker gene amplicons across multiple host populations. Host mitochondrial sequence data confirmed morphology-based identification of six species and revealed two divergent clades of Philaenus spumarius. All of them hosted the primary symbiont Sulcia that was almost always accompanied by Sodalis. Interestingly, populations and individuals often differed in the presence of Sodalis sequence variants, suggestive of intra-genome 16S rRNA variant polymorphism combined with rapid genome evolution and/or recent additional infections or replacements of the co-primary symbiont. The prevalence of facultative endosymbionts, including Wolbachia, Rickettsia, and Spiroplasma, varied among populations. Notably, cytochrome I oxidase (COI) amplicon data also showed that nearly a quarter of P. spumarius were infected by parasitoid flies (Verralia aucta). One of the Wolbachia operational taxonomic units (OTUs) was exclusively present in Verralia-parasitized specimens, suggestive of parasitoids as their source and highlighting the utility of host gene amplicon sequencing in microbiome studies.}, } @article {pmid37524915, year = {2023}, author = {Lee, H and Seo, P and Teklay, S and Yuguchi, E and Benetta, ED and Werren, JH and Ferree, PM}, title = {Ability of a selfish B chromosome to evade genome elimination in the jewel wasp, Nasonia vitripennis.}, journal = {Heredity}, volume = {131}, number = {3}, pages = {230-237}, pmid = {37524915}, issn = {1365-2540}, mesh = {*Genome, Insect ; Animals ; *Chromosomes, Insect ; Protamines/genetics/metabolism ; Histones/genetics/metabolism ; Male ; Female ; Genes, rRNA ; DNA, Ribosomal/genetics/metabolism ; Genetic Loci ; }, abstract = {B chromosomes are non-essential, extra chromosomes that can exhibit transmission-enhancing behaviors, including meiotic drive, mitotic drive, and induction of genome elimination, in plants and animals. A fundamental but poorly understood question is what characteristics allow B chromosomes to exhibit these extraordinary behaviors. The jewel wasp, Nasonia vitripennis, harbors a heterochromatic, paternally transmitted B chromosome known as paternal sex ratio (PSR), which causes complete elimination of the sperm-contributed half of the genome during the first mitotic division of fertilized embryos. This genome elimination event may result from specific, previously observed alterations of the paternal chromatin. Due to the haplo-diploid reproduction of the wasp, genome elimination by PSR causes female-destined embryos to develop as haploid males that transmit PSR. PSR does not undergo self-elimination despite its presence with the paternal chromatin until the elimination event. Here we performed fluorescence microscopic analyses aimed at understanding this unexplained property. Our results show that PSR, like the rest of the genome, participates in the histone-to-protamine transition, arguing that PSR does not avoid this transition to escape self-elimination. In addition, PSR partially escapes the chromatin-altering activity of the intracellular bacterium, Wolbachia, demonstrating that this ability to evade chromatin alteration is not limited to PSR's own activity. Finally, we observed that the rDNA locus and other unidentified heterochromatic regions of the wasp's genome also seem to evade chromatin disruption by PSR, suggesting that PSR's genome-eliminating activity does not affect heterochromatin. Thus, PSR may target an aspect of euchromatin to cause genome elimination.}, } @article {pmid37491306, year = {2023}, author = {Moorhead, AR and Evans, CC and Sakamoto, K and Dzimianski, MT and Mansour, A and DiCosty, U and Fricks, C and McCall, S and Carson, B and Nelson, CT and McCall, JW}, title = {Effects of doxycycline dose rate and pre-adulticide wait period on heartworm-associated pathology and adult worm mass.}, journal = {Parasites & vectors}, volume = {16}, number = {1}, pages = {251}, pmid = {37491306}, issn = {1756-3305}, mesh = {Animals ; Dogs ; *Dirofilaria immitis ; Doxycycline ; *Dirofilariasis/drug therapy ; *Filaricides ; *Dog Diseases/drug therapy ; *Wolbachia ; }, abstract = {BACKGROUND: The American Heartworm Society canine guidelines recommend treatment with doxycycline prior to adulticide administration to reduce levels of Wolbachia and its associated metabolites, which are known to be a leading cause of pulmonary pathology. Studies have determined that doxycycline administered at 10 mg/kg BID for 28 days is an effective dose for eliminating Wolbachia, but what has not been determined is the clinical relevance of this elimination. The current guidelines also recommend a 30-day wait period following administration of doxycycline to allow for clearance of metabolites, such as Wolbachia surface protein, and for further reduction in heartworm biomass before administration of adulticide. Reducing the doxycycline dose and eliminating the wait period may carry practical benefits for the animal, client, and practitioner.

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

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

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

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

EXPERT OPINION: We recommend that human infections with Mansonella species be treated, regardless of any apparent clinical manifestations. We argue that mansonellosis, despite being widely considered a benign infection, may represent a direct or indirect cause of significant morbidity that remains poorly characterized at present.}, } @article {pmid37474590, year = {2023}, author = {Rainey, SM and Geoghegan, V and Lefteri, DA and Ant, TH and Martinez, J and McNamara, CJ and Kamel, W and de Laurent, ZR and Castello, A and Sinkins, SP}, title = {Differences in proteome perturbations caused by the Wolbachia strain wAu suggest multiple mechanisms of Wolbachia-mediated antiviral activity.}, journal = {Scientific reports}, volume = {13}, number = {1}, pages = {11737}, pmid = {37474590}, issn = {2045-2322}, support = {108508/A/15/Z/WT_/Wellcome Trust/United Kingdom ; 202888/Z/16/Z/WT_/Wellcome Trust/United Kingdom ; MC_UU_12014/8/MRC_/Medical Research Council/United Kingdom ; 108508/WT_/Wellcome Trust/United Kingdom ; /WT_/Wellcome Trust/United Kingdom ; 202888/WT_/Wellcome Trust/United Kingdom ; MC_UU_00034/4/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Animals ; *Dengue Virus/physiology ; Proteome ; *Wolbachia/physiology ; Antiviral Agents ; Proteomics ; Lipids ; *Aedes ; }, abstract = {Some strains of the inherited bacterium Wolbachia have been shown to be effective at reducing the transmission of dengue virus (DENV) and other RNA viruses by Aedes aegypti in both laboratory and field settings and are being deployed for DENV control. The degree of virus inhibition varies between Wolbachia strains. Density and tissue tropism can contribute to these differences but there are also indications that this is not the only factor involved: for example, strains wAu and wAlbA are maintained at similar intracellular densities but only wAu produces strong DENV inhibition. We previously reported perturbations in lipid transport dynamics, including sequestration of cholesterol in lipid droplets, with strains wMel/wMelPop in Ae. aegypti. To further investigate the cellular basis underlying these differences, proteomic analysis of midguts was carried out on Ae. aegypti lines carrying strains wAu and wAlbA: with the hypothesis that differences in perturbations may underline Wolbachia-mediated antiviral activity. Surprisingly, wAu-carrying midguts not only showed distinct proteome perturbations when compared to non-Wolbachia carrying and wAlbA-carrying midguts but also wMel-carrying midguts. There are changes in RNA processing pathways and upregulation of a specific set of RNA-binding proteins in the wAu-carrying line, including genes with known antiviral activity. Lipid transport and metabolism proteome changes also differ between strains, and we show that strain wAu does not produce the same cholesterol sequestration phenotype as wMel. Moreover, in contrast to wMel, wAu antiviral activity was not rescued by cyclodextrin treatment. Together these results suggest that wAu could show unique features in its inhibition of arboviruses compared to previously characterized Wolbachia strains.}, } @article {pmid37464886, year = {2023}, author = {Shen, R and Wenzel, M and Messer, PW and Aquadro, CF}, title = {Evolution under a model of functionally buffered deleterious mutations can lead to positive selection in protein-coding genes.}, journal = {Evolution; international journal of organic evolution}, volume = {77}, number = {10}, pages = {2200-2212}, pmid = {37464886}, issn = {1558-5646}, support = {R01 GM095793/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; *Drosophila melanogaster/genetics ; Evolution, Molecular ; Drosophila/genetics ; Selection, Genetic ; Mutation ; Amino Acids/genetics ; RNA-Binding Proteins/genetics ; *Drosophila Proteins/genetics ; }, abstract = {Selective pressures on DNA sequences often result in departures from neutral evolution that can be captured by the McDonald-Kreitman (MK) test. However, the nature of such selective forces often remains unknown to experimentalists. Amino acid fixations driven by natural selection in protein-coding genes are commonly associated with a genetic arms race or changing biological purposes, leading to proteins with new functionality. Here, we evaluate the expectations of population genetic patterns under a buffering mechanism driving selective amino acids to fixation, which is motivated by an observed phenotypic rescue of otherwise deleterious nonsynonymous substitutions at bag of marbles (bam) and Sex lethal (Sxl) in Drosophila melanogaster. These two genes were shown to experience strong episodic bursts of natural selection potentially due to infections of the endosymbiotic bacteria Wolbachia observed among multiple Drosophila species. Using simulations to implement and evaluate the evolutionary dynamics of a Wolbachia buffering model, we demonstrate that selectively fixed amino acid replacements will occur, but that the proportion of adaptive amino acid fixations and the statistical power of the MK test to detect the departure from an equilibrium neutral model are both significantly lower than seen for an arms race/change-in-function model that favors proteins with diversified amino acids. We find that the observed selection pattern at bam in a natural population of D. melanogaster is more consistent with an arms race model than with the buffering model.}, } @article {pmid37443389, year = {2023}, author = {Wybouw, N and Van Reempts, E and Zarka, J and Zélé, F and Bonte, D}, title = {Egg provisioning explains the penetrance of symbiont-mediated sex allocation distortion in haplodiploids.}, journal = {Heredity}, volume = {131}, number = {3}, pages = {221-229}, pmid = {37443389}, issn = {1365-2540}, mesh = {Animals ; Male ; Female ; Reproduction/physiology ; Penetrance ; *Tetranychidae/genetics ; Bacteroidetes ; Cytoplasm ; *Wolbachia/genetics ; Symbiosis/genetics ; Sex Ratio ; }, abstract = {Maternally transmitted symbionts such as Wolbachia can alter sex allocation in haplodiploid arthropods. By biasing population sex ratios towards females, these changes in sex allocation may facilitate the spread of symbionts. In contrast to symbiont-induced cytoplasmic incompatibility (CI), the mechanisms that underpin sex allocation distortion remain poorly understood. Using a nuclear genotype reference panel of the haplodiploid mite Tetranychus urticae and a single Wolbachia variant that is able to simultaneously induce sex allocation distortion and CI, we unraveled the mechanistic basis of Wolbachia-mediated sex allocation distortion. Host genotype was an important determinant for the strength of sex allocation distortion. We further show that sex allocation distortion by Wolbachia in haplodiploid mites is driven by increasing egg size, hereby promoting egg fertilization. This change in reproductive physiology was also coupled to increased male and female adult size. Our results echo previous work on Cardinium symbionts, suggesting that sex allocation distortion by regulating host investment in egg size is a common strategy among symbionts that infect haplodiploids. To better understand the relevance that sex allocation distortion may have for the spread of Wolbachia in natural haplodiploid populations, we parametrized a model based on generated phenotypic data. Our simulations show that empirically derived levels of sex allocation distortion can be sufficient to remove invasion thresholds, allowing CI to drive the spread of Wolbachia independently of the initial infection frequency. Our findings help elucidate the mechanisms that underlie the widespread occurrence of symbionts in haplodiploid arthropods and the evolution of sex allocation.}, } @article {pmid37432953, year = {2023}, author = {Medina, P and Russell, SL and Corbett-Detig, R}, title = {Deep data mining reveals variable abundance and distribution of microbial reproductive manipulators within and among diverse host species.}, journal = {PloS one}, volume = {18}, number = {7}, pages = {e0288261}, pmid = {37432953}, issn = {1932-6203}, support = {R35 GM128932/GM/NIGMS NIH HHS/United States ; T32 HG008345/HG/NHGRI NIH HHS/United States ; }, mesh = {Animals ; Drosophila melanogaster/genetics ; *Arthropods ; Data Mining ; Ecology ; Reproduction ; *Wolbachia/genetics ; }, abstract = {Bacterial symbionts that manipulate the reproduction of their hosts are important factors in invertebrate ecology and evolution, and are being leveraged for host biological control. Infection prevalence restricts which biological control strategies are possible and is thought to be strongly influenced by the density of symbiont infection within hosts, termed titer. Current methods to estimate infection prevalence and symbiont titers are low-throughput, biased towards sampling infected species, and rarely measure titer. Here we develop a data mining approach to estimate symbiont infection frequencies within host species and titers within host tissues. We applied this approach to screen ~32,000 publicly available sequence samples from the most common symbiont host taxa, discovering 2,083 arthropod and 119 nematode infected samples. From these data, we estimated that Wolbachia infects approximately 44% of all arthropod and 34% of all nematode species, while other reproductive manipulators only infect 1-8% of arthropod and nematode species. Although relative titers within hosts were highly variable within and between arthropod species, a combination of arthropod host species and Wolbachia strain explained approximately 36% of variation in Wolbachia titer across the dataset. To explore potential mechanisms for host control of symbiont titer, we leveraged population genomic data from the model system Drosophila melanogaster. In this host, we found a number of SNPs associated with titer in candidate genes potentially relevant to host interactions with Wolbachia. Our study demonstrates that data mining is a powerful tool to detect bacterial infections and quantify infection intensities, thus opening an array of previously inaccessible data for further analysis in host-symbiont evolution.}, } @article {pmid37430172, year = {2023}, author = {Porter, J and Sullivan, W}, title = {The cellular lives of Wolbachia.}, journal = {Nature reviews. Microbiology}, volume = {21}, number = {11}, pages = {750-766}, pmid = {37430172}, issn = {1740-1534}, mesh = {*Wolbachia/physiology ; Animals ; *Symbiosis ; Arthropods/microbiology ; Host Microbial Interactions/physiology ; Filarioidea/microbiology/physiology ; Insecta/microbiology ; Host-Pathogen Interactions ; }, abstract = {Wolbachia are successful Gram-negative bacterial endosymbionts, globally infecting a large fraction of arthropod species and filarial nematodes. Efficient vertical transmission, the capacity for horizontal transmission, manipulation of host reproduction and enhancement of host fitness can promote the spread both within and between species. Wolbachia are abundant and can occupy extraordinary diverse and evolutionary distant host species, suggesting that they have evolved to engage and manipulate highly conserved core cellular processes. Here, we review recent studies identifying Wolbachia-host interactions at the molecular and cellular levels. We explore how Wolbachia interact with a wide array of host cytoplasmic and nuclear components in order to thrive in a diversity of cell types and cellular environments. This endosymbiont has also evolved the ability to precisely target and manipulate specific phases of the host cell cycle. The remarkable diversity of cellular interactions distinguishes Wolbachia from other endosymbionts and is largely responsible for facilitating its global propagation through host populations. Finally, we describe how insights into Wolbachia-host cellular interactions have led to promising applications in controlling insect-borne and filarial nematode-based diseases.}, } @article {pmid37428804, year = {2023}, author = {Alami, N and Carter, DC and Kwatra, NV and Zhao, W and Snodgrass, L and Porcalla, AR and Klein, CE and Cohen, DE and Gallenberg, L and Neenan, M and Carr, RA and Marsh, KC and Kempf, DJ}, title = {A Phase-I pharmacokinetic, safety and food-effect study on flubentylosin, a novel analog of Tylosin-A having potent anti-Wolbachia and antifilarial activity.}, journal = {PLoS neglected tropical diseases}, volume = {17}, number = {7}, pages = {e0011392}, pmid = {37428804}, issn = {1935-2735}, mesh = {Adult ; Humans ; Tylosin ; *Wolbachia ; *Onchocerciasis ; Double-Blind Method ; Anti-Bacterial Agents/pharmacokinetics ; Macrolides ; Area Under Curve ; Dose-Response Relationship, Drug ; Administration, Oral ; }, abstract = {BACKGROUND: The parasitic filariae responsible for onchocerciasis and lymphatic filariasis are host to an endosymbiotic bacterium, Wolbachia, which is essential to the fertility and development of the parasites. We performed a Phase-I pharmacokinetic, safety and food-effect study on single and multiple ascending doses of flubentylosin (ABBV-4083), a macrolide antibacterial with activity against Wolbachia, intended to sterilize and eliminate the parasites.

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

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

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

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

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

DISCUSSION: Future adoption of these mouse models will benefit end-user laboratories conducting research and development of novel heartworm preventatives via increased access, rapid turnaround, and reduced costs and may simultaneously decrease the need for experimental cat or dog use.}, } @article {pmid37425681, year = {2023}, author = {Boehm, EC and Jaeger, AS and Ries, HJ and Castañeda, D and Weiler, AM and Valencia, CC and Weger-Lucarelli, J and Ebel, GD and O'Connor, SL and Friedrich, TC and Zamanian, M and Aliota, MT}, title = {Wolbachia -mediated resistance to Zika virus infection in Aedes aegypti is dominated by diverse transcriptional regulation and weak evolutionary pressures.}, journal = {bioRxiv : the preprint server for biology}, volume = {}, number = {}, pages = {}, pmid = {37425681}, issn = {2692-8205}, support = {P51 RR000167/RR/NCRR NIH HHS/United States ; T32 AI083196/AI/NIAID NIH HHS/United States ; R01 AI132563/AI/NIAID NIH HHS/United States ; R21 AI131454/AI/NIAID NIH HHS/United States ; P51 OD011106/OD/NIH HHS/United States ; }, abstract = {UNLABELLED: A promising candidate for arbovirus control and prevention relies on replacing arbovirus-susceptible Aedes aegypti populations with mosquitoes that have been colonized by the intracellular bacterium Wolbachia and thus have a reduced capacity to transmit arboviruses. This reduced capacity to transmit arboviruses is mediated through a phenomenon referred to as pathogen blocking. Pathogen blocking has primarily been proposed as a tool to control dengue virus (DENV) transmission, however it works against a range of viruses, including Zika virus (ZIKV). Despite years of research, the molecular mechanisms underlying pathogen blocking still need to be better understood. Here, we used RNA-seq to characterize mosquito gene transcription dynamics in Ae. aegypti infected with the w Mel strain of Wolbachia that are being released by the World Mosquito Program in Medellín, Colombia. Comparative analyses using ZIKV-infected, uninfected tissues, and mosquitoes without Wolbachia revealed that the influence of w Mel on mosquito gene transcription is multifactorial. Importantly, because Wolbachia limits, but does not completely prevent, replication of ZIKV and other viruses in coinfected mosquitoes, there is a possibility that these viruses could evolve resistance to pathogen blocking. Therefore, to understand the influence of Wolbachia on within-host ZIKV evolution, we characterized the genetic diversity of molecularly barcoded ZIKV virus populations replicating in Wolbachia -infected mosquitoes and found that within-host ZIKV evolution was subject to weak purifying selection and, unexpectedly, loose anatomical bottlenecks in the presence and absence of Wolbachia . Together, these findings suggest that there is no clear transcriptional profile associated with Wolbachia -mediated ZIKV restriction, and that there is no evidence for ZIKV escape from this restriction in our system.

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

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

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

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

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

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

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

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

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

DISCUSSION: The findings of the study have advanced our understanding of the potential pathogen groups of vectors in marmot (Marmota himalayana) in the Qinghai-Tibet Plateau.}, } @article {pmid37399133, year = {2023}, author = {Beliavskaia, A and Tan, KK and Sinha, A and Husin, NA and Lim, FS and Loong, SK and Bell-Sakyi, L and Carlow, CKS and AbuBakar, S and Darby, AC and Makepeace, BL and Khoo, JJ}, title = {Metagenomics of culture isolates and insect tissue illuminate the evolution of Wolbachia, Rickettsia and Bartonella symbionts in Ctenocephalides spp. fleas.}, journal = {Microbial genomics}, volume = {9}, number = {7}, pages = {}, pmid = {37399133}, issn = {2057-5858}, support = {BB/P024378/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; BB/P024270/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; 223743/Z/21/Z/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Animals ; *Rickettsia/genetics ; *Bartonella/genetics ; *Siphonaptera/microbiology ; *Ctenocephalides/microbiology ; *Wolbachia/genetics ; Metagenomics ; Insecta ; }, abstract = {While fleas are often perceived simply as a biting nuisance and a cause of allergic dermatitis, they represent important disease vectors worldwide, especially for bacterial zoonoses such as plague (transmitted by rodent fleas) and some of the rickettsioses and bartonelloses. The cosmopolitan cat (Ctenocephalides felis) and dog (Ctenocephalides canis) fleas, as well as Ctenocephalides orientis (restricted to tropical and subtropical Asia), breed in human dwellings and are vectors of cat-scratch fever (caused by Bartonella spp.) and Rickettsia spp., including Rickettsia felis (agent of flea-borne spotted fever) and Rickettsia asembonensis , a suspected pathogen. These Rickettsia spp. are members of a phylogenetic clade known as the ‘transitional group’, which includes both human pathogens and arthropod-specific endosymbionts. The relatively depauperate flea microbiome can also contain other endosymbionts, including a diverse range of Wolbachia strains. Here, we present circularized genome assemblies for two C. orientis-derived pathogens (Bartonella clarridgeiae and R. asembonensis) from Malaysia, a novel Wolbachia strain (wCori), and the C. orientis mitochondrion; all were obtained by direct metagenomic sequencing of flea tissues. Moreover, we isolated two Wolbachia strains from Malaysian C. felis into tick cell culture and recovered circularized genome assemblies for both, one of which (wCfeF) is newly sequenced. We demonstrate that the three Wolbachia strains are representatives of different major clades (‘supergroups’), two of which appear to be flea-specific. These Wolbachia genomes exhibit unique combinations of features associated with reproductive parasitism or mutualism, including prophage WO, cytoplasmic incompatibility factors and the biotin operon of obligate intracellular microbes. The first circularized assembly for R. asembonensis includes a plasmid with a markedly different structure and gene content compared to the published plasmid; moreover, this novel plasmid was also detected in cat flea metagenomes from the USA. Analysis of loci under positive selection in the transitional group revealed genes involved in host–pathogen interactions that may facilitate host switching. Finally, the first B. clarridgeiae genome from Asia exhibited large-scale genome stability compared to isolates from other continents, except for SNPs in regions predicted to mediate interactions with the vertebrate host. These findings highlight the paucity of data on the genomic diversity of Ctenocephalides-associated bacteria and raise questions regarding how interactions between members of the flea microbiome might influence vector competence.}, } @article {pmid37397989, year = {2023}, author = {Pujhari, S and Hughes, GL and Pakpour, N and Suzuki, Y and Rasgon, JL}, title = {Wolbachia-induced inhibition of O'nyong nyong virus in Anopheles mosquitoes is mediated by Toll signaling and modulated by cholesterol.}, journal = {bioRxiv : the preprint server for biology}, volume = {}, number = {}, pages = {}, pmid = {37397989}, issn = {2692-8205}, support = {R01 AI116636/AI/NIAID NIH HHS/United States ; }, abstract = {Enhanced host immunity and competition for metabolic resources are two main competing hypotheses for the mechanism of Wolbachia-mediated pathogen inhibition in arthropods. Using an Anopheles mosquito - somatic Wolbachia infection - O'nyong nyong virus (ONNV) model, we demonstrate that the mechanism underpinning Wolbachia-mediated virus inhibition is up-regulation of the Toll innate immune pathway. However, the viral inhibitory properties of Wolbachia were abolished by cholesterol supplementation. This result was due to Wolbachia-dependent cholesterol-mediated suppression of Toll signaling rather than competition for cholesterol between Wolbachia and virus. The inhibitory effect of cholesterol was specific to Wolbachia-infected Anopheles mosquitoes and cells. These data indicate that both Wolbachia and cholesterol influence Toll immune signaling in Anopheles mosquitoes in a complex manner and provide a functional link between the host immunity and metabolic competition hypotheses for explaining Wolbachia-mediated pathogen interference in mosquitoes. In addition, these results provide a mechanistic understanding of the mode of action of Wolbachia-induced pathogen blocking in Anophelines, which is critical to evaluate the long-term efficacy of control strategies for malaria and Anopheles-transmitted arboviruses.}, } @article {pmid37391513, year = {2023}, author = {Abel, SM and Hong, Z and Williams, D and Ireri, S and Brown, MQ and Su, T and Hung, KY and Henke, JA and Barton, JP and Le Roch, KG}, title = {Small RNA sequencing of field Culex mosquitoes identifies patterns of viral infection and the mosquito immune response.}, journal = {Scientific reports}, volume = {13}, number = {1}, pages = {10598}, pmid = {37391513}, issn = {2045-2322}, support = {U01CK000516/ACL/ACL HHS/United States ; S10 OD026929/OD/NIH HHS/United States ; U01 CK000516/CK/NCEZID CDC HHS/United States ; }, mesh = {Humans ; Animals ; *Culex ; *Culicidae ; Mosquito Vectors ; *Virus Diseases ; Antiviral Agents ; }, abstract = {Mosquito-borne disease remains a significant burden on global health. In the United States, the major threat posed by mosquitoes is transmission of arboviruses, including West Nile virus by mosquitoes of the Culex genus. Virus metagenomic analysis of mosquito small RNA using deep sequencing and advanced bioinformatic tools enables the rapid detection of viruses and other infecting organisms, both pathogenic and non-pathogenic to humans, without any precedent knowledge. In this study, we sequenced small RNA samples from over 60 pools of Culex mosquitoes from two major areas of Southern California from 2017 to 2019 to elucidate the virome and immune responses of Culex. Our results demonstrated that small RNAs not only allowed the detection of viruses but also revealed distinct patterns of viral infection based on location, Culex species, and time. We also identified miRNAs that are most likely involved in Culex immune responses to viruses and Wolbachia bacteria, and show the utility of using small RNA to detect antiviral immune pathways including piRNAs against some pathogens. Collectively, these findings show that deep sequencing of small RNA can be used for virus discovery and surveillance. One could also conceive that such work could be accomplished in various locations across the world and over time to better understand patterns of mosquito infection and immune response to many vector-borne diseases in field samples.}, } @article {pmid37381006, year = {2023}, author = {Zeb, J and Song, B and Khan, MA and Senbill, H and Aziz, MU and Hussain, S and Waris, A and E-Tabor, A and Sparagano, OA}, title = {Genetic diversity of vector-borne pathogens in ixodid ticks infesting dogs from Pakistan with notes on Ehrlichia canis, Rickettsia raoultii and Dirofilaria immitis detection.}, journal = {Parasites & vectors}, volume = {16}, number = {1}, pages = {214}, pmid = {37381006}, issn = {1756-3305}, support = {9380108//Olivier Andre Sparagano/ ; }, mesh = {Humans ; Dogs ; Animals ; Ehrlichia canis/genetics ; *Dirofilaria immitis ; Pakistan/epidemiology ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; *Ixodidae ; *Rickettsia/genetics ; Ehrlichia/genetics ; *Canidae ; Dirofilaria ; Genetic Variation ; }, abstract = {BACKGROUND: Vector-/tick-borne pathogens (V/TBPs) pose a potential threat to human and animal health globally. Information regarding canine V/TBPs is scarce and no specific study has been conducted so far to explore the microbial diversity within ticks infesting dogs from Pakistan. Herein, this knowledge gap is addressed by assessing the genetic diversity and prevalence pattern of V/TBPs in ixodid ticks with special implications for public and canine health.

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

CONCLUSION: Overall, we found that targeting high burden cities with Wolbachia deployments would be a cost-effective intervention in Vietnam and generate notable broader benefits besides health gains.}, } @article {pmid37250803, year = {2023}, author = {Arai, H and Anbutsu, H and Nishikawa, Y and Kogawa, M and Ishii, K and Hosokawa, M and Lin, SR and Ueda, M and Nakai, M and Kunimi, Y and Harumoto, T and Kageyama, D and Takeyama, H and Inoue, MN}, title = {Combined actions of bacteriophage-encoded genes in Wolbachia-induced male lethality.}, journal = {iScience}, volume = {26}, number = {6}, pages = {106842}, pmid = {37250803}, issn = {2589-0042}, abstract = {Some Wolbachia endosymbionts induce male killing, whereby male offspring of infected females are killed during development; however, the origin and diversity of the underlying mechanisms remain unclear. In this study, we identified a 76 kbp prophage region specific to male-killing Wolbachia hosted by the moth Homona magnanima. The prophage encoded a homolog of the male-killing gene oscar in Ostrinia moths and the wmk gene that induces various toxicities in Drosophila melanogaster. Upon overexpressing these genes in D. melanogaster, wmk-1 and wmk-3 killed all males and most females, whereas Hm-oscar, wmk-2, and wmk-4 had no impact on insect survival. Strikingly, co-expression of tandemly arrayed wmk-3 and wmk-4 killed 90% of males and restored 70% of females, suggesting their conjugated functions for male-specific lethality. While the male-killing gene in the native host remains unknown, our findings highlight the role of bacteriophages in male-killing evolution and differences in male-killing mechanisms among insects.}, } @article {pmid37247378, year = {2023}, author = {Oladipupo, SO and Laidoudi, Y and Beckmann, JF and Hu, XP and Appel, AG}, title = {The prevalence of Wolbachia in multiple cockroach species and its implication for urban insect management.}, journal = {Journal of economic entomology}, volume = {116}, number = {4}, pages = {1307-1316}, doi = {10.1093/jee/toad098}, pmid = {37247378}, issn = {1938-291X}, mesh = {Animals ; *Wolbachia/genetics ; Biotin/genetics ; Phylogeny ; Prevalence ; Insecta ; *Bedbugs ; *Blattellidae ; Symbiosis ; }, abstract = {Cockroach management relies heavily on the use of conventional insecticides in urban settings, which no longer provide the anticipated level of control. Knowledge of cockroach endosymbionts, like Wolbachia, might provide novel avenues for control. Therefore, we screened 16 cockroach species belonging to 3 families (Ectobiidae, Blattidae, and Blaberidae) for the presence of Wolbachia. We mapped the evolution of Wolbachia-cockroach relationships based on maximum likelihood phylogeny and phylogenetic species clustering on a multi-loci sequence dataset (i.e., coxA, virD4, hcpA, and gatB) of Wolbachia genes. We confirmed the previous report of Wolbachia in 1 Ectobiid species; Supella longipalpa (Fab.), and detected the presence of Wolbachia in 2 Ectobiid species; Balta notulata (Stål) and Pseudomops septentrionalis Hebard, and 1 Blaberid species; Gromphadorhina portentosa (Schaum). All cockroach-associated Wolbachia herein detected were clustered with the ancestor of F clade Wolbachia of Cimex lectularius L. (bed bugs). Since Wolbachia provision C. lectularius with biotin vitamins that confer reproductive fitness, we screened the cockroach-associated Wolbachia for the presence of biotin genes. In toto, our results reveal 2 important findings: (i) Wolbachia is relatively uncommon among cockroach species infecting about 25% of species investigated, and (ii) cockroach-associated Wolbachia have biotin genes that likely provide nutritional benefits to their hosts. Thus, we discuss the potential of exploring Wolbachia as a tool for urban insect management.}, } @article {pmid37231184, year = {2023}, author = {Zheng, R and Wang, Q and Wu, R and Paradkar, PN and Hoffmann, AA and Wang, GH}, title = {Holobiont perspectives on tripartite interactions among microbiota, mosquitoes, and pathogens.}, journal = {The ISME journal}, volume = {17}, number = {8}, pages = {1143-1152}, pmid = {37231184}, issn = {1751-7370}, mesh = {Animals ; Humans ; *Culicidae ; Mosquito Vectors ; *Microbiota/physiology ; *Malaria ; *Wolbachia/physiology ; }, abstract = {Mosquito-borne diseases like dengue and malaria cause a significant global health burden. Unfortunately, current insecticides and environmental control strategies aimed at the vectors of these diseases are only moderately effective in decreasing disease burden. Understanding and manipulating the interaction between the mosquito holobiont (i.e., mosquitoes and their resident microbiota) and the pathogens transmitted by these mosquitoes to humans and animals could help in developing new disease control strategies. Different microorganisms found in the mosquito's microbiota affect traits related to mosquito survival, development, and reproduction. Here, we review the physiological effects of essential microbes on their mosquito hosts; the interactions between the mosquito holobiont and mosquito-borne pathogen (MBP) infections, including microbiota-induced host immune activation and Wolbachia-mediated pathogen blocking (PB); and the effects of environmental factors and host regulation on the composition of the microbiota. Finally, we briefly overview future directions in holobiont studies, and how these may lead to new effective control strategies against mosquitoes and their transmitted diseases.}, } @article {pmid37231093, year = {2023}, author = {Bruner-Montero, G and Jiggins, FM}, title = {Wolbachia protects Drosophila melanogaster against two naturally occurring and virulent viral pathogens.}, journal = {Scientific reports}, volume = {13}, number = {1}, pages = {8518}, pmid = {37231093}, issn = {2045-2322}, mesh = {Animals ; Female ; Drosophila melanogaster ; *Wolbachia ; *Viruses ; *RNA Viruses ; Antiviral Agents/pharmacology ; Symbiosis ; }, abstract = {Wolbachia is a common endosymbiont that can protect insects against viral pathogens. However, whether the antiviral effects of Wolbachia have a significant effect on fitness remains unclear. We have investigated the interaction between Drosophila melanogaster, Wolbachia and two viruses that we recently isolated from wild flies, La Jolla virus (LJV; Iflaviridae) and Newfield virus (NFV; Permutotetraviridae). Flies infected with these viruses have increased mortality rates, and NFV partially sterilizes females. These effects on fitness were reduced in Wolbachia-infected flies, and this was associated with reduced viral titres. However, Wolbachia alone also reduces survival, and under our experimental conditions these costs of the symbiont can outweigh the benefits of antiviral protection. In contrast, protection against the sterilizing effect of NFV leads to a net benefit of Wolbachia infection after exposure to the virus. These results support the hypothesis that Wolbachia is an important defense against the natural pathogens of D. melanogaster. Furthermore, by reducing the cost of Wolbachia infection, the antiviral effects of Wolbachia may aid its invasion into populations and help explain why it is so common in nature.}, } @article {pmid37214959, year = {2023}, author = {Moldovan, OT and Carrell, AA and Bulzu, PA and Levei, E and Bucur, R and Sitar, C and Faur, L and Mirea, IC and Enilă, M and Cadar, O and Podar, M}, title = {The gut microbiome mediates adaptation to scarce food in Coleoptera.}, journal = {bioRxiv : the preprint server for biology}, volume = {}, number = {}, pages = {}, doi = {10.1101/2023.05.12.540564}, pmid = {37214959}, issn = {2692-8205}, abstract = {Beetles are ubiquitous cave invertebrates worldwide that adapted to scarce subterranean resources when they colonized caves. Here, we investigated the potential role of gut microbiota in the adaptation of beetles to caves from different climatic regions of the Carpathians. The beetles' microbiota was host-specific, reflecting phylogenetic and nutritional adaptation. The microbial community structure further resolved conspecific beetles by caves suggesting microbiota-host coevolution and influences by local environmental factors. The detritivore species hosted a variety of bacteria known to decompose and ferment organic matter, suggesting turnover and host cooperative digestion of the sedimentary microbiota and allochthonous-derived nutrients. The cave Carabidae, with strong mandibulae adapted to predation and scavenging of animal and plant remains, had distinct microbiota dominated by symbiotic lineages Spiroplasma or Wolbachia . All beetles had relatively high levels of fermentative Carnobacterium and Vagococcus involved in lipid accumulation and a reduction of metabolic activity, and both features characterize adaptation to caves.}, } @article {pmid37214831, year = {2023}, author = {Mfopit, YM and Weber, JS and Chechet, GD and Ibrahim, MAM and Signaboubo, D and Achukwi, DM and Mamman, M and Balogun, EO and Shuaibu, MN and Kabir, J and Kelm, S}, title = {Molecular detection of Sodalis glossinidius, Spiroplasma and Wolbachia endosymbionts in wild population of tsetse flies collected in Cameroon, Chad and Nigeria.}, journal = {Research square}, volume = {}, number = {}, pages = {}, pmid = {37214831}, issn = {2693-5015}, support = {K43 TW012015/TW/FIC NIH HHS/United States ; }, abstract = {Background Tsetse flies are cyclical vectors of African trypanosomiasis. They have established symbiotic associations with different bacteria, which influence certain aspects of their physiology. The vector competence of tsetse flies for different trypanosome species is highly variable and is suggested to be affected by various factors, amongst which are bacterial endosymbionts. Symbiotic interactions may provide an avenue for the disease control. The current study provided the prevalence of 3 tsetse symbionts in Glossina species from Cameroon, Chad and Nigeria. Results Tsetse flies were collected from five different locations and dissected. DNA was extracted and polymerase chain reaction PCR was used to detect the presence of Sodalis glossinidius , Spiroplasma sp and Wolbachia using specific primers. A total of 848 tsetse samples were analysed: Glossina morsitans submorsitans (47.52%), Glossina palpalis palpalis (37.26%), Glossina fuscipes fuscipes (9.08%) and Glossina tachinoides (6.13%). Only 95 (11.20%) were infected with at least one of the 3 symbionts. Among the infected, 6 (6.31%) were carrying mixed infection (Wolbachia and Spiroplasma). The overall symbiont prevalence was 0.88%, 3.66% and 11.00% respectively, for Sodalis , Spiroplasma and Wolbachia . Prevalence varied between countries and tsetse species. No Spiroplasma was detected in samples from Cameroon and no Sodalis was found in samples from Nigeria. Conclusion The present study revealed for the first time, the presence of infection by Spiroplasma in tsetse in Chad and Nigeria. These findings provide useful information to the repertoire of bacterial flora of tsetse flies and incite to more investigations to understand their implication in the vector competence of tsetse flies.}, } @article {pmid37213490, year = {2023}, author = {Jackson, R and Patapiou, PA and Golding, G and Helanterä, H and Economou, CK and Chapuisat, M and Henry, LM}, title = {Evidence of phylosymbiosis in Formica ants.}, journal = {Frontiers in microbiology}, volume = {14}, number = {}, pages = {1044286}, pmid = {37213490}, issn = {1664-302X}, abstract = {INTRODUCTION: Insects share intimate relationships with microbes that play important roles in their biology. Yet our understanding of how host-bound microbial communities assemble and perpetuate over evolutionary time is limited. Ants host a wide range of microbes with diverse functions and are an emerging model for studying the evolution of insect microbiomes. Here, we ask whether phylogenetically related ant species have formed distinct and stable microbiomes.

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

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

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

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

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

DISCUSSION: The results presented indicate that Mansonella sp "DEUX" is a new Mansonella species. These findings reflect the neglect of this research topic. And the availability of whole genome data will allow further investigations of these species.}, } @article {pmid37121168, year = {2023}, author = {Biney, C and Graham, GE and Asiedu, E and Sakyi, SA and Kwarteng, A}, title = {Wolbachia Ferrochelatase as a potential drug target against filarial infections.}, journal = {Journal of molecular graphics & modelling}, volume = {122}, number = {}, pages = {108490}, doi = {10.1016/j.jmgm.2023.108490}, pmid = {37121168}, issn = {1873-4243}, mesh = {Animals ; *Wolbachia/metabolism ; Ferrochelatase/metabolism/therapeutic use ; *Filariasis/drug therapy/parasitology ; *Brugia malayi ; Heme/metabolism ; }, abstract = {Filarial infections are among the world's most disturbing diseases caused by 3 major parasitic worms; Onchocerca volvulus, Wuchereria bancrofti, and Brugia malayi, affecting more than 500 million people worldwide. Currently used drugs for mass drug administration (MDA) have been met with several challenges including the development of complications in individuals with filaria co-infections and parasitic drug resistance. The filarial endosymbiont, Wolbachia, has emerged as an attractive therapeutic target for filariasis elimination, due to the dependence of the filaria on this endosymbiont for survival. Here, we target an important enzyme in the Wolbachia heme biosynthetic pathway (ferrochelatase), using high-throughput virtual screening and molecular dynamics with MM-PBSA calculations. We identified four drug candidates; Nilotinib, Ledipasvir, 3-benzhydryloxy-8-methyl-8-azabicyclo[3.2.1]octane, and 2-(4-Amino-piperidin-1-yl)-ethanol as potential small molecules inhibitors as they could compete with the enzyme's natural substrate (Protoporphyrin IX) for active pocket binding. This prevents the worm from receiving the heme molecule from Wolbachia for their growth and survival, resulting in their death. This study which involved targeting enzymes in biosynthetic pathways of the parasitic worms' endosymbiont (Wolbachia), has proven to be an alternative therapeutic option leading to the discovery of new drugs, which will help facilitate the elimination of parasitic infections.}, } @article {pmid37117399, year = {2023}, author = {Ghousein, A and Tutagata, J and Schrieke, H and Etienne, M and Chaumeau, V and Boyer, S and Pages, N and Roiz, D and Eren, AM and Cambray, G and Reveillaud, J}, title = {pWCP is a widely distributed and highly conserved Wolbachia plasmid in Culex pipiens and Culex quinquefasciatus mosquitoes worldwide.}, journal = {ISME communications}, volume = {3}, number = {1}, pages = {40}, pmid = {37117399}, issn = {2730-6151}, support = {948135//EC | EC Seventh Framework Programm | FP7 Ideas: European Research Council (FP7-IDEAS-ERC - Specific Programme: "Ideas" Implementing the Seventh Framework Programme of the European Community for Research, Technological Development and Demonstration Activities (2007 to 2013))/ ; 220211//Wellcome Trust (Wellcome)/ ; }, abstract = {Mosquitoes represent the most important pathogen vectors and are responsible for the spread of a wide variety of poorly treatable diseases. Wolbachia are obligate intracellular bacteria that are widely distributed among arthropods and collectively represents one of the most promising solutions for vector control. In particular, Wolbachia has been shown to limit the transmission of pathogens, and to dramatically affect the reproductive behavior of their host through its phage WO. While much research has focused on deciphering and exploring the biocontrol applications of these WO-related phenotypes, the extent and potential impact of the Wolbachia mobilome remain poorly appreciated. Notably, several Wolbachia plasmids, carrying WO-like genes and Insertion Sequences (IS), thus possibly interrelated to other genetic units of the endosymbiont, have been recently discovered. Here we investigated the diversity and biogeography of the first described plasmid of Wolbachia in Culex pipiens (pWCP) in several islands and continental countries around the world-including Cambodia, Guadeloupe, Martinique, Thailand, and Mexico-together with mosquito strains from colonies that evolved for 2 to 30 years in the laboratory. We used PCR and qPCR to determine the presence and copy number of pWCP in individual mosquitoes, and highly accurate Sanger sequencing to evaluate potential variations. Together with earlier observation, our results show that pWCP is omnipresent and strikingly conserved among Wolbachia populations within mosquitoes from distant geographies and environmental conditions. These data suggest a critical role for the plasmid in Wolbachia ecology and evolution, and the potential of a great tool for further genetic dissection and possible manipulation of this endosymbiont.}, } @article {pmid37117271, year = {2023}, author = {Řezáč, M and Řezáčová, V and Gloríková, N and Némethová, E and Heneberg, P}, title = {Food provisioning to Pardosa spiders decreases the levels of tissue-resident endosymbiotic bacteria.}, journal = {Scientific reports}, volume = {13}, number = {1}, pages = {6943}, pmid = {37117271}, issn = {2045-2322}, mesh = {Animals ; *Spiders/microbiology ; Symbiosis ; *Coxiellaceae ; Host Specificity ; Drosophila ; *Rickettsia ; }, abstract = {The diversity, host specificity, and physiological effects of endosymbiotic bacteria in spiders (Araneae) are poorly characterized. We used 16S rDNA sequencing to evaluate endosymbionts in the cephalothorax and legs of a wolf spider Pardosa agrestis. We tested the effects of feeding once or twice daily with fruit flies, aphids, or starved and compared them to those of syntopically occurring Pardosa palustris. The feeding increased traveled distance up to five times in some of the groups provisioned with food relative to the starved control. The Shannon diversity t-test revealed significant differences between these component communities of the two spider species. The increased frequency of feeding with fruit flies, but not aphids, increased the dominance and decreased the alpha diversity of OTUs. The obligate or facultative endosymbionts were present in all analyzed spider individuals and were represented mostly by Rickettsiella, Rhabdochlamydia, Spiroplasma, and the facultative intracellular parasite Legionella. Vertically transmitted endosymbionts were less common, represented by Wolbachia pipientis and Rickettsia sp. H820. The relative abundance of Mycoplasma spp. was negatively correlated with provisioned or killed aphids. In conclusion, the tissues of Pardosa spiders host tremendously diverse assemblages of bacteria, including obligate or facultative endosymbionts, with yet unknown phenotypic effects.}, } @article {pmid37112932, year = {2023}, author = {Petersen, MT and Couto-Lima, D and Garcia, GA and Pavan, MG and David, MR and Maciel-de-Freitas, R}, title = {Dengue Exposure and Wolbachia wMel Strain Affects the Fertility of Quiescent Eggs of Aedes aegypti.}, journal = {Viruses}, volume = {15}, number = {4}, pages = {}, pmid = {37112932}, issn = {1999-4915}, mesh = {Humans ; Animals ; Female ; *Dengue Virus ; *Aedes ; *Wolbachia ; Fertility ; *Dengue ; }, abstract = {(1) Background: The deployment of the bacterium Wolbachia to reduce arbovirus transmission is ongoing in several countries worldwide. When Wolbachia-carrying Aedes aegypti are released and established in the field, females may feed on dengue-infected hosts. The effects of simultaneous exposure on life-history traits of Ae. aegypti to Wolbachia wMel strain and dengue-1 virus DENV-1 remain unclear. (2) Methods: We monitored 4 groups (mosquitoes with either DENV-1 or Wolbachia, coinfected with DENV-1 and Wolbachia, as well as negative controls) to estimate Ae. aegypti survival, oviposition success, fecundity, collapsing and fertility of quiescent eggs for 12 weeks. (3) Results: Neither DENV-1 nor Wolbachia had a significant impact on mosquito survival nor on mosquito fecundity, although the last parameter showed a tendency to decrease with ageing. There was a significant decrease in oviposition success in individuals carrying Wolbachia. Wolbachia infection and storage time significantly increased egg collapse parameter on the egg viability assay, while DENV-1 had a slight protective effect on the first four weeks of storage. (4) Conclusions: Despite limitations, our results contribute to better understanding of the tripartite interaction of virus, bacteria and mosquito that may take place in field conditions and aid in guaranteeing the Wolbachia strategy success.}, } @article {pmid37105498, year = {2023}, author = {Gong, JT and Li, TP and Wang, MK and Hong, XY}, title = {Wolbachia-based strategies for control of agricultural pests.}, journal = {Current opinion in insect science}, volume = {57}, number = {}, pages = {101039}, doi = {10.1016/j.cois.2023.101039}, pmid = {37105498}, issn = {2214-5753}, mesh = {Humans ; Animals ; *Aedes ; *Wolbachia ; Population Dynamics ; }, abstract = {Wolbachia-based incompatible insect technique (IIT) and pathogen blocking technique (PBT) have been shown to be effective at protecting humans from mosquito-borne diseases in the past decades. Population suppression based on IIT and population replacement based on PBT have become major field application strategies that have continuously been improved by the translational research on Wolbachia-transinfected mosquitoes. Similarly, Wolbachia-based approaches have been proposed for the protection of plants from agricultural pests and their associated diseases. However, a bottleneck in Wolbachia-based strategies for the control of agricultural pests is the need for methods to establish Wolbachia-transinfected insect lines. As a first step in this direction, we compare field control strategies for mosquitos with the potential strategies for agricultural pests based on Wolbachia. Our results show that there is a critical need for establishing productive insect lines and accumulating field test data.}, } @article {pmid37103495, year = {2023}, author = {Towett-Kirui, S and Morrow, JL and Close, S and Royer, JE and Riegler, M}, title = {Bacterial Communities Are Less Diverse in a Strepsipteran Endoparasitoid than in Its Fruit Fly Hosts and Dominated by Wolbachia.}, journal = {Microbial ecology}, volume = {86}, number = {3}, pages = {2120-2132}, pmid = {37103495}, issn = {1432-184X}, support = {IC150100026//Australian Research Council (ARC) Industrial Transformation Training Centre (ITTC) Fruit Fly Biosecurity Innovation/ ; }, mesh = {*Wolbachia/genetics ; RNA, Ribosomal, 16S/genetics ; Bacteria/genetics ; *Microbiota ; High-Throughput Nucleotide Sequencing ; }, abstract = {Microbiomes play vital roles in insect fitness and health and can be influenced by interactions between insects and their parasites. Many studies investigate the microbiome of free-living insects, whereas microbiomes of endoparasitoids and their interactions with parasitised insects are less explored. Due to their development in the constrained environment within a host, endoparasitoids are expected to have less diverse yet distinct microbiomes. We used high-throughput 16S rRNA gene amplicon sequencing to characterise the bacterial communities of Dipterophagus daci (Strepsiptera) and seven of its tephritid fruit fly host species. Bacterial communities of D. daci were less diverse and contained fewer taxa relative to the bacterial communities of the tephritid hosts. The strepsipteran's microbiome was dominated by Pseudomonadota (formerly Proteobacteria) (> 96%), mainly attributed to the presence of Wolbachia, with few other bacterial community members, indicative of an overall less diverse microbiome in D. daci. In contrast, a dominance of Wolbachia was not found in flies parasitised by early stages of D. daci nor unparasitised flies. Yet, early stages of D. daci parasitisation resulted in structural changes in the bacterial communities of parasitised flies. Furthermore, parasitisation with early stages of D. daci with Wolbachia was associated with a change in the relative abundance of some bacterial taxa relative to parasitisation with early stages of D. daci lacking Wolbachia. Our study is a first comprehensive characterisation of bacterial communities in a Strepsiptera species together with the more diverse bacterial communities of its hosts and reveals effects of concealed stages of parasitisation on host bacterial communities.}, } @article {pmid37103216, year = {2023}, author = {Lv, N and Peng, J and He, ZQ and Wen, Q and Su, ZQ and Ali, S and Liu, CZ and Qiu, BL}, title = {The Dynamic Distribution of Wolbachia and Rickettsia in AsiaII1 Bemisia tabaci.}, journal = {Insects}, volume = {14}, number = {4}, pages = {}, pmid = {37103216}, issn = {2075-4450}, support = {32060250//the National Science Foundation of China/ ; }, abstract = {Wolbachia and Rickettsia are bacterial endosymbionts that can induce a number of reproductive abnormalities in their arthropod hosts. We screened and established the co-infection of Wolbachia and Rickettsia in Bemisia tabaci and compared the spatial and temporal distribution of Wolbachia and Rickettsia in eggs (3-120 h after spawning), nymphs, and adults of B. tabaci by qPCR quantification and fluorescent in situ hybridization (FISH). The results show that the titer of Wolbachia and Rickettsia in the 3-120 h old eggs showed a "w" patterned fluctuation, while the titers of Wolbachia and Rickettsia had a "descending-ascending descending-ascending" change process. The titers of Rickettsia and Wolbachia nymphal and the adult life stages of Asia II1 B. tabaci generally increased with the development of whiteflies. However, the location of Wolbachia and Rickettsia in the egg changed from egg stalk to egg base, and then from egg base to egg posterior, and finally back to the middle of the egg. These results will provide basic information on the quantity and localization of Wolbachia and Rickettsia within different life stages of B. tabaci. These findings help to understand the dynamics of the vertical transmission of symbiotic bacteria.}, } @article {pmid37103189, year = {2023}, author = {Cheong, YL and Nazni, WA and Lee, HL and NoorAfizah, A and MohdKhairuddin, IC and Kamarul, GMR and Nizam, NMN and Arif, MAK and NurZatilAqmar, ZM and Irwan, SM and Khadijah, K and Paid, YM and Topek, O and Hasnor, AH and AbuBakar, R and Singh Gill, B and Fadzilah, K and Tahir, A and Sinkins, SP and Hoffmann, AA}, title = {Spatial Distribution and Long-Term Persistence of Wolbachia-Infected Aedes aegypti in the Mentari Court, Malaysia.}, journal = {Insects}, volume = {14}, number = {4}, pages = {}, pmid = {37103189}, issn = {2075-4450}, support = {MC_UU_00034/4/MRC_/Medical Research Council/United Kingdom ; 108508/A/15/Z/WT_/Wellcome Trust/United Kingdom ; Wellcome Trust Award 108508 and Wellcome Trust Award 108508/WT_/Wellcome Trust/United Kingdom ; 202888/Z/16/Z/WT_/Wellcome Trust/United Kingdom ; MC_UU_12014/8/MRC_/Medical Research Council/United Kingdom ; /WT_/Wellcome Trust/United Kingdom ; }, abstract = {Dengue is endemic in Malaysia, and vector control strategies are vital to reduce dengue transmission. The Wolbachia strain wAlbB carried by both sexes of Ae. aegypti was released in Mentari Court, a high-rise residential site, in October 2017 and stopped after 20 weeks. Wolbachia frequencies are still being monitored at multiple traps across this site, providing an opportunity to examine the spatiotemporal distribution of Wolbachia and mosquito density with respect to year, residential block, and floor, using spatial interpolation in ArcGIS, GLMs, and contingency analyses. In just 12 weeks, Wolbachia-infected mosquitoes were established right across the Mentari Court site with an overall infection frequency of >90%. To date, the Wolbachia frequency of Ae. aegypti has remained high in all areas across the site despite releases finishing four years ago. Nevertheless, the Wolbachia invaded more rapidly in some residential blocks than others, and also showed a relatively higher frequency on the eighth floor. The Ae. aegypti index tended to differ somewhat between residential blocks, whilst the Ae. albopictus index was relatively higher at the top and bottom floors of buildings. In Mentari Court, only a short release period was required to infiltrate Wolbachia completely and stably into the natural population. The results inform future releases in comparable sites in a dengue control programme.}, } @article {pmid37103172, year = {2023}, author = {Karpova, EK and Bobrovskikh, MA and Deryuzhenko, MA and Shishkina, OD and Gruntenko, NE}, title = {Wolbachia Effect on Drosophila melanogaster Lipid and Carbohydrate Metabolism.}, journal = {Insects}, volume = {14}, number = {4}, pages = {}, pmid = {37103172}, issn = {2075-4450}, support = {19-04-00458//Russian Foundation for Basic Research/ ; }, abstract = {The effect of maternally inherited endosymbiotic bacteria Wolbachia on triglyceride and carbohydrate metabolism, starvation resistance and feeding behavior of Drosophila melanogaster females was studied. Eight D. melanogaster lines of the same nuclear background were investigated; one had no infection and served as the control, and seven others were infected with different Wolbachia strains pertaining to wMel and wMelCS groups of genotypes. Most of the infected lines had a higher overall lipid content and triglyceride level than the control line and their expression of the bmm gene regulating triglyceride catabolism was reduced. The glucose content was higher in the infected lines compared to that in the control, while their trehalose levels were similar. It was also found that the Wolbachia infection reduced the level of tps1 gene expression (coding for enzyme for trehalose synthesis from glucose) and had no effect on treh gene expression (coding for trehalose degradation enzyme). The infected lines exhibited lower appetite but higher survival under starvation compared to the control. The data obtained may indicate that Wolbachia foster their hosts' energy exchange through increasing its lipid storage and glucose content to ensure the host's competitive advantage over uninfected individuals. The scheme of carbohydrate and lipid metabolism regulation under Wolbachia's influence was suggested.}, } @article {pmid37103136, year = {2023}, author = {Heffernan, E and Markee, A and Truglio, MR and Barkdull, M and Steele Cabrera, S and Daniels, J}, title = {Population Genetic Structure of a Rare Butterfly in a Fragmented South Florida Ecosystem.}, journal = {Insects}, volume = {14}, number = {4}, pages = {}, pmid = {37103136}, issn = {2075-4450}, support = {NA//Disney Conservation Fund/ ; }, abstract = {We investigated the genetic structure and diversity between populations of a rare butterfly, the Florida duskywing (Ephyriades brunnea floridensis E. Bell and W. Comstock, 1948) (Lepidoptera: Hesperiidae) across a network of South Florida pine rockland habitat fragments. Based on 81 individuals from seven populations and using multiple polymorphic microsatellite loci, our analyses support the presence of mainland Florida (peninsular) and Florida Keys (island) population groupings, with a moderate, asymmetrical gene flow connecting them, and the presence of private alleles providing unique identities to each. We additionally found that despite a prevalence in many Lepidoptera, the presence of Wolbachia was not identified in any of the samples screened. Our findings can be used to inform conservation and recovery decisions, including population monitoring, organism translocation, and priority areas for management, restoration or stepping-stone creation to help maintain the complex genetic structure of separate populations.}, } @article {pmid37098937, year = {2023}, author = {Arai, H and Takamatsu, T and Lin, SR and Mizutani, T and Omatsu, T and Katayama, Y and Nakai, M and Kunimi, Y and Inoue, MN}, title = {Diverse Molecular Mechanisms Underlying Microbe-Inducing Male Killing in the Moth Homona magnanima.}, journal = {Applied and environmental microbiology}, volume = {89}, number = {5}, pages = {e0209522}, pmid = {37098937}, issn = {1098-5336}, mesh = {Animals ; Female ; Male ; *Moths ; Symbiosis ; Larva/microbiology ; Reproduction ; Apoptosis ; *Wolbachia/genetics ; *Spiroplasma/genetics ; }, abstract = {Male killing (MK) is a type of reproductive manipulation induced by microbes, where sons of infected mothers are killed during development. MK is a strategy that enhances the fitness of the microbes, and the underlying mechanisms and the process of their evolution have attracted substantial attention. Homona magnanima, a moth, harbors two embryonic MK bacteria, namely, Wolbachia (Alphaproteobacteria) and Spiroplasma (Mollicutes), and a larval MK virus, Osugoroshi virus (OGV; Partitiviridae). However, whether the three distantly related male killers employ similar or different mechanisms to accomplish MK remains unknown. Here, we clarified the differential effects of the three male killers on the sex-determination cascades and development of H. magnanima males. Reverse transcription-PCR demonstrated that Wolbachia and Spiroplasma, but not OGVs, disrupted the sex-determination cascade of males by inducing female-type splice variants of doublesex (dsx), a downstream regulator of the sex-determining gene cascade. We also found that MK microbes altered host transcriptomes in different manners; Wolbachia impaired the host dosage compensation system, whereas Spiroplasma and OGVs did not. Moreover, Wolbachia and Spiroplasma, but not OGVs, triggered abnormal apoptosis in male embryos. These findings suggest that distantly related microbes employ distinct machineries to kill males of the identical host species, which would be the outcome of the convergent evolution. IMPORTANCE Many microbes induce male killing (MK) in various insect species. However, it is not well understood whether microbes adopt similar or different MK mechanisms. This gap in our knowledge is partly because different insect models have been examined for each MK microbe. Here, we compared three taxonomically distinct male killers (i.e., Wolbachia, Spiroplasma, and a partiti-like virus) that infect the same host. We provided evidence that microbes can cause MK through distinct mechanisms that differ in the expression of genes involved in sex determination, dosage compensation, and apoptosis. These results imply independent evolutionary scenarios for the acquisition of their MK ability.}, } @article {pmid37098535, year = {2023}, author = {da Moura, AJF and Valadas, V and Da Veiga Leal, S and Montalvo Sabino, E and Sousa, CA and Pinto, J}, title = {Screening of natural Wolbachia infection in mosquitoes (Diptera: Culicidae) from the Cape Verde islands.}, journal = {Parasites & vectors}, volume = {16}, number = {1}, pages = {142}, pmid = {37098535}, issn = {1756-3305}, support = {PhD fellowship//Camões I.P/ ; PTDC/BIA-OUT/29477/2017//Fundação para a Ciência e a Tecnologia/ ; }, mesh = {Animals ; *Culicidae/genetics ; *Wolbachia/genetics ; Multilocus Sequence Typing ; Cabo Verde ; Mosquito Vectors/microbiology ; *Culex/genetics ; *Aedes/genetics ; }, abstract = {BACKGROUND: Wolbachia pipientis is an endosymbiont bacterium that induces cytoplasmic incompatibility and inhibits arboviral replication in mosquitoes. This study aimed to assess Wolbachia prevalence and genetic diversity in different mosquito species from Cape Verde.

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

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

CONCLUSIONS: A high prevalence and diversity of Wolbachia was found in species from the Cx. pipiens complex. This diversity may be related to the mosquito's colonization history on the Cape Verde islands. To the best of our knowledge, this is the first study to detect Wolbachia in Cx. tigripes, which may provide an additional opportunity for biocontrol initiatives.}, } @article {pmid37094148, year = {2023}, author = {Gu, X and Ross, PA and Gill, A and Yang, Q and Ansermin, E and Sharma, S and Soleimannejad, S and Sharma, K and Callahan, A and Brown, C and Umina, PA and Kristensen, TN and Hoffmann, AA}, title = {A rapidly spreading deleterious aphid endosymbiont that uses horizontal as well as vertical transmission.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {120}, number = {18}, pages = {e2217278120}, pmid = {37094148}, issn = {1091-6490}, mesh = {Animals ; *Aphids/microbiology ; *Coxiellaceae/genetics ; Bacteria ; Phenotype ; Reproduction ; Symbiosis ; }, abstract = {Endosymbiotic bacteria that live inside the cells of insects are typically only transmitted maternally and can spread by increasing host fitness and/or modifying reproduction in sexual hosts. Transinfections of Wolbachia endosymbionts are now being used to introduce useful phenotypes into sexual host populations, but there has been limited progress on applications using other endosymbionts and in asexual populations. Here, we develop a unique pathway to application in aphids by transferring the endosymbiont Rickettsiella viridis to the major crop pest Myzus persicae. Rickettsiella infection greatly reduced aphid fecundity, decreased heat tolerance, and modified aphid body color, from light to dark green. Despite inducing host fitness costs, Rickettsiella spread rapidly through caged aphid populations via plant-mediated horizontal transmission. The phenotypic effects of Rickettsiella were sensitive to temperature, with spread only occurring at 19 °C and not 25 °C. Body color modification was also lost at high temperatures despite Rickettsiella maintaining a high density. Rickettsiella shows the potential to spread through natural M. persicae populations by horizontal transmission and subsequent vertical transmission. Establishment of Rickettsiella in natural populations could reduce crop damage by modifying population age structure, reducing population growth and providing context-dependent effects on host fitness. Our results highlight the importance of plant-mediated horizontal transmission and interactions with temperature as drivers of endosymbiont spread in asexual insect populations.}, } @article {pmid37079598, year = {2023}, author = {Choubdar, N and Karimian, F and Koosha, M and Nejati, J and Shabani Kordshouli, R and Azarm, A and Oshaghi, MA}, title = {Wolbachia infection in native populations of Blattella germanica and Periplaneta americana.}, journal = {PloS one}, volume = {18}, number = {4}, pages = {e0284704}, pmid = {37079598}, issn = {1932-6203}, mesh = {Animals ; *Periplaneta/microbiology ; *Blattellidae/genetics/microbiology ; *Wolbachia/genetics ; Phylogeny ; *Cockroaches/microbiology ; Allergens ; }, abstract = {Cockroaches are significant pests worldwide, being important in medical, veterinary, and public health fields. Control of cockroaches is difficult because they have robust reproductive ability and high adaptability and are resistant to many insecticides. Wolbachia is an endosymbiont bacterium that infects the reproductive organs of approximately 70% of insect species and has become a promising biological agent for controlling insect pests. However, limited data on the presence or strain typing of Wolbachia in cockroaches are available. PCR amplification and sequencing of the wsp and gltA genes were used to study the presence, prevalence and molecular typing of Wolbachia in two main cockroach species, Blattella germanica (German cockroach) and Periplaneta americana (American cockroach), from different geographical locations of Iran. The Wolbachia endosymbiont was found only in 20.6% of German cockroaches while it was absent in American cockroach samples. Blast search and phylogenetic analysis revealed that the Wolbachia strain found in the German cockroach belongs to Wolbachia supergroup F. Further studies should investigate the symbiotic role of Wolbachia in cockroaches and determine whether lack of Wolbachia infection may increase this insect's ability to tolerate or acquire various pathogens. Results of our study provide a foundation for continued work on interactions between cockroaches, bacterial endosymbionts, and pathogens.}, } @article {pmid37075471, year = {2023}, author = {Che Lah, EF and Ahamad, M and Dmitry, A and Md-Zain, BM and Yaakop, S}, title = {Metagenomic profile of the bacterial communities associated with Ixodes granulatus (Acari: Ixodidae): a potential vector of tick-borne diseases.}, journal = {Journal of medical entomology}, volume = {60}, number = {4}, pages = {753-768}, doi = {10.1093/jme/tjad044}, pmid = {37075471}, issn = {1938-2928}, mesh = {Humans ; Female ; Animals ; *Ixodes/microbiology ; *Ixodidae/microbiology ; Rickettsiales ; Ehrlichia ; *Rickettsia/genetics ; *Borrelia/genetics ; *Tick-Borne Diseases/microbiology ; }, abstract = {Ixodes granulatus Supino, 1897 (Acari: Ixodida) is one of Malaysia's most common hard ticks and is a potential vector for tick-borne diseases (TBDs). Despite its great public health importance, research on I. granulatus microbial communities remains largely unexplored. Therefore, this study aimed to investigate the bacterial communities of on-host I. granulatus collected from three different recreational areas on the East Coast of Peninsular Malaysia using high throughput Next Generation Sequencing (NGS). A total of 9 females on-host I. granulatus were subjected to metabarcoding analysis targeting V3-V4 regions of 16S ribosomal RNA (rRNA) using the Illumina MiSeq platform. This study identified 15 bacterial phyla corresponding to 19 classes, 54 orders, and 90 families from 435 amplicon sequence variants (ASVs), revealing a diverse bacterial community profile. Together with 130 genera assigned, local I. granulatus harbored 4 genera of pathogens, i.e., Rickettsia da Rocha Lima, 1916 (Rickettsiales: Rickettsiaceae) (58.6%), Borrelia Swellengrebel 1907 (Spirochaetales: Borreliaceae) (31.6%), Borreliella Adeolu and Gupta 2015 (Spirochaetales: Borreliaceae) (0.6%), and Ehrlichia Cowdria Moshkovski 1947 (Rickettsiales: Ehrlichiaceae) (39.9%). Some endosymbiont bacteria, such as Coxiella (Philip, 1943) (Legionellales: Coxiellaceae), Wolbachia Hertig 1936 (Rickettsiales: Ehrlichiaceae), and Rickettsiella Philip, 1956 (Legionellales: Coxiellaceae), were also detected at very low abundance. Interestingly, this study reported the co-infection of Borrelia and Ehrlichia for the first time, instilling potential health concerns in the context of co-transmission to humans, especially in areas with a high population of I. granulatus. This study successfully characterized the tick microbiome and provided the first baseline data of I. granulatus bacterial communities in Malaysia. These results support the need for way-forward research on tick-associated bacteria using NGS, focusing on medically important species toward TBD prevention.}, } @article {pmid37073332, year = {2023}, author = {Ye, Z and Bishop, T and Wang, Y and Shahriari, R and Lynch, M}, title = {Evolution of sex determination in crustaceans.}, journal = {Marine life science & technology}, volume = {5}, number = {1}, pages = {1-11}, pmid = {37073332}, issn = {2662-1746}, support = {R35 GM122566/GM/NIGMS NIH HHS/United States ; }, abstract = {Sex determination (SD) involves mechanisms that determine whether an individual will develop into a male, female, or in rare cases, hermaphrodite. Crustaceans harbor extremely diverse SD systems, including hermaphroditism, environmental sex determination (ESD), genetic sex determination (GSD), and cytoplasmic sex determination (e.g., Wolbachia controlled SD systems). Such diversity lays the groundwork for researching the evolution of SD in crustaceans, i.e., transitions among different SD systems. However, most previous research has focused on understanding the mechanism of SD within a single lineage or species, overlooking the transition across different SD systems. To help bridge this gap, we summarize the understanding of SD in various clades of crustaceans, and discuss how different SD systems might evolve from one another. Furthermore, we review the genetic basis for transitions between different SD systems (i.e., Dmrt genes) and propose the microcrustacean Daphnia (clade Branchiopoda) as a model to study the transition from ESD to GSD.}, } @article {pmid37060070, year = {2023}, author = {Li, Y and Sun, Y and Zou, J and Zhong, D and Liu, R and Zhu, C and Li, W and Zhou, Y and Cui, L and Zhou, G and Lu, G and Li, T}, title = {Characterizing the Wolbachia infection in field-collected Culicidae mosquitoes from Hainan Province, China.}, journal = {Parasites & vectors}, volume = {16}, number = {1}, pages = {128}, pmid = {37060070}, issn = {1756-3305}, support = {YSPTZX202004//Research project of Hainan academician innovation platform/ ; 82060379//National Natural Science Foundation of China/ ; 2022NHCTDCKFKT31002//Open Foundation of NHC Key Laboratory of Tropical Disease Control, Hainan Medical University/ ; ZDKJ202003//Major Science and Technology Program of Hainan Province/ ; U19 AI089672/AI/NIAID NIH HHS/United States ; XRC220012//Talent Introduction Fund of Hainan Medical University/ ; 820RC653//Hainan Provincial Natural Science Foundation/ ; }, mesh = {Animals ; Humans ; *Culicidae ; *Wolbachia/genetics ; RNA, Ribosomal, 16S/genetics ; Phylogeny ; Mosquito Vectors/genetics ; *Aedes/genetics ; *Culex/genetics ; China/epidemiology ; }, abstract = {BACKGROUND: Mosquitoes are vectors of many pathogens, such as malaria, dengue virus, yellow fever virus, filaria and Japanese encephalitis virus. Wolbachia are capable of inducing a wide range of reproductive abnormalities in their hosts, such as cytoplasmic incompatibility. Wolbachia has been proposed as a tool to modify mosquitoes that are resistant to pathogen infection as an alternative vector control strategy. This study aimed to determine natural Wolbachia infections in different mosquito species across Hainan Province, China.

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

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

CONCLUSION: Our study revealed the prevalence and distribution of Wolbachia in mosquitoes from Hainan Province, China. Knowledge of the prevalence and diversity of Wolbachia strains in local mosquito populations will provide part of the baseline information required for current and future Wolbachia-based vector control approaches to be conducted in Hainan Province.}, } @article {pmid37055642, year = {2023}, author = {Lozano-Sardaneta, YN and Marina, CF and Torres-Monzón, JA and Sánchez-Cordero, V and Becker, I}, title = {Molecular detection of Wolbachia and Bartonella as part of the microbiome of phlebotomine sand flies from Chiapas, Mexico.}, journal = {Parasitology research}, volume = {122}, number = {6}, pages = {1293-1301}, pmid = {37055642}, issn = {1432-1955}, support = {6682//Consejo Nacional de Ciencia y Tecnología/ ; CRISP-CONACYT-FOSSIS-69530//Consejo Nacional de Ciencia y Tecnología/ ; IG201221//Programa de Apoyo a Proyectos de Investigación e Innovación Tecnológica (PAPIIT)/ ; }, mesh = {Animals ; *Psychodidae/parasitology ; *Wolbachia/genetics ; *Bartonella/genetics ; Mexico ; Phylogeny ; Retrospective Studies ; *Phlebotomus ; *Leishmania ; *Microbiota ; Bacteria ; }, abstract = {Phlebotomine sand flies are dipterans of relevance due to their role as vectors of several pathogens worldwide. Bacteria in the gut of sand flies possibly affect their vectorial capacity and competence to transmit parasites. A retrospective study was performed in sand fly specimens that had previously been collected in four localities of the state of Chiapas during the period 2009-2011 to detect Wolbachia and Bartonella and their possible coinfection with Leishmania. For the molecular detection of bacteria, we used primers and conditions that had previously been reported. A total of 531 sand fly specimens of 10 species were analyzed. Four Wolbachia strains were detected in five sand fly species, showing a prevalence of 8.6%. All the Wolbachia strains had previously been reported in other taxa. In one sand fly species, we also detected a new lineage of Bartonella evidenced by a phylogenetic analysis. No sand fly specimens showed coinfections of these bacteria and Leishmania. The bacteria found in the phlebotomine sand flies are possibly transmitted by plant-mediated horizontal transmission and during blood meal feeding.}, } @article {pmid37052365, year = {2023}, author = {Xu, J and Tan, JB and Li, YD and Xu, YH and Tang, A and Zhou, HK and Shi, PQ}, title = {Diversity and dynamics of endosymbionts in a single population of sweet potato weevil, Cylas formicarius (Coleoptera: Brentidae): a preliminary study.}, journal = {Journal of insect science (Online)}, volume = {23}, number = {2}, pages = {}, pmid = {37052365}, issn = {1536-2442}, mesh = {Animals ; *Weevils ; *Coleoptera ; *Ipomoea batatas ; Reproduction ; }, abstract = {Endosymbionts live symbiotically with insect hosts and play important roles in the evolution, growth, development, reproduction, and environmental fitness of hosts. Weevils are one of the most abundant insect groups that can be infected by various endosymbionts, such as Sodalis, Nardonella, and Wolbachia. The sweet potato weevil, Cylas formicarius (Coleoptera: Brentidae), is a notorious pest in sweet potato (Ipomoea batatas L.) cultivation. Currently, little is known about the presence of endosymbionts in C. formicarius. Herein, we assessed the endosymbiont load of a single geographic population of C. formicarius. The results showed that Nardonella and Rickettsia could infect C. formicarius at different rates, which also varied according to the developmental stages of C. formicarius. The relative titer of Nardonella was significantly related to C. formicarius developmental stages. The Nardonella-infecting sweet potato weevils were most closely related to the Nardonella in Sphenophorus levis (Coleoptera, Curculionidae). The Rickettsia be identified in bellii group. These results preliminarily revealed the endosymbionts in C. formicarius and helped to explore the diversity of endosymbionts in weevils and uncover the physiological roles of endosymbionts in weevils.}, } @article {pmid37043605, year = {2023}, author = {Dobson, KL and Blore, K and Henke, JA and Hung, KY and Morgan, T and Posey, T and Sun, S and Sypes, O and Tremblay, NP and Dobson, SL}, title = {Satellite Rearing of Aedes Mosquito Eggs: Synchronized Empirical Test of a Novel Mass Rearing Model.}, journal = {Journal of the American Mosquito Control Association}, volume = {39}, number = {1}, pages = {12-17}, doi = {10.2987/22-7104}, pmid = {37043605}, issn = {1943-6270}, mesh = {Animals ; Male ; *Aedes ; Humidity ; Mosquito Control/methods ; }, abstract = {Mosquito suppression strategies based on "rear and release" of male mosquitoes are attracting renewed interest from governments, municipalities, and private businesses. These include irradiation-based sterile insect technique, Wolbachia-based technologies, and genetic modification. Each of these approaches requires the mass rearing and release of adult male mosquitoes, which typically is accomplished via a rearing facility near the release site. Although some release programs have relied on centralized rearing and shipment of adult males, adult male mosquitoes are relatively fragile, and their fitness can be diminished by temperature fluctuations, humidity, nutritional deficiencies, and other stresses that occur during shipment. Furthermore, expensive, expedited shipment is typically used to maximize the amount of adult lifetime in the field following the release. In contrast, Aedes aegypti and Ae. albopictus eggs can be desiccated and stored for long periods. They are small, and many millions of eggs can be shipped without specialized environmental conditions and using less expensive means. Here we examine a model in which mosquito eggs are centrally produced and then mailed to satellite rearing facilities. As a control, a replicate set of eggs was reared at the factory of origin. At each of the rearing sites, cloud-based software was used to track and compare rearing at the different locations. The results demonstrate similar rearing outcomes (i.e., egg hatch, immature development, and number of adult males) at each of the different sites for both species. We discuss the outcome in relation to downstream applications and potential future studies.}, } @article {pmid37043515, year = {2023}, author = {Novelo, M and Dutra, HL and Metz, HC and Jones, MJ and Sigle, LT and Frentiu, FD and Allen, SL and Chenoweth, SF and McGraw, EA}, title = {Dengue and chikungunya virus loads in the mosquito Aedes aegypti are determined by distinct genetic architectures.}, journal = {PLoS pathogens}, volume = {19}, number = {4}, pages = {e1011307}, pmid = {37043515}, issn = {1553-7374}, support = {R01 AI143758/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; *Chikungunya virus/physiology ; *Aedes ; Mosquito Vectors ; *Chikungunya Fever ; *Dengue ; }, abstract = {Aedes aegypti is the primary vector of the arboviruses dengue (DENV) and chikungunya (CHIKV). These viruses exhibit key differences in their vector interactions, the latter moving more quicky through the mosquito and triggering fewer standard antiviral pathways. As the global footprint of CHIKV continues to expand, we seek to better understand the mosquito's natural response to CHIKV-both to compare it to DENV:vector coevolutionary history and to identify potential targets in the mosquito for genetic modification. We used a modified full-sibling design to estimate the contribution of mosquito genetic variation to viral loads of both DENV and CHIKV. Heritabilities were significant, but higher for DENV (40%) than CHIKV (18%). Interestingly, there was no genetic correlation between DENV and CHIKV loads between siblings. These data suggest Ae. aegypti mosquitoes respond to the two viruses using distinct genetic mechanisms. We also examined genome-wide patterns of gene expression between High and Low CHIKV families representing the phenotypic extremes of viral load. Using RNAseq, we identified only two loci that consistently differentiated High and Low families: a long non-coding RNA that has been identified in mosquito screens post-infection and a distant member of a family of Salivary Gland Specific (SGS) genes. Interestingly, the latter gene is also associated with horizontal gene transfer between mosquitoes and the endosymbiotic bacterium Wolbachia. This work is the first to link the SGS gene to a mosquito phenotype. Understanding the molecular details of how this gene contributes to viral control in mosquitoes may, therefore, also shed light on its role in Wolbachia.}, } @article {pmid37035661, year = {2023}, author = {Michalik, A and Franco, DC and Deng, J and Szklarzewicz, T and Stroiński, A and Kobiałka, M and Łukasik, P}, title = {Variable organization of symbiont-containing tissue across planthoppers hosting different heritable endosymbionts.}, journal = {Frontiers in physiology}, volume = {14}, number = {}, pages = {1135346}, pmid = {37035661}, issn = {1664-042X}, abstract = {Sap-feeding hemipteran insects live in associations with diverse heritable symbiotic microorganisms (bacteria and fungi) that provide essential nutrients deficient in their hosts' diets. These symbionts typically reside in highly specialized organs called bacteriomes (with bacterial symbionts) or mycetomes (with fungal symbionts). The organization of these organs varies between insect clades that are ancestrally associated with different microbes. As these symbioses evolve and additional microorganisms complement or replace the ancient associates, the organization of the symbiont-containing tissue becomes even more variable. Planthoppers (Hemiptera: Fulgoromorpha) are ancestrally associated with bacterial symbionts Sulcia and Vidania, but in many of the planthopper lineages, these symbionts are now accompanied or have been replaced by other heritable bacteria (e.g., Sodalis, Arsenophonus, Purcelliella) or fungi. We know the identity of many of these microbes, but the symbiont distribution within the host tissues and the bacteriome organization have not been systematically studied using modern microscopy techniques. Here, we combine light, fluorescence, and transmission electron microscopy with phylogenomic data to compare symbiont tissue distributions and the bacteriome organization across planthoppers representing 15 families. We identify and describe seven primary types of symbiont localization and seven types of the organization of the bacteriome. We show that Sulcia and Vidania, when present, usually occupy distinct bacteriomes distributed within the body cavity. The more recently acquired gammaproteobacterial and fungal symbionts generally occupy separate groups of cells organized into distinct bacteriomes or mycetomes, distinct from those with Sulcia and Vidania. They can also be localized in the cytoplasm of fat body cells. Alphaproteobacterial symbionts colonize a wider range of host body habitats: Asaia-like symbionts often colonize the host gut lumen, whereas Wolbachia and Rickettsia are usually scattered across insect tissues and cell types, including cells containing other symbionts, bacteriome sheath, fat body cells, gut epithelium, as well as hemolymph. However, there are exceptions, including Gammaproteobacteria that share bacteriome with Vidania, or Alphaproteobacteria that colonize Sulcia cells. We discuss how planthopper symbiont localization correlates with their acquisition and replacement patterns and the symbionts' likely functions. We also discuss the evolutionary consequences, constraints, and significance of these findings.}, } @article {pmid37032902, year = {2023}, author = {Wangwiwatsin, A and Kulwong, S and Phetcharaburanin, J and Namwat, N and Klanrit, P and Loilome, W and Maleewong, W and Reid, AJ}, title = {Toward novel treatment against filariasis: Insight into genome-wide co-evolutionary analysis of filarial nematodes and Wolbachia.}, journal = {Frontiers in microbiology}, volume = {14}, number = {}, pages = {1052352}, pmid = {37032902}, issn = {1664-302X}, support = {/WT_/Wellcome Trust/United Kingdom ; }, abstract = {Infectious diseases caused by filarial nematodes are major health problems for humans and animals globally. Current treatment using anti-helminthic drugs requires a long treatment period and is only effective against the microfilarial stage. Most species of filarial nematodes harbor a specific strain of Wolbachia bacteria, which are essential for the survival, development, and reproduction of the nematodes. This parasite-bacteria obligate symbiosis offers a new angle for the cure of filariasis. In this study, we utilized publicly available genome data and putative protein sequences from seven filarial nematode species and their symbiotic Wolbachia to screen for protein-protein interactions that could be a novel target against multiple filarial nematode species. Genome-wide in silico screening was performed to predict molecular interactions based on co-evolutionary signals. We identified over 8,000 pairs of gene families that show evidence of co-evolution based on high correlation score and low false discovery rate (FDR) between gene families and obtained a candidate list that may be keys in filarial nematode-Wolbachia interactions. Functional analysis was conducted on these top-scoring pairs, revealing biological processes related to various signaling processes, adult lifespan, developmental control, lipid and nucleotide metabolism, and RNA modification. Furthermore, network analysis of the top-scoring genes with multiple co-evolving pairs suggests candidate genes in both Wolbachia and the nematode that may play crucial roles at the center of multi-gene networks. A number of the top-scoring genes matched well to known drug targets, suggesting a promising drug-repurposing strategy that could be applicable against multiple filarial nematode species.}, } @article {pmid37027941, year = {2023}, author = {Liu, W and Nasir, M and Yan, M and Zhu, X and Iqbal, MS and Wang, L and Zhang, K and Li, D and Ji, J and Gao, X and Luo, J and Cui, J}, title = {Response of the Pardosa astrigera bacterial community to Cry1B protein.}, journal = {Ecotoxicology and environmental safety}, volume = {256}, number = {}, pages = {114855}, doi = {10.1016/j.ecoenv.2023.114855}, pmid = {37027941}, issn = {1090-2414}, mesh = {Female ; Male ; Humans ; Animals ; RNA, Ribosomal, 16S/metabolism ; *Bacteria ; *Spiders/metabolism ; Symbiosis ; Proteobacteria ; }, abstract = {While genetically modified (GM) crops bring economic benefits to human beings, their impact on non-target organisms has become an important part of environmental safety assessments. Symbiotic bacteria play an important role in eukaryotic biological functions and can adjust host communities to adapt to new environments. Therefore, this study examined the effects of Cry1B protein on the growth and development of non-target natural enemies of Pardosa astrigera (L. Koch) from the perspective of symbiotic bacteria. Cry1B protein had no significant effect on the health indicators of P. astrigera (adults and 2nd instar spiderlings). 16S rRNA sequencing results revealed that Cry1B protein did not change the symbiotic bacteria species composition of P. astrigera, but did reduce the number of OTU and species diversity. In 2nd instar spiderlings, neither the dominant phylum (Proteobacteria) nor the dominant genus (Acinetobacter) changed, but the relative abundance of Corynebacterium-1 decreased significantly; in adult spiders, the dominant bacteria genera of females and males were different. The dominant bacterial genera were Brevibacterium in females and Corynebacterium-1 in males, but Corynebacterium-1 was the dominant bacteria in both females and males feeding on Cry1B. The relative abundance of Wolbachia also increased significantly. In addition, bacteria in other genera varied significantly by sex. KEGG results showed that Cry1B protein only altered the significant enrichment of metabolic pathways in female spiders. In conclusion, the effects of Cry1B protein on symbiotic bacteria vary by growth and development stage and sex.}, } @article {pmid37022600, year = {2023}, author = {Caputo, B and Moretti, R and Virgillito, C and Manica, M and Lampazzi, E and Lombardi, G and Serini, P and Pichler, V and Beebe, NW and Della Torre, A and Calvitti, M}, title = {A bacterium against the tiger: further evidence of the potential of noninundative releases of males with manipulated Wolbachia infection in reducing fertility of Aedes albopictus field populations in Italy.}, journal = {Pest management science}, volume = {79}, number = {9}, pages = {3167-3176}, doi = {10.1002/ps.7495}, pmid = {37022600}, issn = {1526-4998}, support = {MUR PNRR Extended Partnership initiative on Emerging Infectious Diseases (project no. PE00000007, INF-ACT, research node 2).//Ministero della RIcerca/ ; }, mesh = {Animals ; Male ; Female ; *Aedes ; *Wolbachia ; Mosquito Control/methods ; Fertility ; Italy ; }, abstract = {BACKGROUND: Incompatible insect technique (IIT) is a population suppression approach based on the release of males with manipulated Wolbachia infection inducing egg inviability in wild females. We here present results of multiple field releases of incompatible ARwP males carried out in 2019 in a 2.7-ha green area within urban Rome (Italy) to assess the effect on Aedes albopictus egg viability. Data are compared with results obtained in 2018, when the approach was tested for the first time in Europe.

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

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

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

RESULTS AND DISCUSSION: We observed that high competition stress extended development time, decreased the likelihood of eclosion, decreased body size, and increased susceptibility to West Nile virus (WNV) infection. We also observed that Wolbachia infection reduced WNV load under low competition stress, and significantly improved the rate of survival for larval reared under higher competition stress. Consequently, our data suggest that native Wolbachia infection in Cx. quinquefasciatus has differential consequences for host fitness and susceptibility to WNV infection depending on competition stress.}, } @article {pmid37001324, year = {2023}, author = {Romano, DMM and Pereira, TN and Almeida, IB and Coelho, CSG and Duarte, FC and Harakava, R and Cassiano, LL and Mendes, MC}, title = {First molecular evidence of Wolbachia occurrence in Amblyomma sculptum (Acari: Ixodidae).}, journal = {Veterinary parasitology}, volume = {317}, number = {}, pages = {109907}, doi = {10.1016/j.vetpar.2023.109907}, pmid = {37001324}, issn = {1873-2550}, mesh = {Animals ; *Ixodidae/microbiology ; *Rocky Mountain Spotted Fever/epidemiology/microbiology/veterinary ; Amblyomma/genetics ; *Wolbachia/genetics ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Brazil/epidemiology ; *Ticks/microbiology ; *Rickettsia ; }, abstract = {As the main vector for the bacterium Rickettsia rickettsii in Brazil, the tick Amblyomma sculptum is a parasite of great public health importance in this country. Wolbachia is an endosymbiont bacterium highly widespread among invertebrates and because of its impact on its hosts' biology, form a powerful alternative for pests and disease control. The aim of this study was to investigate the occurrence of this bacterium in A. sculptum. For this, 187 adult ticks collected in two municipalities in the interior of the state of São Paulo, Brazil, were analyzed using molecular techniques and bioinformatics tools. A total of 15 ticks were positive for the presence of Wolbachia. Phylogenetic analysis on the 16S rRNA gene indicated that the Wolbachia DNA sequences obtained in this investigation belonged to different clades, probably in supergroups B and F. This was the first study to report the occurrence of Wolbachia in A. sculptum and it enriches knowledge about the susceptibility of ticks to this bacterium. Now that we know that Wolbachia can be found in A. sculptum, the objective for a next study must be to investigate Wolbachia's possible origin in this tick.}, } @article {pmid37000335, year = {2023}, author = {Rohlfing, K and Yue, L and Franke, S and Zeng, C and Podsiadlowski, L and Dobler, S}, title = {When does the female bias arise? Insights from the sex determination cascade of a flea beetle with a strongly skewed sex ratio.}, journal = {Functional & integrative genomics}, volume = {23}, number = {2}, pages = {112}, pmid = {37000335}, issn = {1438-7948}, mesh = {Animals ; Male ; Female ; *Coleoptera/genetics ; Sex Ratio ; *Siphonaptera/genetics ; Larva ; DNA, Mitochondrial ; RNA, Messenger ; }, abstract = {Reproduction-manipulating bacteria like Wolbachia can shift sex ratios in insects towards females, but skewed sex ratios may also arise from genetic conflicts. The flea beetle Altica lythri harbors three main mtDNA strains that are coupled to three different Wolbachia infections. Depending on the mtDNA types, the females produce either offspring with a balanced sex ratio or exclusively daughters. To obtain markers that can monitor when sex bias arises in the beetle's ontogeny, we elucidated the sex determination cascade of A. lythri. We established a RT-PCR method based on length variants of dsx (doublesex) transcripts to determine the sex of morphologically indistinguishable eggs and larvae. In females of one mtDNA type (HT1/HT1*) known to produce only daughters, male offspring were already missing at the egg stage while for females of another type (HT2), the dsx splice variants revealed a balanced sex ratio among eggs and larvae. Our data suggest that the sex determination cascade in A. lythri is initiated by maternally transmitted female-specific tra (transformer) mRNA as primary signal. This tra mRNA seems to be involved in a positive feedback loop that maintains the production of the female splice variant, as known for female offspring in Tribolium castaneum. The translation of the maternally transmitted female tra mRNA must be inhibited in male offspring, but the underlying primary genetic signal remains to be identified. We discuss which differences between the mtDNA types can influence sex determination and lead to the skewed sex ratio of HT1.}, } @article {pmid36996252, year = {2023}, author = {Hornett, EA and Hurst, GDD}, title = {One strain may hide another: Cryptic male-killing Wolbachia.}, journal = {PLoS biology}, volume = {21}, number = {3}, pages = {e3002076}, pmid = {36996252}, issn = {1545-7885}, mesh = {Animals ; Male ; *Wolbachia ; Insecta ; Host-Pathogen Interactions ; Host Microbial Interactions ; }, abstract = {While heritable symbionts are common in insects, strains that act as male-killers are considered rare. A new study in PLOS Biology identifies a novel male-killer hidden by coinfection and host resistance, highlighting the complexity of host-microbial interactions in natural systems.}, } @article {pmid36993585, year = {2023}, author = {Beckmann, J and Gillespie, J and Tauritz, D}, title = {Modelling Emergence of Wolbachia Toxin-Antidote Protein Functions with an Evolutionary Algorithm.}, journal = {bioRxiv : the preprint server for biology}, volume = {}, number = {}, pages = {}, pmid = {36993585}, issn = {2692-8205}, support = {R21 AI146773/AI/NIAID NIH HHS/United States ; R21 AI156762/AI/NIAID NIH HHS/United States ; R21 AI166832/AI/NIAID NIH HHS/United States ; }, abstract = {Evolutionary algorithms (EAs) simulate Darwinian evolution and adeptly mimic natural evolution. Most EA applications in biology encode high levels of abstraction in top-down ecological population models. In contrast, our research merges protein alignment algorithms from bioinformatics into codon based EAs that simulate molecular protein string evolution from the bottom up. We apply our EA to reconcile a problem in the field of Wolbachia induced cytoplasmic incompatibility (CI). Wolbachia is a microbial endosymbiont that lives inside insect cells. CI is conditional insect sterility that operates as a toxin antidote (TA) system. Although, CI exhibits complex phenotypes not fully explained under a single discrete model. We instantiate in-silico genes that control CI, CI factors (cifs), as strings within the EA chromosome. We monitor the evolution of their enzymatic activity, binding, and cellular localization by applying selective pressure on their primary amino acid strings. Our model helps rationalize why two distinct mechanisms of CI induction might coexist in nature. We find that nuclear localization signals (NLS) and Type IV secretion system signals (T4SS) are of low complexity and evolve fast, whereas binding interactions have intermediate complexity, and enzymatic activity is the most complex. Our model predicts that as ancestral TA systems evolve into eukaryotic CI systems, the placement of NLS or T4SS signals can stochastically vary, imparting effects that might impact CI induction mechanics. Our model highlights how preconditions, genetic diversity, and sequence length can bias evolution of cifs towards one mechanism or another.}, } @article {pmid36992487, year = {2023}, author = {Ferreira, QR and Lemos, FFB and Moura, MN and Nascimento, JOS and Novaes, AF and Barcelos, IS and Fernandes, LA and Amaral, LSB and Barreto, FK and Melo, FF}, title = {Role of the Microbiome in Aedes spp. Vector Competence: What Do We Know?.}, journal = {Viruses}, volume = {15}, number = {3}, pages = {}, pmid = {36992487}, issn = {1999-4915}, mesh = {Animals ; Female ; Humans ; *Aedes ; *Chikungunya Fever ; *Arboviruses/genetics ; *Yellow Fever ; *Zika Virus Infection ; *Zika Virus ; *Microbiota ; }, abstract = {Aedes aegypti and Aedes albopictus are the vectors of important arboviruses: dengue fever, chikungunya, Zika, and yellow fever. Female mosquitoes acquire arboviruses by feeding on the infected host blood, thus being able to transmit it to their offspring. The intrinsic ability of a vector to infect itself and transmit a pathogen is known as vector competence. Several factors influence the susceptibility of these females to be infected by these arboviruses, such as the activation of the innate immune system through the Toll, immunodeficiency (Imd), JAK-STAT pathways, and the interference of specific antiviral response pathways of RNAi. It is also believed that the presence of non-pathogenic microorganisms in the microbiota of these arthropods could influence this immune response, as it provides a baseline activation of the innate immune system, which may generate resistance against arboviruses. In addition, this microbiome has direct action against arboviruses, mainly due to the ability of Wolbachia spp. to block viral genome replication, added to the competition for resources within the mosquito organism. Despite major advances in the area, studies are still needed to evaluate the microbiota profiles of Aedes spp. and their vector competence, as well as further exploration of the individual roles of microbiome components in activating the innate immune system.}, } @article {pmid36989319, year = {2023}, author = {Wimalasiri-Yapa, BMCR and Huang, B and Ross, PA and Hoffmann, AA and Ritchie, SA and Frentiu, FD and Warrilow, D and van den Hurk, AF}, title = {Differences in gene expression in field populations of Wolbachia-infected Aedes aegypti mosquitoes with varying release histories in northern Australia.}, journal = {PLoS neglected tropical diseases}, volume = {17}, number = {3}, pages = {e0011222}, pmid = {36989319}, issn = {1935-2735}, mesh = {Animals ; *Aedes ; *Dengue Virus/physiology ; *Wolbachia/genetics ; Mosquito Vectors ; *Zika Virus/genetics ; *Zika Virus Infection ; Australia ; Gene Expression ; }, abstract = {Aedes aegypti is the principal mosquito vector of dengue, yellow fever, Zika and chikungunya viruses. The wMel strain of the endosymbiotic bacteria Wolbachia pipientis was introduced into the vector as a novel biocontrol strategy to stop transmission of these viruses. Mosquitoes with Wolbachia have been released in the field in Northern Queensland, Australia since 2011, at various locations and over several years, with populations remaining stably infected. Wolbachia infection is known to alter gene expression in its mosquito host, but whether (and how) this changes over the long-term in the context of field releases remains unknown. We sampled mosquitoes from Wolbachia-infected populations with three different release histories along a time gradient and performed RNA-seq to investigate gene expression changes in the insect host. We observed a significant impact on gene expression in Wolbachia-infected mosquitoes versus uninfected controls. Fewer genes had significantly upregulated expression in mosquitoes from the older releases (512 and 486 from the 2011 and 2013/14 release years, respectively) versus the more recent releases (1154 from the 2017 release year). Nonetheless, a fundamental signature of Wolbachia infection on host gene expression was observed across all releases, comprising upregulation of immunity (e.g. leucine-rich repeats, CLIPs) and metabolism (e.g. lipid metabolism, iron transport) genes. There was limited downregulation of gene expression in mosquitoes from the older releases (84 and 71 genes from the 2011 and 2013/14 release years, respectively), but significantly more in the most recent release (509 from the 2017 release year). Our findings indicate that at > 8 years post-introgression into field populations, Wolbachia continues to profoundly impact expression of host genes, such as those involved in insect immune response and metabolism. If Wolbachia-mediated virus blocking is underpinned by these differential gene expression changes, our results suggest it may remain stable long-term.}, } @article {pmid36986288, year = {2023}, author = {Moore, C and Lashnits, E and Neupane, P and Herrin, BH and Lappin, M and André, MR and Breitschwerdt, EB}, title = {Feeding on a Bartonella henselae Infected Host Triggers Temporary Changes in the Ctenocephalides felis Microbiome.}, journal = {Pathogens (Basel, Switzerland)}, volume = {12}, number = {3}, pages = {}, pmid = {36986288}, issn = {2076-0817}, support = {FAPESP Process 2019/09464-6//Fundação de Amparo à Pesquisa do Estado de São Paulo/ ; }, abstract = {The effect of Bartonella henselae on the microbiome of its vector, Ctenocephalides felis (the cat flea) is largely unknown, as the majority of C. felis microbiome studies have utilized wild-caught pooled fleas. We surveyed the microbiome of laboratory-origin C. felis fed on B. henselae-infected cats for 24 h or 9 days to identify changes to microbiome diversity and microbe prevalence compared to unfed fleas, and fleas fed on uninfected cats. Utilizing Next Generation Sequencing (NGS) on the Illumina platform, we documented an increase in microbial diversity in C. felis fed on Bartonella-infected cats for 24 h. These changes returned to baseline (unfed fleas or fleas fed on uninfected cats) after 9 days on the host. Increased diversity in the C. felis microbiome when fed on B. henselae-infected cats may be related to the mammalian, flea, or endosymbiont response. Poor B. henselae acquisition was documented with only one of four infected flea pools having B. henselae detected by NGS. We hypothesize this is due to the use of adult fleas, flea genetic variation, or lack of co-feeding with B. henselae-infected fleas. Future studies are necessary to fully characterize the effect of endosymbionts and C. felis diversity on B. henselae acquisition.}, } @article {pmid36985370, year = {2023}, author = {Remmal, I and Bel Mokhtar, N and Maurady, A and Reda Britel, M and El Fakhouri, K and Asimakis, E and Tsiamis, G and Stathopoulou, P}, title = {Characterization of the Bacterial Microbiome in Natural Populations of Barley Stem Gall Midge, Mayetiola hordei, in Morocco.}, journal = {Microorganisms}, volume = {11}, number = {3}, pages = {}, pmid = {36985370}, issn = {2076-2607}, support = {MIS 5010952//Hellenic Ministry of Rural Development and Food/ ; 00//European Union Erasmus+ Programme/ ; }, abstract = {Mayetiola hordei (Kieffer), known as barley stem gall midge, is one of the most destructive barley pests in many areas around the world, inflicting significant qualitative and quantitative damage to crop production. In this study, we investigate the presence of reproductive symbionts, the effect of geographical origin on the bacterial microbiome's structure, and the diversity associated with natural populations of M. hordei located in four barley-producing areas in Morocco. Wolbachia infection was discovered in 9% of the natural populations using a precise 16S rDNA PCR assay. High-throughput sequencing of the V3-V4 region of the bacterial 16S rRNA gene indicated that the native environments of samples had a substantial environmental impact on the microbiota taxonomic assortment. Briefly, 5 phyla, 7 classes, and 42 genera were identified across all the samples. To our knowledge, this is the first report on the bacterial composition of M. hordei natural populations. The presence of Wolbachia infection may assist in the diagnosis of ideal natural populations, providing a new insight into the employment of Wolbachia in the control of barley midge populations, in the context of the sterile insect technique or other biological control methods.}, } @article {pmid36985289, year = {2023}, author = {Huynh, LN and Diarra, AZ and Pham, QL and Berenger, JM and Ho, VH and Nguyen, XQ and Parola, P}, title = {Identification of Vietnamese Flea Species and Their Associated Microorganisms Using Morphological, Molecular, and Protein Profiling.}, journal = {Microorganisms}, volume = {11}, number = {3}, pages = {}, pmid = {36985289}, issn = {2076-2607}, abstract = {Fleas are obligatory blood-sucking ectoparasites of medical and veterinary importance. The identification of fleas and associated flea-borne microorganisms, therefore, plays an important role in controlling and managing these vectors. Recently, Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS) has been reported as an innovative and effective approach to the identification of arthropods, including fleas. This study aims to use this technology to identify ethanol-preserved fleas collected in Vietnam and to use molecular biology to search for microorganisms associated with these fleas. A total of 502 fleas were collected from wild and domestic animals in four provinces in Vietnam. Morphological identification led to the recognition of five flea species, namely Xenopsylla cheopis, Xenopsylla astia, Pulex irritans, Ctenocephalides canis, and Ctenocephalides felis. The cephalothoraxes of 300 individual, randomly selected fleas were tested using MALDI-TOF MS and molecular analysis for the identification and detection of microorganisms. A total of 257/300 (85.7%) of the obtained spectra from the cephalothoraxes of each species were of good enough quality to be used for our analyses. Our laboratory MALDI-TOF MS reference database was upgraded with spectra achieved from five randomly selected fleas for every species of Ctenocephalides canis and Ctenocephalides felis. The remaining spectra were then queried against the upgraded MALDI-TOF MS database, which showed 100% correspondence between morphology and MALDI-TOF MS identification for two flea species (Ctenocephalides canis and Ctenocephalides felis). The MS spectra of the remaining species (three P. irritans, five X. astia, and two X. cheopis) were visually generated low-intensity MS profiles with high background noise that could not be used to update our database. Bartonella and Wolbachia spp. were detected in 300 fleas from Vietnam using PCR and sequencing with primers derived from the gltA gene for Bartonella and the 16S rRNA gene for Wolbachia, including 3 Bartonella clarridgeiae (1%), 3 Bartonella rochalimae (1%), 1 Bartonella coopersplainsensis (0.3%), and 174 Wolbachia spp. endosymbionts (58%).}, } @article {pmid36977163, year = {2023}, author = {Florez, D and Young, AJ and Bernabé, KJ and Hyman, JM and Qu, Z}, title = {Modeling Sustained Transmission of Wolbachia among Anopheles Mosquitoes: Implications for Malaria Control in Haiti.}, journal = {Tropical medicine and infectious disease}, volume = {8}, number = {3}, pages = {}, pmid = {36977163}, issn = {2414-6366}, support = {New Faculty Startup Funds//The University of Texas at San Antonio/ ; }, abstract = {Wolbachia infection in Anopheles albimanus mosquitoes can render mosquitoes less capable of spreading malaria. We developed and analyzed a mechanistic compartmental ordinary differential equation model to evaluate the effectiveness of Wolbachia-based vector control strategies among wild Anopheles mosquitoes in Haiti. The model tracks the mosquito life stages, including egg, larva, and adult (male and female). It also accounts for critical biological effects, such as the maternal transmission of Wolbachia through infected females and cytoplasmic incompatibility, which effectively sterilizes uninfected females when they mate with infected males. We derive and interpret dimensionless numbers, including the basic reproductive number and next-generation numbers. The proposed system presents a backward bifurcation, which indicates a threshold infection that needs to be exceeded to establish a stable Wolbachia infection. The sensitivity analysis ranks the relative importance of the epidemiological parameters at baseline. We simulate different intervention scenarios, including prerelease mitigation using larviciding and thermal fogging before the release, multiple releases of infected populations, and different release times of the year. Our simulations show that the most efficient approach to establishing Wolbachia is to release all the infected mosquitoes immediately after the prerelease mitigation process. Moreover, the model predicts that it is more efficient to release during the dry season than the wet season.}, } @article {pmid36975996, year = {2023}, author = {Jiang, RX and Shang, F and Jiang, HB and Dou, W and Cernava, T and Wang, JJ}, title = {Environmental Factors Affect the Bacterial Community in Diaphorina citri, an Important Vector of "Candidatus Liberibacter asiaticus".}, journal = {Microbiology spectrum}, volume = {11}, number = {2}, pages = {e0529822}, pmid = {36975996}, issn = {2165-0497}, abstract = {Insects are associated with diverse microbial communities that can have substantial effects on hosts. Here, we characterized the bacterial communities in the Asian citrus psyllid (ACP), Diaphorina citri (Hemiptera: Psyllidae), a major vector of the devastating pathogen "Candidatus Liberibacter asiaticus," which causes citrus Huanglongbing (HLB). In total, 256 ACP individuals across 15 field sites and one laboratory population in China were sequenced. The results showed that the bacterial community diversity was the highest in the Guilin population (average Shannon index, 1.27), and the highest value for richness was found in the Chenzhou population (average Chao1 index, 298). The bacterial community structures of the field-collected populations were significantly different, and all of them harbored Wolbachia, which was assigned to strain ST-173. Structural equation models revealed that the dominant Wolbachia strain had a significantly negative correlation with the annual mean temperature. In addition, the results obtained with populations infected with "Ca. Liberibacter asiaticus" indicated that in total, 140 bacteria could be involved in interactions with this bacterium. The ACP field populations harbored a more diverse bacterial community than the laboratory population, and the relative occurrences of some symbionts differed significantly. However, the bacterial community of the ACP laboratory colony was connected in a more complex network structure (average degree, 54.83) than that of the field populations (average degree, 10.62). Our results provide evidence that environmental factors can influence the bacterial community structure and bacterial relative abundance in ACP populations. This is likely due to the adaptation of ACPs to local environments. IMPORTANCE The Asian citrus psyllid (ACP) is an important vector of the HLB pathogen, which is a major threat to citrus production around the world. Bacterial communities harbored by insects could be affected by different environmental factors. Understanding these factors that affect the bacterial community of the ACP could be important for the better management of HLB transmission. This work surveyed ACP field populations in mainland China in order to explore the bacterial community diversity of different populations and the potential relationships between environmental factors and predominant symbionts. We have assessed the differences in ACP bacterial communities and identified the prevalent Wolbachia strains in the field. In addition, we compared the bacterial communities of ACP field-collected and laboratory populations. Comparing populations subjected to contrasting conditions could help us to better understand how the ACP adapts to local environmental conditions. Our study provides new insights into how environmental factors influence the bacterial community of the ACP.}, } @article {pmid36958642, year = {2023}, author = {Detcharoen, M and Jiggins, FM and Schlick-Steiner, BC and Steiner, FM}, title = {Wolbachia endosymbiotic bacteria alter the gut microbiome in the fly Drosophila nigrosparsa.}, journal = {Journal of invertebrate pathology}, volume = {198}, number = {}, pages = {107915}, doi = {10.1016/j.jip.2023.107915}, pmid = {36958642}, issn = {1096-0805}, mesh = {Animals ; Drosophila/microbiology ; *Wolbachia ; *Gastrointestinal Microbiome ; Reproduction ; Bacteria ; Symbiosis ; Drosophila melanogaster/microbiology ; }, abstract = {Wolbachia are known to cause reproductive manipulations and in some arthropod species, Wolbachia were reported to cause changes in gut microbiome. However, the effects of Wolbachia bacteria on the microbiomes of their hosts, including Drosophila flies, have not been fully accessed. Here, we checked the bacterial microbiome in guts of Wolbachia-uninfected and of Wolbachia-infected Drosophila nigrosparsa, both separated into a bleach-only (embryos bleached) and a gnotobiotic (embryos bleached and inoculated with bacteria) treatment. We observed a clear separation between the Wolbachia-infected and the Wolbachia-uninfected samples, and the infected samples had higher variation in alpha diversity than the uninfected ones. There were reductions in the abundances of Proteobacteria (Pseudomonadota), especially Acetobacter, in the infected samples of both treatments. These findings highlight that Wolbachia change the gut microbiome in D. nigrosparsa as well as that the interactions between Wolbachia and bacteria like Acetobacter need to be investigated.}, } @article {pmid36958587, year = {2023}, author = {Verhulst, EC and Pannebakker, BA and Geuverink, E}, title = {Variation in sex determination mechanisms may constrain parthenogenesis-induction by endosymbionts in haplodiploid systems.}, journal = {Current opinion in insect science}, volume = {56}, number = {}, pages = {101023}, doi = {10.1016/j.cois.2023.101023}, pmid = {36958587}, issn = {2214-5753}, mesh = {Animals ; Female ; Parthenogenesis ; *Wolbachia ; *Arthropods ; Sex Determination Processes ; }, abstract = {Endosymbionts are maternally transmitted, and therefore benefit from maximizing female offspring numbers. Parthenogenesis-induction (PI) is the most effective type of manipulation for transmission, but has solely been detected in haplodiploid species, whereas cytoplasmic incompatibility (CI) is detected frequently across the arthropod phylum, including haplodiploids. This puzzling observation led us to hypothesize that the molecular sex-determination mechanism of the haplodiploid host may be a constraining factor in the ability of endosymbionts to induce parthenogenesis. Recent insights indicate that PI-endosymbionts may be able to directly manipulate sex-determination genes to induce the necessary steps required for PI in haplodiploids. However, sex-determination cascades vary extensively, so PI-induction would require a specialized and host-dependent tool set. Contrastingly, CI-related genes target conserved cell-cycle mechanisms, are located on mobile elements, and spread easily. Finally, endosymbiont-manipulations may have a strong impact on the effectiveness of haplodiploid biocontrol agents, but can also be used to enhance their efficacy.}, } @article {pmid36950155, year = {2023}, author = {Moore, C and Breitschwerdt, EB and Kim, L and Li, Y and Ferris, K and Maggi, R and Lashnits, E}, title = {The association of host and vector characteristics with Ctenocephalides felis pathogen and endosymbiont infection.}, journal = {Frontiers in microbiology}, volume = {14}, number = {}, pages = {1137059}, pmid = {36950155}, issn = {1664-302X}, abstract = {Surveillance of the fleas and flea-borne pathogens infecting cats is important for both human and animal health. Multiple zoonotic Bartonella and Rickettsia species are known to infect the most common flea infesting cats and dogs worldwide: Ctenocephalides felis, the cat flea. The ability of other flea species to transmit pathogens is relatively unexplored. We aimed to determine cat host and flea factors independently associated with flea Bartonella and Rickettsia infection. We also assessed flea and cat infection by flea-host pair and location. To accomplish these aims, we performed qPCR for the detection of Bartonella, hemotropic Mycoplasma, Rickettsia, and Wolbachia DNA using paired cat and flea samples obtained from free-roaming cats presenting for spay or neuter across four locations in the United States. A logistic regression model was employed to identify the effect of cat (sex, body weight, geographic location, and Bartonella, hemotropic Mycoplasma, and Rickettsia spp., infection) and flea (clade and Rickettsia and Wolbachia infection) factors on C. felis Bartonella clarridgeiae infection. From 189 free roaming cats, we collected 84 fleas: Ctenocephalides felis (78/84), Cediopsylla simplex (4/84), Orchopeas howardi (1/84), and Nosopsyllus fasciatus (1/84). Ctenocephalides felis were phylogenetically assigned to Clades 1, 4, and 6 by cox1 gene amplification. Rickettsia asembonensis (52/84) and B. clarridgeiae (16/84) were the most common pathogenic bacteria detected in fleas. Our model identified host cat sex and weight as independently associated with B. clarridgeiae infection in fleas. Rickettsia asembonensis (52/84), Rickettsia felis (7/84) and Bartonella henselae (7/84) were detected in specific clades: R. felis was detected only in Clades 1 and 6 while B. henselae and R. asembonensis were detected only in Clade 4. Wolbachia spp., also displayed clade specificity with strains other than Wolbachia wCfeT only infecting fleas from Clade 6. There was poor flea and host agreement for Bartonella spp., infection; however, there was agreement in the Bartonella species detected in cats and fleas by geographic location. These findings reinforce the importance of considering reservoir host attributes and vector phylogenetic diversity in epidemiological studies of flea-borne pathogens. Widespread sampling is necessary to identify the factors driving flea-borne pathogen presence and transmission.}, } @article {pmid36949814, year = {2023}, author = {Ou, D and Qiu, JH and Su, ZQ and Wang, L and Qiu, BL}, title = {The phylogeny and distribution of Wolbachia in two pathogen vector insects, Asian citrus psyllid and Longan psyllid.}, journal = {Frontiers in cellular and infection microbiology}, volume = {13}, number = {}, pages = {1121186}, pmid = {36949814}, issn = {2235-2988}, mesh = {Animals ; *Hemiptera/microbiology ; *Wolbachia/genetics ; Phylogeny ; *Citrus/microbiology ; }, abstract = {BACKGROUND: Wolbachia is the most abundant bacterial endosymbiont among insects. It can play a prominent role in the development, reproduction and immunity of its given insect host. To date, Wolbachia presence is well studied within aphids, whiteflies and planthoppers, but relatively few studies have investigated its presence in psyllids.

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

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

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

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

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

CONCLUSIONS: We describe the time and temperature limits for maintaining viability of Wolbachia-infected Ae. aegypti eggs when encapsulated or exposed to extreme temperatures. These findings could improve the efficiency of mass releases by providing transport and storage constraints to ensure only high-quality material is utilised during field releases.}, } @article {pmid36933571, year = {2023}, author = {Oladipupo, SO and Carroll, JD and Beckmann, JF}, title = {Convergent Aedes and Drosophila CidB interactomes suggest cytoplasmic incompatibility targets are conserved.}, journal = {Insect biochemistry and molecular biology}, volume = {155}, number = {}, pages = {103931}, doi = {10.1016/j.ibmb.2023.103931}, pmid = {36933571}, issn = {1879-0240}, mesh = {Animals ; Male ; Drosophila ; Drosophila melanogaster ; *Aedes ; Semen ; Cytoplasm/metabolism ; *Wolbachia ; }, abstract = {Wolbachia-mediated cytoplasmic incompatibility (CI) is a conditional embryonic lethality induced when Wolbachia-modified sperm fertilizes an uninfected egg. The Wolbachia proteins, CidA and CidB control CI. CidA is a rescue factor that reverses lethality. CidA binds to CidB. CidB contains a deubiquitinating enzyme and induces CI. Precisely how CidB induces CI and what it targets are unknown. Likewise, how CidA prevents sterilization by CidB is not clear. To identify CidB substrates in mosquitos we conducted pull-down assays using recombinant CidA and CidB mixed with Aedes aegypti lysates to identify the protein interactomes of CidB and the CidB/CidA protein complex. Our data allow us to cross compare CidB interactomes across taxa for Aedes and Drosophila. Our data replicate several convergent interactions, suggesting that CI targets conserved substrates across insects. Our data support a hypothesis that CidA rescues CI by tethering CidB away from its substrates. Specifically, we identify ten convergent candidate substrates including P32 (protamine-histone exchange factor), karyopherin alpha, ubiquitin-conjugating enzyme, and bicoid stabilizing factor. Future analysis on how these candidates contribute to CI will clarify mechanisms.}, } @article {pmid36929176, year = {2023}, author = {Eugénio, AT and Marialva, MSP and Beldade, P}, title = {Effects of Wolbachia on Transposable Element Expression Vary Between Drosophila melanogaster Host Genotypes.}, journal = {Genome biology and evolution}, volume = {15}, number = {3}, pages = {}, pmid = {36929176}, issn = {1759-6653}, mesh = {Animals ; *Drosophila melanogaster/genetics ; DNA Transposable Elements ; *Wolbachia/genetics ; Evolution, Molecular ; Genotype ; }, abstract = {Transposable elements (TEs) are repetitive DNA sequences capable of changing position in host genomes, thereby causing mutations. TE insertions typically have deleterious effects but they can also be beneficial. Increasing evidence of the contribution of TEs to adaptive evolution further raises interest in understanding what factors impact TE activity. Based on previous studies associating the bacterial endosymbiont Wolbachia with changes in the abundance of piRNAs, a mechanism for TE repression, and to transposition of specific TEs, we hypothesized that Wolbachia infection would interfere with TE activity. We tested this hypothesis by studying the expression of 14 TEs in a panel of 25 Drosophila melanogaster host genotypes, naturally infected with Wolbachia and annotated for TE insertions. The host genotypes differed significantly in Wolbachia titers inside individual flies, with broad-sense heritability around 20%, and in the number of TE insertions, which depended greatly on TE identity. By removing Wolbachia from the target host genotypes, we generated a panel of 25 pairs of Wolbachia-positive and Wolbachia-negative lines in which we quantified transcription levels for our target TEs. We found variation in TE expression that was dependent on Wolbachia status, TE identity, and host genotype. Comparing between pairs of Wolbachia-positive and Wolbachia-negative flies, we found that Wolbachia removal affected TE expression in 21.1% of the TE-genotype combinations tested, with up to 2.3 times differences in the median level of transcript. Our data show that Wolbachia can impact TE activity in host genomes, underscoring the importance this endosymbiont can have in the generation of genetic novelty in hosts.}, } @article {pmid36928089, year = {2023}, author = {Terretaz, K and Horard, B and Weill, M and Loppin, B and Landmann, F}, title = {Functional analysis of Wolbachia Cid effectors unravels cooperative interactions to target host chromatin during replication.}, journal = {PLoS pathogens}, volume = {19}, number = {3}, pages = {e1011211}, pmid = {36928089}, issn = {1553-7374}, mesh = {Animals ; Male ; Chromatin/metabolism ; *Wolbachia/physiology ; Semen/metabolism ; Animals, Genetically Modified ; Drosophila/metabolism ; Cytoplasm/metabolism ; Centromere Protein A/metabolism ; *Drosophila Proteins/metabolism ; }, abstract = {Wolbachia are common bacteria among terrestrial arthropods. These endosymbionts transmitted through the female germline manipulate their host reproduction through several mechanisms whose most prevalent form called Cytoplasmic Incompatibility -CI- is a conditional sterility syndrome eventually favoring the infected progeny. Upon fertilization, the sperm derived from an infected male is only compatible with an egg harboring a compatible Wolbachia strain, this sperm leading otherwise to embryonic death. The Wolbachia Cif factors CidA and CidB responsible for CI and its neutralization function as a Toxin-Antitoxin system in the mosquito host Culex pipiens. However, the mechanism of CidB toxicity and its neutralization by the CidA antitoxin remain unexplored. Using transfected insect cell lines to perform a structure-function analysis of these effectors, we show that both CidA and CidB are chromatin interactors and CidA anchors CidB to the chromatin in a cell-cycle dependent-manner. In absence of CidA, the CidB toxin localizes to its own chromatin microenvironment and acts by preventing S-phase completion, independently of its deubiquitylase -DUB- domain. Experiments with transgenic Drosophila show that CidB DUB domain is required together with CidA during spermatogenesis to stabilize the CidA-CidB complex. Our study defines CidB functional regions and paves the way to elucidate the mechanism of its toxicity.}, } @article {pmid36914655, year = {2023}, author = {Kageyama, D and Harumoto, T and Nagamine, K and Fujiwara, A and Sugimoto, TN and Jouraku, A and Tamura, M and Katoh, TK and Watada, M}, title = {A male-killing gene encoded by a symbiotic virus of Drosophila.}, journal = {Nature communications}, volume = {14}, number = {1}, pages = {1357}, pmid = {36914655}, issn = {2041-1723}, support = {P40 OD018537/OD/NIH HHS/United States ; }, mesh = {*Drosophila melanogaster/embryology/virology ; *Insect Viruses/genetics ; *Genes, Viral/physiology ; *Symbiosis ; Male ; Animals ; Embryonic Development ; RNA, Viral/physiology ; RNA, Double-Stranded/physiology ; Evolution, Molecular ; Open Reading Frames/genetics ; *Sex Ratio ; Sex Characteristics ; Female ; }, abstract = {In most eukaryotes, biparentally inherited nuclear genomes and maternally inherited cytoplasmic genomes have different evolutionary interests. Strongly female-biased sex ratios that are repeatedly observed in various arthropods often result from the male-specific lethality (male-killing) induced by maternally inherited symbiotic bacteria such as Spiroplasma and Wolbachia. However, despite some plausible case reports wherein viruses are raised as male-killers, it is not well understood how viruses, having much smaller genomes than bacteria, are capable of inducing male-killing. Here we show that a maternally inherited double-stranded RNA (dsRNA) virus belonging to the family Partitiviridae (designated DbMKPV1) induces male-killing in Drosophila. DbMKPV1 localizes in the cytoplasm and possesses only four genes, i.e., one gene in each of the four genomic segments (dsRNA1-dsRNA4), in contrast to ca. 1000 or more genes possessed by Spiroplasma or Wolbachia. We also show that a protein (designated PVMKp1; 330 amino acids in size), encoded by a gene on the dsRNA4 segment, is necessary and sufficient for inducing male-killing. Our results imply that male-killing genes can be easily acquired by symbiotic viruses through reassortment and that symbiotic viruses are hidden players in arthropod evolution. We anticipate that host-manipulating genes possessed by symbiotic viruses can be utilized for controlling arthropods.}, } @article {pmid36913994, year = {2023}, author = {Junsiri, W and Kamkong, P and Chinkangsadarn, T and Ouisuwan, S and Taweethavonsawat, P}, title = {Molecular identification and genetic diversity of equine ocular setariasis in Thailand based on the COI, 12S rDNA, and ITS1 regions.}, journal = {Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases}, volume = {110}, number = {}, pages = {105425}, doi = {10.1016/j.meegid.2023.105425}, pmid = {36913994}, issn = {1567-7257}, mesh = {Animals ; Horses/genetics ; *Setariasis/diagnosis ; DNA, Ribosomal/genetics ; Thailand/epidemiology ; Phylogeny ; *Genetic Variation ; }, abstract = {Equine ocular setariasis is mainly caused by Setaria digitata, and the identification of this filarial nematode is based on morphology. However, morphological characterization alone is insufficient for the detection and differentiation of S. digitata from its congeners. In Thailand, the molecular detection of S. digitata is lacking and its genetic diversity is still unknown. This study aimed to phylogenetically characterize equine S. digitata from Thailand based on sequences derived from the mitochondrial cytochrome c oxidase subunit 1 (COI), the mitochondrial small subunit ribosomal DNA (12S rDNA), the nuclear internal transcribed spacer 1 (ITS1) and Wolbachia surface protein (wsp). Five samples of S. digitata were characterized, submitted to the NCBI database, and used for phylogenetic analysis as well as the assessment of similarity, entropy, and haplotype diversity. Phylogenetic analyses revealed that the S. digitata Thai strain was similar to S. digitata from China and Sri Lanka, with 99 to 100% similarity. The entropy and haplotype diversity indicated that the S. digitata Thai isolate was conserved and closely related to S. digitata worldwide. This is the first report on the molecular detection of equine ocular setariasis caused by S. digitata in Thailand.}, } @article {pmid36911919, year = {2023}, author = {Radousky, YA and Hague, MTJ and Fowler, S and Paneru, E and Codina, A and Rugamas, C and Hartzog, G and Cooper, BS and Sullivan, W}, title = {Distinct Wolbachia localization patterns in oocytes of diverse host species reveal multiple strategies of maternal transmission.}, journal = {Genetics}, volume = {224}, number = {1}, pages = {}, pmid = {36911919}, issn = {1943-2631}, support = {R35 GM124701/GM/NIGMS NIH HHS/United States ; R35 GM139595/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; *Wolbachia/genetics ; Drosophila melanogaster ; Oocytes ; Oogenesis ; Drosophila/genetics ; }, abstract = {A broad array of endosymbionts radiate through host populations via vertical transmission, yet much remains unknown concerning the cellular basis, diversity, and routes underlying this transmission strategy. Here, we address these issues, by examining the cellular distributions of Wolbachia strains that diverged up to 50 million years ago in the oocytes of 18 divergent Drosophila species. This analysis revealed 3 Wolbachia distribution patterns: (1) a tight clustering at the posterior pole plasm (the site of germline formation); (2) a concentration at the posterior pole plasm, but with a significant bacteria population distributed throughout the oocyte; and (3) a distribution throughout the oocyte, with none or very few located at the posterior pole plasm. Examination of this latter class indicates Wolbachia accesses the posterior pole plasm during the interval between late oogenesis and the blastoderm formation. We also find that 1 Wolbachia strain in this class concentrates in the posterior somatic follicle cells that encompass the pole plasm of the developing oocyte. In contrast, strains in which Wolbachia concentrate at the posterior pole plasm generally exhibit no or few Wolbachia in the follicle cells associated with the pole plasm. Taken together, these studies suggest that for some Drosophila species, Wolbachia invade the germline from neighboring somatic follicle cells. Phylogenomic analysis indicates that closely related Wolbachia strains tend to exhibit similar patterns of posterior localization, suggesting that specific localization strategies are a function of Wolbachia-associated factors. Previous studies revealed that endosymbionts rely on 1 of 2 distinct routes of vertical transmission: continuous maintenance in the germline (germline-to-germline) or a more circuitous route via the soma (germline-to-soma-to-germline). Here, we provide compelling evidence that Wolbachia strains infecting Drosophila species maintain the diverse arrays of cellular mechanisms necessary for both of these distinct transmission routes. This characteristic may account for its ability to infect and spread globally through a vast range of host insect species.}, } @article {pmid36909520, year = {2023}, author = {Martin, M and Newton, ILG}, title = {The Wolbachia WalE1 effector alters Drosophila endocytosis.}, journal = {bioRxiv : the preprint server for biology}, volume = {}, number = {}, pages = {}, pmid = {36909520}, issn = {2692-8205}, support = {R01 AI144430/AI/NIAID NIH HHS/United States ; }, abstract = {The most common intracellular bacterial infection is Wolbachia pipientis, a microbe that manipulates host reproduction and is used in control of insect vectors. Phenotypes induced by Wolbachia have been studied for decades and range from sperm-egg incompatibility to male killing. How Wolbachia alters host biology is less well understood. Previously, we characterized the first Wolbachia effector - WalE1, which encodes a synuclein domain at the N terminus. Purified WalE1 sediments with and bundles actin and when heterologously expressed in flies, increases Wolbachia titer in the developing oocyte. In this work, we first identify the native expression WalE1 by Wolbachia infecting both fly cells and whole animals. WalE1 appears as aggregates, separate from Wolbachia cells. We next show that WalE1 co-immunoprecipitates with the host protein Past1 and that WalE1 manipulates host endocytosis. Yeast expressing WalE1 show deficiency in uptake of FM4-64 dye, and flies harboring mutations in Past1 or overexpressing WalE1 are sensitive to AgNO3, a hallmark of endocytosis defects. Finally, we also show that Past1 null flies harbor more Wolbachia overall and in late egg chambers. Our results identify interactions between a Wolbachia secreted effector and a host protein and point to yet another important host cell process impinged upon by Wolbachia.}, } @article {pmid36907292, year = {2023}, author = {Ruiz, A and Gutiérrez-Bugallo, G and Rodríguez-Roche, R and Pérez, L and González-Broche, R and Piedra, LA and Martínez, LC and Menéndez, Z and Vega-Rúa, A and Bisset, JA}, title = {First report of natural Wolbachia infections in mosquitoes from Cuba.}, journal = {Acta tropica}, volume = {242}, number = {}, pages = {106891}, doi = {10.1016/j.actatropica.2023.106891}, pmid = {36907292}, issn = {1873-6254}, mesh = {Animals ; *Wolbachia/genetics ; Phylogeny ; Cuba ; Mosquito Vectors/microbiology ; *Aedes/microbiology ; }, abstract = {Mosquitoes are extensively responsible for the transmission of pathogens. Novel strategies using Wolbachia could transform that scenario, since these bacteria manipulate mosquito reproduction, and can confer a pathogen transmission-blocking phenotype in culicids. Here, we screened the Wolbachia surface protein region by PCR in eight Cuban mosquito species. We confirmed the natural infections by sequencing and assessed the phylogenetic relationships among the Wolbachia strains detected. We identified four Wolbachia hosts: Aedes albopictus, Culex quinquefasciatus, Mansonia titillans, and Aedes mediovittatus (first report worldwide). Knowledge of Wolbachia strains and their natural hosts is essential for future operationalization of this vector control strategy in Cuba.}, } @article {pmid36897930, year = {2023}, author = {Davison, HR and Hurst, GDD and Siozios, S}, title = {'Candidatus Megaira' are diverse symbionts of algae and ciliates with the potential for defensive symbiosis.}, journal = {Microbial genomics}, volume = {9}, number = {3}, pages = {}, pmid = {36897930}, issn = {2057-5858}, mesh = {*Symbiosis ; *Ciliophora ; Phylogeny ; }, abstract = {Symbiotic microbes from the genus 'Candidatus Megaira' (Rickettsiales) are known to be common associates of algae and ciliates. However, genomic resources for these bacteria are scarce, limiting our understanding of their diversity and biology. We therefore utilize Sequence Read Archive and metagenomic assemblies to explore the diversity of this genus. We successfully extract four draft 'Ca. Megaira' genomes including one complete scaffold for a 'Ca. Megaira' and identify an additional 14 draft genomes from uncategorized environmental metagenome-assembled genomes. We use this information to resolve the phylogeny for the hyper-diverse 'Ca. Megaira', with hosts broadly spanning ciliates, and micro- and macro-algae, and find that the current single genus designation 'Ca. Megaira' significantly underestimates their diversity. We also evaluate the metabolic potential and diversity of ''Ca. Megaira' from this new genomic data and find no clear evidence of nutritional symbiosis. In contrast, we hypothesize a potential for defensive symbiosis in 'Ca. Megaira'. Intriguingly, one symbiont genome revealed a proliferation of ORFs with ankyrin, tetratricopeptide and leucine-rich repeats such as those observed in the genus Wolbachia where they are considered important for host–symbiont protein–protein interactions. Onward research should investigate the phenotypic interactions between 'Ca. Megaira' and their various potential hosts, including the economically important Nemacystus decipiens, and target acquisition of genomic information to reflect the diversity of this massively variable group.}, } @article {pmid36889655, year = {2023}, author = {Zhu, X and Liu, T and He, A and Zhang, L and Li, J and Li, T and Miao, X and You, M and You, S}, title = {Diversity of Wolbachia infection and its influence on mitochondrial DNA variation in the diamondback moth, Plutella xylostella.}, journal = {Molecular phylogenetics and evolution}, volume = {182}, number = {}, pages = {107751}, doi = {10.1016/j.ympev.2023.107751}, pmid = {36889655}, issn = {1095-9513}, mesh = {Animals ; *Moths/genetics ; *Wolbachia/genetics ; Phylogeny ; DNA, Mitochondrial/genetics ; Mitochondria/genetics ; }, abstract = {Plutella xylostella is a pest that severely damages cruciferous vegetables worldwide and has been shown to be infected with the maternally inherited bacteria Wolbachia, with the main infected strain was plutWB1. In this study, we performed a large-scale global sampling of P. xylostella and amplified 3 mtDNA genes of P. xylostella and 6 Wolbachia genes to analyze the infection status, diversity of Wolbachia in P. xylostella, and its effect on mtDNA variation in P. xylostella. This study provides a conservative estimate of Wolbachia infection rates in P. xylostella, which was found to be 7% (104/1440). The ST 108 (plutWB1) was shared among butterfly species and the moth species P. xylostella, revealing that Wolbachia strain plutWB1 acquisition in P. xylostella may be through horizontal transmission. The Parafit analyses indicated a significant association between Wolbachia and Wolbachia-infected P. xylostella individuals, and individuals infected with plutWB1 tended to cluster in the basal positions of the phylogenetic tree based on the mtDNA data. Additionally, Wolbachia infections were associated with increased mtDNA polymorphism in the infected P. xylostella population. These data suggest that Wolbachia endosymbionts may have a potential effect on mtDNA variation of P. xylostella.}, } @article {pmid36889142, year = {2023}, author = {Hussain, M and Etebari, K and Asgari, S}, title = {Analysing inhibition of dengue virus in Wolbachia-infected mosquito cells following the removal of Wolbachia.}, journal = {Virology}, volume = {581}, number = {}, pages = {48-55}, doi = {10.1016/j.virol.2023.02.017}, pmid = {36889142}, issn = {1096-0341}, mesh = {Animals ; *Dengue Virus/physiology ; *Wolbachia/physiology ; Virus Replication ; *RNA Viruses/genetics ; *Aedes ; *Dengue ; }, abstract = {Wolbachia pipientis is known to block replication of positive sense RNA viruses. Previously, we created an Aedes aegypti Aag2 cell line (Aag2.wAlbB) transinfected with the wAlbB strain of Wolbachia and a matching tetracycline-cured Aag2.tet cell line. While dengue virus (DENV) was blocked in Aag2.wAlbB cells, we found significant inhibition of DENV in Aag2.tet cells. RNA-Seq analysis of the cells confirmed removal of Wolbachia and lack of expression of Wolbachia genes that could have been due to lateral gene transfer in Aag2.tet cells. However, we noticed a substantial increase in the abundance of phasi charoen-like virus (PCLV) in Aag2.tet cells. When RNAi was used to reduce the PCLV levels, DENV replication was significantly increased. Further, we found significant changes in the expression of antiviral and proviral genes in Aag2.tet cells. Overall, the results reveal an antagonistic interaction between DENV and PCLV and how PCLV-induced changes could contribute to DENV inhibition.}, } @article {pmid36883556, year = {2023}, author = {Bueno-Marí, R and Domínguez-Santos, R and Trelis, M and Garrote-Sánchez, E and Cholvi, M and Quero de Lera, F and Khoubbane, M and Marcilla, A and Gil, R}, title = {[Wolbachia pipientis infections in populations of Aedes albopictus in the city of València (Spain): implications for mosquito control].}, journal = {Revista espanola de salud publica}, volume = {97}, number = {}, pages = {}, pmid = {36883556}, issn = {2173-9110}, mesh = {Adult ; Male ; Animals ; Humans ; *Aedes ; Mosquito Control ; Spain/epidemiology ; *Wolbachia ; }, abstract = {OBJECTIVE: The presence of Aedes albopictus, of high sanitary and social impact, was first reported in Valencia (Eastern Spain) in 2015. Innovative tools for its control include the use of the endosymbiotic bacterium Wolbachia pipientis. The release of mosquito males infected with the wPip strain, has proven very promising for large-scale Incompatible Insect Technique (IIT) applications. Before this strategy can be implemented in Valencia, it is important to know whether the natural local mosquito populations are Wolbachia-infected and, if so, identifying the infecting strains/supergroups, these being the objectives of the present work.

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

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

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

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

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

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

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

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

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

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

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

CONCLUSION: Nodule analysis by experienced microscopists was reproducible with regard to important parameters such as identification of living female worms or detection of normal embryogenesis. Assessments varied more for other parameters, and we recommend continued use of two independent readers for detailed analyzes. Analysis of scanned images provided similar results to direct microscopy. This facilitates training and comparison of nodule findings by readers in different locations. Analysis of high quality digital images that can be viewed remotely should improve the quality and availability of nodule assessments that are primary endpoints for onchocerciasis clinical trials.}, } @article {pmid36810669, year = {2023}, author = {Manoj, RRS and Latrofa, MS and Louni, M and Laidoudi, Y and Fenollar, F and Otranto, D and Mediannikov, O}, title = {In vitro maintenance of the endosymbiont Wolbachia of Dirofilaria immitis.}, journal = {Parasitology research}, volume = {122}, number = {4}, pages = {939-943}, pmid = {36810669}, issn = {1432-1955}, mesh = {Animals ; Dogs ; *Dirofilaria immitis ; *Dirofilariasis ; *Wolbachia ; Microfilariae ; *Dog Diseases/microbiology ; }, abstract = {Wolbachia has an obligatory mutualistic relationship with many onchocercid nematodes of the subfamilies Dirofilariinae and Onchocercinae. Till date, no attempts have been made for the in vitro cultivation of this intracellular bacterium from the filarioid host. Hence, the current study attempted cell co-culture method using embryonic Drosophila S2 and the LD cell lines to cultivate Wolbachia from Dirofilaria immitis microfilariae (mfs) harvested from infected dogs. Microfilariae (mfs = 1500) were inoculated in shell vials supplemented with Schneider medium using both cell lines. The establishment and multiplication of the bacterium were observed during the initial inoculation, at day 0 and before every medium change (from days 14 to 115). An aliquot (50 µl) from each time point was tested by quantitative real-time PCR (qPCR). Comparing the average of Ct values, obtained by the tested parameters (i.e., LD/S2 cell lines and mfs with/without treatment), the S2 cell line without mechanical disruption of mfs provided the highest Wolbachia cell count by qPCR. Despite the maintenance of Wolbachia within both S2 and LD-based cell co-culture models for up to 115 days, a definitive conclusion is still far. Further trials using fluorescent microscopy and viable staining will help to demonstrate the cell line infection and viability of Wolbachia. Use of considerable amount of untreated mfs to inoculate the Drosophilia S2 cell lines, as well as the supplementation of the culture media with growth stimulants or pre-treated cells to increase their susceptibility for the infection and development of a filarioid-based cell line system are recommended for the future trials.}, } @article {pmid36810610, year = {2023}, author = {Muro, T and Hikida, H and Fujii, T and Kiuchi, T and Katsuma, S}, title = {Two Complete Genomes of Male-Killing Wolbachia Infecting Ostrinia Moth Species Illuminate Their Evolutionary Dynamics and Association with Hosts.}, journal = {Microbial ecology}, volume = {86}, number = {3}, pages = {1740-1754}, pmid = {36810610}, issn = {1432-184X}, support = {17H06431//Japan Society for the Promotion of Science/ ; 22H00366//Japan Society for the Promotion of Science/ ; }, mesh = {Animals ; Male ; *Moths/genetics ; *Wolbachia/genetics ; Phylogeny ; Sex Ratio ; Genomics ; }, abstract = {Wolbachia is an extremely widespread intracellular symbiont which causes reproductive manipulation on various arthropod hosts. Male progenies are killed in Wolbachia-infected lineages of the Japanese Ostrinia moth population. While the mechanism of male killing and the evolutionary interaction between host and symbiont are significant concerns for this system, the absence of Wolbachia genomic information has limited approaches to these issues. We determined the complete genome sequences of wFur and wSca, the male-killing Wolbachia of Ostrinia furnacalis and Ostrinia scapulalis. The two genomes shared an extremely high degree of homology, with over 95% of the predicted protein sequences being identical. A comparison of these two genomes revealed nearly minimal genome evolution, with a strong emphasis on the frequent genome rearrangements and the rapid evolution of ankyrin repeat-containing proteins. Additionally, we determined the mitochondrial genomes of both species' infected lineages and performed phylogenetic analyses to deduce the evolutionary dynamics of Wolbachia infection in the Ostrinia clade. According to the inferred phylogenetic relationship, two possible scenarios were proposed: (1) Wolbachia infection was established in the Ostrinia clade prior to the speciation of related species such as O. furnacalis and O. scapulalis, or (2) Wolbachia infection in these species was introgressively transferred from a currently unidentified relative. Simultaneously, the relatively high homology of mitochondrial genomes suggested recent Wolbachia introgression between infected Ostrinia species. The findings of this study collectively shed light on the host-symbiont interaction from an evolutionary standpoint.}, } @article {pmid36800397, year = {2023}, author = {Mills, MK and McCabe, LG and Rodrigue, EM and Lechtreck, KF and Starai, VJ}, title = {Wbm0076, a candidate effector protein of the Wolbachia endosymbiont of Brugia malayi, disrupts eukaryotic actin dynamics.}, journal = {PLoS pathogens}, volume = {19}, number = {2}, pages = {e1010777}, pmid = {36800397}, issn = {1553-7374}, support = {R01 AI100913/AI/NIAID NIH HHS/United States ; R01 GM110413/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Humans ; Actins/metabolism ; *Brugia malayi/genetics ; Eukaryotic Cells ; Saccharomyces cerevisiae/genetics ; Symbiosis/genetics ; *Wolbachia/physiology ; Bacterial Proteins ; }, abstract = {Brugia malayi, a parasitic roundworm of humans, is colonized by the obligate intracellular bacterium, Wolbachia pipientis. The symbiosis between this nematode and bacterium is essential for nematode reproduction and long-term survival in a human host. Therefore, identifying molecular mechanisms required by Wolbachia to persist in and colonize B. malayi tissues will provide new essential information regarding the basic biology of this endosymbiosis. Wolbachia utilize a Type IV secretion system to translocate so-called "effector" proteins into the cytosol of B. malayi cells to promote colonization of the eukaryotic host. However, the characterization of these Wolbachia secreted proteins has remained elusive due to the genetic intractability of both organisms. Strikingly, expression of the candidate Wolbachia Type IV-secreted effector protein, Wbm0076, in the surrogate eukaryotic cell model, Saccharomyces cerevisiae, resulted in the disruption of the yeast actin cytoskeleton and inhibition of endocytosis. Genetic analyses show that Wbm0076 is a member of the family of Wiskott-Aldrich syndrome proteins (WAS [p]), a well-conserved eukaryotic protein family required for the organization of actin skeletal structures. Thus, Wbm0076 likely plays a central role in the active cell-to-cell movement of Wolbachia throughout B. malayi tissues during nematode development. As most Wolbachia isolates sequenced to date encode at least partial orthologs of wBm0076, we find it likely that the ability of Wolbachia to directly manipulate host actin dynamics is an essential requirement of all Wolbachia endosymbioses, independent of host cell species.}, } @article {pmid36786616, year = {2023}, author = {Li, C and Liu, S and Zhou, H and Zhu, W and Cui, M and Li, J and Wang, J and Liu, J and Zhu, J and Li, W and Bi, Y and Carr, MJ and Holmes, EC and Shi, W}, title = {Metatranscriptomic Sequencing Reveals Host Species as an Important Factor Shaping the Mosquito Virome.}, journal = {Microbiology spectrum}, volume = {11}, number = {2}, pages = {e0465522}, pmid = {36786616}, issn = {2165-0497}, abstract = {Mosquitoes are important vector hosts for numerous viral pathogens and harbor a large number of mosquito-specific viruses as well as human-infecting viruses. Previous studies have mainly focused on the discovery of mosquito viruses, and our understanding of major ecological factors associated with virome structure in mosquitoes remains limited. We utilized metatranscriptomic sequencing to characterize the viromes of five mosquito species sampled across eight locations in Yunnan Province, China. This revealed the presence of 52 viral species, of which 19 were novel, belonging to 15 viral families/clades. Of particular note was Culex hepacivirus 1, clustering within the avian clade of hepaciviruses. Notably, both the viromic diversity and abundance of Aedes genus mosquitoes were significantly higher than those of the Culex genus, while Aedes albopictus mosquitoes harbored a higher diversity than Aedes aegypti mosquitoes. Our findings thus point to discernible differences in viromic structure between mosquito genera and even between mosquito species within the same genus. Importantly, such differences were not attributable to differences in sampling between geographical location. Our study also revealed the ubiquitous presence of the endosymbiont bacterium Wolbachia, with the genetic diversity and abundance also varying between mosquito species. In conclusion, our results suggested that the mosquito host species play an important role in shaping the virome's structure. IMPORTANCE This study revealed the huge capability of mosquitoes in harboring a rich diversity of RNA viruses, although relevant studies have characterized the intensively unparalleled diversity of RNA viruses previously. Furthermore, our findings showed discernible differences not only in viromic structure between mosquito genera and even between mosquito species within the same genus but also in the genetic diversity and abundance of Wolbachia between different mosquito populations. These findings emphasize the importance of host genetic background in shaping the virome composition of mosquitoes.}, } @article {pmid36785954, year = {2023}, author = {Sanaei, E and Albery, GF and Yeoh, YK and Lin, YP and Cook, LG and Engelstädter, J}, title = {Host phylogeny and ecological associations best explain Wolbachia host shifts in scale insects.}, journal = {Molecular ecology}, volume = {32}, number = {9}, pages = {2351-2363}, doi = {10.1111/mec.16883}, pmid = {36785954}, issn = {1365-294X}, mesh = {Animals ; *Hemiptera/microbiology ; Insecta/genetics ; Phylogeny ; Symbiosis/genetics ; Wasps/genetics ; *Wolbachia/genetics ; }, abstract = {Wolbachia are among the most prevalent and widespread endosymbiotic bacteria on Earth. Wolbachia's success in infecting an enormous number of arthropod species is attributed to two features: the range of phenotypes they induce in their hosts, and their ability to switch between host species. Whilst much progress has been made in elucidating their induced phenotypes, our understanding of Wolbachia host-shifting is still very limited: we lack answers to even fundamental questions concerning Wolbachia's routes of transfer and the importance of factors influencing host shifts. Here, we investigate the diversity and host-shift patterns of Wolbachia in scale insects, a group of arthropods with intimate associations with other insects that make them well suited to studying host shifts. Using Illumina multitarget amplicon sequencing of Wolbachia-infected scale insects and their direct associates we determined the identity of all Wolbachia strains. We then fitted a generalized additive mixed model to our data to estimate the influence of host phylogeny and the geographical distribution on Wolbachia strain sharing among scale insect species. The model predicts no significant contribution of host geography but strong effects of host phylogeny, with high rates of Wolbachia sharing among closely related species and a sudden drop-off in sharing with increasing phylogenetic distance. We also detected the same Wolbachia strain in scale insects and several intimately associated species (ants, wasps and flies). This indicates putative host shifts and potential routes of transfers via these associates and highlights the importance of ecological connectivity in Wolbachia host-shifting.}, } @article {pmid36781724, year = {2023}, author = {Takasuka, K and Arakawa, K}, title = {The Method of Eliminating the Wolbachia Endosymbiont Genomes from Insect Samples Prior to a Long-Read Sequencing.}, journal = {Methods in molecular biology (Clifton, N.J.)}, volume = {2632}, number = {}, pages = {101-112}, pmid = {36781724}, issn = {1940-6029}, mesh = {Animals ; *Wolbachia/genetics ; *Wasps/genetics ; Genome ; Insecta/genetics ; Rifampin ; Symbiosis/genetics ; }, abstract = {When extracting DNA of invertebrates for long-read sequencing, not only enough quantity and size of the DNA but, depending on the species, elimination of contamination of endosymbiotic Wolbachia genome also has to be achieved. These requirements become troublesome, especially in small-sized species with a limited number of individuals available for the experiment. In this chapter, using tiny parasitoid wasps (Reclinervellus nielseni) parasitizing spiders as hosts, we developed a method of eliminating the Wolbachia genomes by means of an antibiotic administration to adult wasps via honey solution. Twenty days of rifampicin treatment since their emergence from cocoons resulted in a significant decrease in the Wolbachia genomes while keeping good DNA conditions for nanopore sequencing. An adequate quantity of DNA was then gained by pooling several individuals. The method could be applied to other insects or invertebrates that can be maintained by laboratory feeding with liquid food.}, } @article {pmid36778977, year = {2022}, author = {Sinha, DK and Gupta, A and Padmakumari, AP and Bentur, JS and Nair, S}, title = {Infestation of Rice by Gall Midge Influences Density and Diversity of Pseudomonas and Wolbachia in the Host Plant Microbiome.}, journal = {Current genomics}, volume = {23}, number = {2}, pages = {126-136}, pmid = {36778977}, issn = {1389-2029}, abstract = {Background: The virulence of phytophagous insects is predominantly determined by their ability to evade or suppress host defense for their survival. The rice gall midge (GM, Orseolia oryzae), a monophagous pest of rice, elicits a host defense similar to the one elicited upon pathogen attack. This could be due to the GM feeding behaviour, wherein the GM endosymbionts are transferred to the host plant via oral secretions, and as a result, the host mounts an appropriate defense response(s) (i.e., up-regulation of the salicylic acid pathway) against these endosymbionts. Methods: The current study aimed to analyze the microbiome present at the feeding site of GM maggots to determine the exchange of bacterial species between GM and its host and to elucidate their role in rice-GM interaction using a next-generation sequencing approach. Results: Our results revealed differential representation of the phylum Proteobacteria in the GM-infested and -uninfested rice tissues. Furthermore, analysis of the species diversity of Pseudomonas and Wolbachia supergroups at the feeding sites indicated the exchange of bacterial species between GM and its host upon infestation. Conclusion: As rice-GM microbial associations remain relatively unstudied, these findings not only add to our current understanding of microbe-assisted insect-plant interactions but also provide valuable insights into how these bacteria drive insect-plant coevolution. Moreover, to the best of our knowledge, this is the first report analyzing the microbiome of a host plant (rice) at the feeding site of its insect pest (GM).}, } @article {pmid36761111, year = {2022}, author = {Pang, Y and Su, CY and Zhu, JQ and Yang, XH and Zhong, JL and Zhu, DH and Liu, Z}, title = {A new species of Andricus Hartig, 1840 (Hymenoptera, Cynipidae) from China, with references to DNA taxonomy and Wolbachia infection.}, journal = {ZooKeys}, volume = {1134}, number = {}, pages = {52-73}, pmid = {36761111}, issn = {1313-2989}, abstract = {In the present paper, a new species of cynipid gall wasp, Andricuselodeoides Liu & Pang, is described from several provinces in southern China. The new species is closely related to the recently redescribed A.mairei (Kieffer, 1906). In addition to differences in adult and gall morphology, the new species is also readily separated by COI sequences, with a 6.2-8.9% genetic distance between populations of the new species and those of A.mairei. A contrasting difference in sex ratios was also observed between the two species, with A.elodeoides extremely female-biased (95.5-97.8% female) while A.mairei male-biased to more balanced (5.4-43.5% female). PCR screening for Wolbachia infection further revealed contrasting infection rates between populations of A.elodeoides and A.mairei: the Wolbachia infection rate was 0% in A.elodeoides and 100% in A.mairei. Cytoplasmic incompatibility induced by Wolbachia is proposed as a potential mechanism of speciation of the sympatric A.elodeoides and A.mairei.}, } @article {pmid36760975, year = {2022}, author = {Zarka, J and Parmentier, T and Wybouw, N}, title = {Intersexuality in a natural population of the terrestrial isopod Porcellioscaber.}, journal = {ZooKeys}, volume = {1101}, number = {}, pages = {183-190}, pmid = {36760975}, issn = {1313-2989}, abstract = {Intersex phenotypes are rarely observed in natural isopod populations and their expression is typically associated with infection of Wolbachia, a reproductive parasite that manipulates arthropod reproduction. During an intensive sampling effort of a natural population of the isopod Porcellioscaber, an adult individual was isolated that expressed both male and female traits. The intersex individual exhibited clearly developed external male genitalia and carried multiple eggs in its brood pouch. No Wolbachia infection could be identified in this individual, a result that needs to be approached with caution due to suboptimal DNA preservation for diagnostic PCR assays. Wolbachia were, however, detected in two adult females of the same population, and appear closely related to isolates that infect other terrestrial isopod species. This is the first demonstration that intersex phenotypes can arise under natural conditions in P.scaber.}, } @article {pmid36757767, year = {2023}, author = {Halter, T and Köstlbacher, S and Rattei, T and Hendrickx, F and Manzano-Marín, A and Horn, M}, title = {One to host them all: genomics of the diverse bacterial endosymbionts of the spider Oedothorax gibbosus.}, journal = {Microbial genomics}, volume = {9}, number = {2}, pages = {}, pmid = {36757767}, issn = {2057-5858}, mesh = {Animals ; *Spiders/genetics/microbiology ; DNA Transposable Elements ; Bacteroidetes/genetics ; Genomics ; *Rickettsia/genetics ; *Wolbachia/genetics ; }, abstract = {Bacterial endosymbionts of the groups Wolbachia, Cardinium and Rickettsiaceae are well known for their diverse effects on their arthropod hosts, ranging from mutualistic relationships to reproductive phenotypes. Here, we analysed a unique system in which the dwarf spider Oedothorax gibbosus is co-infected with up to five different endosymbionts affiliated with Wolbachia, 'Candidatus Tisiphia' (formerly Torix group Rickettsia), Cardinium and Rhabdochlamydia. Using short-read genome sequencing data, we show that the endosymbionts are heterogeneously distributed among O. gibbosus populations and are frequently found co-infecting spider individuals. To study this intricate host-endosymbiont system on a genome-resolved level, we used long-read sequencing to reconstruct closed genomes of the Wolbachia, 'Ca. Tisiphia' and Cardinium endosymbionts. We provide insights into the ecology and evolution of the endosymbionts and shed light on the interactions with their spider host. We detected high quantities of transposable elements in all endosymbiont genomes and provide evidence that ancestors of the Cardinium, 'Ca. Tisiphia' and Wolbachia endosymbionts have co-infected the same hosts in the past. Our findings contribute to broadening our knowledge about endosymbionts infecting one of the largest animal phyla on Earth and show the usefulness of transposable elements as an evolutionary 'contact-tracing' tool.}, } @article {pmid36754115, year = {2023}, author = {Prigot-Maurice, C and Lheraud, B and Guéritault, S and Beltran-Bech, S and Cordaux, R and Peccoud, J and Braquart-Varnier, C}, title = {Investigating Wolbachia symbiont-mediated host protection against a bacterial pathogen using a natural Wolbachia nuclear insert.}, journal = {Journal of invertebrate pathology}, volume = {197}, number = {}, pages = {107893}, doi = {10.1016/j.jip.2023.107893}, pmid = {36754115}, issn = {1096-0805}, mesh = {Female ; Animals ; *Symbiosis ; *Wolbachia ; Bacteria ; }, abstract = {Wolbachia bacterial endosymbionts provide protection against pathogens in various arthropod species but the underlying mechanisms remain misunderstood. By using a natural Wolbachia nuclear insert (f-element) in the isopod Armadillidium vulgare, we explored whether Wolbachia presence is mandatory to observe protection in this species or the presence of its genes is sufficient. We assessed survival of closely related females carrying or lacking the f-element (and lacking Wolbachia) challenged with the bacterial pathogen Salmonella enterica. Despite marginal significant effects, the f-element alone did not appear to confer survival benefits to its host, suggesting that Wolbachia presence in cells is crucial for protection.}, } @article {pmid36750860, year = {2023}, author = {Mahmood, S and Nováková, E and Martinů, J and Sychra, O and Hypša, V}, title = {Supergroup F Wolbachia with extremely reduced genome: transition to obligate insect symbionts.}, journal = {Microbiome}, volume = {11}, number = {1}, pages = {22}, pmid = {36750860}, issn = {2049-2618}, mesh = {Animals ; Phylogeny ; *Wolbachia/genetics ; Biological Evolution ; *Nematoda ; Insecta ; Symbiosis/physiology ; }, abstract = {BACKGROUND: Wolbachia belong to highly abundant bacteria which are frequently found in invertebrate microbiomes and manifest by a broad spectrum of lifestyles from parasitism to mutualism. Wolbachia supergroup F is a particularly interesting clade as it gave rise to symbionts of both arthropods and nematodes, and some of its members are obligate mutualists. Investigations on evolutionary transitions among the different symbiotic stages have been hampered by a lack of the known diversity and genomic data for the supergroup F members.

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

CONCLUSIONS: The genome of wMeur1 strain, from the M. eurysternus microbiome, represents the smallest currently known Wolbachia genome and the first example of Wolbachia which has completed genomic streamlining as known from the typical obligate symbionts. This points out that despite the large amount and great diversity of the known Wolbachia strains, evolutionary potential of these bacteria still remains underexplored. The diversity of the four chewing lice microbiomes indicates that this vast parasitic group may provide suitable models for further investigations. Video Abstract.}, } @article {pmid36748430, year = {2022}, author = {Izraeli, Y and Lepetit, D and Atias, S and Mozes-Daube, N and Wodowski, G and Lachman, O and Luria, N and Steinberg, S and Varaldi, J and Zchori-Fein, E and Chiel, E}, title = {Genomic characterization of viruses associated with the parasitoid Anagyrus vladimiri (Hymenoptera: Encyrtidae).}, journal = {The Journal of general virology}, volume = {103}, number = {12}, pages = {}, doi = {10.1099/jgv.0.001810}, pmid = {36748430}, issn = {1465-2099}, mesh = {Humans ; Female ; Animals ; Phylogeny ; *Wasps ; *Viruses ; Genomics ; *Reoviridae/genetics ; }, abstract = {Knowledge on symbiotic microorganisms of insects has increased dramatically in recent years, yet relatively little data are available regarding non-pathogenic viruses. Here we studied the virome of the parasitoid wasp Anagyrus vladimiri Triapitsyn (Hymenoptera: Encyrtidae), a biocontrol agent of mealybugs. By high-throughput sequencing of viral nucleic acids, we revealed three novel viruses, belonging to the families Reoviridae [provisionally termed AnvRV (Anagyrus vladimiri reovirus)], Iflaviridae (AnvIFV) and Dicistroviridae (AnvDV). Phylogenetic analysis further classified AnvRV in the genus Idnoreovirus, and AnvDV in the genus Triatovirus. The genome of AnvRV comprises 10 distinct genomic segments ranging in length from 1.5 to 4.2 kb, but only two out of the 10 ORFs have a known function. AnvIFV and AnvDV each have one polypeptide ORF, which is typical of iflaviruses but very un-common among dicistroviruses. Five conserved domains were found along both the ORFs of those two viruses. AnvRV was found to be fixed in an A. vladimiri population that was obtained from a mass rearing facility, whereas its prevalence in field-collected A. vladimiri was ~15 %. Similarly, the prevalence of AnvIFV and AnvDV was much higher in the mass rearing population than in the field population. The presence of AnvDV was positively correlated with the presence of Wolbachia in the same individuals. Transmission electron micrographs of females' ovaries revealed clusters and viroplasms of reovirus-like particles in follicle cells, suggesting that AnvRV is vertically transmitted from mother to offspring. AnvRV was not detected in the mealybugs, supporting the assumption that this virus is truly associated with the wasps. The possible effects of these viruses on A. vladimiri's biology, and on biocontrol agents in general, are discussed. Our findings identify RNA viruses as potentially involved in the multitrophic system of mealybugs, their parasitoids and other members of the holobiont.}, } @article {pmid36744754, year = {2023}, author = {Chen, J and Wang, MK and Xie, QX and Bing, XL and Li, TP and Hong, XY}, title = {NDUFA8 potentially rescues Wolbachia-induced cytoplasmic incompatibility in Laodelphax striatellus.}, journal = {Insect science}, volume = {30}, number = {6}, pages = {1689-1700}, doi = {10.1111/1744-7917.13182}, pmid = {36744754}, issn = {1744-7917}, support = {32020103011//National Natural Science Foundation of China/ ; 32001905//National Natural Science Foundation of China/ ; 31871976//National Natural Science Foundation of China/ ; }, mesh = {Female ; Male ; Animals ; *Wolbachia/genetics ; *Hemiptera/genetics/metabolism ; Fertility ; Reproduction ; Gene Expression Profiling ; }, abstract = {The endosymbiont Wolbachia manipulates host reproduction by several strategies, one of the most important of which is cytoplasmic incompatibility (CI). CI can be rescued when Wolbachia-infected males mate with females infected with the same Wolbachia strain. However, the potential rescue mechanism of CI in the small brown planthopper Laodelphax striatellus is unclear. In this study, comparative transcriptome analysis was applied to explore the effect of Wolbachia on L. striatellus eggs. A total of 1387 differentially expressed genes were identified. RNA interference of 7 Wolbachia-upregulated key planthopper genes reduced egg reproduction, suggesting that Wolbachia might improve fecundity in L. striatellus by affecting these 7 genes. Suppressing the expression of another upregulated gene, NDUFA8 (encoding NADH dehydrogenase [ubiquinone] 1 α subcomplex subunit 8-like) by RNA interference significantly increased the mortality of early embryos without affecting the number of deposited eggs. Wolbachia infection upregulated the mRNA level of NDUFA8, and dsNDUFA8 treatment of Wolbachia-infected females recreated CI-like symptoms, suggesting that NDUFA8 is associated with the rescue phenotype. Because all L. striatellus populations worldwide are infected with Wolbachia, NDUFA8 is a potential pest control target.}, } @article {pmid36719928, year = {2023}, author = {Ross, PA and Elfekih, S and Collier, S and Klein, MJ and Lee, SS and Dunn, M and Jackson, S and Zhang, Y and Axford, JK and Gu, X and Home, JL and Nassar, MS and Paradkar, PN and Tawfik, EA and Jiggins, FM and Almalik, AM and Al-Fageeh, MB and Hoffmann, AA}, title = {Developing Wolbachia-based disease interventions for an extreme environment.}, journal = {PLoS pathogens}, volume = {19}, number = {1}, pages = {e1011117}, pmid = {36719928}, issn = {1553-7374}, mesh = {Animals ; *Wolbachia ; Saudi Arabia ; Australia ; Extreme Environments ; *Aedes ; *Dengue ; }, abstract = {Aedes aegypti mosquitoes carrying self-spreading, virus-blocking Wolbachia bacteria are being deployed to suppress dengue transmission. However, there are challenges in applying this technology in extreme environments. We introduced two Wolbachia strains into Ae. aegypti from Saudi Arabia for a release program in the hot coastal city of Jeddah. Wolbachia reduced infection and dissemination of dengue virus (DENV2) in Saudi Arabian mosquitoes and showed complete maternal transmission and cytoplasmic incompatibility. Wolbachia reduced egg hatch under a range of environmental conditions, with the Wolbachia strains showing differential thermal stability. Wolbachia effects were similar across mosquito genetic backgrounds but we found evidence of local adaptation, with Saudi Arabian mosquitoes having lower egg viability but higher adult desiccation tolerance than Australian mosquitoes. Genetic background effects will influence Wolbachia invasion dynamics, reinforcing the need to use local genotypes for mosquito release programs, particularly in extreme environments like Jeddah. Our comprehensive characterization of Wolbachia strains provides a foundation for Wolbachia-based disease interventions in harsh climates.}, } @article {pmid36715911, year = {2023}, author = {Nevalainen, LBM and Newton, ILG}, title = {Detection and Assessment of Wolbachia pipientis Infection.}, journal = {Methods in molecular biology (Clifton, N.J.)}, volume = {2626}, number = {}, pages = {291-307}, pmid = {36715911}, issn = {1940-6029}, support = {R01 AI144430/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; *Drosophila melanogaster/genetics ; *Wolbachia/genetics ; Multilocus Sequence Typing ; In Situ Hybridization, Fluorescence ; Drosophila/genetics ; }, abstract = {Wolbachia pipientis is a widespread vertically transmitted intracellular bacterium naturally present in the model organism Drosophila melanogaster. As Wolbachia is present in a large number of Drosophila lines, it is critical for researchers to be able to identify which of their stocks maintain this infection to avoid any potential confounding variables. Here, we describe methods for detecting the bacterium and assessing the infection, including polymerase chain reaction (PCR) of DNA, multi-locus sequence typing (MLST) to identify strains, western blotting for protein detection, and immunohistochemistry and fluorescence in situ hybridization (FISH) of Drosophila ovaries to visually detect Wolbachia by fluorescence microscopy.}, } @article {pmid36714832, year = {2022}, author = {Liang, X and Tan, CH and Sun, Q and Zhang, M and Wong, PSJ and Li, MI and Mak, KW and Martín-Park, A and Contreras-Perera, Y and Puerta-Guardo, H and Manrique-Saide, P and Ng, LC and Xi, Z}, title = {Wolbachia wAlbB remains stable in Aedes aegypti over 15 years but exhibits genetic background-dependent variation in virus blocking.}, journal = {PNAS nexus}, volume = {1}, number = {4}, pages = {pgac203}, pmid = {36714832}, issn = {2752-6542}, abstract = {The ability of the maternally transmitted endosymbiotic bacterium Wolbachia to induce cytoplasmic incompatibility (CI) and virus blocking makes it a promising weapon for combatting mosquito-borne diseases through either suppression or replacement of wild-type populations. Recent field trials show that both approaches significantly reduce the incidence of dengue fever in humans. However, new questions emerge about how Wolbachia-mosquito associations will co-evolve over time and whether Wolbachia-mediated virus blocking will be affected by the genetic diversity of mosquitoes and arboviruses in the real world. Here, we have compared the Wolbachia density and CI expression of two wAlbB-infected Aedes aegypti lines transinfected 15 years apart. We have also assessed wAlbB-mediated virus blocking against dengue (DENV), Zika (ZIKV), and Chikungunya (CHIKV) viruses and examined whether host genetic backgrounds modulate viral blocking effects by comparing ZIKV infection in mosquitoes with a Mexican genetic background to those with a Singaporean background. Our results show that over 15 years, wAlbB maintained the capacity to form a stable association with Ae. aegypti in terms of both density and CI expression. There were variations in wAlbB-induced virus blocking against CHIKV, DENV, and ZIKV, and higher inhibitory effects on ZIKV in mosquitoes on the Singaporean genetic background than on the Mexican background. These results provide important information concerning the robustness and long-term stability of Wolbachia as a biocontrol agent for arbovirus disease control.}, } @article {pmid36714306, year = {2022}, author = {Barman, M and Samanta, S and Ahmed, B and Dey, S and Chakraborty, S and Deeksha, MG and Dutta, S and Samanta, A and Tarafdar, J and Roy, D}, title = {Transcription dynamics of heat-shock proteins (Hsps) and endosymbiont titres in response to thermal stress in whitefly, Bemisia tabaci (Asia-I).}, journal = {Frontiers in physiology}, volume = {13}, number = {}, pages = {1097459}, pmid = {36714306}, issn = {1664-042X}, abstract = {The sweet potato whitefly, Bemisia tabaci (Gennadius), is one of the several species complexes of whitefly that are currently significant agricultural pests. Bemisia tabaci infests more than 600 plant species and thrives under a wide range of temperature conditions. In addition to the direct damage caused by sucking plant sap, it vectors several plant viruses. Heat-shock proteins play a pivotal role in enabling the insect to extend its geographical location, survival, and reproduction under different stress conditions. B. tabaci harbours several endosymbionts under the genera Portiera, Rickettsia, Hamiltonella, Wolbachia, Arsenophonus, Cardinium, and Fritschea that directly or indirectly affect its fitness. By accelerating cuticle biosynthesis and sclerotisation, symbiotic microbes can reduce or enhance tolerance to extreme temperatures and detoxify heavy metals. Thus, symbionts or microbial communities can expand or constrain the abiotic niche space of their host and affect its ability to adapt to changing conditions. The present study delineates the effect of thermal stress on the expression of heat-shock genes and endosymbionts in B. tabaci. Studies of the expression level of heat-shock proteins with the help of quantitative real-time polymerase chain reaction (qRT-PCR) showed that heat- and cold-shock treatment fuels the increased expression of heat-shock proteins (Hsp40 and Hsp70). However, Hsp90 was not induced by a heat- and cold-shock treatment. A significant decrease in the relative titre of secondary endosymbionts, such as Rickettsia, Arsenophonus, and Wolbachia, were recorded in B. tabaci upon heat treatment. However, the titre of the primary symbiont, C. Portiera, was relatively unaffected by both cold and heat treatments. These results are indicative of the fact that Hsp genes and endosymbionts in B. tabaci are modulated in response to thermal stress, and this might be responsible for the adaptation of whitefly under changing climatic scenario.}, } @article {pmid36712932, year = {2023}, author = {Herran, B and Sugimoto, TN and Watanabe, K and Imanishi, S and Tsuchida, T and Matsuo, T and Ishikawa, Y and Kageyama, D}, title = {Cell-based analysis reveals that sex-determining gene signals in Ostrinia are pivotally changed by male-killing Wolbachia.}, journal = {PNAS nexus}, volume = {2}, number = {1}, pages = {pgac293}, pmid = {36712932}, issn = {2752-6542}, abstract = {Wolbachia, a maternally transmitted bacterium, shows male-killing, an adaptive phenotype for cytoplasmic elements, in various arthropod species during the early developmental stages. In lepidopteran insects, lethality of males is accounted for by improper dosage compensation in sex-linked genes owing to Wolbachia-induced feminization. Herein, we established Ostrinia scapulalis cell lines that retained sex specificity per the splicing pattern of the sex-determining gene doublesex (Osdsx). We found that Wolbachia transinfection in male cell lines enhanced the female-specific splice variant of Osdsx (Osdsx[F]) while suppressing the male-specific variant (Osdsx[M]), indicating that Wolbachia affects sex-determining gene signals even in vitro. Comparative transcriptome analysis isolated only two genes that behave differently upon Wolbachia infection. The two genes were respectively homologous to Masculinizer (BmMasc) and zinc finger-2 (Bmznf-2), male-specifically expressed sex-determining genes of the silkworm Bombyx mori that encode CCCH-type zinc finger motif proteins. By using cultured cells and organismal samples, OsMasc and Osznf-2 were found to be sex-determining genes of O. scapulalis that are subjected to sex-specific alternative splicing depending upon the chromosomal sex, developmental stage, and infection status. Overall, our findings expound the cellular autonomy in insect sex determination and the mechanism through which sex is manipulated by intracellular selfish microbes.}, } @article {pmid36711723, year = {2023}, author = {Dodson, BL and Pujhari, S and Brustolin, M and Metz, HC and Rasgon, JL}, title = {Variable effects of Wolbachia on alphavirus infection in Aedes aegypti.}, journal = {bioRxiv : the preprint server for biology}, volume = {}, number = {}, pages = {}, pmid = {36711723}, issn = {2692-8205}, support = {R01 AI116636/AI/NIAID NIH HHS/United States ; R01 AI150251/AI/NIAID NIH HHS/United States ; }, abstract = {Wolbachia pipientis (=Wolbachia) has promise as a tool to suppress virus transmission by Aedes aegypti mosquitoes. However, Wolbachia can have variable effects on mosquito-borne viruses. This variation remains poorly characterized, yet the multimodal effects of Wolbachia on diverse pathogens could have important implications for public health. Here, we examine the effects of somatic infection with two strains of Wolbachia (wAlbB and wMel) on the alphaviruses Sindbis virus (SINV), O'nyong-nyong virus (ONNV), and Mayaro virus (MAYV) in Ae. aegypti. We found variable effects of Wolbachia including enhancement and suppression of viral infections, with some effects depending on Wolbachia strain. Both wAlbB- and wMel-infected mosquitoes showed enhancement of SINV infection rates one week post-infection, with wAlbB-infected mosquitoes also having higher viral titers than controls. Infection rates with ONNV were low across all treatments and no significant effects of Wolbachia were observed. The effects of Wolbachia on MAYV infections were strikingly strain-specific; wMel strongly blocked MAYV infections and suppressed viral titers, while wAlbB did not influence MAYV infection. The variable effects of Wolbachia on vector competence underscore the importance of further research into how this bacterium impacts the virome of wild mosquitoes including the emergent human pathogens they transmit.}, } @article {pmid36711506, year = {2024}, author = {Lindsey, AR and Tennessen, JM and Gelaw, MA and Jones, MW and Parish, AJ and Newton, IL and Nemkov, T and D'Alessandro, A and Rai, M and Stark, N}, title = {The intracellular symbiont Wolbachia alters Drosophila development and metabolism to buffer against nutritional stress.}, journal = {bioRxiv : the preprint server for biology}, volume = {}, number = {}, pages = {}, doi = {10.1101/2023.01.20.524972}, pmid = {36711506}, issn = {2692-8205}, support = {R21 AI175957/AI/NIAID NIH HHS/United States ; R35 GM150991/GM/NIGMS NIH HHS/United States ; }, abstract = {UNLABELLED: The intracellular bacterium Wolbachia is a common symbiont of many arthropods and nematodes, well studied for its impacts on host reproductive biology. However, its broad success as a vertically transmitted infection cannot be attributed to manipulations of host reproduction alone. Using the Drosophila melanogaster model and their natively associated Wolbachia strain " w Mel", we show that Wolbachia infection supports fly development and buffers against nutritional stress. Wolbachia infection across several fly genotypes and a range of nutrient conditions resulted in reduced pupal mortality, increased adult emergence, and larger size. We determined that the exogenous supplementation of pyrimidines partially rescued developmental phenotypes in the Wolbachia -free flies, and that Wolbachia titers were responsive to reduced gene expression of the fly's de novo pyrimidine synthesis pathway. In parallel, transcriptomic and metabolomic analyses indicated that Wolbachia impacts larval biology far beyond pyrimidine metabolism. Wolbachia -infected larvae had strong signatures of shifts in glutathione and mitochondrial metabolism, plus significant changes in the expression of key developmental regulators including Notch , the insulin receptor (lnR), and the juvenile hormone receptor Methoprene-tolerant (Met). We propose that Wolbachia acts as a beneficial symbiont to support fly development and enhance host fitness, especially during periods of nutrient stress.

SIGNIFICANCE: Wolbachia is a bacterial symbiont of arthropods and nematodes, well described for its manipulations of arthropod reproduction. However, many have theorized there must be more to this symbiosis, even in well-studied Wolbachia- host relationships such as with Drosophila . Reproductive impacts alone cannot explain the success and ubiquity of this bacterium. Here, we use Drosophila melanogaster and their native Wolbachia infections to show that Wolbachia supports fly development and significantly buffers flies against nutritional stress. These developmental advantages might help explain the ubiquity of Wolbachia infections.}, } @article {pmid36708161, year = {2023}, author = {Hubert, J and Nesvorna, M and Sopko, B and Green, SJ}, title = {Diet modulation of the microbiome of the pest storage mite Tyrophagus putrescentiae.}, journal = {FEMS microbiology ecology}, volume = {99}, number = {3}, pages = {}, doi = {10.1093/femsec/fiad011}, pmid = {36708161}, issn = {1574-6941}, mesh = {Animals ; *Acaridae/genetics/microbiology ; RNA, Ribosomal, 16S/genetics ; *Microbiota ; *Mites ; Diet ; Bacteria/genetics ; Bacteroidetes/genetics ; Enterobacteriaceae/genetics ; }, abstract = {Storage mites colonize a wide spectrum of food commodities and adaptations to diets have been suggested as mechanisms enabling successful colonization. We characterized the response of seven unique Tyrophagus putrescentiae cultures (5K, 5L, 5N, 5P, 5Pi, 5S, and 5Tk) with different baseline microbiomes to different diets. The offered diets included a rearing diet, protein-enriched diet, oat flakes, and sunflower seeds. Microbiome characterization was performed using 16S ribosomal RNA (rRNA) gene amplicon sequencing and 16S rRNA gene quantitative PCR. The mite culture microbiomes were classified into four groups: (i) Sodalis-dominated (5Pi), (ii) Wolbachia-dominated (5N and 5P), (iii) Cardinium-dominated (5L and 5S), and (iv) asymbiontic (5K and 5Tk) mites dominated by Bacillus and Bartonella. Mite growth rates were most strongly affected by nutrients in the diet, while respiration and microbial community profiles were largely influenced by mite culture. While growth rate was not directly explained by microbiome composition, microbiomes strongly influenced mite fitness as measured by respiration. While diet significantly influenced microbial profiles in all cultures, the effect of diet differed in impact between cultures (5Pi > 5S > 5N > 5K > 5Tk > 5L > 5P). Furthermore, no new bacterial taxa were acquired by mites after dietary changes. Bacteria from the taxa Bacillus, Bartonella-like, Solitalea-like, Kocuria, and Sodalis-like contributed most strongly to differentiating mite-associated microbiomes.}, } @article {pmid36706527, year = {2023}, author = {Wei, X and Peng, H and Li, Y and Meng, B and Wang, S and Bi, S and Zhao, X}, title = {Pyrethroids exposure alters the community and function of the internal microbiota in Aedes albopictus.}, journal = {Ecotoxicology and environmental safety}, volume = {252}, number = {}, pages = {114579}, doi = {10.1016/j.ecoenv.2023.114579}, pmid = {36706527}, issn = {1090-2414}, mesh = {Animals ; *Insecticides/pharmacology ; Permethrin/toxicity ; *Aedes ; Mosquito Vectors ; *Pyrethrins/pharmacology ; Insecticide Resistance/genetics ; *Microbiota ; }, abstract = {Large amounts of insecticides bring selection pressure and then develop insecticide resistance in Aedes albopictus. This study demonstrated for the first time the effect of pyrethroid exposure on the internal microbiota in Ae. albopictus. 36, 48, 57 strains of virgin adult Ae. albopictus were exposed to the pyrethroids deltamethrin (Dme group), β-cypermethrin (Bcy group), and cis-permethrin (Cper group), respectively, with n-hexane exposure (Hex group) as the controls (n = 36). The internal microbiota community and functions were analyzed based on the metagenomic analysis. The analysis of similarity (ANOSIM) results showed that the Hex/Bcy (p = 0.001), Hex/Cper (p = 0.006), Hex/Dme (p = 0.001) groups were well separated, and the internal microbes of Ae. albopictus vary in the composition and functions depending on the type of pyrethroid insecticide they are applied. Four short chain fatty acid-producing genera, Butyricimonas, Prevotellaceae, Anaerococcus, Pseudorhodobacter were specifically absent in the pyrethroid-exposed mosquitoes. Morganella and Streptomyces were significantly enriched in cis-permethrin-exposed mosquitoes. Wolbachia and Chryseobacterium showed significant enrichment in β-cypermethrin-exposed mosquitoes. Pseudomonas was significantly abundant in deltamethrin-exposed mosquitoes. The significant proliferation of these bacteria may be closely related to insecticide metabolism. Our study recapitulated a specifically enhanced metabolic networks relevant to the exposure to cis-permethrin and β-cypermethrin, respectively. Benzaldehyde dehydrogenase (EC 1.2.1.28), key enzyme in aromatic compounds metabolism, was detected enhanced in cis-permethrin and β-cypermethrin exposed mosquitoes. The internal microbiota metabolism of aromatic compounds may be important influencing factors for pyrethroid resistance. Future work will be needed to elucidate the specific mechanisms by which mosquito microbiota influences host resistance and vector ability.}, } @article {pmid36700745, year = {2023}, author = {Indriani, C and Tanamas, SK and Khasanah, U and Ansari, MR and Rubangi, and Tantowijoyo, W and Ahmad, RA and Dufault, SM and Jewell, NP and Utarini, A and Simmons, CP and Anders, KL}, title = {Impact of randomised wmel Wolbachia deployments on notified dengue cases and insecticide fogging for dengue control in Yogyakarta City.}, journal = {Global health action}, volume = {16}, number = {1}, pages = {2166650}, pmid = {36700745}, issn = {1654-9880}, support = {R01 AI148127/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Humans ; *Aedes ; *Dengue/epidemiology/prevention & control ; *Dengue Virus ; *Insecticides ; *Wolbachia ; }, abstract = {BACKGROUND: Releases of Wolbachia (wMel)-infected Aedes aegypti mosquitoes significantly reduced the incidence of virologically confirmed dengue in a previous cluster randomised trial in Yogyakarta City, Indonesia. Following the trial, wMel releases were extended to the untreated control areas, to achieve city-wide coverage of Wolbachia.

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

CONCLUSIONS: Overall, this study provided important information regarding the Parascaris spp. microbiota and provides a first step towards determining whether the microbiota may be a viable target for future parasite control options.}, } @article {pmid36318064, year = {2022}, author = {Martinez, J and Ross, PA and Gu, X and Ant, TH and Murdochy, SM and Tong, L and da Silva Filipe, A and Hoffmann, AA and Sinkins, SP}, title = {Genomic and Phenotypic Comparisons Reveal Distinct Variants of Wolbachia Strain wAlbB.}, journal = {Applied and environmental microbiology}, volume = {88}, number = {22}, pages = {e0141222}, pmid = {36318064}, issn = {1098-5336}, support = {/WT_/Wellcome Trust/United Kingdom ; 108508/A/15/Z/WT_/Wellcome Trust/United Kingdom ; 202888/Z/16/Z/WT_/Wellcome Trust/United Kingdom ; MC_UU_12014/8/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Animals ; Humans ; *Wolbachia/genetics ; *Dengue Virus ; Mosquito Vectors ; *Aedes/microbiology ; *Dengue ; Genomics ; }, abstract = {The intracellular bacterium Wolbachia inhibits virus replication and is being harnessed around the world to fight mosquito-borne diseases through releases of mosquitoes carrying the symbiont. Wolbachia strains vary in their ability to invade mosquito populations and suppress viruses in part due to differences in their density within the insect and associated fitness costs. Using whole-genome sequencing, we demonstrate the existence of two variants in wAlbB, a Wolbachia strain being released in natural populations of Aedes aegypti mosquitoes. The two variants display striking differences in genome architecture and gene content. Differences in the presence/absence of 52 genes between variants include genes located in prophage regions and others potentially involved in controlling the symbiont's density. Importantly, we show that these genetic differences correlate with variation in wAlbB density and its tolerance to heat stress, suggesting that different wAlbB variants may be better suited for field deployment depending on local environmental conditions. Finally, we found that the wAlbB genome remained stable following its introduction in a Malaysian mosquito population. Our results highlight the need for further genomic and phenotypic characterization of Wolbachia strains in order to inform ongoing Wolbachia-based programs and improve the selection of optimal strains in future field interventions. IMPORTANCE Dengue is a viral disease transmitted by Aedes mosquitoes that threatens around half of the world population. Recent advances in dengue control involve the introduction of Wolbachia bacterial symbionts with antiviral properties into mosquito populations, which can lead to dramatic decreases in the incidence of the disease. In light of these promising results, there is a crucial need to better understand the factors affecting the success of such strategies, in particular the choice of Wolbachia strain for field releases and the potential for evolutionary changes. Here, we characterized two variants of a Wolbachia strain used for dengue control that differ at the genomic level and in their ability to replicate within the mosquito. We also found no evidence for the evolution of the symbiont within the 2 years following its deployment in Malaysia. Our results have implications for current and future Wolbachia-based health interventions.}, } @article {pmid36317957, year = {2023}, author = {Zhou, JC and Shang, D and Liu, SM and Zhang, C and Huo, LX and Zhang, LS and Dong, H}, title = {Wolbachia-infected Trichogramma dendrolimi is outcompeted by its uninfected counterpart in superparasitism but does not have developmental delay.}, journal = {Pest management science}, volume = {79}, number = {3}, pages = {1005-1017}, doi = {10.1002/ps.7269}, pmid = {36317957}, issn = {1526-4998}, support = {CAAS-ZDRW202108//Agricultural Science and Technology Innovation Program/ ; //Major Projects of China National Tobacco Corporation/ ; 32102275//National Natural Science Foundation of China/ ; 32172476//National Natural Science Foundation of China/ ; //Projects of Guizhou Tobacco Corporation/ ; 201936//Guizhou Tobacco Corporation/ ; 201937/WT_/Wellcome Trust/United Kingdom ; 201941/WT_/Wellcome Trust/United Kingdom ; 110202001032 (LS-01)//China National Tobacco Corporation/ ; 201937/WT_/Wellcome Trust/United Kingdom ; 201941/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Animals ; Female ; Oviposition ; *Wasps/microbiology ; *Wolbachia ; }, abstract = {BACKGROUND: Wolbachia infection increases the superparasitism frequency of Trichogramma females and provides an opportunity for horizontal intraspecific transmission. However, superparasitism may lead to interstrain competition between Wolbachia-infected Trichogramma offspring and their uninfected counterparts. This study investigated the outcome of interstrain intrinsic competition between Wolbachia-infected thelytokous strain (W) and uninfected bisexual strain (B) of Trichogramma dendrolimi. To determine the developmental rate of both strains, the sizes of immature stages of T. dendrolimi offspring at different times after parasitisation were measured in single parasitism and superparasitism conditions.

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

CONCLUSION: The results reconfirmed that W females presented a higher tendency for superparasitism than B females, and showed that B offspring outcompeted W offspring even when the latter was deposited 8 h earlier. The inferiority of Wolbachia-infected Trichogramma compared with their uninfected counterparts is not due to the developmental delay. This study provides insights into the effects of intrinsic competition on the control efficacy of Wolbachia-infected Trichogramma against pests in biological control programs. © 2022 Society of Chemical Industry.}, } @article {pmid36316689, year = {2022}, author = {Manvell, C and Berman, H and Callahan, B and Breitschwerdt, E and Swain, W and Ferris, K and Maggi, R and Lashnits, E}, title = {Identification of microbial taxa present in Ctenocephalides felis (cat flea) reveals widespread co-infection and associations with vector phylogeny.}, journal = {Parasites & vectors}, volume = {15}, number = {1}, pages = {398}, pmid = {36316689}, issn = {1756-3305}, support = {T32 GM133366/GM/NIGMS NIH HHS/United States ; T32 OD011130/OD/NIH HHS/United States ; 1T32GM133366/NH/NIH HHS/United States ; T32OD011130/NH/NIH HHS/United States ; }, mesh = {Animals ; Cats ; *Bartonella/genetics ; Cat Diseases/parasitology ; *Coinfection ; *Ctenocephalides/microbiology ; Flea Infestations/epidemiology/veterinary ; Phylogeny ; *Rickettsia/genetics ; }, abstract = {BACKGROUND: Ctenocephalides felis, the cat flea, is the most common ectoparasite of cats and dogs worldwide. As a cause of flea allergy dermatitis and a vector for two genera of zoonotic pathogens (Bartonella and Rickettsia spp.), the effect of the C. felis microbiome on pathogen transmission and vector survival is of substantial medical importance to both human and veterinary medicine. The aim of this study was to assay the pathogenic and commensal eubacterial microbial communities of individual C. felis from multiple geographic locations and analyze these findings by location, qPCR pathogen prevalence, and flea genetic diversity.

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

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

CONCLUSIONS: Our findings confirm the widespread coinfection of fleas with multiple bacterial genera and strains, proposing three additional microbiome members. The presence of minor Bartonella, Rickettsia, and Wolbachia ASVs was found to vary by location and flea haplotype. These findings have important implications for flea-borne pathogen transmission and control.}, } @article {pmid36314668, year = {2022}, author = {Roslan, MA and Ngui, R and Vythilingam, I and Wan Sulaiman, WY}, title = {Community surveillance of Aedes albopictus associated with Wolbachia detection in low-rise residential areas in Selangor, Malaysia.}, journal = {Journal of vector ecology : journal of the Society for Vector Ecology}, volume = {47}, number = {2}, pages = {142-152}, doi = {10.52707/1081-1710-47.2.142}, pmid = {36314668}, issn = {1948-7134}, mesh = {Animals ; *Wolbachia ; *Aedes ; Malaysia ; Temperature ; Polymerase Chain Reaction ; }, abstract = {The study assessed the distribution of Malaysian Ae. albopictus adults associated with Wolbachia detection in low-rise residential areas using a modified sticky ovitrap (MSO). The relationship between Ae. albopictus and climatological parameters were also determined. Fifty-two weeks of surveillance using 273 MSOs were conducted in four installation areas of eleven sampling sites. Specimens were subjected to PCR using wsp-specific primers for Wolbachia detection. The relationship between climatological parameters and Ae. albopictus captured were analyzed using Spearman rank correlation coefficient test. The majority of Ae. albopictus were captured in residential houses (87%), followed by playgrounds or parks (11.5%), guardhouses (1%), and community halls (0.5%). Most of the specimens (92%) were superinfected with wAlbA and wAlbB strains. A positive correlation with no significant association was found for rainfall (r = 0.015, P = 0.072), relative humidity (r = 0.005, P = 0.526), minimum temperature (r = 0.005, P = 0.516), and mean temperature (r = 0.003, P = 0.689). MSO effectively captured a high number of Ae. albopictus that was determined to be the predominant mosquito species found in low-rise residential areas. The adult collection is not only influenced by climatological parameters but also by other factors, including environmental conditions and general sanitation status.}, } @article {pmid36313820, year = {2022}, author = {Ryabinin, AS and Shishkina, OD and Ilinsky, YY and Bykov, RA}, title = {Rare Wolbachia genotypes in laboratory Drosophila melanogaster strains.}, journal = {Vavilovskii zhurnal genetiki i selektsii}, volume = {26}, number = {6}, pages = {553-559}, doi = {10.18699/VJGB-22-67}, pmid = {36313820}, issn = {2500-0462}, abstract = {Symbiotic bacteria of the genus Wolbachia are widespread in Drosophila melanogaster populations. Based on the polymorphism of the Wolbachia genome, the symbionts' diversity in D. melanogaster is presented by two groups: MEL (wMel, wMel2, wMel3 and wMel4) and CS (wMelCS and wMelCS2). The wMel genotype is predominant in natural D. melanogaster populations and is distributed all over the world. The CS genotypes, on the other hand, are of particular interest because it is unclear how they are maintained in the fruit f ly populations since they should have been eliminated from them due to their low frequency and genetic drift or been replaced by the wMel genotype. However, this is not what is really observed, which means these genotypes are supported by selection. It is known that the wMelPlus strain of the wMelCS genotype can increase the lifespan of infected f lies at high temperatures. The same genotype also increases the intensity of dopamine metabolism in Drosophila compared to the MEL-group genotypes. In the present study, we searched for the rare Wolbachia wMelCS and wMelCS2 genotypes, as well as for new genotypes in wild-type D. melanogaster strains and in several mutant laboratory strains. The symbiont was found in all populations, in 200 out of 385 wild-type strains and in 83 out of 170 mutant strains. Wolbachia diversity in D. melanogaster wild-type strains was represented by the wMel, wMelCS and wMelCS2 genotypes. More than 90 % of the infected strains carried wMel; 9 %, wMelCS2; and only two strains were found to carry wMelCS. No new Wolbachia genotypes were found. The northernmost point reported for the wMelCS2 genotype was Izhevsk city (Udmurtia, Russia). For the f irst time the wMelCS2 genotype was detected in D. melanogaster from the Sakhalin Island, and wMelCS, in the f lies from Nalchik (the North Caucasus). A comparison of Wolbachia genetic diversity between the wild-type laboratory strains and previously obtained data on mutant laboratory strains demonstrated differences in the frequencies of rare CS genotypes, which were more prevalent in mutant strains, apparently due to the breeding history of these Drosophila strains.}, } @article {pmid36304111, year = {2022}, author = {Ritchie, IT and Needles, KT and Leigh, BA and Kaur, R and Bordenstein, SR}, title = {Transgenic cytoplasmic incompatibility persists across age and temperature variation in Drosophila melanogaster.}, journal = {iScience}, volume = {25}, number = {11}, pages = {105327}, pmid = {36304111}, issn = {2589-0042}, support = {R01 AI132581/AI/NIAID NIH HHS/United States ; R01 AI143725/AI/NIAID NIH HHS/United States ; }, abstract = {Environmental stressors can impact the basic biology and applications of host-microbe symbioses. For example, Wolbachia symbiont densities and cytoplasmic incompatibility (CI) levels can decline in response to extreme temperatures and host aging. To investigate whether transgenic expression of CI-causing cif genes overcomes the environmental sensitivity of CI, we exposed transgenic male flies to low and high temperatures as well as aging treatments. Our results indicate that transgenic cif expression induces nearly complete CI regardless of temperature and aging, despite severe weakening of Wolbachia-based wild-type CI. Strong CI levels correlate with higher levels of cif transgene expression in young males. Altogether, our results highlight that transgenic CI persists against common environmental pressures and may be relevant for future control applications involving the cifA and cifB transgenes.}, } @article {pmid36303000, year = {2022}, author = {Kache, PA and Santos-Vega, M and Stewart-Ibarra, AM and Cook, EM and Seto, KC and Diuk-Wasser, MA}, title = {Bridging landscape ecology and urban science to respond to the rising threat of mosquito-borne diseases.}, journal = {Nature ecology & evolution}, volume = {6}, number = {11}, pages = {1601-1616}, pmid = {36303000}, issn = {2397-334X}, support = {U01 CK000509/CK/NCEZID CDC HHS/United States ; }, mesh = {Animals ; Humans ; Mosquito Vectors ; *Aedes ; Ecology ; Urbanization ; *Vector Borne Diseases/prevention & control ; }, abstract = {The prevalence of diseases borne by mosquitoes, particularly in the genus Aedes, is rising worldwide. This has been attributed, in part, to the dramatic rates of contemporary urbanization. While Aedes-borne disease risk varies within and between cities, few investigations use urban science-based approaches to examine how city structure and function contribute to vector or pathogen introduction and maintenance. Here, we integrate theories from complex adaptive systems, landscape ecology and urban geography to develop an urban systems framework for understanding Aedes-borne diseases. The framework establishes that cities comprise hierarchically structured patches of different land uses and characteristics. Properties of the patches (that is, composition) determine localized disease risk, while configuration and connectivity drive emergent patterns of pathogen spread. Complexity is added by incorporating individual and collective human social structures, considering how feedbacks among social actors and with the landscape drive risk and transmission. We discuss how these concepts apply to case studies of Aedes-borne disease from around the world. Ultimately, the framework strengthens existing theoretical and mixed qualitative-quantitative approaches, and advances considerations of how interventions including urban planning (for example, piped water provisioning) and emerging vector control strategies (for example, Wolbachia-infected mosquitoes) can be implemented to prevent and control the rising threat of Aedes-borne diseases.}, } @article {pmid36299486, year = {2022}, author = {Hargitai, D and Kenéz, L and Al-Lami, M and Szenczi, G and Lőrincz, P and Juhász, G}, title = {Autophagy controls Wolbachia infection upon bacterial damage and in aging Drosophila.}, journal = {Frontiers in cell and developmental biology}, volume = {10}, number = {}, pages = {976882}, pmid = {36299486}, issn = {2296-634X}, abstract = {Autophagy is a conserved catabolic process in eukaryotic cells that degrades intracellular components in lysosomes, often in an organelle-specific selective manner (mitophagy, ERphagy, etc). Cells also use autophagy as a defense mechanism, eliminating intracellular pathogens via selective degradation known as xenophagy. Wolbachia pipientis is a Gram-negative intracellular bacterium, which is one of the most common parasites on Earth affecting approximately half of terrestrial arthropods. Interestingly, infection grants the host resistance against other pathogens and modulates lifespan, so this bacterium resembles an endosymbiont. Here we demonstrate that Drosophila somatic cells normally degrade a subset of these bacterial cells, and autophagy is required for selective elimination of Wolbachia upon antibiotic damage. In line with these, Wolbachia overpopulates in autophagy-compromised animals during aging while its presence fails to affect host lifespan unlike in case of control flies. The autophagic degradation of Wolbachia thus represents a novel antibacterial mechanism that controls the propagation of this unique bacterium, behaving both as parasite and endosymbiont at the same time.}, } @article {pmid36296267, year = {2022}, author = {Ibrahim, YE and Paredes-Montero, JR and Al-Saleh, MA and Widyawan, A and He, R and El Komy, MH and Al Dhafer, HM and Kitchen, N and Gang, DR and Brown, JK}, title = {Characterization of the Asian Citrus Psyllid-'Candidatus Liberibacter Asiaticus' Pathosystem in Saudi Arabia Reveals Two Predominant CLas Lineages and One Asian Citrus Psyllid Vector Haplotype.}, journal = {Microorganisms}, volume = {10}, number = {10}, pages = {}, pmid = {36296267}, issn = {2076-2607}, support = {2018-70016-27411//USDA-NIFA CDRE/ ; 14-BIO-627-02//National Plan for Science, Technology, and Innovation (MAARIFAH), King Abdul-Aziz City for Science and Technology, Kingdom of Saudi Arabia/ ; }, abstract = {In Saudi Arabia (SA), the citrus greening disease is caused by 'Candidatus Liberibacter asiaticus' (CLas) transmitted by the Asian citrus psyllid (ACP) Diaphorina citri. The origin and route(s) of the ACP-CLas pathosystem invasion in SA have not been studied. Adult ACP were collected from citrus trees in SA and differentiated by analysis of the mitochondrial cytochrome oxidase I (mtCOI) and nuclear copper transporting protein (atox1) genes. A phylogenetic analysis of the Wolbachia spp. surface protein (wsp) gene was used to identify the ACP-associated Wolbachia spp. A phylogenetic analysis of the atox1 and mtCOI gene sequences revealed one predominant ACP haplotype most closely related to the Indian subcontinent founder populations. The detection and identification of CLas in citrus trees were carried out by polymerase chain reaction (PCR) amplification and sequencing of the 16S rDNA gene. The CLas-integrated prophage genomes were sequenced, annotated, and used to differentiate CLas populations. The ML and ASTRAL trees reconstructed with prophages type 1 and 2 genome sequences, separately and concatenated, resolved two major lineages, CLas-1 and -2. The CLas-1 clade, reported here for the first time, consisted of isolates from SA isolates and Pakistan. The CLas-2 sequences formed two groups, CLas-2-1 and -2-2, previously the 'Asiatic' and 'Floridian' strains, respectively. Members of CLas-2-1 originated from Southeast Asia, the USA, and other worldwide locations, while CLas-2-2 was identified only in Florida. This study provides the first snapshot into the status of the ACP-CLas pathosystem in SA. In addition, the results provide new insights into the pathosystem coevolution and global invasion histories of two ACP-CLas lineages with a predicted center of origin in South and Southeast Asia, respectively.}, } @article {pmid36296233, year = {2022}, author = {Ngnindji-Youdje, Y and Diarra, AZ and Lontsi-Demano, M and Tchuinkam, T and Parola, P}, title = {Detection of Tick-Borne Pathogens in Ticks from Cattle in Western Highlands of Cameroon.}, journal = {Microorganisms}, volume = {10}, number = {10}, pages = {}, pmid = {36296233}, issn = {2076-2607}, support = {ANR-10-IAHU-03//Méditerranée Infection Foundation/ ; }, abstract = {This study aimed to detect and identify microorganisms in ticks collected in the Western Highlands of Cameroon. Quantitative real-time and standard PCR assays, coupled with sequencing, were used. A total of 944 ticks collected from cattle in five distinct sites in Cameroon were selected for the analyses. They belonged to five genera (Amblyomma, Hyalomma, Rhipicephalus, Haemaphysalis, and Ixodes) and twelve species. Real-time PCR revealed that 23% (n = 218) of the ticks were positive for Rickettsia spp., 15% (n = 141) for bacteria of the Anaplasmataceae family, 3% (n = 29) for Piroplasmida, 0.5% (n = 5) for Coxiella burnetii, 0.4% (n = 4) for Borrelia spp., and 0.2% (n = 2) for Bartonella spp. The co-infection rate (3.4%, n = 32) involved mainly Rickettsia spp. and Anaplasmataceae. Of the Rickettsia spp. positive ticks, the targeted PCR and sequencing yielded Rickettsia africae (78.9%), Rickettsia aeschlimannii (6.4%), Rickettsia massiliae (7.8%), Candidatus Rickettsia barbariae (0.9%), and Rickettsia sp. (0.9%). Anaplasmataceae included Anaplasma marginale (4.3%), Anaplasma platys (1.4%), Anaplasma centrale (0.7%), Ehrlichia ruminantium (0.7%), Wolbachia sp., Candidatus Ehrlichia rustica (13.5%), Candidatus Ehrlichia urmitei (7%), and an uncultured Ehrlichia sp. (4.2%). Borrelia theileri was identified in one Rhipicephalus microplus tick. Unfortunately, Piroplasmida could not be identified to the species level. This study demonstrates that in Cameroon, ticks harbour a wide variety of microorganisms and present a risk of zoonotic diseases.}, } @article {pmid36292897, year = {2022}, author = {Pagendam, D and Elfekih, S and Nassar, MS and Nelson, S and Almalik, AM and Tawfik, EA and Al-Fageeh, MB and Hoffmann, AA}, title = {Spatio-Temporal Modelling Informing Wolbachia Replacement Releases in a Low Rainfall Climate.}, journal = {Insects}, volume = {13}, number = {10}, pages = {}, pmid = {36292897}, issn = {2075-4450}, support = {ETSC&KACST-CSIRO-2018-12-30-21//KACST-CSIRO collaborative research agreement/ ; 132412//National Health and Medical Research Council/ ; 1118640//National Health and Medical Research Council/ ; }, abstract = {Releases of Aedes aegypti carrying Wolbachia bacteria are known to suppress arbovirus transmission and reduce the incidence of vector-borne diseases. In planning for Wolbachia releases in the arid environment of Jeddah, Saudi Arabia, we collected entomological data with ovitraps across a 7-month period in four locations. Herein, we show that mosquito presence in basements does not differ from that of non-basement areas of buildings. In modelling mosquito presence across the study sites, we found the spatial structure to be statistically significant in one of the four sites, while a significant spatial structure was found for egg production data across three of the four sites. The length scales of the spatial covariance functions fitted to the egg production data ranged from 143 m to 574 m, indicating that high productivity regions can be extensive in size. Rank-correlation analyses indicated that mosquito presence tended to persist from the dry to wet season, but that egg production ranks at locations could reverse. The data suggest that, in Jeddah, the quality of the local environment for breeding can vary over time. The data support the feasibility of dry season releases but with release numbers needing to be flexible depending on local rates of invasion.}, } @article {pmid36292872, year = {2022}, author = {Buchori, D and Mawan, A and Nurhayati, I and Aryati, A and Kusnanto, H and Hadi, UK}, title = {Risk Assessment on the Release of Wolbachia-Infected Aedes aegypti in Yogyakarta, Indonesia.}, journal = {Insects}, volume = {13}, number = {10}, pages = {}, pmid = {36292872}, issn = {2075-4450}, support = {28-Mei-2016//Yayasan Tahija/ ; }, abstract = {Wolbachia-infected Aedes aegypti is the latest technology that was developed to eliminate dengue fever. The Ministry of Research and Technology of the Republic of Indonesia (Kemenristekdikti) established an expert group to identify future potential risks that may occur over a period of 30 years associated with the release of Wolbachia-infected Ae. aegypti. The risk assessment consisted of identifying different hazards that may have impacts on humans and the environment. From the consensus among the experts, there were 56 hazards identified and categorized into 4 components, namely, ecological matters, efficacy in mosquito management, economic and sociocultural issues, and public health standards. There were 19 hazards in the ecological group. The overall likelihood in the ecology of the mosquito is very low (0.05), with moderate consequence (0.74), which resulted in negligible risk. For the efficacy in mosquito management group, there were 12 hazards that resulted in very low likelihood (0.11) with high consequence (0.85). The overall risk for mosquito management efficacy was very low (0.09). There were 14 hazards identified in the public health standard with very low likelihood (0.07), moderate consequence (0.50) and negligible risk (0.04). Lastly, 13 hazards were identified in the economic and sociocultural group with low likelihood (0.01) but of moderate consequence (0.5), which resulted in a very low risk (0.09). The risk severity level of the four components leading to the endpoint risk of "cause more harm" due to releasing Wolbachia-infected Ae. aegypti is negligible (0.01).}, } @article {pmid36271174, year = {2022}, author = {Fallon, AM}, title = {Mitotically inactivated mosquito cells support robust Wolbachia infection and replication.}, journal = {In vitro cellular & developmental biology. Animal}, volume = {58}, number = {9}, pages = {780-787}, pmid = {36271174}, issn = {1543-706X}, mesh = {Animals ; *Wolbachia ; *Aedes ; Cell Line ; Mammals ; }, abstract = {Wolbachia is an obligate intracellular bacterium that infects many species of insects, and has been of particular interest in recent efforts to reduce disease transmission by mosquitoes. Two aspects of Wolbachia biology underlie its applications for insect control: first, the bacterium behaves as a natural gene drive agent and, second, when introduced into mosquitoes that do not harbor Wolbachia in nature, infection reduces survival of pathogens. These properties support efforts to explore the basic biology of Wolbachia in insect cell lines, which can produce sufficient infectious material for microbiological studies and microinjection into novel hosts. When introduced into naïve C7-10 Aedes albopictus mosquito cells, the yield of Wolbachia strain wStri improves, roughly in proportion to the size of the inoculum, as exponential growth of the host cell ceases. Wolbachia yields also increase when persistently infected C/wStri1 cells or naive, newly infected cells are treated with 20-hydroxyecdysone (20E), which inhibits growth in the G1 phase of the cell cycle. These observations suggest that Wolbachia infection and replication are independent of exponential growth and mitosis of host cells. To explore yields of infectious bacteria in cells arrested prior to infection, I tested host cells pre-treated with mitomycin C, an agent that crosslinks DNA and prevents cell division that is used to produce "feeder layers" with mammalian cells. Yields of wStri per plate increased by about 50-fold relative to exponentially growing cells, and the multiplicity of infection necessary for a robust infection was reduced to a single bacterium per cell. These results suggest that Wolbachia infection and replication are supported by mitotically arrested cells and provide new insights into biological processes that influence maintenance of a widespread obligate intracellular bacterium.}, } @article {pmid36270115, year = {2022}, author = {Araújo, IM and Cordeiro, MD and Soares, RFP and Guterres, A and Sanavria, A and Baêta, BA and da Fonseca, AH}, title = {Survey of bacterial and protozoan agents in ticks and fleas found on wild animals in the state of Rio de Janeiro, Brazil.}, journal = {Ticks and tick-borne diseases}, volume = {13}, number = {6}, pages = {102037}, doi = {10.1016/j.ttbdis.2022.102037}, pmid = {36270115}, issn = {1877-9603}, abstract = {This study evaluates the presence of bacterial and protozoan agents in ticks and fleas found on wild animals in the state of Rio de Janeiro, Brazil. These ectoparasites were collected on mammal species Hydrochoerus hydrochaeris, Tapirus terrestris, Dicotyles tajacu, Didelphis aurita, Cuniculus paca, Cerdocyon thous, and Coendou prehensilis, and on the terrestrial bird Dromaius novaehollandiae. Ticks and fleas were identified morphologically using specific taxonomic keys. A total of 396 ticks and 54 fleas were tested via polymerase chain reaction (PCR) for the presence of Rickettsia spp., Borrelia spp., microorganisms of the order Piroplasmida and Anaplasmataceae family. This total is distributed among nine tick species of the genus Amblyomma and one flea species. Rickettsia bellii was detected in Amblyomma dubitatum and Amblyomma pacae; Rickettsia sp. strain AL was found in Amblyomma longirostre; Rickettsia parkeri strain Atlantic rainforest was found in Amblyomma ovale; and "Candidatus Rickettsia senegalensis" and Rickettsia felis were detected in Ctenocephalides felis felis. Wolbachia sp. was detected in C. f. felis, and Borrelia sp. was detected in Amblyomma calcaratum (here named Borrelia sp. strain Acalc110). All tested samples were negative for Ehrlichia spp. and microorganisms of the Piroplasmida order. This study detected a new bacterial strain, Borrelia sp. strain Acalc 110 (which is genetically close to B. miyamotoi and B. venezuelensis) and the Rickettsia sp. strain 19P, which is 100% similar to "Ca. R. senegalensis", a bacterium recently discovered and now being reported for the first time in Brazil.}, } @article {pmid36250862, year = {2022}, author = {Huggins, LG and Colella, V and Atapattu, U and Koehler, AV and Traub, RJ}, title = {Nanopore Sequencing Using the Full-Length 16S rRNA Gene for Detection of Blood-Borne Bacteria in Dogs Reveals a Novel Species of Hemotropic Mycoplasma.}, journal = {Microbiology spectrum}, volume = {10}, number = {6}, pages = {e0308822}, pmid = {36250862}, issn = {2165-0497}, mesh = {Animals ; Dogs ; *Dog Diseases/diagnosis/epidemiology/microbiology ; Genes, rRNA ; High-Throughput Nucleotide Sequencing ; *Mycoplasma/classification/genetics ; *Nanopore Sequencing ; RNA, Ribosomal, 16S/genetics ; *Blood-Borne Pathogens/classification ; }, abstract = {Dogs across the globe are afflicted by diverse blood- and vector-borne bacteria (VBB), many of which cause severe disease and can be fatal. Diagnosis of VBB infections can be challenging due to the low concentration of bacteria in the blood, the frequent occurrence of coinfections, and the wide range of known, emerging, and potentially novel VBB species encounterable. Therefore, there is a need for diagnostics that address these challenges by being both sensitive and capable of detecting all VBB simultaneously. We detail the first employment of a nanopore-based sequencing methodology conducted on the Oxford Nanopore Technologies (ONT) MinION device to accurately elucidate the "hemobacteriome" from canine blood through sequencing of the full-length 16S rRNA gene. We detected a diverse range of important canine VBB, including Ehrlichia canis, Anaplasma platys, Mycoplasma haemocanis, Bartonella clarridgeiae, "Candidatus Mycoplasma haematoparvum", a novel species of hemotropic mycoplasma, and Wolbachia endosymbionts of filarial worms, indicative of filariasis. Our nanopore-based protocol was equivalent in sensitivity to both quantitative PCR (qPCR) and Illumina sequencing when benchmarked against these methods, achieving high agreement as defined by the kappa statistics (k > 0.81) for three key VBB. Utilizing the ability of the ONT' MinION device to sequence long read lengths provides an excellent alternative diagnostic method by which the hemobacteriome can be accurately characterized to the species level in a way previously unachievable using short reads. We envision our method to be translatable to multiple contexts, such as the detection of VBB in other vertebrate hosts, including humans, while the small size of the MinION device is highly amenable to field use. IMPORTANCE Blood- and vector-borne bacteria (VBB) can cause severe pathology and even be lethal for dogs in many regions across the globe. Accurate characterization of all the bacterial pathogens infecting a canine host is critical, as coinfections are common and emerging and novel pathogens that may go undetected by traditional diagnostics frequently arise. Deep sequencing using devices from Oxford Nanopore Technologies (ONT) provides a solution, as the long read lengths achievable provide species-level taxonomic identification of pathogens that previous short-read technologies could not accomplish. We developed a protocol using ONT' MinION sequencer to accurately detect and classify a wide spectrum of VBB from canine blood at a sensitivity comparable to that of regularly used diagnostics, such as qPCR. This protocol demonstrates great potential for use in biosurveillance and biosecurity operations for the detection of VBB in a range of vertebrate hosts, while the MinION sequencer's portability allows this method to be used easily in the field.}, } @article {pmid36247321, year = {2022}, author = {Tomaru, M and Takano-Shimizu-Kouno, T and Wakada, H}, title = {No Wolbachia infection was detected in Drosophila elegans collected from the wild in the Ryukyu Islands, Japan.}, journal = {microPublication biology}, volume = {2022}, number = {}, pages = {}, pmid = {36247321}, issn = {2578-9430}, abstract = {Flower breeding, tropical and subtropical Drosophila elegans is distributed in the Ryukyu Islands and Taiwan (black morph) and in southern China, Philippines, Indonesia, and New Guinea (brown morph). Although reproductive and behavioral manipulations by Wolbachia are reported in many insect taxa, Wolbachia infection in D. elegans is unclear. There is only a report of no Wolbachia detected in a laboratory strain of brown morph. This PCR diagnosis study revealed no Wolbachia infection in D. elegans males collected from the wild in the Ryukyu Islands. We concluded that D. elegans black morph in the Ryukyu Islands is not infected with Wolbachia .}, } @article {pmid36244506, year = {2022}, author = {Djoukzoumka, S and Mahamat Hassane, H and Khan Payne, V and Ibrahim, MAM and Tagueu Kanté, S and Mouliom Mfopit, Y and Berger, P and Kelm, S and Simo, G}, title = {Sodalis glossinidius and Wolbachia infections in wild population of Glossina morsitans submorsitans caught in the area of Lake Iro in the south of Chad.}, journal = {Journal of invertebrate pathology}, volume = {195}, number = {}, pages = {107835}, doi = {10.1016/j.jip.2022.107835}, pmid = {36244506}, issn = {1096-0805}, mesh = {Animals ; *Tsetse Flies/microbiology ; *Wolbachia ; Lakes ; Chad ; *Trypanosoma/genetics ; Symbiosis ; }, abstract = {Investigations on the bacterial fauna and their association with trypanosome infections in tsetse fly have revealed contrasting results. This study aimed to detect Wolbachia and S. glossinidius in wild populations of G. m. submorsistans and subsequently, understand the influence that these bacteria may have on the vectorial competence of this tsetse species. Tsetse flies were captured in the area of Lake Iro in the south of Chad using biconical traps. After DNA extraction from each tsetse fly, Sodalis glossinidius and Wolbachia were detected using specific primers. Sodalis glossinidius and Wolbachia infection rates were compared and association studies involving trypanosome infections and S. glossinidius or Wolbachia were performed. From 345 G. m. submorsitans analyzed, 9.0% and 14.5% were respectively infected with S. glossinidius and Wolbachia. Only 2.31% of all tsetse flies were co-infected by the 2 bacteria. Of all trypanosome-infected flies, 7.1% and 9.8% harbored, respectively, S. glossinidius and Wolbachia. No association was observed between Wolbachia and trypanosome infections while a significant association (r = 4.992; P = 0.025) was found between S. glossinidius and the presence of trypanosomes. A significant association (r = 3.147; P = 0.043) was also observed between S. glossinidius and T. simiae; and none with T. congolense or T. godfreyi. This study revealed S. glossinidius and Wolbachia in G. m. submorsitans of the area of lake Iro. It showed that co-infections between Wolbachia and S. glossinidius are rare in wild populations of G. m. submorsitans and that the tripartite associations vary according to trypanosome species as well as symbiotic mricroorganisms.}, } @article {pmid36227923, year = {2022}, author = {Hugo, LE and Rašić, G and Maynard, AJ and Ambrose, L and Liddington, C and Thomas, CJE and Nath, NS and Graham, M and Winterford, C and Wimalasiri-Yapa, BMCR and Xi, Z and Beebe, NW and Devine, GJ}, title = {Wolbachia wAlbB inhibit dengue and Zika infection in the mosquito Aedes aegypti with an Australian background.}, journal = {PLoS neglected tropical diseases}, volume = {16}, number = {10}, pages = {e0010786}, pmid = {36227923}, issn = {1935-2735}, mesh = {*Aedes ; Animals ; Australia ; DNA ; *Dengue/prevention & control ; *Dengue Virus/physiology ; Humans ; *Insecticides ; Male ; Mosquito Vectors ; *Wolbachia/physiology ; *Zika Virus/genetics ; *Zika Virus Infection/prevention & control ; }, abstract = {Biological control of mosquito vectors using the endosymbiotic bacteria Wolbachia is an emerging strategy for the management of human arboviral diseases. We recently described the development of a strain of Aedes aegypti infected with the Wolbachia strain wAlbB (referred to as the wAlbB2-F4 strain) through simple backcrossing of wild type Australian mosquitoes with a wAlbB infected Ae. aegypti strain from the USA. Field releases of male wAlbB2-F4 mosquitoes resulted in the successful suppression of wild populations of mosquitoes in the trial sites by exploiting the strain's Wolbachia-induced cytoplasmic incompatibility. We now demonstrate that the strain is resistant to infection by dengue and Zika viruses and is genetically similar to endemic Queensland populations. There was a fourfold reduction in the proportion of wAlbB2-F4 mosquitoes that became infected following a blood meal containing dengue 2 virus (16.7%) compared to wild type mosquitoes (69.2%) and a 6-7 fold reduction in the proportion of wAlbB2-F4 mosquitoes producing virus in saliva following a blood meal containing an epidemic strain of Zika virus (8.7% in comparison to 58.3% in wild type mosquitoes). Restriction-site Associated DNA (RAD) sequencing revealed that wAlbB2-F4 mosquitoes have > 98% Australian ancestry, confirming the successful introduction of the wAlbB2 infection into the Australian genomic background through backcrossing. Genotypic and phenotypic analyses showed the wAlbB2-F4 strain retains the insecticide susceptible phenotype and genotype of native Australian mosquitoes. We demonstrate that the Wolbachia wAlbB2-F4, in addition to being suitable for population suppression programs, can also be effective in population replacement programs given its inhibition of virus infection in mosquitoes. The ease at which a target mosquito population can be transfected with wAlbB2, while retaining the genotypes and phenotypes of the target population, shows the utility of this strain for controlling the Ae. aegypti mosquitoes and the pathogens they transmit.}, } @article {pmid36214563, year = {2022}, author = {Zhou, JC and Zhao, X and Huo, LX and Shang, D and Dong, H and Zhang, LS}, title = {Wolbachia-Driven Memory Loss in a Parasitic Wasp Increases Superparasitism to Enhance Horizontal Transmission.}, journal = {mBio}, volume = {13}, number = {6}, pages = {e0236222}, pmid = {36214563}, issn = {2150-7511}, mesh = {Humans ; Animals ; Female ; *Wasps/microbiology ; *Wolbachia/genetics ; In Situ Hybridization, Fluorescence ; Larva ; Memory Disorders ; }, abstract = {Horizontal transmission of the endosymbiont, Wolbachia, may occur during superparasitism when parasitoid females deposit a second clutch of eggs on a host. Wolbachia may increase the superparasitism tendency of Trichogramma wasps by depriving their memory. To test this hypothesis, we investigated the effects of conditioning experience and memory inhibitors (actinomycin D [ACD] and anisomycin [ANI]) on memory capacity, and expressions of memory-related genes (CREB1 and PKA), and superparasitism frequency of Wolbachia-infected (TDW) and uninfected (TD) lines of Trichogramma dendrolimi after conditioning with lemon or peppermint odor. We detected the presence of Wolbachia in eggs, larvae, pre-pupae, pupae, and adults of Trichogramma by using fluorescence in situ hybridization. The results showed that TDW females had a more reduced memory capacity than TD females after conditioning. Compared with TD females, TDW females showed a higher proportion of superparasitism and a downregulation of CREB1 and PKA genes after conditioning. TD females fed ACD or ANI showed a higher tendency for superparasitism and a downregulation of CREB1 and PKA, along with memory loss after conditioning than TD females fed honey solution only. The presence of Wolbachia was detected in the anterior region of the larva, pre-pupa, and pupa, but was not found in the head of the adult. The results provide evidence of host behavioral manipulation of Wolbachia by depriving memory of host Trichogramma wasps based on Poulin' s criteria. These host behavioral changes led by Wolbachia may be caused by the virulence of Wolbachia on the nervous system of the host. IMPORTANCE The endosymbiotic bacteria, Wolbachia, live widely within cells of arthropods. Wolbachia are not only transmitted vertically from host mother to offspring, but are also transmitted horizontally among host individuals. Horizontal transmission is expected to occur during superparasitism when host parasitoid females deposit a clutch of eggs on a host previously parasitized by the same parasitoid species. Thus, a question is proposed regarding whether superparasitism behavior is a behavior modification induced by the symbiont to favor symbiont transmission. This study highlights behavioral mechanisms of Wolbachia-induced superparasitism in Trichogramma wasps and the manipulation of symbionts on host parasitoids.}, } @article {pmid36205550, year = {2023}, author = {Ant, TH and Mancini, MV and McNamara, CJ and Rainey, SM and Sinkins, SP}, title = {Wolbachia-Virus interactions and arbovirus control through population replacement in mosquitoes.}, journal = {Pathogens and global health}, volume = {117}, number = {3}, pages = {245-258}, pmid = {36205550}, issn = {2047-7732}, support = {/WT_/Wellcome Trust/United Kingdom ; 108508/A/15/Z/WT_/Wellcome Trust/United Kingdom ; 202888/Z/16/Z/WT_/Wellcome Trust/United Kingdom ; MC_UU_12014/8/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Animals ; Humans ; *Arboviruses/physiology ; *Wolbachia ; *Dengue Virus/physiology ; Mosquito Vectors ; *Aedes ; *Zika Virus ; *Zika Virus Infection ; *Dengue/prevention & control ; Population Dynamics ; }, abstract = {Following transfer into the primary arbovirus vector Aedes aegypti, several strains of the intracellular bacterium Wolbachia have been shown to inhibit the transmission of dengue, Zika, and chikungunya viruses, important human pathogens that cause significant morbidity and mortality worldwide. In addition to pathogen inhibition, many Wolbachia strains manipulate host reproduction, resulting in an invasive capacity of the bacterium in insect populations. This has led to the deployment of Wolbachia as a dengue control tool, and trials have reported significant reductions in transmission in release areas. Here, we discuss the possible mechanisms of Wolbachia-virus inhibition and the implications for long-term success of dengue control. We also consider the evidence presented in several reports that Wolbachia may cause an enhancement of replication of certain viruses under particular conditions, and conclude that these should not cause any concerns with respect to the application of Wolbachia to arbovirus control.}, } @article {pmid36201377, year = {2022}, author = {Kiefer, JST and Schmidt, G and Krüsemer, R and Kaltenpoth, M and Engl, T}, title = {Wolbachia causes cytoplasmic incompatibility but not male-killing in a grain pest beetle.}, journal = {Molecular ecology}, volume = {31}, number = {24}, pages = {6570-6587}, doi = {10.1111/mec.16717}, pmid = {36201377}, issn = {1365-294X}, mesh = {Animals ; Male ; Female ; *Wolbachia/genetics ; *Coleoptera/genetics/microbiology ; Drosophila melanogaster/genetics ; Phylogeny ; Cytoplasm/genetics/microbiology ; Symbiosis/genetics ; }, abstract = {The endosymbiotic Wolbachia is one of the most common intracellular bacteria known in arthropods and nematodes. Its ability for reproductive manipulation can cause unequal inheritance to male and female offspring, allowing the manipulator to spread, but potentially also impact the evolutionary dynamics of infected hosts. Estimated to be present in up to 66% of insect species, little is known about the phenotypic impact of Wolbachia within the order Coleoptera. Here, we describe the reproductive manipulation by the Wolbachia strain wSur harboured by the sawtoothed grain beetle Oryzaephilus surinamensis (Coleoptera, Silvanidae), through a combination of genomics approaches and bioassays. The Wolbachia strain wSur belongs to supergroup B that contains well-described reproductive manipulators of insects and encodes a pair of cytoplasmic incompatibility factor (cif) genes, as well as multiple homologues of the WO-mediated killing (wmk) gene. A phylogenetic comparison with wmk homologues of wMel of Drosophila melanogaster identified 18 wmk copies in wSur, including one that is closely related to the wMel male-killing homologue. However, further analysis of this particular wmk gene revealed an eight-nucleotide deletion leading to a stop-codon and subsequent reading frame shift midsequence, probably rendering it nonfunctional. Concordantly, utilizing a Wolbachia-deprived O. surinamensis population and controlled mating pairs of wSur-infected and noninfected partners, we found no experimental evidence for male-killing. However, a significant ~50% reduction of hatching rates in hybrid crosses of uninfected females with infected males indicates that wSur is causing cytoplasmic incompatibility. Thus, Wolbachia also represents an important determinant of host fitness in Coleoptera.}, } @article {pmid36200325, year = {2022}, author = {Ross, PA and Hoffmann, AA}, title = {Fitness costs of Wolbachia shift in locally-adapted Aedes aegypti mosquitoes.}, journal = {Environmental microbiology}, volume = {24}, number = {12}, pages = {5749-5759}, pmid = {36200325}, issn = {1462-2920}, mesh = {Animals ; Female ; *Wolbachia/genetics ; *Aedes/genetics ; }, abstract = {Aedes aegypti mosquito eggs can remain quiescent for many months before hatching, allowing populations to persist through unfavourable conditions. A. aegypti infected with the Wolbachia strain wMel have been released in tropical and subtropical regions for dengue control. wMel reduces the viability of quiescent eggs, but this physiological cost might be expected to evolve in natural mosquito populations that frequently experience stressful conditions. We found that the cost of wMel infection differed consistently between mosquitoes collected from different locations and became weaker across laboratory generations, suggesting environment-specific adaptation of mosquitoes to the wMel infection. Reciprocal crossing experiments show that differences in the cost of wMel to quiescent egg viability were mainly due to mosquito genetic background and not Wolbachia origin. wMel-infected mosquitoes hatching from long-term quiescent eggs showed partial loss of cytoplasmic incompatibility and female infertility, highlighting additional costs of long-term quiescence. Our study provides the first evidence for a shift in Wolbachia phenotypic effects following deliberate field release and establishment and it highlights interactions between Wolbachia infections and mosquito genetic backgrounds. The unexpected changes in fitness costs observed here suggest potential tradeoffs with undescribed fitness benefits of the wMel infection.}, } @article {pmid36192576, year = {2022}, author = {Štarhová Serbina, L and Gajski, D and Malenovský, I and Corretto, E and Schuler, H and Dittmer, J}, title = {Wolbachia infection dynamics in a natural population of the pear psyllid Cacopsylla pyri (Hemiptera: Psylloidea) across its seasonal generations.}, journal = {Scientific reports}, volume = {12}, number = {1}, pages = {16502}, pmid = {36192576}, issn = {2045-2322}, mesh = {Animals ; Female ; *Hemiptera/genetics ; Male ; Phylogeny ; *Pyrus ; Seasons ; *Wolbachia/genetics ; }, abstract = {Wolbachia is one of the most abundant intracellular symbionts of arthropods and has profound effects on host biology. Wolbachia transmission and host phenotypes often depend on its density within the host, which can be affected by multiple biotic and abiotic factors. However, very few studies measured Wolbachia density in natural host populations. Here, we describe Wolbachia in the pear psyllid Cacopsylla pyri from three populations in the Czech Republic. Using phylogenetic analyses based on wsp and multilocus sequence typing genes, we demonstrate that C. pyri harbours three new Wolbachia strains from supergroup B. A fourth Wolbachia strain from supergroup A was also detected in parasitised immatures of C. pyri, but likely came from a hymenopteran parasitoid. To obtain insights into natural Wolbachia infection dynamics, we quantified Wolbachia in psyllid individuals from the locality with the highest prevalence across an entire year, spanning several seasonal generations of the host. All tested females were infected and Wolbachia density remained stable across the entire period, suggesting a highly efficient vertical transmission and little influence from the environment and different host generations. In contrast, we observed a tendency towards reduced Wolbachia density in males which may suggest sex-related differences in Wolbachia-psyllid interactions.}, } @article {pmid36182681, year = {2022}, author = {Ooi, EE and Wilder-Smith, A}, title = {Externalities modulate the effectiveness of the Wolbachia release programme.}, journal = {The Lancet. Infectious diseases}, volume = {22}, number = {11}, pages = {1518-1519}, doi = {10.1016/S1473-3099(22)00497-2}, pmid = {36182681}, issn = {1474-4457}, mesh = {Animals ; Humans ; *Wolbachia ; *Aedes ; *Dengue ; }, } @article {pmid36182679, year = {2022}, author = {Ribeiro Dos Santos, G and Durovni, B and Saraceni, V and Souza Riback, TI and Pinto, SB and Anders, KL and Moreira, LA and Salje, H}, title = {Estimating the effect of the wMel release programme on the incidence of dengue and chikungunya in Rio de Janeiro, Brazil: a spatiotemporal modelling study.}, journal = {The Lancet. Infectious diseases}, volume = {22}, number = {11}, pages = {1587-1595}, pmid = {36182679}, issn = {1474-4457}, mesh = {Humans ; Animals ; *Chikungunya Fever/epidemiology/prevention & control ; *Dengue Virus ; Incidence ; Brazil/epidemiology ; *Wolbachia ; *Aedes ; *Dengue/epidemiology/prevention & control ; Mosquito Vectors ; }, abstract = {BACKGROUND: Introgression of genetic material from species of the insect bacteria Wolbachia into populations of Aedes aegypti mosquitoes has been shown in randomised and non-randomised trials to reduce the incidence of dengue; however, evidence for the real-world effectiveness of large-scale deployments of Wolbachia-infected mosquitoes for arboviral disease control in endemic settings is still scarce. A large Wolbachia (wMel strain) release programme was implemented in 2017 in Rio de Janeiro, Brazil. We aimed to assess the effect of this programme on the incidence of dengue and chikungunya in the city.

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

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

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

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

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

CONCLUSION: Together, these results indicate that SHOT1s are positively correlated with Wolbachia density and account for Wolbachia-mediated phenotypes. Our results provide new evidence that herbivore salivary proteins are related to Wolbachia-mediated manipulations of host performance on plants. © 2022 Society of Chemical Industry.}, } @article {pmid36149408, year = {2022}, author = {Warecki, B and Titen, SWA and Alam, MS and Vega, G and Lemseffer, N and Hug, K and Minden, JS and Sullivan, W}, title = {Wolbachia action in the sperm produces developmentally deferred chromosome segregation defects during the Drosophila mid-blastula transition.}, journal = {eLife}, volume = {11}, number = {}, pages = {}, pmid = {36149408}, issn = {2050-084X}, support = {R35 GM139595/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Blastula ; Chromatin ; Chromosome Segregation ; Cytoplasm ; Drosophila/genetics ; Female ; In Situ Hybridization, Fluorescence ; Male ; Semen ; Spermatozoa ; *Wolbachia/genetics ; }, abstract = {Wolbachia, a vertically transmitted endosymbiont infecting many insects, spreads rapidly through uninfected populations by a mechanism known as cytoplasmic incompatibility (CI). In CI, a paternally delivered modification of the sperm leads to chromatin defects and lethality during and after the first mitosis of embryonic development in multiple species. However, whether CI-induced defects in later stage embryos are a consequence of the first division errors or caused by independent defects remains unresolved. To address this question, we focused on ~1/3 of embryos from CI crosses in Drosophila simulans that develop apparently normally through the first and subsequent pre-blastoderm divisions before exhibiting mitotic errors during the mid-blastula transition and gastrulation. We performed single embryo PCR and whole genome sequencing to find a large percentage of these developed CI-derived embryos bypass the first division defect. Using fluorescence in situ hybridization, we find increased chromosome segregation errors in gastrulating CI-derived embryos that had avoided the first division defect. Thus, Wolbachia action in the sperm induces developmentally deferred defects that are not a consequence of the first division errors. Like the immediate defect, the delayed defect is rescued through crosses to infected females. These studies inform current models on the molecular and cellular basis of CI.}, } @article {pmid36147860, year = {2022}, author = {Guo, L and Tang, C and Gao, C and Li, Z and Cheng, Y and Chen, J and Wang, T and Xu, J}, title = {Bacterial and fungal communities within and among geographic samples of the hemp pest Psylliodes attenuata from China.}, journal = {Frontiers in microbiology}, volume = {13}, number = {}, pages = {964735}, pmid = {36147860}, issn = {1664-302X}, abstract = {The hemp flea beetle Psylliodes attenuata (Coleoptera: Chrysomelidae: Psylliodes) is a common pest of Cannabis sativa, including cultivars of both medicinal marijuana and industrial hemp. Both the larval and adult stages of this beetle can cause significant damages to C. sativa, resulting in substantial crop losses. At present, little is known about the bacterial and fungal community diversity among populations of this pest insect. In the present study, we obtained P. attenuata samples from nine field sites representing broad industrial hemp productions in China and analyzed their microbial communities using DNA metabarcoding. Bacterial sequences of all the samples were assigned to 3728 OTUs, which belonged to 45 phyla, 1058 genera and 1960 known species. The most common genera were Rickettsia, Wolbachia, and Candidatus_Brownia. Fungal sequences of all the samples were assigned to 910 OTUs, which belonged to 9 phyla, 308 genera and 464 known species. The most common fungal genera were Cladosporium, Cutaneotrichosporon, and Aspergillus. Principal coordinate analysis revealed a significant difference in the bacterial and fungal community structure among the nine P. attenuata populations. Understanding the microbial symbionts may provide clues to help develop potential biocontrol techniques against this pest.}, } @article {pmid36147838, year = {2022}, author = {Liu, YH and Ma, YM and Tian, HO and Yang, B and Han, WX and Zhao, WH and Chai, HL and Zhang, ZS and Wang, LF and Chen, L and Xing, Y and Ding, YL and Zhao, L}, title = {First determination of DNA virus and some additional bacteria from Melophagus ovinus (sheep ked) in Tibet, China.}, journal = {Frontiers in microbiology}, volume = {13}, number = {}, pages = {988136}, pmid = {36147838}, issn = {1664-302X}, abstract = {Melophagus ovinus (sheep ked) is one of the common ectoparasites in sheep. In addition to causing direct damage to the host through biting and sucking blood, sheep ked is a potential vector of helminths, protozoa, bacteria, and viruses. Sheep M. ovinus samples from three regions in Tibet were selected for DNA extraction. The 16S rDNA V3-V4 hypervariable region was amplified, after genomic DNA fragmentation, Illumina Hiseq libraries were constructed. The 16S rRNA sequencing and viral metagenomics sequencing were separately conducted on the Illumina Novaseq 6000 platform and molecular biology software and platforms were employed to analyze the sequencing data. Illumina PE250 sequencing results demonstrated that the dominant bacteria phylum in M. ovinus from Tibet, China was Proteobacteria, where 29 bacteria genera were annotated. The dominant bacterial genera were Bartonella, Wolbachia, and Arsenophonus; Bartonella chomelii, Wolbachia spp., and Arsenophonus spp. were the dominant bacterial species in M. ovinus from Tibet, China. We also detected Kluyvera intermedia, Corynebacterium maris DSM 45190, Planomicrobium okeanokoites, and Rhodococcus erythropolis, of which the relative abundance of Kluyvera intermedia was high. Illumina Hiseq sequencing results demonstrated that 4 virus orders were detected in M. ovinus from Tibet, China, and 3 samples were annotated into 29 families, 30 families, and 28 families of viruses, respectively. Virus families related to vertebrates and insects mainly included Mimiviridae, Marseilleviridae, Poxviridae, Ascoviridae, Iridoviridae, Baculoviridae, Hytrosaviridae, Nudiviridae, Polydnaviridae, Adomaviridae, Asfarviridae, Hepeviridae, Herpesviridae, and Retroviridae; at the species level, the relative abundance of Tupanvirus_soda_lake, Klosneuvirus_KNV1, and Indivirus_ILV1 was higher. African swine fever virus and many poxviruses from the family Poxviridae were detected, albeit their relative abundance was low. The dominant bacterial phylum of M. ovinus from Tibet, China was Proteobacteria, and the dominant bacterial genera were Bartonella, Wolbachia, and Arsenophonus, where 23 out of 29 annotated bacteria genera were first reported in M. ovinus. Kluyvera intermedia, Corynebacterium maris DSM 45190, Planomicrobium okeanokoites, and Rhodococcus erythropolis were detected for the first time. All DNA viruses detected in this study have been reported in M. ovinus for the first time.}, } @article {pmid36138209, year = {2023}, author = {Liu, Q and Zhang, H and Huang, X}, title = {Strong Linkage Between Symbiotic Bacterial Community and Host Age and Morph in a Hemipteran Social Insect.}, journal = {Microbial ecology}, volume = {86}, number = {2}, pages = {1213-1225}, pmid = {36138209}, issn = {1432-184X}, support = {31970446//National Natural Science Foundation of China/ ; }, mesh = {Animals ; Female ; Biological Evolution ; *Aphids/microbiology ; Insecta ; Symbiosis/physiology ; *Buchnera/genetics ; }, abstract = {The relationships between symbionts and insects are complex, and symbionts usually have diverse ecological and evolutionary effects on their hosts. The phloem sap-sucking aphids are good models to study the interactions between insects and symbiotic microorganisms. Although aphids usually exhibit remarkable life cycle complexity, most previous studies on symbiotic diversity sampled only apterous viviparous adult females or very few morphs. In this study, high-throughput 16S rDNA amplicon sequencing was used to assess the symbiotic bacterial communities of eleven morphs or developmental stages of the social aphid Pseudoregma bambucicola. We found there were significant differences in bacterial composition in response to different morphs and developmental stages, and for the first time, we revealed male aphids hosted very different symbiotic composition featured with low abundance of dominant symbionts but high diversity of total symbionts. The relative abundance of Pectobacterium showed relatively stable across different types of samples, while that of Wolbachia fluctuated greatly, indicating the former may have a consistent function in this species and the latter may provide specific function for certain morphs or developmental stages. Our study presents new evidence of complexity of symbiotic associations and indicates strong linkage between symbiotic bacterial community and host age and morph.}, } @article {pmid36135489, year = {2022}, author = {Andrianto, E and Kasai, A}, title = {Wolbachia in Black Spiny Whiteflies and Their New Parasitoid Wasp in Japan: Evidence of the Distinct Infection Status on Aleurocanthus camelliae Cryptic Species Complex.}, journal = {Insects}, volume = {13}, number = {9}, pages = {}, pmid = {36135489}, issn = {2075-4450}, abstract = {Wolbachia, an alphaproteobacterial reproductive parasite, can cause profound mitochondrial divergence in insects, which might eventually be a part of cryptic speciation. Aleurocanthus camelliae is a cryptic species complex consisting of several morphospecies and/or haplotypes that are genetically different but morphologically indistinctive. However, little is known about the Wolbachia infection status in these tea and Citrus pests. Thus, this study aimed to profile the diversity and phenotypic characteristics of Wolbachia natural infections in the A. camelliae cryptic species complex. A monophyletic strain of Wolbachia that infected the A. camelliae cryptic species complex (wAlec) with different patterns was discovered. Whiteflies that are morphologically identical to Aleurocanthus spiniferus (Aleurocanthus cf. A. spiniferus in Eurya japonica and A. spiniferus in Citrus) were grouped into uninfected populations, whereas the fixed infection was detected in A. camelliae B1 from Theaceae. The rapid evolution of wAlec was also found to occur through a high recombination event, which produced subgroups A and B in wAlec. It may also be associated with the non-cytoplasmic incompatibility (CI) phenotype of wAlec due to undetectable CI-related genes from phage WO (WOAlec). The current discovery of a novel cryptic species of A. camelliae led to a discussion about the oscillation hypothesis, which may provide insights on cryptic speciation, particularly on how specialization and host expansion have been recorded among these species. This study also identified a parasitoid wasp belonging to the genus Eretmocerus in A. camelliae, for the first time in Japan.}, } @article {pmid36125236, year = {2023}, author = {Brinker, P and Chen, F and Chehida, YB and Beukeboom, LW and Fontaine, MC and Salles, JF}, title = {Microbiome composition is shaped by geography and population structure in the parasitic wasp Asobara japonica, but not in the presence of the endosymbiont Wolbachia.}, journal = {Molecular ecology}, volume = {32}, number = {23}, pages = {6644-6658}, doi = {10.1111/mec.16699}, pmid = {36125236}, issn = {1365-294X}, mesh = {Animals ; *Wasps/genetics/microbiology ; *Wolbachia/genetics ; RNA, Ribosomal, 16S/genetics ; *Microbiota/genetics ; Bacteria/genetics ; Geography ; }, abstract = {The microbial community composition is crucial for diverse life-history traits in many organisms. However, we still lack a sufficient understanding of how the host microbiome is acquired and maintained, a pressing issue in times of global environmental change. Here we investigated to what extent host genotype, environmental conditions, and the endosymbiont Wolbachia influence the bacterial communities in the parasitic wasp Asobara japonica. We sampled multiple wasp populations across 10 locations in their natural distribution range in Japan and sequenced the host genome (whole genome sequencing) and microbiome (16S rRNA gene). We compared the host population structure and bacterial community composition of wasps that reproduce sexually and are uninfected with Wolbachia with wasps that reproduce asexually and carry Wolbachia. The bacterial communities in asexual wasps were highly similar due to a strong effect of Wolbachia rather than host genomic structure. In contrast, in sexual wasps, bacterial communities appear primarily shaped by a combination of population structure and environmental conditions. Our research highlights that multiple factors shape the bacterial communities of an organism and that the presence of a single endosymbiont can strongly alter their compositions. This information is crucial to understanding how organisms and their associated microbiome will react in the face of environmental change.}, } @article {pmid36124585, year = {2022}, author = {Chinnathambi, R and Rihan, FA}, title = {Analysis and control of Aedes Aegypti mosquitoes using sterile-insect techniques with Wolbachia.}, journal = {Mathematical biosciences and engineering : MBE}, volume = {19}, number = {11}, pages = {11154-11171}, doi = {10.3934/mbe.2022520}, pmid = {36124585}, issn = {1551-0018}, mesh = {*Aedes ; Animals ; Humans ; *Infertility ; Male ; Mosquito Control/methods ; Mosquito Vectors ; *Wolbachia ; }, abstract = {Combining Sterile and Incompatible Insect techniques can significantly reduce mosquito populations and prevent the transmission of diseases between insects and humans. This paper describes impulsive differential equations for the control of a mosquito with Wolbachia. Several interesting conditions are created when sterile male mosquitoes are released impulsively, ensuring both open- and closed-loop control. To determine the wild mosquito population size in real-time, we propose an open-loop control system, which uses impulsive and constant releases of sterile male mosquitoes. A closed-loop control scheme is also being investigated, which specifies the release of sterile mosquitoes according to the size of the wild mosquito population. To eliminate or reduce a mosquito population below a certain threshold, the Sterile insect technique involves mass releases of sterile insects. Numerical simulations verify the theoretical results.}, } @article {pmid36121852, year = {2022}, author = {Martinez, J and Ant, TH and Murdochy, SM and Tong, L and da Silva Filipe, A and Sinkins, SP}, title = {Genome sequencing and comparative analysis of Wolbachia strain wAlbA reveals Wolbachia-associated plasmids are common.}, journal = {PLoS genetics}, volume = {18}, number = {9}, pages = {e1010406}, pmid = {36121852}, issn = {1553-7404}, support = {/WT_/Wellcome Trust/United Kingdom ; 108508/A/15/Z/WT_/Wellcome Trust/United Kingdom ; 202888/Z/16/Z/WT_/Wellcome Trust/United Kingdom ; MC_UU_12014/8/MRC_/Medical Research Council/United Kingdom ; }, mesh = {*Aedes/genetics ; Animals ; Plasmids/genetics ; Prophages/genetics ; Symbiosis/genetics ; *Wolbachia/genetics ; }, abstract = {Wolbachia are widespread maternally-transmitted bacteria of arthropods that often spread by manipulating their host's reproduction through cytoplasmic incompatibility (CI). Their invasive potential is currently being harnessed in field trials aiming to control mosquito-borne diseases. Wolbachia genomes commonly harbour prophage regions encoding the cif genes which confer their ability to induce CI. Recently, a plasmid-like element was discovered in wPip, a Wolbachia strain infecting Culex mosquitoes; however, it is unclear how common such extra-chromosomal elements are in Wolbachia. Here we sequenced the complete genome of wAlbA, a strain of the symbiont found in Aedes albopictus, after eliminating the co-infecting and higher density wAlbB strain that previously made sequencing of wAlbA challenging. We show that wAlbA is associated with two new plasmids and identified additional Wolbachia plasmids and related chromosomal islands in over 20% of publicly available Wolbachia genome datasets. These plasmids encode a variety of accessory genes, including several phage-like DNA packaging genes as well as genes potentially contributing to host-symbiont interactions. In particular, we recovered divergent homologues of the cif genes in both Wolbachia- and Rickettsia-associated plasmids. Our results indicate that plasmids are common in Wolbachia and raise fundamental questions around their role in symbiosis. In addition, our comparative analysis provides useful information for the future development of genetic tools to manipulate and study Wolbachia symbionts.}, } @article {pmid36117539, year = {2022}, author = {Power, NR and Rugman-Jones, PF and Stouthamer, R and Ganjisaffar, F and Perring, TM}, title = {High temperature mortality of Wolbachia impacts the sex ratio of the parasitoid Ooencyrtus mirus (Hymenoptera: Encyrtidae).}, journal = {PeerJ}, volume = {10}, number = {}, pages = {e13912}, pmid = {36117539}, issn = {2167-8359}, mesh = {Animals ; Male ; Female ; Temperature ; *Wolbachia/genetics ; Sex Ratio ; *Wasps/microbiology ; Reproduction ; *Heteroptera ; }, abstract = {BACKGROUND: Wolbachia bacteria are estimated to occur in more than half of all insect species. In Hymenoptera, Wolbachia often manipulates its host's reproduction to its own advantage. Wolbachia is likely the reason that males are rare in the uniparental Ooencyrtus mirus Triapitsyn & Power (Hymenoptera: Encyrtidae). The likelihood of producing male offspring can be increased by giving mothers a continuous supply of Bagrada hilaris (Burmeister) (Heteroptera: Pentatomidae) host eggs to parasitize for 2-3 weeks, by feeding the parents antibiotics, or by rearing parent wasps at high temperatures; all variables that have been shown to correlate with depleting Wolbachia titers in other organisms. The purpose of the current study was to determine whether thelytoky in O. mirus is due to Wolbachia, and if so, at what time in development the sex change occurs. We also wished to determine if Wolbachia removal results in the production of intersexes, as in some other hymenopterans. Finally, mating behavior was observed to see if and where it breaks down as a result of the species becoming thelytokous.

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

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

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

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

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

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

CONCLUSIONS: Our results indicate that many of the host-symbiont associations remain largely unexplored, with the majority of associations we identify never being recorded before. This highlights the usefulness of public databases to explore the hidden diversity of symbiotic entities, allowing the development of hypotheses regarding host-symbiont associations. The ever-expanding genomic databases provide a diverse databank from which one can characterize and explore the true diversity of symbiotic entities.}, } @article {pmid36040904, year = {2022}, author = {Conjard, S and Meyer, DF and Aprelon, R and Pagès, N and Gros, O}, title = {Evidence of new strains of Wolbachia symbiont colonising semiaquatic bugs (Hemiptera: Gerroidea) in mangrove environment of the Lesser Antilles.}, journal = {PloS one}, volume = {17}, number = {8}, pages = {e0273668}, pmid = {36040904}, issn = {1932-6203}, mesh = {Animals ; *Arthropods/microbiology ; DNA, Bacterial ; *Heteroptera ; Phylogeny ; *Wolbachia/genetics ; }, abstract = {Wolbachia Hertig, 1936 is an intracellular bacterial symbiont colonizing many arthropods. Of the studies done on the bacteria present in the superfamily Gerroidea Leach, 1815, no report of Wolbachia infection had yet been made. Thus, we checked the presence of Wolbachia in six Gerroidea species which colonize tropical aquatic environments by PCR using wsp primer set before sequencing and phylogenetic analyses. Insects were collected in the marine fringe of mangroves, in river estuaries, in swampy mangroves, and in ponds from Guadeloupe islands (Caribbean). Two new strains of Wolbachia were detected in these Gerroidea. They were named wLfran and wRmang. The wsp sequences suggest that the strains belong to the already described E supergroup or similar. wLfran is present in Limnogonus franciscanus Stål, 1859 and Rheumatobates trinitatis (China, 1943) while wRmang appears to be present exclusively in R. mangrovensis (China, 1943). Three other species were analysed, but did not appear to be infected: Brachymetra albinerva (Amyot & Serville, 1843), Halobates micans Eschscheltz, 1822, and Microvelia pulchella Westwood, 1834. The results presented here highlight for the first time the presence of new intracellular Wolbachia strains in Gerroidea colonising tropical aquatic environments like mangrove habitats from inlands to sea shore.}, } @article {pmid36037217, year = {2022}, author = {Formisano, G and Iodice, L and Cascone, P and Sacco, A and Quarto, R and Cavalieri, V and Bosco, D and Guerrieri, E and Giorgini, M}, title = {Wolbachia infection and genetic diversity of Italian populations of Philaenus spumarius, the main vector of Xylella fastidiosa in Europe.}, journal = {PloS one}, volume = {17}, number = {8}, pages = {e0272028}, pmid = {36037217}, issn = {1932-6203}, mesh = {Animals ; Europe ; Genetic Variation ; *Hemiptera/genetics/microbiology ; Insect Vectors/microbiology ; Italy ; Multilocus Sequence Typing ; Plant Diseases/microbiology ; *Wolbachia/genetics ; *Xylella/genetics ; }, abstract = {Philaenus spumarius is a cosmopolitan species that has become a major threat to European agriculture being recognized as the main vector of the introduced plant pathogen Xylella fastidiosa, the agent of the "olive quick decline syndrome", a disease which is devastating olive orchards in southern Italy. Wolbachia are bacterial symbionts of many insects, frequently as reproductive parasites, sometime by establishing mutualistic relationships, able to spread within host populations. Philaenus spumarius harbors Wolbachia, but the role played by this symbiont is unknown and data on the infection prevalence within host populations are limited. Here, the Wolbachia infection rate was analyzed in relation to the geographic distribution and the genetic diversity of the Italian populations of P. spumarius. Analysis of the COI gene sequences revealed a geographically structured distribution of the three main mitochondrial lineages of P. spumarius. Wolbachia was detected in half of the populations sampled in northern Italy where most individuals belonged to the western-Mediterranean lineage. All populations sampled in southern and central Italy, where the individuals of the eastern-Mediterranean lineage were largely prevalent, were uninfected. Individuals of the north-eastern lineage were found only in populations from the Alps in the northernmost part of Italy, at high altitudes. In this area, Wolbachia infection reached the highest prevalence, with no difference between north-eastern and western-Mediterranean lineage. Analysis of molecular diversity of COI sequences suggested no significant effect of Wolbachia on population genetics of P. spumarius. Using the MLST approach, six new Wolbachia sequence types were identified. Using FISH, Wolbachia were observed within the host's reproductive tissues and salivary glands. Results obtained led us to discuss the role of Wolbachia in P. spumarius, the factors influencing the geographic distribution of the infection, and the exploitation of Wolbachia for the control of the vector insect to reduce the spread of X. fastidiosa.}, } @article {pmid36012723, year = {2022}, author = {Zong, Q and Mao, B and Zhang, HB and Wang, B and Yu, WJ and Wang, ZW and Wang, YF}, title = {Comparative Ubiquitome Analysis Reveals Deubiquitinating Effects Induced by Wolbachia Infection in Drosophila melanogaster.}, journal = {International journal of molecular sciences}, volume = {23}, number = {16}, pages = {}, pmid = {36012723}, issn = {1422-0067}, support = {31872288//National Natural Science Foundation of China/ ; }, mesh = {Animals ; Cytoplasm/metabolism ; *Drosophila melanogaster/genetics ; Female ; Male ; Proteasome Endopeptidase Complex/metabolism ; Semen ; Testis/metabolism ; *Wolbachia ; }, abstract = {The endosymbiotic Wolbachia bacteria frequently cause cytoplasmic incompatibility (CI) in their insect hosts, where Wolbachia-infected males cross with uninfected females, leading to no or fewer progenies, indicating a paternal modification by Wolbachia. Recent studies have identified a Wolbachia protein, CidB, containing a DUB (deubiquitylating enzyme) domain, which can be loaded into host sperm nuclei and involved in CI, though the DUB activity is not necessary for CI in Drosophila melanogaster. To investigate whether and how Wolbachia affect protein ubiquitination in testes of male hosts and are thus involved in male fertility, we compared the protein and ubiquitinated protein expressions in D. melanogaster testes with and without Wolbachia. A total of 643 differentially expressed proteins (DEPs) and 309 differentially expressed ubiquitinated proteins (DEUPs) were identified to have at least a 1.5-fold change with a p-value of <0.05. Many DEPs were enriched in metabolic pathway, ribosome, RNA transport, and post-translational protein modification pathways. Many DEUPs were involved in metabolism, ribosome, and proteasome pathways. Notably, 98.1% DEUPs were downregulated in the presence of Wolbachia. Four genes coding for DEUPs in ubiquitin proteasome pathways were knocked down, respectively, in Wolbachia-free fly testes. Among them, Rpn6 and Rpn7 knockdown caused male sterility, with no mature sperm in seminal vesicles. These results reveal deubiquitylating effects induced by Wolbachia infection, suggesting that Wolbachia can widely deubiquitinate proteins that have crucial functions in male fertility of their hosts, but are not involved in CI. Our data provide new insights into the regulatory mechanisms of endosymbiont/host interactions and male fertility.}, } @article {pmid36009794, year = {2022}, author = {Suo, P and Wang, K and Yu, H and Fu, X and An, L and Bhowmick, B and Zhang, J and Han, Q}, title = {Seasonal Variation of Midgut Bacterial Diversity in Culexquinquefasciatus Populations in Haikou City, Hainan Province, China.}, journal = {Biology}, volume = {11}, number = {8}, pages = {}, pmid = {36009794}, issn = {2079-7737}, support = {ZDYF2021xx//Key Science and Technology Program of Hainan Province (ZDYF2021xx)/ ; ZDYF2019073//Key Research and Development Program of Hainan Province (ZDYF2019073)/ ; }, abstract = {Culex quinquefasciatus, one of the most significant mosquito vectors in the world, is widespread in most parts of southern China. A variety of diseases including Bancroft's filariasis, West Nile disease, and St. Louis encephalitis could be transmitted by the vector. Mosquitoes have been shown to host diverse bacterial communities that vary depending on environmental factors such as temperature and rainfall. In this work, 16S rDNA sequencing was used to analyze the seasonal variation of midgut bacterial diversity of Cx. Quinquefasciatus in Haikou City, Hainan Province, China. Proteobacteria was the dominant phylum, accounting for 79.7% (autumn), 73% (winter), 80.4% (spring), and 84.5% (summer). The abundance of Bacteroidetes in autumn and winter was higher than in others. Interestingly, Epsilonbacteraeota, which only exists in autumn and winter, was discovered accidentally in the midgut. We speculated that this might participate in the nutritional supply of adult mosquitoes when temperatures drop. Wolbachia is the most abundant in autumn, accounting for 31.6% of bacteria. The content of Pantoea was highest in the summer group, which might be related to the enhancement of the ability of mosquitoes as temperatures increased. Pseudomonas is carried out as the highest level in winter. On the contrary, in spring and summer, the genus in highest abundance is Enterobacter. Acinetobacter enriches in the spring when it turns from cold to hot. By studying the diversity of midgut bacteria of Cx. quinquefasciatus, we can further understand the co-evolution of mosquitoes and their symbiotic microbes. This is necessary to discuss the seasonal variation of microorganisms and ultimately provide a new perspective for the control of Cx. quinquefasciatus to reduce the spread of the diseases which have notably vital practical significance for the effective prevention of Cx. quinquefasciatus.}, } @article {pmid36005381, year = {2022}, author = {Fallon, AM}, title = {From Mosquito Ovaries to Ecdysone; from Ecdysone to Wolbachia: One Woman's Career in Insect Biology.}, journal = {Insects}, volume = {13}, number = {8}, pages = {}, pmid = {36005381}, issn = {2075-4450}, abstract = {In anautogenous mosquitoes, synchronous development of terminal ovarian follicles after a blood meal provides an important model for studies on insect reproduction. Removal and implantation of ovaries, in vitro culture of dissected tissues and immunological assays for vitellogenin synthesis by the fat body showed that the Aedes aegypti (L.) (Diptera, Culicidae) mosquito ovary produces a factor essential for egg production. The discovery that the ovarian factor was the insect steroid hormone, ecdysone, provided a model for co-option of the larval hormones as reproductive hormones in adult insects. In later work on cultured mosquito cells, ecdysone was shown to arrest the cell cycle, resulting in an accumulation of diploid cells in G1, prior to initiation of DNA synthesis. Some mosquito species, such as Culex pipiens L. (Diptera, Culicidae), harbor the obligate intracellular bacterium, Wolbachia pipientis Hertig (Rickettsiales, Anaplasmataceae), in their reproductive tissues. When maintained in mosquito cell lines, Wolbachia abundance increases in ecdysone-arrested cells. This observation facilitated the recovery of high levels of Wolbachia from cultured cells for microinjection and genetic manipulation. In female Culex pipiens, it will be of interest to explore how hormonal cues that support initiation and progression of the vitellogenic cycle influence Wolbachia replication and transmission to subsequent generations via infected eggs.}, } @article {pmid36005362, year = {2022}, author = {Kyritsis, GA and Koskinioti, P and Bourtzis, K and Papadopoulos, NT}, title = {Effect of Wolbachia Infection and Adult Food on the Sexual Signaling of Males of the Mediterranean Fruit Fly Ceratitis capitata.}, journal = {Insects}, volume = {13}, number = {8}, pages = {}, pmid = {36005362}, issn = {2075-4450}, support = {511 (Coordinated Research Project (CRP) D41025)//International Atomic Energy Agency/ ; HFRI-FM17-4289//Hellenic Foundation for Research and Innovation/ ; }, abstract = {Sexual signaling is a fundamental component of sexual behavior of Ceratitis capitata that highly determines males' mating success. Nutritional status and age are dominant factors known to affect males' signaling performance and define the female decision to accept a male as a sexual partner. Wolbachia pipientis, a widespread endosymbiotic bacterium of insects and other arthropods, exerts several biological effects on its hosts. However, the effects of Wolbachia infection on the sexual behavior of medfly and the interaction between Wolbachia infection and adult food remain unexplored. This study was conducted to determine the effects of Wolbachia on sexual signaling of protein-fed and protein-deprived males. Our findings demonstrate that: (a) Wolbachia infection reduced male sexual signaling rates in both food regimes; (b) the negative effect of Wolbachia infection was more pronounced on protein-fed than protein-deprived males, and it was higher at younger ages, indicating that the bacterium regulates male sexual maturity; (c) Wolbachia infection alters the daily pattern of sexual signaling; and (d) protein deprivation bears significant descent on sexual signaling frequency of the uninfected males, whereas no difference was observed for the Wolbachia-infected males. The impact of our findings on the implementation of Incompatible Insect Technique (IIT) or the combined SIT/IIT towards controlling insect pests is discussed.}, } @article {pmid36003274, year = {2022}, author = {Sharma, M and Kumar, V}, title = {Mosquito-larvicidal Binary (BinA/B) proteins for mosquito control programs -advancements, challenges, and possibilities.}, journal = {Current research in insect science}, volume = {2}, number = {}, pages = {100028}, pmid = {36003274}, issn = {2666-5158}, abstract = {The increasing global burden of mosquito-borne diseases require targeted, environmentally friendly, and sustainable approaches for effective vector control without endangering the non-target beneficial insect population. Biological interventions such as biopesticides, Wolbachia-mediated biological controls, or sterile insect techniques are used worldwide. Here we review Binary or BinAB toxin-the mosquito-larvicidal component of WHO-recognized Lysinibacillus sphaericus bacterium employed in mosquito control programs. Binary (BinAB) toxin is primarily responsible for the larvicidal effect of the bacterium. BinAB is a single-receptor-specific toxin and is effective against larvae of Culex and Anopheles, but not against Aedes aegypti. The receptor in Culex, the Cqm1 protein, has been extensively studied. It is a GPI-anchored amylomaltase and is located apically in the lipid rafts of the larval-midgut epithelium. The interaction of the toxin components with the receptor is crucial for the mosquito larvicidal activity of the BinAB toxin. Here we extend support for the pore formation model of BinAB toxin internalization and the role of toxin-glycan interactions in the endoplasmic reticulum in mediating larval death. BinAB is phylogenetically safe for humans, as Cqm1-like protein is not expected in the human proteome. This review aims to initiate targeted R&D efforts, such as applying fusion technologies (chimera of BinA, chemical modification of BinA), for efficient mosquito control interventions. In addition, the review also examines other areas such as bioremediation and cancer therapeutics, in which L. sphaericus is proving useful and showing potential for further development.}, } @article {pmid36003268, year = {2022}, author = {Davies, OK and Dorey, JB and Stevens, MI and Gardner, MG and Bradford, TM and Schwarz, MP}, title = {Unparalleled mitochondrial heteroplasmy and Wolbachia co-infection in the non-model bee, Amphylaeus morosus.}, journal = {Current research in insect science}, volume = {2}, number = {}, pages = {100036}, pmid = {36003268}, issn = {2666-5158}, abstract = {Mitochondrial heteroplasmy is the occurrence of more than one type of mitochondrial DNA within a single individual. Although generally reported to occur in a small subset of individuals within a species, there are some instances of widespread heteroplasmy across entire populations. Amphylaeus morosus is an Australian native bee species in the diverse and cosmopolitan bee family Colletidae. This species has an extensive geographical range along the eastern Australian coast, from southern Queensland to western Victoria, covering approximately 2,000 km. Seventy individuals were collected from five localities across this geographical range and sequenced using Sanger sequencing for the mitochondrial cytochrome c oxidase subunit I (COI) gene. These data indicate that every individual had the same consistent heteroplasmic sites but no other nucleotide variation, suggesting two conserved and widespread heteroplasmic mitogenomes. Ion Torrent shotgun sequencing revealed that heteroplasmy occurred across multiple mitochondrial protein-coding genes and is unlikely explained by transposition of mitochondrial genes into the nuclear genome (NUMTs). DNA sequence data also demonstrated a consistent co-infection of Wolbachia across the A. morosus distribution with every individual infected with both bacterial strains. Our data are consistent with the presence of two mitogenomes within all individuals examined in this species and suggest a major divergence from standard patterns of mitochondrial inheritance. Because the host's mitogenome and the Wolbachia genome are genetically linked through maternal inheritance, we propose three possible hypotheses that could explain maintenance of the widespread and conserved co-occurring bacterial and mitochondrial genomes in this species.}, } @article {pmid35992686, year = {2022}, author = {Ma, TC and Guo, WJ and Wen, JB}, title = {Effects of feeding on different parts of Ailanthus altissima on the intestinal microbiota of Eucryptorrhynchus scrobiculatus and Eucryptorrhynchus brandti (Coleoptera: Curculionidae).}, journal = {Frontiers in microbiology}, volume = {13}, number = {}, pages = {899313}, pmid = {35992686}, issn = {1664-302X}, abstract = {Eucryptorrhynchus brandti and Eucryptorrhynchus scrobiculatus (Coleoptera: Curculionidae) are two monophagous weevil pests that feed on Ailanthus altissima (Mill.) Swingle but differ in their diet niche. In the field, adults of E. brandti prefer to feed on the trunk of A. altissima, whereas adults of E. scrobiculatus prefer to feed on the tender parts. We conducted Illumina sequencing of 16S rRNA to examine changes in bacterial diversity in the adults of these two weevil species after they fed on different parts of A. altissima (trunk, 2-3-year-old branches, annual branches, and petioles). Proteobacteria, Tenericutes, and Firmicutes were the dominant phyla in E. brandti (relative abundance was 50.64, 41.56, and 5.63%, respectively) and E. scrobiculatus (relative abundance was 78.63, 11.91, and 7.41%, respectively). At the genus level, Spiroplasma, endosymbionts2, Unclassified Enterobacteriaceae, and Lactococcus were dominant in E. brandti, and Unclassified Enterobacteriaceae, Wolbachia and Spiroplasma, and endosymbionts2 were dominant in E. scrobiculatus. Linear discriminant analysis effect size analysis revealed microbial biomarkers in the different treatment group of adults of both weevil species. Adults of E. brandti may require the trunk, and adults of E. scrobiculatus may require the petioles and annual branches to maintain the high diversity of their gut microbes. The results of this study indicate that feeding on different parts of A. altissima affects the composition and function of the microbes of E. brandti and the microbial composition of E. scrobiculatus. Variation in the abundance of Wolbachia and Spiroplasma in E. brandti and E. scrobiculatus is associated with dietary niche changes, and this might explain the evolution of reproductive isolation between these two sibling weevil species.}, } @article {pmid35992676, year = {2022}, author = {Sadanandane, C and Gunasekaran, K and Panneer, D and Subbarao, SK and Rahi, M and Vijayakumar, B and Athithan, V and Sakthivel, A and Dinesh, S and Jambulingam, P}, title = {Studies on the fitness characteristics of wMel- and wAlbB-introgressed Aedes aegypti (Pud) lines in comparison with wMel- and wAlbB-transinfected Aedes aegypti (Aus) and wild-type Aedes aegypti (Pud) lines.}, journal = {Frontiers in microbiology}, volume = {13}, number = {}, pages = {947857}, pmid = {35992676}, issn = {1664-302X}, abstract = {Wolbachia, an intracellular maternally transmitted endosymbiont, has been shown to interfere with the replication of dengue virus in Aedes aegypti mosquitoes. The Wolbachia-transinfected Ae. aegypti has been currently released in many countries to test its effectiveness in preventing the transmission of dengue virus. ICMR-Vector Control Research Centre in collaboration with World Mosquito Program Monash University, Australia, has generated two new Wolbachia-introgressed Ae. aegypti Puducherry (Pud) lines via backcrossing Ae. aegypti females of Australian (Aus) strains, infected with wMel and wAlbB Wolbachia with wild-type Ae. aegypti Puducherry (Pud) males. Wolbachia infections are known to induce a fitness cost and confer benefit on the host mosquito populations that will influence spread of the Wolbachia into native wild mosquito populations during the field release. Hence, the induced fitness cost or benefit/advantage in the two newly generated Ae. aegypti (Pud) lines was assessed in the laboratory in comparison with the wild-type Ae. aegypti (Pud) strain. In addition, maternal transmission (MT) efficiency, induced cytoplasmic incompatibility (CI), and insecticide resistance status of the two (Pud) lines were determined to assess the likely frequency of wMel and wAlbB infections in the native wild population after field invasion. The study shows that wMel and wAlbB infections did not induce any fitness cost on the two newly generated (Pud) lines. Rather, in terms of wing length, fecundity, egg hatch rate, and adult survival, the Wolbachia introgression conferred fitness benefits on the (Pud) lines compared to uninfected Wolbachia free wild Ae. aegypti population. wMel and wAlbB exhibited a high maternal transmission (99-100%) and induced nearly complete (98-100%) cytoplasmic incompatibility. Both the (Pud) lines were resistant to deltamethrin, malathion, DDT, and temephos, and the level of resistance was almost the same between the two lines as in the wild type. Overall, the stable association of wMel and wAlbB established with Ae. aegypti and the reproductive advantages of the (Pud) lines encourage a pilot release in the field for population replacement potential.}, } @article {pmid35976120, year = {2022}, author = {Shastry, V and Bell, KL and Buerkle, CA and Fordyce, JA and Forister, ML and Gompert, Z and Lebeis, SL and Lucas, LK and Marion, ZH and Nice, CC}, title = {A continental-scale survey of Wolbachia infections in blue butterflies reveals evidence of interspecific transfer and invasion dynamics.}, journal = {G3 (Bethesda, Md.)}, volume = {12}, number = {10}, pages = {}, pmid = {35976120}, issn = {2160-1836}, mesh = {Animals ; *Butterflies/genetics/microbiology ; DNA, Mitochondrial/genetics ; Haplotypes/genetics ; Phylogeny ; *Wolbachia/genetics ; }, abstract = {Infections by maternally inherited bacterial endosymbionts, especially Wolbachia, are common in insects and other invertebrates but infection dynamics across species ranges are largely under studied. Specifically, we lack a broad understanding of the origin of Wolbachia infections in novel hosts, and the historical and geographical dynamics of infections that are critical for identifying the factors governing their spread. We used Genotype-by-Sequencing data from previous population genomics studies for range-wide surveys of Wolbachia presence and genetic diversity in North American butterflies of the genus Lycaeides. As few as one sequence read identified by assembly to a Wolbachia reference genome provided high accuracy in detecting infections in host butterflies as determined by confirmatory PCR tests, and maximum accuracy was achieved with a threshold of only 5 sequence reads per host individual. Using this threshold, we detected Wolbachia in all but 2 of the 107 sampling localities spanning the continent, with infection frequencies within populations ranging from 0% to 100% of individuals, but with most localities having high infection frequencies (mean = 91% infection rate). Three major lineages of Wolbachia were identified as separate strains that appear to represent 3 separate invasions of Lycaeides butterflies by Wolbachia. Overall, we found extensive evidence for acquisition of Wolbachia through interspecific transfer between host lineages. Strain wLycC was confined to a single butterfly taxon, hybrid lineages derived from it, and closely adjacent populations in other taxa. While the other 2 strains were detected throughout the rest of the continent, strain wLycB almost always co-occurred with wLycA. Our demographic modeling suggests wLycB is a recent invasion. Within strain wLycA, the 2 most frequent haplotypes are confined almost exclusively to separate butterfly taxa with haplotype A1 observed largely in Lycaeides melissa and haplotype A2 observed most often in Lycaeides idas localities, consistent with either cladogenic mode of infection acquisition from a common ancestor or by hybridization and accompanying mutation. More than 1 major Wolbachia strain was observed in 15 localities. These results demonstrate the utility of using resequencing data from hosts to quantify Wolbachia genetic variation and infection frequency and provide evidence of multiple colonizations of novel hosts through hybridization between butterfly lineages and complex dynamics between Wolbachia strains.}, } @article {pmid35967981, year = {2022}, author = {Shropshire, JD and Hamant, E and Conner, WR and Cooper, BS}, title = {cifB-transcript levels largely explain cytoplasmic incompatibility variation across divergent Wolbachia.}, journal = {PNAS nexus}, volume = {1}, number = {3}, pages = {pgac099}, pmid = {35967981}, issn = {2752-6542}, support = {R35 GM124701/GM/NIGMS NIH HHS/United States ; }, abstract = {Divergent hosts often associate with intracellular microbes that influence their fitness. Maternally transmitted Wolbachia bacteria are the most common of these endosymbionts, due largely to cytoplasmic incompatibility (CI) that kills uninfected embryos fertilized by Wolbachia-infected males. Closely related infections in females rescue CI, providing a relative fitness advantage that drives Wolbachia to high frequencies. One prophage-associated gene (cifA) governs rescue, and two contribute to CI (cifA and cifB), but CI strength ranges from very strong to very weak for unknown reasons. Here, we investigate CI-strength variation and its mechanistic underpinnings in a phylogenetic context across 20 million years (MY) of Wolbachia evolution in Drosophila hosts diverged up to 50 MY. These Wolbachia encode diverse Cif proteins (100% to 7.4% pairwise similarity), and AlphaFold structural analyses suggest that CifB sequence similarities do not predict structural similarities. We demonstrate that cifB-transcript levels in testes explain CI strength across all but two focal systems. Despite phylogenetic discordance among cifs and the bulk of the Wolbachia genome, closely related Wolbachia tend to cause similar CI strengths and transcribe cifB at similar levels. This indicates that other non-cif regions of the Wolbachia genome modulate cif-transcript levels. CI strength also increases with the length of the host's larval life stage, presumably due to prolonged cif action. Our findings reveal that cifB-transcript levels largely explain CI strength, while highlighting other covariates. Elucidating CI's mechanism contributes to our understanding of Wolbachia spread in natural systems and to improving the efficacy of CI-based biocontrol of arboviruses and agricultural pests globally.}, } @article {pmid35939432, year = {2022}, author = {, }, title = {Retraction: Wolbachia Transcription Elongation Factor "Wol GreA" Interacts with α2ββ'σ Subunits of RNA Polymerase through Its Dimeric C-Terminal Domain.}, journal = {PLoS neglected tropical diseases}, volume = {16}, number = {8}, pages = {e0010694}, pmid = {35939432}, issn = {1935-2735}, } @article {pmid35925631, year = {2022}, author = {Denton, JA and Joubert, DA and Goundar, AA and Gilles, JRL}, title = {International shipments of Wolbachia-infected mosquito eggs: towards the scaling-up of World Mosquito Program operations.}, journal = {Revue scientifique et technique (International Office of Epizootics)}, volume = {41}, number = {1}, pages = {91-99}, doi = {10.20506/rst.41.1.3306}, pmid = {35925631}, issn = {0253-1933}, support = {212914/Z/18/B/WT_/Wellcome Trust/United Kingdom ; }, mesh = {*Aedes ; Animals ; Australia ; *Dengue/epidemiology/veterinary ; Mosquito Vectors ; Ovum ; Pest Control, Biological ; *Wolbachia ; }, abstract = {The Wolbachia insect control method, employed by the World Mosquito Program (WMP), relies on introgressing Wolbachia through target Aedes aegypti populations to reduce the incidence of dengue. Since 2010, the WMP has been producing Wolbachia-infected mosquitoes at numerous sites across the globe for release in 11 countries. As the technology has matured, greater focus has been placed on mosquito production at larger central facilities for transport to remote release sites, both domestically and internationally. Of particular note is the production of Wolbachia-infected mosquitoes at the WMP's Australian production facility for successful international deployments in Fiji, Vanuatu, Kiribati and Sri Lanka. This requires careful management of both production and supply-chain processes to ensure that the quality of the mosquito eggs, specifically the hatch rate and Wolbachia infection rate, is maintained. To ensure the cost-effectiveness and scalability of the Wolbachia method, these processes will be further refined to facilitate deployment from large centralised production facilities.}, } @article {pmid35923389, year = {2022}, author = {Tibbs-Cortes, LE and Tibbs-Cortes, BW and Schmitz-Esser, S}, title = {Tardigrade Community Microbiomes in North American Orchards Include Putative Endosymbionts and Plant Pathogens.}, journal = {Frontiers in microbiology}, volume = {13}, number = {}, pages = {866930}, pmid = {35923389}, issn = {1664-302X}, abstract = {The microbiome of tardigrades, a phylum of microscopic animals best known for their ability to survive extreme conditions, is poorly studied worldwide and completely unknown in North America. An improved understanding of tardigrade-associated bacteria is particularly important because tardigrades have been shown to act as vectors of the plant pathogen Xanthomonas campestris in the laboratory. However, the potential role of tardigrades as reservoirs and vectors of phytopathogens has not been investigated further. This study analyzed the microbiota of tardigrades from six apple orchards in central Iowa, United States, and is the first analysis of the microbiota of North American tardigrades. It is also the first ever study of the tardigrade microbiome in an agricultural setting. We utilized 16S rRNA gene amplicon sequencing to characterize the tardigrade community microbiome across four contrasts: location, substrate type (moss or lichen), collection year, and tardigrades vs. their substrate. Alpha diversity of the tardigrade community microbiome differed significantly by location and year of collection but not by substrate type. Our work also corroborated earlier findings, demonstrating that tardigrades harbor a distinct microbiota from their environment. We also identified tardigrade-associated taxa that belong to genera known to contain phytopathogens (Pseudomonas, Ralstonia, and the Pantoea/Erwinia complex). Finally, we observed members of the genera Rickettsia and Wolbachia in the tardigrade microbiome; because these are obligate intracellular genera, we consider these taxa to be putative endosymbionts of tardigrades. These results suggest the presence of putative endosymbionts and phytopathogens in the microbiota of wild tardigrades in North America.}, } @article {pmid35916725, year = {2022}, author = {Dereeper, A and Summo, M and Meyer, DF}, title = {PanExplorer: a web-based tool for exploratory analysis and visualization of bacterial pan-genomes.}, journal = {Bioinformatics (Oxford, England)}, volume = {38}, number = {18}, pages = {4412-4414}, pmid = {35916725}, issn = {1367-4811}, support = {//European Union on the Guadeloupe Region/ ; 2018-FED-1084//European Research and Development Funds/ ; }, mesh = {*Genome, Bacterial ; Genomics ; Software ; *Libraries ; Internet ; }, abstract = {MOTIVATION: As pan-genome approaches are largely employed for bacterial comparative genomics and evolution analyses, but still difficult to be carried out by non-bioinformatician biologists, there is a need for an innovative tool facilitating the exploration of bacterial pan-genomes.

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

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

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

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

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

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

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

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

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

CONCLUSIONS: Exposure of larvae to higher rearing temperatures in the laboratory and simulated-field conditions reduced the densities of wMel and wAlbB strains particularly in males, but the impact was more pronounced for wMel strain. The actual effect of heat stress on the stability of these two Wolbachia strains needs to be tested under natural field conditions.}, } @article {pmid35726500, year = {2022}, author = {Dzul-Rosado, K and Maldonado-Borges, JI and Puerto-Manzano, FI and Lammoglia-Villagómez, MA and Becker, I and Sánchez-Montes, S}, title = {First exploratory study of bacterial pathogens and endosymbionts in head lice from a Mayan community in southern Mexico.}, journal = {Zoonoses and public health}, volume = {69}, number = {6}, pages = {729-736}, doi = {10.1111/zph.12982}, pmid = {35726500}, issn = {1863-2378}, mesh = {*Acinetobacter/genetics ; Animals ; Bacteria/genetics ; DNA ; Humans ; *Lice Infestations/epidemiology/veterinary ; Mexico/epidemiology ; *Pediculus/genetics/microbiology ; Phylogeny ; }, abstract = {Lice represent one of the most neglected group of vectors worldwide, particularly in Latin America. Records of bacterial agents related to head lice are non-existent in this region of the continent. Many of these communities often do not have adequate access to public services and/or health protection. The normalization of this condition prevents them from manifesting discomfort, such as bites and itching, which further aggravates the situation, as they can be vectors of important diseases. For this reason, the aim of this work was to identify the richness of bacterial pathogens (Acinetobacter, Bartonella, and Rickettsia) and endosymbionts (Wolbachia) in head lice of paediatric patients from the indigenous municipality of Hoctun, Yucatan, Mexico. DNA extraction was performed using the QIAamp DNA Mini Kit. For the detection of bacterial pathogens, fragments of the gltA, rpoB, and 16S rDNA genes were amplified. For the detection of Wolbachia, the wsp gene was amplified. Of the 28 lice analysed, the presence of two genera of bacterial pathogens was detected Acinetobacter (42.9% = 12/28) and Bartonella (7.14% = 2/28). We also detected the endosymbiont Wolbachia (71.42% = 20/28). Our results showed that DNA from three bacteria species (Acinetobacter baumannii, Bartonella quintana, and Wolbachia pipientis) was present with frequencies ranging from 3.57% to 71.42%. This work represents the first exploratory study of the diversity of agents associated with head lice (Pediculus humanus capitis) in Mexico and Latin America. Due to the findings generated in the present study, it is important to perform surveillance of head lice populations to identify the degree of spread of these pathogens and their impact on populations in the region.}, } @article {pmid35724982, year = {2023}, author = {Faulk, C}, title = {De novo sequencing, diploid assembly, and annotation of the black carpenter ant, Camponotus pennsylvanicus, and its symbionts by one person for $1000, using nanopore sequencing.}, journal = {Nucleic acids research}, volume = {51}, number = {1}, pages = {17-28}, pmid = {35724982}, issn = {1362-4962}, mesh = {Animals ; Humans ; *Ants/genetics/microbiology ; Diploidy ; Genome, Mitochondrial ; *High-Throughput Nucleotide Sequencing/economics/methods ; Nanopore Sequencing ; *Sequence Analysis, DNA/economics/methods ; Symbiosis ; Wolbachia/genetics/physiology ; Enterobacteriaceae/classification/genetics/physiology ; *Genome, Insect ; }, abstract = {The black carpenter ant (Camponotus pennsylvanicus) is a pest species found widely throughout North America. From a single individual I used long-read nanopore sequencing to assemble a phased diploid genome of 306 Mb and 60X coverage, with quality assessed by a 97.0% BUSCO score, improving upon other ant assemblies. The mitochondrial genome reveals minor rearrangements from other ants. The reads also allowed assembly of parasitic and symbiont genomes. I include a complete Wolbachia bacterial assembly with a size of 1.2 Mb, as well as a commensal symbiont Blochmannia pennsylvanicus, at 791 kb. DNA methylation and hydroxymethylation were measured at base-pair resolution level from the same reads and confirmed extremely low levels seen in the Formicidae family. There was moderate heterozygosity, with 0.16% of bases being biallelic from the parental haplotypes. Protein prediction yielded 14 415 amino acid sequences with 95.8% BUSCO score and 86% matching to previously known proteins. All assemblies were derived from a single MinION flow cell generating 20 Gb of sequence for a cost of $1047 including consumable reagents. Adding fixed costs for equipment brings the total for an ant-sized genome to less than $5000. All analyses were performed in 1 week on a single desktop computer.}, } @article {pmid35723456, year = {2022}, author = {Chen, L and Xiao, Q and Shi, M and Cheng, J and Wu, J}, title = {Detecting Wolbachia Strain wAlbB in Aedes albopictus Cell Lines.}, journal = {Journal of visualized experiments : JoVE}, volume = {}, number = {184}, pages = {}, doi = {10.3791/63662}, pmid = {35723456}, issn = {1940-087X}, mesh = {*Aedes ; Animals ; Cell Line ; Mosquito Vectors ; *RNA Viruses ; *Wolbachia/physiology ; }, abstract = {As a maternally harbored endosymbiont, Wolbachia infects large proportions of insect populations. Studies have recently reported the successful regulation of RNA virus transmission using Wolbachia-transfected mosquitoes. Key strategies to control viruses include the manipulation of host reproduction via cytoplasmic incompatibility and the inhibition of viral transcripts via immune priming and competition for host-derived resources. However, the underlying mechanisms of the responses of Wolbachia-transfected mosquitoes to viral infection are poorly understood. This paper presents a protocol for the in vitro identification of Wolbachia infection at the nucleic acid and protein levels in Aedes albopictus (Diptera: Culicidae) Aa23 cells to enhance the understanding of the interactions between Wolbachia and its insect vectors. Through the combined use of polymerase chain reaction (PCR), quantitative PCR, western blot, and immunological analytical methods, a standard morphologic protocol has been described for the detection of Wolbachia-infected cells that is more accurate than the use of a single method. This approach may also be applied to the detection of Wolbachia infection in other insect taxa.}, } @article {pmid35715692, year = {2022}, author = {Hickin, ML and Kakumanu, ML and Schal, C}, title = {Effects of Wolbachia elimination and B-vitamin supplementation on bed bug development and reproduction.}, journal = {Scientific reports}, volume = {12}, number = {1}, pages = {10270}, pmid = {35715692}, issn = {2045-2322}, support = {P30 ES025128/ES/NIEHS NIH HHS/United States ; }, mesh = {Animals ; *Bedbugs ; Dietary Supplements ; Female ; Nymph ; Reproduction ; *Vitamin B Complex/pharmacology ; *Wolbachia ; }, abstract = {Obligate blood feeders, such as Cimex lectularius (common bed bug), have symbiotic associations with nutritional endosymbionts that produce B-vitamins. To quantify the symbiont's contribution to host fitness in these obligate mutualisms, the symbiont must be eliminated and its absence rigorously confirmed. We developed and validated procedures for complete elimination of Wolbachia (Wb) in bed bugs and quantified development and reproduction in bed bugs with and without Wb and with and without B-vitamins supplementation. Aposymbiotic bed bugs had slower nymphal development, reduced adult survivorship, smaller adult size, fewer eggs per female, and lower hatch rate than bed bugs that harbored Wb. In aposymbiotic bed bugs that were fed B-vitamins-supplemented blood, nymph development time, adult survivorship and hatch rate recovered to control levels, but adult size and egg number only partially recovered. These results underscore the nutritional dependence of bed bugs on their Wb symbiont and suggest that Wb may provide additional nutritional benefits beyond the B-vitamin mix that we investigated.}, } @article {pmid35708345, year = {2022}, author = {Ettinger, CL and Byrne, FJ and Redak, RA and Stajich, JE}, title = {Metagenome-Assembled Genomes of Bacterial Symbionts Associated with Insecticide-Resistant and -Susceptible Individuals of the Glassy-Winged Sharpshooter (Homalodisca vitripennis).}, journal = {Microbiology resource announcements}, volume = {11}, number = {7}, pages = {e0050622}, pmid = {35708345}, issn = {2576-098X}, support = {14-0379-000-SA-2//Pierce's Disease Control Program/ ; 01170-002//California Department of Food and Agriculture (CDFA)/ ; 007011-003//California Department of Food and Agriculture (CDFA)/ ; //Canadian Institute for Advanced Research (ICRA)/ ; CA-R-PPA-5062-H//U.S. Department of Agriculture (USDA)/ ; 012604-002//USDA | Animal and Plant Health Inspection Service (APHIS)/ ; }, abstract = {The role of microbes in insecticide resistance is an emerging question. Here, we describe six metagenome-assembled genomes (MAGs) associated with the glassy-winged sharpshooter (Homalodisca vitripennis [Germar, 1821]) (Hemiptera, Cicadellidae). MAGs representing the obligate symbionts Candidatus Sulcia muelleri and Candidatus Baumannia cicadellinicola and the facultative symbiont Wolbachia were obtained from imidacloprid-resistant and imidacloprid-susceptible sharpshooters.}, } @article {pmid35701454, year = {2022}, author = {Dufault, SM and Tanamas, SK and Indriani, C and Utarini, A and Ahmad, RA and Jewell, NP and Simmons, CP and Anders, KL}, title = {Disruption of spatiotemporal clustering in dengue cases by wMel Wolbachia in Yogyakarta, Indonesia.}, journal = {Scientific reports}, volume = {12}, number = {1}, pages = {9890}, pmid = {35701454}, issn = {2045-2322}, support = {/WT_/Wellcome Trust/United Kingdom ; R01 AI148127/AI/NIAID NIH HHS/United States ; R01AI148127/AI/NIAID NIH HHS/United States ; }, mesh = {*Aedes ; Animals ; Cluster Analysis ; *Dengue ; *Dengue Virus/genetics ; Humans ; Indonesia/epidemiology ; Pest Control, Biological ; *Wolbachia/genetics ; }, abstract = {Dengue exhibits focal clustering in households and neighborhoods, driven by local mosquito population dynamics, human population immunity, and fine scale human and mosquito movement. We tested the hypothesis that spatiotemporal clustering of homotypic dengue cases is disrupted by introduction of the arbovirus-blocking bacterium Wolbachia (wMel-strain) into the Aedes aegypti mosquito population. We analysed 318 serotyped and geolocated dengue cases (and 5921 test-negative controls) from a randomized controlled trial in Yogyakarta, Indonesia of wMel deployments. We find evidence of spatial clustering up to 300 m among the 265 dengue cases (3083 controls) in the untreated trial arm. Participant pairs enrolled within 30 days and 50 m had a 4.7-fold increase (compared to 95% CI on permutation-based null distribution: 0.1, 1.2) in the odds of being homotypic (i.e. potentially transmission-related) as compared to pairs occurring at any distance. In contrast, we find no evidence of spatiotemporal clustering among the 53 dengue cases (2838 controls) resident in the wMel-treated arm. Introgression of wMel Wolbachia into Aedes aegypti mosquito populations interrupts focal dengue virus transmission leading to reduced case incidence; the true intervention effect may be greater than the 77% efficacy measured in the primary analysis of the Yogyakarta trial.}, } @article {pmid35699129, year = {2022}, author = {Mulenga, GM and Namangala, B and Gummow, B}, title = {Prevalence of trypanosomes and selected symbionts in tsetse species of eastern Zambia.}, journal = {Parasitology}, volume = {149}, number = {11}, pages = {1406-1410}, pmid = {35699129}, issn = {1469-8161}, mesh = {Animals ; Enterobacteriaceae/genetics ; Insect Vectors/parasitology ; Prevalence ; *Trypanosoma/genetics ; *Trypanosomiasis, African/parasitology ; *Tsetse Flies/parasitology ; *Wolbachia/genetics ; Zambia/epidemiology ; }, abstract = {Insect symbionts have attracted attention for their potential use as anti-parasitic gene products in arthropod disease vectors. While tsetse species of the Luangwa valley have been extensively studied, less is known about the prevalence of symbionts and their interactions with the trypanosome parasite. Polymerase chain reaction was used to investigate the presence of Wolbachia and Sodalis bacteria, in tsetse flies infected with trypanosomes (Trypanosoma vivax, Trypanosoma congolense and Trypanosoma brucei). Out of 278 captured tsetse flies in eastern Zambia, 95.3% (n = 265, 95% CI = 92.8–97.8) carried endosymbionts: Wolbachia (79.1%, 95% CI 73.9–83.8) and Sodalis (86.3%, 95% CI 81.7–90.1). Overall, trypanosome prevalence was 25.5% (n = 71, 95% CI = 20.4–30.7), 10.8% (n = 30, 95% CI 7.1–14.4) for T. brucei, 1.4% (n = 4, 95% CI = 0.4–3.6) for both T. congolense and T. vivax, and 0.7% (n = 2, 95% CI 0.1–2.6) for T. b. rhodesiense. Out of 240 tsetse flies that were infected with Sodalis, trypanosome infection was reported in 40 tsetse flies (16.7%, 95% CI = 12.0–21.4) while 37 (16.8%, 95% CI 11.9–21.8) of the 220 Wolbachia infected tsetse flies were infected with trypanosomes. There was 1.3 times likelihood of T. brucei infection to be present when Wolbachia was present and 1.7 likelihood of T. brucei infection when Sodalis was present. Overall findings suggest absence of correlation between the presence of tsetse endosymbionts and tsetse with trypanosome infection. Lastly, the presence of pathogenic trypanosomes in tsetse species examined provided insights into the risk communities face, and the importance of African trypanosomiasis in the area.}, } @article {pmid35687421, year = {2022}, author = {Vörös, K and Becker, Z and Kónya, R and Arany-Tóth, A and Farkas, R}, title = {Application of Moxidectin and Ultrasound-Aided Injection of Melarsomine During the American Heartworm Society Recommended Treatment Protocol in Dirofilaria immitis Infected Dogs.}, journal = {Vector borne and zoonotic diseases (Larchmont, N.Y.)}, volume = {22}, number = {7}, pages = {382-390}, doi = {10.1089/vbz.2021.0108}, pmid = {35687421}, issn = {1557-7759}, mesh = {Animals ; Arsenicals ; Clinical Protocols ; Dexamethasone/pharmacology/therapeutic use ; *Dirofilaria immitis ; *Dirofilariasis/diagnosis/drug therapy ; *Dog Diseases/drug therapy ; Dogs ; Doxycycline/therapeutic use ; *Filaricides/therapeutic use ; Macrolides/pharmacology/therapeutic use ; Triazines ; }, abstract = {The American Heartworm Society (AHS) recommends the three-dose alternate melarsomine therapeutic regimen, together with a macrocyclic lactone (ML) to reduce new infections and eliminate susceptible larvae and doxycycline against Wolbachia bacteria. Till now, only reports on ivermectin as an ML exist in the frame of this protocol. Between 2014 and 2020, the AHS protocol was used in 44 heartworm-positive dogs. Microfilaremic dogs were pretreated with prednisolone and clopidogrel for 1 week before the first moxidectin application. Moxidectin was applied on the 1st, 30th, 60th, and 90th therapeutic days. On the first day, dexamethasone and chloropyramine were used to avoid potential adverse effects caused by the destroyed microfilariae. During the 1st-28th days, doxycycline 10 mg/kg BID was given with probiotics. Adult heartworms were destroyed with melarsomine on the 60th, 90[th], and 91st days. Butorphanol and dexamethasone were given just before melarsomine injections. The depth of the intramuscular injection site was determined by ultrasound examination of the lumbar muscles. From the 60th day, dalteparin was applied for 10 days to decrease the chance of pulmonary thromboembolism. Moxidectin did not cause adverse reactions, even in microfilaremic dogs. Gastrointestinal side effects of doxycycline were observed in three (6%) dogs, they recovered after symptomatic therapy and by lowering the initial dose to 5 mg/kg BID. Transient anorexia and diarrhea were found in one (2%), and coughing and mild dyspnea in one (2%) animal as systemic post-therapeutic complications of melarsomine. No local side effects were observed in 13 (30%) dogs, mild local side effects occurred in 29 (66%) patients, and severe local swelling in 2 (4%) cases. All dogs recovered clinically by the 120th day and no microfilaremia was seen that time. An antigen test performed in 37/44 animals on the 271st day was also negative in all cases.}, } @article {pmid35686927, year = {2022}, author = {Kamiyama, T and Shimada-Niwa, Y and Tanaka, H and Katayama, M and Kuwabara, T and Mori, H and Kunihisa, A and Itoh, T and Toyoda, A and Niwa, R}, title = {Whole-genome sequencing analysis and protocol for RNA interference of the endoparasitoid wasp Asobara japonica.}, journal = {DNA research : an international journal for rapid publication of reports on genes and genomes}, volume = {29}, number = {4}, pages = {}, pmid = {35686927}, issn = {1756-1663}, support = {//JSPS KAKENHI/ ; //Japan Science and Technology Agency/ ; }, mesh = {Animals ; Drosophila/genetics ; Female ; Larva/parasitology ; Molecular Sequence Annotation ; RNA Interference ; RNA, Double-Stranded/genetics ; *Wasps/genetics ; }, abstract = {Asobara japonica is an endoparasitic wasp that parasitizes Drosophila flies. It synthesizes various toxic components in the venom gland and injects them into host larvae during oviposition. To identify and characterize these toxic components for enabling parasitism, we performed the whole-genome sequencing (WGS) and devised a protocol for RNA interference (RNAi) with A. japonica. Because it has a parthenogenetic lineage due to Wolbachia infection, we generated a clonal strain from a single wasp to obtain highly homogenous genomic DNA. The WGS analysis revealed that the estimated genome size was 322 Mb with a heterozygosity of 0.132%. We also performed RNA-seq analyses for gene annotation. Based on the qualified WGS platform, we cloned ebony-Aj, which encodes the enzyme N-β-alanyl dopamine synthetase, which is involved in melanin production. The microinjection of double-stranded RNA (dsRNA) targeting ebony-Aj led to body colour changes in adult wasps, phenocopying ebony-Dm mutants. Furthermore, we identified putative venom genes as a target of RNAi, confirming that dsRNA injection-based RNAi specifically suppressed the expression of the target gene in wasp adults. Taken together, our results provide a powerful genetic toolkit for studying the molecular mechanisms of parasitism.}, } @article {pmid35686658, year = {2022}, author = {Szklarzewicz, T and Kalandyk-Kołodziejczyk, M and Michalik, A}, title = {Ovary structure and symbiotic associates of a ground mealybug, Rhizoecus albidus (Hemiptera, Coccomorpha: Rhizoecidae) and their phylogenetic implications.}, journal = {Journal of anatomy}, volume = {241}, number = {3}, pages = {860-872}, pmid = {35686658}, issn = {1469-7580}, mesh = {Animals ; Female ; *Hemiptera/anatomy & histology/chemistry/microbiology ; In Situ Hybridization, Fluorescence ; Ovary/anatomy & histology ; Phylogeny ; Symbiosis ; }, abstract = {The ovary structure and the organization of its symbiotic system of the ground mealybug, Rhizoecus albidus (Rhizoecidae), were examined by means of microscopic and molecular methods. Each of the paired elongated ovaries of R. albidus is composed of circa one hundred short telotrophic-meroistic ovarioles, which are radially arranged along the distal part of the lateral oviduct. Analysis of serial sections revealed that each ovariole contains four germ cells: three trophocytes (nurse cells) occupying the tropharium and a single oocyte in the vitellarium. The ovaries are accompanied by giant cells termed bacteriocytes which are tightly packed with large pleomorphic bacteria. Their identity as Brownia rhizoecola (Bacteroidetes) was confirmed by means of amplicon sequencing and fluorescence in situ hybridization techniques. Moreover, to our knowledge, this is the first report on the morphology and ultrastructure of the Brownia rhizoecola bacterium. In the bacteriocyte cytoplasm bacteria Brownia co-reside with sporadic rod-shaped smaller bacteria, namely Wolbachia (Proteobacteria: Alphaproteobacteria). Both symbionts are transmitted to the next generation vertically (maternally), that is, via female germline cells. We documented that, at the time when ovarioles contain oocytes at the vitellogenic stage, these symbionts leave the bacteriocytes and move toward the neck region of ovarioles (i.e. the region between tropharium and vitellarium). Next, the bacteria enter the cytoplasm of follicular cells surrounding the basal part of the tropharium, leave them and enter the space between the follicular epithelium and surface of the nutritive cord connecting the tropharium and vitellarium. Finally, they gather in the deep depression of the oolemma at the anterior pole of the oocyte in the form of a 'symbiont ball'. Our results provide further arguments strongly supporting the validity of the recent changes in the classification of mealybugs, which involved excluding ground mealybugs from the Pseudococcidae family and raising them to the rank of their own family Rhizoecidae.}, } @article {pmid35678877, year = {2022}, author = {Araújo, NJS and Macêdo, MJF and de Morais, LP and da Cunha, FAB and de Matos, YMLS and de Almeida, RS and Braga, MFBM and Coutinho, HDM}, title = {Control of arboviruses vectors using biological control by Wolbachia pipientis: a short review.}, journal = {Archives of microbiology}, volume = {204}, number = {7}, pages = {376}, pmid = {35678877}, issn = {1432-072X}, mesh = {Animals ; *Arboviruses ; Insect Vectors/microbiology ; *Wolbachia ; }, abstract = {The number of arbovirus cases has increased in recent years, demonstrating a need for investing in effective control actions. Among these actions, are strategies using biological control vectors, a field where Wolbachia pipientis has shown itself as useful. Wolbachia pipientis, an obligatory intracellular Gram-negative bacteria, which parasites arthropods naturally or through laboratory-induced infections, is capable of manipulating the reproduction of its host. A systematic literature review gathering studies on this bacteria over last 10 years (2007-2021) was performed given its important role in the reduction of insect disease vectors. 111 articles were found, from which 78 were used in this study. Information on the Wolbachia biology, mechanism of action and potential for the biological control of insect disease vectors was gathered. The present study may contribute to the knowledge surrounding the bacterium, as well as stimulate the production of other studies with the same theme.}, } @article {pmid35678697, year = {2022}, author = {Withers, AJ and Rice, A and de Boer, J and Donkersley, P and Pearson, AJ and Chipabika, G and Karangwa, P and Uzayisenga, B and Mensah, BA and Mensah, SA and Nkunika, POY and Kachigamba, D and Smith, JA and Jones, CM and Wilson, K}, title = {The distribution of covert microbial natural enemies of a globally invasive crop pest, fall armyworm, in Africa: Enemy release and spillover events.}, journal = {The Journal of animal ecology}, volume = {91}, number = {9}, pages = {1826-1841}, pmid = {35678697}, issn = {1365-2656}, support = {BB/P023444/1//Global Challenges Research Fund/ ; NE/L002604/1//Natural Environment Research Council/ ; }, mesh = {Animals ; Introduced Species ; Kenya ; *Nucleopolyhedroviruses ; Spodoptera ; *Wolbachia ; }, abstract = {Invasive species pose a significant threat to biodiversity and agriculture world-wide. Natural enemies play an important part in controlling pest populations, yet we understand very little about the presence and prevalence of natural enemies during the early invasion stages. Microbial natural enemies of fall armyworm Spodoptera frugiperda are known in its native region, however, they have not yet been identified in Africa where fall armyworm has been an invasive crop pest since 2016. Larval samples were screened from Malawi, Rwanda, Kenya, Zambia, Sudan and Ghana for the presence of four different microbial natural enemies; two nucleopolyhedroviruses, Spodoptera frugiperda NPV (SfMNPV) and Spodoptera exempta NPV (SpexNPV); the fungal pathogen Metarhizium rileyi; and the bacterium Wolbachia. This study aimed to identify which microbial pathogens are present in invasive fall armyworm, and determine the geographical, meteorological and temporal variables that influence prevalence. Within 3 years of arrival, fall armyworm was exposed to all four microbial natural enemies. SfMNPV probably arrived with fall armyworm from the Americas, but this is the first putative evidence of host spillover from Spodoptera exempta (African armyworm) to fall armyworm for the endemic pathogen SpexNPV and for Wolbachia. It is also the first confirmed incidence of M. rileyi infecting fall armyworm in Africa. Natural enemies were localised, with variation being observed both nationally and temporally. The prevalence of SfMNPV (the most common natural enemy) was predominantly explained by variables associated with the weather; declining with increasing rainfall and increasing with temperature. However, virus prevalence also increased as the growing season progressed. The infection of an invasive species with a natural enemy from its native range and novel pathogens specific to its new range has important consequences for understanding the population ecology of invasive species and insect-pathogen interactions. Additionally, while it is widely known that temporal and geographic factors affect insect populations, this study reveals that these are important in understanding the distribution of microbial natural enemies associated with invasive pests during the early stages of invasion, and provide baseline data for future studies.}, } @article {pmid35671755, year = {2022}, author = {Tvedte, ES and Gasser, M and Zhao, X and Tallon, LJ and Sadzewicz, L and Bromley, RE and Chung, M and Mattick, J and Sparklin, BC and Dunning Hotopp, JC}, title = {Accumulation of endosymbiont genomes in an insect autosome followed by endosymbiont replacement.}, journal = {Current biology : CB}, volume = {32}, number = {12}, pages = {2786-2795.e5}, pmid = {35671755}, issn = {1879-0445}, support = {R01 CA206188/CA/NCI NIH HHS/United States ; U19 AI110820/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Chromosomes ; Drosophila/genetics/microbiology ; Gene Transfer, Horizontal ; Genome ; Symbiosis/genetics ; *Wolbachia/genetics ; }, abstract = {Eukaryotic genomes can acquire bacterial DNA via lateral gene transfer (LGT).[1] A prominent source of LGT is Wolbachia,[2] a widespread endosymbiont of arthropods and nematodes that is transmitted maternally through female germline cells.[3,4] The DNA transfer from the Wolbachia endosymbiont wAna to Drosophila ananassae is extensive[5-7] and has been localized to chromosome 4, contributing to chromosome expansion in this lineage.[6] As has happened frequently with claims of bacteria-to-eukaryote LGT, the contribution of wAna transfers to the expanded size of D. ananassae chromosome 4 has been specifically contested[8] owing to an assembly where Wolbachia sequences were classified as contaminants and removed.[9] Here, long-read sequencing with DNA from a Wolbachia-cured line enabled assembly of 4.9 Mbp of nuclear Wolbachia transfers (nuwts) in D. ananassae and a 24-kbp nuclear mitochondrial transfer. The nuwts are <8,000 years old in at least two locations in chromosome 4 with at least one whole-genome integration followed by rapid extensive duplication of most of the genome with regions that have up to 10 copies. The genes in nuwts are accumulating small indels and mobile element insertions. Among the highly duplicated genes are cifA and cifB, two genes associated with Wolbachia-mediated Drosophila cytoplasmic incompatibility. The wAna strain that was the source of nuwts was subsequently replaced by a different wAna endosymbiont. Direct RNA Nanopore sequencing of Wolbachia-cured lines identified nuwt transcripts, including spliced transcripts, but functionality, if any, remains elusive.}, } @article {pmid35671324, year = {2022}, author = {Marriott, AE and Furlong Silva, J and Pionnier, N and Sjoberg, H and Archer, J and Steven, A and Kempf, D and Taylor, MJ and Turner, JD}, title = {A mouse infection model and long-term lymphatic endothelium co-culture system to evaluate drugs against adult Brugia malayi.}, journal = {PLoS neglected tropical diseases}, volume = {16}, number = {6}, pages = {e0010474}, pmid = {35671324}, issn = {1935-2735}, support = {NC/M00175X/1/NC3RS_/National Centre for the Replacement, Refinement and Reduction of Animals in Research/United Kingdom ; NC/S001131/1/NC3RS_/National Centre for the Replacement, Refinement and Reduction of Animals in Research/United Kingdom ; }, mesh = {Animals ; *Brugia malayi ; Coculture Techniques ; Disease Models, Animal ; Endothelial Cells ; Endothelium, Lymphatic ; Female ; Interleukin-5 ; Male ; Mice ; Mice, Inbred BALB C ; *Wolbachia ; }, abstract = {The development of new drugs targeting adult-stage lymphatic filarial nematodes is hindered by the lack of a robust long-term in vitro culture model. Testing potential direct-acting and anti-Wolbachia therapeutic candidates against adult lymphatic filariae in vitro requires their propagation via chronic infection of gerbils. We evaluated Brugia malayi parasite burden data from male Mongolian gerbils compared with two immune-deficient mouse strains highly susceptible to B. malayi: CB.17 Severe-Combined Immmuno-Deficient (SCID) and interleukin-4 receptor alpha, interleukin-5 double knockout (IL-4Rα-/-IL-5-/-) mice. Adult worms generated in IL-4Rα-/-IL-5-/- mice were tested with different feeder cells (human embryonic kidney cells, human adult dermal lymphatic endothelial cells and human THP-1 monocyte differentiated macrophages) and comparative cell-free conditions to optimise and validate a long-term in vitro culture system. Cultured parasites were compared against those isolated from mice using motility scoring, metabolic viability assay (MTT), ex vivo microfilariae release assay and Wolbachia content by qPCR. A selected culture system was validated as a drug screen using reference anti-Wolbachia (doxycycline, ABBV-4083 / flubentylosin) or direct-acting compounds (flubendazole, suramin). BALB/c IL-4Rα-/-IL-5-/- or CB.17 SCID mice were superior to Mongolian gerbils in generating adult worms and supporting in vivo persistence for periods of up to 52 weeks. Adult females retrieved from BALB/c IL-4Rα-/-IL-5-/- mice could be cultured for up to 21 days in the presence of a lymphatic endothelial cell co-culture system with comparable motility, metabolic activity and Wolbachia titres to those maintained in vivo. Drug studies confirmed significant Wolbachia depletions or direct macrofilaricidal activities could be discerned when female B. malayi were cultured for 14 days. We therefore demonstrate a novel methodology to generate adult B. malayi in vivo and accurately evaluate drug efficacy ex vivo which may be adopted for drug screening with the dual benefit of reducing overall animal use and improving anti-filarial drug development.}, } @article {pmid35668540, year = {2022}, author = {Sarwar, MS and Jahan, N and Ali, A and Yousaf, HK and Munzoor, I}, title = {Establishment of Wolbachia infection in Aedes aegypti from Pakistan via embryonic microinjection and semi-field evaluation of general fitness of resultant mosquito population.}, journal = {Parasites & vectors}, volume = {15}, number = {1}, pages = {191}, pmid = {35668540}, issn = {1756-3305}, mesh = {*Aedes/microbiology ; Animals ; *Dengue/prevention & control ; Female ; Male ; Microinjections ; Mosquito Vectors/microbiology ; Pakistan ; Pest Control, Biological ; *Wolbachia ; }, abstract = {BACKGROUND: Dengue is a mosquito-borne viral disease that is mainly spread by Aedes aegypti. It is prevalent on five continents, predominantly in tropical and sub-tropical zones across the world. Wolbachia bacteria have been extensively used in vector control strategies worldwide. The focus of the current study was to obtain a natural population of Ae. aegypti harbouring Wolbachia and to determine the impact of this bacteria on the new host in a semi-field environment.

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

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

CONCLUSIONS: Wolbachia strain wAlbB has a great potential to control the dengue vector in Ae. aegypti populations by producing 100% CI with a limited burden on its host in natural field conditions. This strain can be used as a biological tool against vector-borne diseases.}, } @article {pmid35666732, year = {2022}, author = {Bordenstein, SR and Bordenstein, SR}, title = {Widespread phages of endosymbionts: Phage WO genomics and the proposed taxonomic classification of Symbioviridae.}, journal = {PLoS genetics}, volume = {18}, number = {6}, pages = {e1010227}, pmid = {35666732}, issn = {1553-7404}, support = {R01 AI132581/AI/NIAID NIH HHS/United States ; R01 AI143725/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; *Arthropods ; *Bacteriophages/genetics ; Eukaryota ; Genomics ; Symbiosis/genetics ; *Wolbachia/genetics ; }, abstract = {Wolbachia are the most common obligate, intracellular bacteria in animals. They exist worldwide in arthropod and nematode hosts in which they commonly act as reproductive parasites or mutualists, respectively. Bacteriophage WO, the largest of Wolbachia's mobile elements, includes reproductive parasitism genes, serves as a hotspot for genetic divergence and genomic rearrangement of the bacterial chromosome, and uniquely encodes a Eukaryotic Association Module with eukaryotic-like genes and an ensemble of putative host interaction genes. Despite WO's relevance to genome evolution, selfish genetics, and symbiotic applications, relatively little is known about its origin, host range, diversification, and taxonomic classification. Here we analyze the most comprehensive set of 150 Wolbachia and phage WO assemblies to provide a framework for discretely organizing and naming integrated phage WO genomes. We demonstrate that WO is principally in arthropod Wolbachia with relatives in diverse endosymbionts and metagenomes, organized into four variants related by gene synteny, often oriented opposite the putative origin of replication in the Wolbachia chromosome, and the large serine recombinase is an ideal typing tool to distinguish the four variants. We identify a novel, putative lytic cassette and WO's association with a conserved eleven gene island, termed Undecim Cluster, that is enriched with virulence-like genes. Finally, we evaluate WO-like Islands in the Wolbachia genome and discuss a new model in which Octomom, a notable WO-like Island, arose from a split with WO. Together, these findings establish the first comprehensive Linnaean taxonomic classification of endosymbiont phages, including non-Wolbachia phages from aquatic environments, that includes a new family and two new genera to capture the collective relatedness of these viruses.}, } @article {pmid35664841, year = {2022}, author = {Montoya-Alonso, JA and García Rodríguez, SN and Carretón, E and Rodríguez Escolar, I and Costa-Rodríguez, N and Matos, JI and Morchón, R}, title = {Seroprevalence of Feline Heartworm in Spain: Completing the Epidemiological Puzzle of a Neglected Disease in the Cat.}, journal = {Frontiers in veterinary science}, volume = {9}, number = {}, pages = {900371}, pmid = {35664841}, issn = {2297-1769}, abstract = {Feline heartworm is a vector-borne zoonotic disease caused by Dirofilaria immitis. It is a cosmopolitan disease that is continuously expanding. Spain is considered an endemic country; however, although there are many published studies in dogs, feline heartworm has been poorly studied in this country. Thus, the objective was to analyze the exposure to D. immitis throughout Spain to complete the epidemiological map in the feline species. For this, 6,588 feline serum samples were analyzed for the presence of D. immitis antigens and antibodies against D. immitis and Wolbachia. The results were analyzed according to sex, age, breed, habitat, origin (owned or shelter cats), presence of clinical signs, use of preventive, location and climatology. The results showed a prevalence of 0.5% and a seroprevalence of 9.4%. The highest antibody seroprevalences were reported in the Canary Islands and the Balearic Islands (19.2 and 16%, respectively), as well as in the autonomous communities located on the Mediterranean coast (9.2-11.2%). Seropositive cats were found in both indoor and outdoor cats, and from 6 months of age. Furthermore, only 5.8% of cats received regular prophylactic treatment. The results show that feline dirofilariasis is widely distributed throughout the national territory and corroborate that, where infected dogs are present, there are cats exposed to the parasite. It is necessary to implement efficient awareness and prophylaxis measures to control the incidence and expansion of feline heartworm in Spain.}, } @article {pmid38468766, year = {2022}, author = {Fernández, MB and Bleidorn, C and Calcaterra, LA}, title = {Wolbachia Infection in Native Populations of the Invasive Tawny Crazy Ant Nylanderia fulva.}, journal = {Frontiers in insect science}, volume = {2}, number = {}, pages = {905803}, pmid = {38468766}, issn = {2673-8600}, abstract = {Antagonistic interactions can affect population growth and dispersal of an invasive species. Wolbachia are intracellular endosymbiont bacteria that infect arthropod and nematode hosts and are able to manipulate reproduction, which in some cases leads to cocladogenesis. Moreover, the presence of the strictly maternally transferred Wolbachia in a population can indirectly induce selective sweeps on the hosts' mitochondria. Ants have a Wolbachia infection rate of about 34%, which makes phylogenetic studies using mitochondrial markers vulnerable of being confounded by the effect of the endosymbiont. Nylanderia fulva is an invasive ant native to South America, considered a pest in the United States. Its distribution and biology are poorly known in its native range, and the taxonomic identity of this and its closely related species, Nylanderia pubens, has only recently been understood with the aid of molecular phylogenies. Aiming at estimating robust phylogenetic relationships of N. fulva in its native range, we investigated the presence and pattern of Wolbachia infection in populations of N. fulva from Argentina, part of its native range, to account for its possible effect on the host population structure. Using the ftsZ gene, 30 nests of N. fulva and four from sympatric Nylanderia species were screened for the presence of Wolbachia. We sequenced the MLST genes, the highly variable gene wsp, as well as glyQ, a novel target gene for which new primers were designed. Phylogeny of the ants was estimated using mtDNA (COI). We found supergroup A Wolbachia strains infecting 73% of N. fulva nests and two nests of Nylanderia sp. 1. Wolbachia phylogenetic tree inferred with MLST genes is partially congruent with the host phylogeny topology, with the exception of a lineage of strains shared by ants from different N. fulva clades. Furthermore, by comparing with Wolbachia sequences infecting other ants, we found that the strains infecting different N. fulva clades are not monophyletic. Our findings suggest there are three recent independent horizontally transmitted Wolbachia infections in N. fulva, and we found no evidence of influence of Wolbachia in the host mtDNA based phylogeny.}, } @article {pmid35652239, year = {2022}, author = {Powell, JR}, title = {Modifying mosquitoes to suppress disease transmission: Is the long wait over?.}, journal = {Genetics}, volume = {221}, number = {3}, pages = {}, pmid = {35652239}, issn = {1943-2631}, mesh = {*Aedes/genetics ; Animals ; *Dengue/epidemiology/prevention & control ; *Dengue Virus ; Humans ; Mosquito Vectors/genetics ; *Wolbachia/genetics ; }, abstract = {For more than 50 years it has been a dream of medical entomologists and public health workers to control diseases like malaria and dengue fever by modifying, through genetics and other methods, the arthropods that transmit them to humans. A brief synopsis of the history of these efforts as applied to mosquitoes is presented; none proved to be effective in reducing disease prevalence. Only in the last few years have novel approaches been developed or proposed that indicate the long wait may be over. Three recent developments are particularly promising: CRISPR-Cas9 driven genetic modification, shifting naturally occurring allele frequencies, and microbe-based modifications. The last is the furthest along in implementation. Dengue fever incidence has been reduced between 40% and 96% in 4 different regions of the world where Wolbachia-infected Aedes aegypti have been established in the field. It is not yet clear how sustainable such control programs will prove to be, but there is good reason for optimism. In light of this, the time is ripe for reinvigorated research on vectors, especially genetics. Vector-borne diseases primarily affect under-developed countries and thus have not received the attention they deserve from wealthier countries with well-developed and funded biomedical research establishments.}, } @article {pmid35644356, year = {2022}, author = {Arguni, E and Indriani, C and Rahayu, A and Supriyati, E and Yohan, B and Hayati, RF and Wardana, S and Tantowijoyo, W and Anshari, MR and Rahayu, E and Rubangi, and Ahmad, RA and Utarini, A and Simmons, CP and Sasmono, RT}, title = {Dengue virus population genetics in Yogyakarta, Indonesia prior to city-wide Wolbachia deployment.}, journal = {Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases}, volume = {102}, number = {}, pages = {105308}, doi = {10.1016/j.meegid.2022.105308}, pmid = {35644356}, issn = {1567-7257}, mesh = {Animals ; Cities ; *Culicidae ; *Dengue/epidemiology ; *Dengue Virus ; Genetics, Population ; Genotype ; Humans ; Indonesia/epidemiology ; Phylogeny ; Serogroup ; *Wolbachia/genetics ; }, abstract = {Dengue has been endemic in Yogyakarta, Indonesia for decades. Here, we report the dengue epidemiology, entomology, and virology in Yogyakarta in 2016-2017, prior to the commencement of the Applying Wolbachia to Eliminate Dengue (AWED) randomized trial. Dengue epidemiological data were compiled and blood samples from dengue-suspected patients were tested for dengue virus (DENV). Ae. aegypti mosquito samples were caught from the field using BG-Sentinel traps and tested for the presence of DENV infection. Sequencing of the DENV E gene was used to determine the phylogeny and genotypes of circulating DENV. Within the last decade, the 2016-2017 dengue incidence was considered very high. Among the 649 plasma samples collected between March 2016-February 2017; and 36,910 mosquito samples collected between December 2016-May 2017, a total of 197 and 38 samples were DENV-positive by qRT-PCR, respectively. All four DENV serotypes were detected, with DENV-3 (n = 88; 44.67%) and DENV-1 (n = 87; 44.16%) as the predominant serotype, followed by DENV-4 (n = 12; 6.09%) and DENV-2 (n = 10; 5.08%). The Yogyakarta DENV-1 isolates were classified into Genotype I and IV, while DENV-2, DENV-3, and DENV-4 isolates were classified into the Cosmopolitan genotype, Genotype I, and Genotype II, respectively. Yogyakarta DENV isolates were closely related to Indonesian strains from neighboring Javanese cities, consistent with the endemic circulation of DENV on this highly populous island. Our study provides comprehensive baseline information on the DENV population genetic characteristics in Yogyakarta, which are useful as baseline data for the AWED trial and the future DENV surveillance in the city in the presence of a Wolbachia-infected Ae. aegypti population.}, } @article {pmid35631057, year = {2022}, author = {Lee, E and Hien Nguyen, T and Yen Nguyen, T and Nam Vu, S and Duong Tran, N and Trung Nghia, L and Mai Vien, Q and Dong Nguyen, T and Kriiger Loterio, R and Iturbe-Ormaetxe, I and Flores, HA and O'Neill, SL and Anh Dang, D and Simmons, CP and Fraser, JE}, title = {Transient Introgression of Wolbachia into Aedes aegypti Populations Does Not Elicit an Antibody Response to Wolbachia Surface Protein in Community Members.}, journal = {Pathogens (Basel, Switzerland)}, volume = {11}, number = {5}, pages = {}, pmid = {35631057}, issn = {2076-0817}, support = {OPP1180815//Bill & Melinda Gates Foundation/ ; OPP1153619//Bill & Melinda Gates Foundation/ ; 102591/WT_/Wellcome Trust/United Kingdom ; N/A//Macquarie Group Foundation/ ; }, abstract = {Wolbachia is an endosymbiotic bacterium that can restrict the transmission of human pathogenic viruses by Aedes aegypti mosquitoes. Recent field trials have shown that dengue incidence is significantly reduced when Wolbachia is introgressed into the local Ae. aegypti population. Female Ae. aegypti are anautogenous and feed on human blood to produce viable eggs. Herein, we tested whether people who reside on Tri Nguyen Island (TNI), Vietnam developed antibodies to Wolbachia Surface Protein (WSP) following release of Wolbachia-infected Ae. aegypti, as a measure of exposure to Wolbachia. Paired blood samples were collected from 105 participants before and after mosquito releases and anti-WSP titres were measured by ELISA. We determined no change in anti-WSP titres after ~30 weeks of high levels of Wolbachia-Ae. aegypti on TNI. These data suggest that humans are not exposed to the major Wolbachia surface antigen, WSP, following introgression of Wolbachia-infected Ae. aegypti mosquitoes.}, } @article {pmid35630383, year = {2022}, author = {Csorba, AB and Fora, CG and Bálint, J and Felföldi, T and Szabó, A and Máthé, I and Loxdale, HD and Kentelky, E and Nyárádi, II and Balog, A}, title = {Endosymbiotic Bacterial Diversity of Corn Leaf Aphid, Rhopalosiphum maidis Fitch (Hemiptera: Aphididae) Associated with Maize Management Systems.}, journal = {Microorganisms}, volume = {10}, number = {5}, pages = {}, pmid = {35630383}, issn = {2076-2607}, abstract = {In this study, different maize fields cultivated under different management systems were sampled to test corn leaf aphid, Rhopalosiphum maidis, populations in terms of total and endosymbiotic bacterial diversity. Corn leaf aphid natural populations were collected from traditionally managed maize fields grown under high agricultural and natural landscape diversity as well as conventionally treated high-input agricultural fields grown in monoculture and with fertilizers use, hence with low natural landscape diversity. Total bacterial community assessment by DNA sequencing was performed using the Illumina MiSeq platform. In total, 365 bacterial genera were identified and 6 endosymbiont taxa. A high abundance of the primary endosymbiont Buchnera and secondary symbionts Serratia and Wolbachia were detected in all maize crops. Their frequency was found to be correlated with the maize management system used, probably with fertilizer input. Three other facultative endosymbionts ("Candidatus Hamiltonella", an uncultured Rickettsiales genus, and Spiroplasma) were also recorded at different frequencies under the two management regimes. Principal components analyses revealed that the relative contribution of the obligate and dominant symbiont Buchnera to the aphid endosymbiotic bacterial community was 72%, whereas for the managed system this was only 16.3%. When facultative symbionts alone were considered, the effect of management system revealed a DNA diversity of 23.3%.}, } @article {pmid35623398, year = {2022}, author = {Agarwal, A and Sarma, DK and Chaurasia, D and Maan, HS}, title = {Novel molecular approaches to combat vectors and vector-borne viruses: Special focus on RNA interference (RNAi) mechanisms.}, journal = {Acta tropica}, volume = {233}, number = {}, pages = {106539}, doi = {10.1016/j.actatropica.2022.106539}, pmid = {35623398}, issn = {1873-6254}, mesh = {*Aedes ; Animals ; Antiviral Agents ; *Chikungunya Fever ; Mosquito Vectors ; RNA Interference ; *Zika Virus/genetics ; *Zika Virus Infection ; }, abstract = {Vector-borne diseases, such as dengue, chikungunya, zika, yellow fever etc pose significant burden among the infectious diseases globally, especially in tropical and sub-tropical regions. Globalization, deforestation, urbanization, climate change, uncontrolled population growth, inadequate waste management and poor vector-management infrastructure have all contributed to the expansion of vector habitats and subsequent increase in vector-borne diseases throughout the world. Conventional vector control methods, such as use of insecticides, have significant negative environmental repercussions in addition to developing resistance in vectors. Till date, a very few vaccines or antiviral therapies have been approved for the treatment of vector borne diseases. In this review, we have discussed emerging molecular approaches like CRISPR (clustered regularly interspaced short palindromic repeats)/Cas-9, sterile insect technique (SIT), release of insects carrying a dominant lethal (RIDL), Wolbachia (virus transmission blocking) and RNA interference (RNAi) to combat vector and vector-borne viruses. Due to the extensive advancements in RNAi research, a special focus has been given on its types, biogenesis, mechanism of action, delivery and experimental studies evaluating their application as anti-mosquito and anti-viral agent. These technologies appear to be highly promising in terms of contributing to vector control and antiviral drug development, and hence can be used to reduce global vector and vector-borne disease burden.}, } @article {pmid35618926, year = {2022}, author = {Bouyer, J and Maiga, H and Vreysen, MJB}, title = {Assessing the efficiency of Verily's automated process for production and release of male Wolbachia-infected mosquitoes.}, journal = {Nature biotechnology}, volume = {40}, number = {10}, pages = {1441-1442}, pmid = {35618926}, issn = {1546-1696}, mesh = {*Aedes ; Animals ; Male ; Mosquito Control ; Mosquito Vectors ; *Wolbachia ; }, } @article {pmid35618925, year = {2022}, author = {Crawford, JE and Hopkins, KC and Buchman, A and Zha, T and Howell, P and Kakani, E and Ohm, JR and Snoad, N and Upson, L and Holeman, J and Massaro, P and Dobson, SL and Mulligan, FS and White, BJ}, title = {Reply to: Assessing the efficiency of Verily's automated process for production and release of male Wolbachia-infected mosquitoes.}, journal = {Nature biotechnology}, volume = {40}, number = {10}, pages = {1443-1446}, pmid = {35618925}, issn = {1546-1696}, mesh = {*Aedes ; Animals ; Male ; Mosquito Control ; *Wolbachia ; }, } @article {pmid35615502, year = {2022}, author = {Wang, L and Li, C and Wang, X and Wang, G and Shang, S and Dou, Z and Luo, Y}, title = {Gut Lignocellulose Activity and Microbiota in Asian Longhorned Beetle and Their Predicted Contribution to Larval Nutrition.}, journal = {Frontiers in microbiology}, volume = {13}, number = {}, pages = {899865}, pmid = {35615502}, issn = {1664-302X}, abstract = {Anoplophora glabripennis (Asian longhorned beetle) is a wood-boring pest that can inhabit a wide range of healthy deciduous host trees in native and invaded areas. The gut microbiota plays important roles in the acquisition of nutrients for the growth and development of A. glabripennis larvae. Herein, we investigated the larval gut structure and studied the lignocellulose activity and microbial communities of the larval gut following feeding on different host trees. The larval gut was divided into foregut, midgut, and hindgut, of which the midgut is the longest, forming a single loop under itself. Microbial community composition and lignocellulose activity in larval gut extracts were correlated with host tree species. A. glabripennis larvae fed on the preferred host (Populus gansuensis) had higher lignocellulose activity and microbial diversity than larvae reared on either a secondary host (Salix babylonica) or a resistant host (Populus alba var. pyramidalis). Wolbachia was the most dominant bacteria in the gut of larvae fed on S. babylonica and P. alba var. pyramidalis, while Enterococcus and Gibbsiella were the most dominant in larvae fed on P. gansuensis, followed by Wolbachia. The lignocellulose-degrading fungus Fusarium solani was dominant in the larval gut fed on different host trees. Functional predictions of microbial communities in the larval gut fed on different resistant host trees suggested that they all play a role in degrading lignocellulose, detoxification, and fixing nitrogen, which likely contribute to the ability of these larvae to thrive in a broad range of host tree species.}, } @article {pmid35613073, year = {2022}, author = {Harumoto, T and Fukatsu, T}, title = {Perplexing dynamics of Wolbachia proteins for cytoplasmic incompatibility.}, journal = {PLoS biology}, volume = {20}, number = {5}, pages = {e3001644}, pmid = {35613073}, issn = {1545-7885}, mesh = {Cytoplasm ; Cytosol ; Humans ; Male ; Prophages ; Spermatozoa ; *Wolbachia ; }, abstract = {The mechanism of symbiont-induced cytoplasmic incompatibility (CI) has been a long-standing mystery. A new study on Wolbachia's Cif proteins in PLOS Biology provides supportive evidence for the "Host modification model," although the alternative "Toxin-antidote model" is still in the running.}, } @article {pmid35610655, year = {2022}, author = {Waymire, E and Duddu, S and Yared, S and Getachew, D and Dengela, D and Bordenstein, SR and Balkew, M and Zohdy, S and Irish, SR and Carter, TE}, title = {Wolbachia 16S rRNA haplotypes detected in wild Anopheles stephensi in eastern Ethiopia.}, journal = {Parasites & vectors}, volume = {15}, number = {1}, pages = {178}, pmid = {35610655}, issn = {1756-3305}, support = {R15 AI151766/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; *Anopheles/genetics ; Ethiopia/epidemiology ; Haplotypes ; Humans ; *Malaria ; *Malaria, Falciparum/epidemiology ; *Malaria, Vivax ; Mosquito Vectors/genetics ; Multilocus Sequence Typing ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; *Wolbachia/genetics ; }, abstract = {BACKGROUND: About two out of three Ethiopians are at risk of malaria, a disease caused by the parasites Plasmodium falciparum and Plasmodium vivax. Anopheles stephensi, an invasive vector typically found in South Asia and the Middle East, was recently found to be distributed across eastern and central Ethiopia and is capable of transmitting both P. falciparum and P. vivax. The detection of this vector in the Horn of Africa (HOA) coupled with widespread insecticide resistance requires that new methods of vector control be investigated in order to control the spread of malaria. Wolbachia, a naturally occurring endosymbiotic bacterium of mosquitoes, has been identified as a potential vector control tool that can be explored for the control of malaria transmission. Wolbachia could be used to control the mosquito population through suppression or potentially decrease malaria transmission through population replacement. However, the presence of Wolbachia in wild An. stephensi in eastern Ethiopia is unknown. This study aimed to identify the presence and diversity of Wolbachia in An. stephensi across eastern Ethiopia.

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

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

CONCLUSIONS: These findings provide the first evidence of natural Wolbachia populations in wild An. stephensi in the HOA. They also identify the need for further research to confirm the endosymbiotic relationship between Wolbachia and An. stephensi and to investigate its utility for malaria control in the HOA.}, } @article {pmid35609042, year = {2022}, author = {Kaur, R and Leigh, BA and Ritchie, IT and Bordenstein, SR}, title = {The Cif proteins from Wolbachia prophage WO modify sperm genome integrity to establish cytoplasmic incompatibility.}, journal = {PLoS biology}, volume = {20}, number = {5}, pages = {e3001584}, pmid = {35609042}, issn = {1545-7885}, support = {F32 AI140694/AI/NIAID NIH HHS/United States ; R01 AI132581/AI/NIAID NIH HHS/United States ; R01 AI143725/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Cytoplasm/metabolism ; Drosophila melanogaster/genetics ; Female ; Male ; Prophages/genetics ; Protamines/metabolism ; Spermatozoa ; *Wolbachia ; }, abstract = {Inherited microorganisms can selfishly manipulate host reproduction to drive through populations. In Drosophila melanogaster, germline expression of the native Wolbachia prophage WO proteins CifA and CifB cause cytoplasmic incompatibility (CI) in which embryos from infected males and uninfected females suffer catastrophic mitotic defects and lethality; however, in infected females, CifA expression rescues the embryonic lethality and thus imparts a fitness advantage to the maternally transmitted Wolbachia. Despite widespread relevance to sex determination, evolution, and vector control, the mechanisms underlying when and how CI impairs male reproduction remain unknown and a topic of debate. Here, we use cytochemical, microscopic, and transgenic assays in D. melanogaster to demonstrate that CifA and CifB proteins of wMel localize to nuclear DNA throughout the process of spermatogenesis. Cif proteins cause abnormal histone retention in elongating spermatids and protamine deficiency in mature sperms that travel to the female reproductive tract with Cif proteins. Notably, protamine gene knockouts enhance wild-type CI. In ovaries, CifA localizes to germ cell nuclei and cytoplasm of early-stage egg chambers; however, Cifs are absent in late-stage oocytes and subsequently in fertilized embryos. Finally, CI and rescue are contingent upon a newly annotated CifA bipartite nuclear localization sequence. Together, our results strongly support the Host modification model of CI in which Cifs initially modify the paternal and maternal gametes to bestow CI-defining embryonic lethality and rescue.}, } @article {pmid35602266, year = {2021}, author = {Hien, NT and Anh, DD and Le, NH and Yen, NT and Phong, TV and Nam, VS and Duong, TN and Nguyen, NB and Huong, DTT and Hung, LQ and Trinh, CNT and Hoang, NV and Mai, VQ and Nghia, LT and Dong, NT and Tho, LH and Kutcher, S and Hurst, TP and Montgomery, JL and Woolfit, M and Rances, E and Kyrylos, P and L Anders, K and Nguyen, L and Brown-Kenyon, J and Caird, A and McLean, BJ and Iturbe-Ormaetxe, I and Ritchie, SA and O'Neill, SL and Ryan, PA}, title = {Environmental factors influence the local establishment of Wolbachia in Aedes aegypti mosquitoes in two small communities in central Vietnam.}, journal = {Gates open research}, volume = {5}, number = {}, pages = {147}, pmid = {35602266}, issn = {2572-4754}, support = {/WT_/Wellcome Trust/United Kingdom ; }, abstract = {Background: The wMel strain of Wolbachia has been successfully introduced into Aedes aegypti mosquitoes and subsequently shown to reduce transmission of dengue and other pathogens, under both laboratory and field conditions. Here we describe the entomological outcomes of wMel Wolbachia mosquito releases in two small communities in Nha Trang City in central Vietnam. Methods: The wMel strain of Wolbachia was backcrossed into local Aedes aegypti genotype and mosquito releases were undertaken by community members or by staff. Field monitoring was undertaken to track Wolbachia establishment in local Ae. aegypti mosquito populations. Ecological studies were undertaken to assess relationships between environmental factors and the spatial and temporal variability in Wolbachia infection prevalence in mosquitoes. Results: Releases of wMel Wolbachia Ae. aegypti mosquitoes in two small communities in Nha Trang City resulted in the initial establishment of Wolbachia in the local Ae. aegypti mosquito populations, followed by seasonal fluctuations in Wolbachia prevalence. There was significant small-scale spatial heterogeneity in Wolbachia infection prevalence in the Tri Nguyen Village site, resulting in the loss of wMel Wolbachia infection in mosquitoes in north and center areas, despite Wolbachia prevalence remaining high in mosquitoes in the south area. In the second site, Vinh Luong Ward, Wolbachia has persisted at a high level in mosquitoes throughout this site despite similar seasonal fluctuations in wMel Wolbachia prevalence. Conclusion: Seasonal variation in Wolbachia infection prevalence in mosquitoes was associated with elevated temperature conditions, and was possibly due to imperfect maternal transmission of Wolbachia. Heterogeneity in Wolbachia infection prevalence was found throughout one site, and indicates additional factors may influence Wolbachia establishment.}, } @article {pmid35604942, year = {2022}, author = {Ørsted, M and Yashiro, E and Hoffmann, AA and Kristensen, TN}, title = {Population bottlenecks constrain host microbiome diversity and genetic variation impeding fitness.}, journal = {PLoS genetics}, volume = {18}, number = {5}, pages = {e1010206}, pmid = {35604942}, issn = {1553-7404}, mesh = {Animals ; Drosophila melanogaster/genetics ; *Genetic Fitness ; Genetic Variation ; *Microbiota/genetics ; RNA, Ribosomal, 16S/genetics ; }, abstract = {It is becoming increasingly clear that microbial symbionts influence key aspects of their host's fitness, and vice versa. This may fundamentally change our thinking about how microbes and hosts interact in influencing fitness and adaptation to changing environments. Here we explore how reductions in population size commonly experienced by threatened species influence microbiome diversity. Consequences of such reductions are normally interpreted in terms of a loss of genetic variation, increased inbreeding and associated inbreeding depression. However, fitness effects of population bottlenecks might also be mediated through microbiome diversity, such as through loss of functionally important microbes. Here we utilise 50 Drosophila melanogaster lines with different histories of population bottlenecks to explore these questions. The lines were phenotyped for egg-to-adult viability and their genomes sequenced to estimate genetic variation. The bacterial 16S rRNA gene was amplified in these lines to investigate microbial diversity. We found that 1) host population bottlenecks constrained microbiome richness and diversity, 2) core microbiomes of hosts with low genetic variation were constituted from subsets of microbiomes found in flies with higher genetic variation, 3) both microbiome diversity and host genetic variation contributed to host population fitness, 4) connectivity and robustness of bacterial networks was low in the inbred lines regardless of host genetic variation, 5) reduced microbial diversity was associated with weaker evolutionary responses of hosts in stressful environments, and 6) these effects were unrelated to Wolbachia density. These findings suggest that population bottlenecks reduce hologenomic variation (combined host and microbial genetic variation). Thus, while the current biodiversity crisis focuses on population sizes and genetic variation of eukaryotes, an additional focal point should be the microbial diversity carried by the eukaryotes, which in turn may influence host fitness and adaptability with consequences for the persistence of populations.}, } @article {pmid35602967, year = {2022}, author = {Pollmann, M and Moore, LD and Krimmer, E and D'Alvise, P and Hasselmann, M and Perlman, SJ and Ballinger, MJ and Steidle, JLM and Gottlieb, Y}, title = {Highly transmissible cytoplasmic incompatibility by the extracellular insect symbiont Spiroplasma.}, journal = {iScience}, volume = {25}, number = {5}, pages = {104335}, pmid = {35602967}, issn = {2589-0042}, support = {P20 GM103646/GM/NIGMS NIH HHS/United States ; }, abstract = {Cytoplasmic incompatibility (CI) is a form of reproductive manipulation caused by maternally inherited endosymbionts infecting arthropods, like Wolbachia, whereby matings between infected males and uninfected females produce few or no offspring. We report the discovery of a new CI symbiont, a strain of Spiroplasma causing CI in the parasitoid wasp Lariophagus distinguendus. Its extracellular occurrence enabled us to establish CI in uninfected adult insects by transferring Spiroplasma-infected hemolymph. We sequenced the CI-Spiroplasma genome and did not find any homologues of any of the cif genes discovered to cause CI in Wolbachia, suggesting independent evolution of CI. Instead, the genome contains other potential CI-causing candidate genes, such as homologues of high-mobility group (HMG) box proteins that are crucial in eukaryotic development but rare in bacterial genomes. Spiroplasma's extracellular nature and broad host range encompassing medically and agriculturally important arthropods make it a promising tool to study CI and its applications.}, } @article {pmid35602212, year = {2021}, author = {Cunha, A}, title = {Trojan mosquitoes control dengue.}, journal = {Communications medicine}, volume = {1}, number = {}, pages = {17}, pmid = {35602212}, issn = {2730-664X}, abstract = {Dengue virus is transmitted by Aedes aegypti mosquitoes and causes the disease known as dengue. In a trial published in The New England Journal of Medicine, Utarini and colleagues report that release of wolbachia-infected A. aegypti populations in a dengue endemic area reduces the number of symptomatic cases and of hospitalisations.}, } @article {pmid35598650, year = {2022}, author = {Louzada-Flores, VN and Kramer, L and Brianti, E and Napoli, E and Mendoza-Roldan, JA and Bezerra-Santos, MA and Latrofa, MS and Otranto, D}, title = {Treatment with doxycycline is associated with complete clearance of circulating Wolbachia DNA in Dirofilaria immitis-naturally infected dogs.}, journal = {Acta tropica}, volume = {232}, number = {}, pages = {106513}, doi = {10.1016/j.actatropica.2022.106513}, pmid = {35598650}, issn = {1873-6254}, mesh = {Animals ; *Cell-Free Nucleic Acids ; *Dirofilaria immitis/genetics ; *Dirofilariasis/drug therapy ; *Dog Diseases/parasitology ; Dogs ; Doxycycline/pharmacology/therapeutic use ; *Wolbachia/genetics ; }, abstract = {Bacteria of the genus Wolbachia are endosymbionts of parasitic filarial nematodes, including Dirofilaria immitis, and are a target for the treatment of canine heartworm disease. In the present study, 53 naturally-infected dogs were divided in three groups, based on their positivity to D. immitis by antigen and Knott tests, to assess the efficacy of doxycycline treatment in eliminating Wolbachia from circulating blood. At T0, dogs that scored positive to both tests (G1) or to antigen only (G2) were submitted to doxycycline (10 mg/kg BID PO) treatment and to 10% Imidacloprid + 2.5% Moxidectin (Advocate®), while those negative to both tests (G3) received only 10% Imidacloprid + 2.5% Moxidectin (Advocate®). All dogs were followed-up for one year, monthly treated with Advocate® and regularly monitored by antigen and Knott tests. During the whole period, all blood samples were screened for Wolbachia-D. immitis DNA load by quantitative real-time PCR (qPCR). At T0, 88.2% of the microfilariemic dogs were positive for Wolbachia DNA, while none of the dogs from G2 or G3 were positive. Wolbachia DNA was no longer detectable in dogs from G1 following 1 month of doxycycline treatment and microfilariae (mfs) were cleared at T2. All dogs from the G1 and G2 were negative for D. immitis antigen at 12 months. Results of this study suggest that successful elimination of mfs by doxycycline is associated with complete clearance of Wolbachia DNA in D. immitis-naturally infected dogs.}, } @article {pmid35581290, year = {2022}, author = {Gomes, TMFF and Wallau, GL and Loreto, ELS}, title = {Multiple long-range host shifts of major Wolbachia supergroups infecting arthropods.}, journal = {Scientific reports}, volume = {12}, number = {1}, pages = {8131}, pmid = {35581290}, issn = {2045-2322}, mesh = {Animals ; *Arthropods/genetics/microbiology ; Female ; Insecta/microbiology ; Phylogeny ; *Wolbachia/genetics ; }, abstract = {Wolbachia is a genus of intracellular bacterial endosymbionts found in 20-66% of all insect species and a range of other invertebrates. It is classified as a single species, Wolbachia pipientis, divided into supergroups A to U, with supergroups A and B infecting arthropods exclusively. Wolbachia is transmitted mainly via vertical transmission through female oocytes, but can also be transmitted across different taxa by host shift (HS): the direct transmission of Wolbachia cells between organisms without involving vertically transmitted gametic cells. To assess the HS contribution, we recovered 50 orthologous genes from over 1000 Wolbachia genomes, reconstructed their phylogeny and calculated gene similarity. Of 15 supergroup A Wolbachia lineages, 10 have similarities ranging from 95 to 99.9%, while their hosts' similarities are around 60 to 80%. For supergroup B, four out of eight lineages, which infect diverse and distantly-related organisms such as Acari, Hemiptera and Diptera, showed similarities from 93 to 97%. These results show that Wolbachia genomes have a much higher similarity when compared to their hosts' genes, which is a major indicator of HS. Our comparative genomic analysis suggests that, at least for supergroups A and B, HS is more frequent than expected, occurring even between distantly-related species.}, } @article {pmid35560029, year = {2022}, author = {Thayanukul, P and Lertanantawong, B and Sirawaraporn, W and Charasmongkolcharoen, S and Chaibun, T and Jittungdee, R and Kittayapong, P}, title = {Simple, sensitive, and cost-effective detection of wAlbB Wolbachia in Aedes mosquitoes, using loop mediated isothermal amplification combined with the electrochemical biosensing method.}, journal = {PLoS neglected tropical diseases}, volume = {16}, number = {5}, pages = {e0009600}, pmid = {35560029}, issn = {1935-2735}, mesh = {*Aedes/genetics ; Animals ; *Arbovirus Infections ; Cost-Benefit Analysis ; Humans ; Molecular Diagnostic Techniques ; Mosquito Vectors ; Nucleic Acid Amplification Techniques ; *Wolbachia/genetics ; }, abstract = {BACKGROUND: Wolbachia is an endosymbiont bacterium generally found in about 40% of insects, including mosquitoes, but it is absent in Aedes aegypti which is an important vector of several arboviral diseases. The evidence that Wolbachia trans-infected Ae. aegypti mosquitoes lost their vectorial competence and became less capable of transmitting arboviruses to human hosts highlights the potential of using Wolbachia-based approaches for prevention and control of arboviral diseases. Recently, release of Wolbachia trans-infected Ae. aegypti has been deployed widely in many countries for the control of mosquito-borne viral diseases. Field surveillance and monitoring of Wolbachia presence in released mosquitoes is important for the success of these control programs. So far, a number of studies have reported the development of loop mediated isothermal amplification (LAMP) assays to detect Wolbachia in mosquitoes, but the methods still have some specificity and cost issues.

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

CONCLUSIONS: Our study revealed that the midgut microbiome of R. microplus is not only composed of a large number of bacteria, but that a portion also comprises eukaryotes and viruses. The data presented here enhance our understanding of this tick's midgut microbiome and provide fundamental information for the control of ticks and tick-borne diseases.}, } @article {pmid35134330, year = {2022}, author = {Horard, B and Terretaz, K and Gosselin-Grenet, AS and Sobry, H and Sicard, M and Landmann, F and Loppin, B}, title = {Paternal transmission of the Wolbachia CidB toxin underlies cytoplasmic incompatibility.}, journal = {Current biology : CB}, volume = {32}, number = {6}, pages = {1319-1331.e5}, doi = {10.1016/j.cub.2022.01.052}, pmid = {35134330}, issn = {1879-0445}, mesh = {Animals ; *Culex/genetics ; Cytoplasm ; Cytosol ; Drosophila ; Male ; *Wolbachia/genetics ; }, abstract = {Wolbachia are widespread endosymbiotic bacteria that manipulate the reproduction of arthropods through a diversity of cellular mechanisms. In cytoplasmic incompatibility (CI), a sterility syndrome originally discovered in the mosquito Culex pipiens, uninfected eggs fertilized by sperm from infected males are selectively killed during embryo development following the abortive segregation of paternal chromosomes in the zygote. Despite the recent discovery of Wolbachia CI factor (cif) genes, the mechanism by which they control the fate of paternal chromosomes at fertilization remains unknown. Here, we have analyzed the cytological distribution and cellular impact of CidA and CidB, a pair of Cif proteins from the Culex-infecting Wolbachia strain wPip. We show that expression of CidB in Drosophila S2R+ cells induces apoptosis unless CidA is co-expressed and associated with its partner. In transgenic Drosophila testes, both effectors colocalize in germ cells until the histone-to-protamine transition in which only CidB is retained in maturing spermatid nuclei. We further show that CidB is similarly targeted to maturing sperm of naturally infected Culex mosquitoes. At fertilization, CidB associates with paternal DNA regions exhibiting DNA replication stress, as a likely cause of incomplete replication of paternal chromosomes at the onset of the first mitosis. Importantly, we demonstrate that inactivation of the deubiquitylase activity of CidB does not abolish its cell toxicity or its ability to induce CI in Drosophila. Our study thus demonstrates that CI functions as a transgenerational toxin-antidote system and suggests that CidB acts by poisoning paternal DNA replication in incompatible crosses.}, } @article {pmid35134226, year = {2022}, author = {Colgan, TJ and Arce, AN and Gill, RJ and Ramos Rodrigues, A and Kanteh, A and Duncan, EJ and Li, L and Chittka, L and Wurm, Y}, title = {Genomic Signatures of Recent Adaptation in a Wild Bumblebee.}, journal = {Molecular biology and evolution}, volume = {39}, number = {2}, pages = {}, pmid = {35134226}, issn = {1537-1719}, support = {BB/K004204/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; BB/T015683/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; }, mesh = {Animals ; Bees/genetics ; *Ecosystem ; *Genomics ; }, abstract = {Environmental changes threaten insect pollinators, creating risks for agriculture and ecosystem stability. Despite their importance, we know little about how wild insects respond to environmental pressures. To understand the genomic bases of adaptation in an ecologically important pollinator, we analyzed genomes of Bombus terrestris bumblebees collected across Great Britain. We reveal extensive genetic diversity within this population, and strong signatures of recent adaptation throughout the genome affecting key processes including neurobiology and wing development. We also discover unusual features of the genome, including a region containing 53 genes that lacks genetic diversity in many bee species, and a horizontal gene transfer from a Wolbachia bacteria. Overall, the genetic diversity we observe and how it is distributed throughout the genome and the population should support the resilience of this important pollinator species to ongoing and future selective pressures. Applying our approach to more species should help understand how they can differ in their adaptive potential, and to develop conservation strategies for those most at risk.}, } @article {pmid35132164, year = {2022}, author = {Xu, W and Wang, Y and Wang, YH and Zhang, YN and Wang, JF}, title = {Diversity and dynamics of bacteria at the Chrysomya megacephala pupal stage revealed by third-generation sequencing.}, journal = {Scientific reports}, volume = {12}, number = {1}, pages = {2006}, pmid = {35132164}, issn = {2045-2322}, mesh = {Animals ; Bacteroidetes/genetics/isolation & purification/physiology ; Calliphoridae/*microbiology ; Erysipelothrix/genetics/isolation & purification ; Firmicutes/genetics/isolation & purification/physiology ; Forensic Entomology ; Gammaproteobacteria/genetics/isolation & purification/physiology ; *Microbiota/genetics/physiology ; Proteobacteria/genetics/isolation & purification/physiology ; Pupa/*microbiology ; RNA, Bacterial/genetics ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, RNA/*methods ; Symbiosis ; Wolbachia/genetics/isolation & purification/physiology ; }, abstract = {Characterization of the microbial community is essential for understanding the symbiotic relationships between microbes and host insects. Chrysomya megacephala is a vital resource, a forensic insect, a pollinator, and a vector for enteric bacteria, protozoa, helminths, and viruses. However, research on its microbial community is incomprehensive, particularly at the pupal stage, which comprises approximately half of the entire larval development stage and is important entomological evidence in forensic medicine. For the first time, this study investigated the bacterial communities of C. megacephala pupae at different ages using third-generation sequencing technology. The results showed that C. megacephala has a diverse and dynamic bacterial community. Cluster analysis at ≥ 97% similarity produced 154 operational taxonomic units (OTUs) that belonged to 10 different phyla and were distributed into 15 classes, 28 orders, 50 families, 88 genera, and 130 species. Overall, the number of bacterial OTUs increased with the development of pupae, and the relative abundance of Wolbachia in the Day5 group was significantly lower than that in the other groups. Within the pupal stage, Proteobacteria, Firmicutes, and Bacteroidetes were the dominant phyla of bacteria. At the genus level, Wolbachia and Ignatzschineria coexisted, a rarely known feature. In addition, we found Erysipelothrix rhusiopathiae, the etiological agent of swine erysipelas, which is rarely identified in insects. This study enriches the understanding of the microbial community of C. megacephala and provides a reference for better utilization and control of C. megacephala.}, } @article {pmid35127567, year = {2021}, author = {Mao, W and Zeng, Q and She, L and Yuan, H and Luo, Y and Wang, R and She, Y and Wang, W and Wang, C and Pan, X}, title = {Wolbachia Utilizes lncRNAs to Activate the Anti-Dengue Toll Pathway and Balance Reactive Oxygen Species Stress in Aedes aegypti Through a Competitive Endogenous RNA Network.}, journal = {Frontiers in cellular and infection microbiology}, volume = {11}, number = {}, pages = {823403}, pmid = {35127567}, issn = {2235-2988}, mesh = {*Aedes/genetics ; Animals ; Gene Regulatory Networks ; *MicroRNAs/genetics ; *RNA, Long Noncoding/genetics/metabolism ; RNA, Messenger/genetics ; Reactive Oxygen Species ; *Wolbachia/genetics ; }, abstract = {Long non-coding RNAs (lncRNA), a class of RNA molecules without protein coding potential, are more than 200 nucleotides in length and widely present in a variety of species. Although increasing progress in regard to the determination of lncRNA function has been made in vertebrates, Aedes aegypti lncRNAs were only identified recently and the functions of few lncRNAs have been annotated so far. Herein, the genome-wide alteration of the lncRNA expression profile trigged by Wolbachia wAlbB infection was investigated by comparing A. aegypti Aag2 cells and W-Aag2 cells infected with Wolbachia wAlbB. Based on lncRNA sequencing, 3035 differentially expressed lncRNAs (DE lncRNAs) in total were identified upon Wolbachia infection, which were further validated by quantitative PCR. The constructed co-expression network of DE lncRNAs and mRNAs revealed that Wolbachia-induced DE lncRNAs were highly enriched in the oxidative phosphorylation pathway via trans-activity, according to the KEGG pathway enrichment analyses. In addition, the established competitive endogenous RNA (ceRNA) network identifies the DE lncRNAs enriched in cellular oxidant detoxification based on GO enrichment analysis. Furthermore, silencing of aae-lnc-7598, the significantly up-regulated lncRNA with the highest fold change induced by Wolbachia, caused a significant reduction of antioxidant catalase 1B (CAT1B) gene expression as well as the enhancement of mitochondrial reactive oxygen species (ROS) production in living cells. These findings indicate that Wolbachia manipulates lncRNA to balance intracellular ROS stress and ensure its endosymbiosis in host A. aegypti. Notably, the function assay demonstrated that aae-lnc-0165 suppressed by Wolbachia could induce expression of the REL1 gene, the key regulator of downstream Toll pathway, through the sequence-specific binding of aae-miR-980-5p, which contributes to the activation of Toll pathway. Moreover, the depletion of aae-lnc-0165 caused the suppression of mitochondrial ROS levels in living cells. Our data reveal that Wolbachia activates the anti-dengue Toll pathway through a lncRNA-ceRNA pattern. Taken together, our finding suggested that Wolbachia utilizes lncRNAs to activate host anti-dengue Toll pathway via a ceRNA network. Moreover, Wolbachia employs lncRNAs to ensure ROS homeostasis for ROS-based anti-dengue defense through either trans-regulation or the ceRNA network. This study identifies novel potential molecular biomarkers for prevention and control of epidemic dengue.}, } @article {pmid35127140, year = {2022}, author = {Turelli, M and Barton, NH}, title = {Why did the Wolbachia transinfection cross the road? drift, deterministic dynamics, and disease control.}, journal = {Evolution letters}, volume = {6}, number = {1}, pages = {92-105}, pmid = {35127140}, issn = {2056-3744}, abstract = {Maternally inherited Wolbachia transinfections are being introduced into natural mosquito populations to reduce the transmission of dengue, Zika, and other arboviruses. Wolbachia-induced cytoplasmic incompatibility provides a frequency-dependent reproductive advantage to infected females that can spread transinfections within and among populations. However, because transinfections generally reduce host fitness, they tend to spread within populations only after their frequency exceeds a critical threshold. This produces bistability with stable equilibrium frequencies at both 0 and 1, analogous to the bistability produced by underdominance between alleles or karyotypes and by population dynamics under Allee effects. Here, we analyze how stochastic frequency variation produced by finite population size can facilitate the local spread of variants with bistable dynamics into areas where invasion is unexpected from deterministic models. Our exemplar is the establishment of wMel Wolbachia in the Aedes aegypti population of Pyramid Estates (PE), a small community in far north Queensland, Australia. In 2011, wMel was stably introduced into Gordonvale, separated from PE by barriers to A. aegypti dispersal. After nearly 6 years during which wMel was observed only at low frequencies in PE, corresponding to an apparent equilibrium between immigration and selection, wMel rose to fixation by 2018. Using analytic approximations and statistical analyses, we demonstrate that the observed fixation of wMel at PE is consistent with both stochastic transition past an unstable threshold frequency and deterministic transformation produced by steady immigration at a rate just above the threshold required for deterministic invasion. The indeterminacy results from a delicate balance of parameters needed to produce the delayed transition observed. Our analyses suggest that once Wolbachia transinfections are established locally through systematic introductions, stochastic "threshold crossing" is likely to only minimally enhance spatial spread, providing a local ratchet that slightly-but systematically-aids area-wide transformation of disease-vector populations in heterogeneous landscapes.}, } @article {pmid35126329, year = {2021}, author = {Zheng, Z and Zhao, M and Zhang, Z and Hu, X and Xu, Y and Wei, C and He, H}, title = {Lactic Acid Bacteria Are Prevalent in the Infrabuccal Pockets and Crops of Ants That Prefer Aphid Honeydew.}, journal = {Frontiers in microbiology}, volume = {12}, number = {}, pages = {785016}, pmid = {35126329}, issn = {1664-302X}, abstract = {Ants are evolutionarily successful species and occupy diverse trophic and habitat niches on the earth. To fulfill dietary requirements, ants have established commensalism with both sap-feeding insects and bacteria. In this study, we used high-throughput sequencing of the bacterial 16S rRNA gene to characterize the bacterial composition and structure of the digestive tracts in three species of Formica ants and Lasius niger (Linnaeus)-species that predominantly feed on honeydew secreted by aphids. We found that bacterial communities displayed species- and colony-level signatures, and that bacterial communities in the infrabuccal pockets and crops were different from those in the midguts and hindguts. Lactobacillus and Wolbachia were dominant in the infrabuccal pockets and crops of workers, whereas Wolbachia was dominant in the midguts, hindguts and brood (larvae, pupae and cocoons). To learn more about the dominant Lactobacillus in ants, we assessed its prevalence in a wide range of aphid-tending ants using diagnostic PCR. We found that Lactobacillus was more prevalent in Formicinae than in Myrmicinae species. We also isolated four strains of lactic acid bacteria (Lactobacillus sanfranciscensis, Lactobacillus lindneri, Weissella cibaria and Fructobacillus sp.) from the infrabuccal pockets and crops of aphid-tending ants using a culture-dependent method. Two predominant lactic acid bacterial isolates, Lactobacillus sanfranciscensis (La2) and Weissella cibaria (La3), exhibited abilities in catabolizing sugars (sucrose, trehalose, melezitose and raffinose) known to be constituents of hemipteran honeydew. These findings contribute to further understanding the association between ants, aphids and bacteria, and provide additional information on the function of lactic acid bacteria in ants.}, } @article {pmid35124699, year = {2022}, author = {Doremus, MR and Stouthamer, CM and Kelly, SE and Schmitz-Esser, S and Hunter, MS}, title = {Quality over quantity: unraveling the contributions to cytoplasmic incompatibility caused by two coinfecting Cardinium symbionts.}, journal = {Heredity}, volume = {128}, number = {3}, pages = {187-195}, pmid = {35124699}, issn = {1365-2540}, mesh = {Animals ; Bacteroidetes/genetics ; Cytoplasm/microbiology ; Female ; Male ; Reproduction ; Symbiosis ; *Wasps/genetics/microbiology ; *Wolbachia ; }, abstract = {Cytoplasmic incompatibility (CI) is a common form of reproductive sabotage caused by maternally inherited bacterial symbionts of arthropods. CI is a two-step manipulation: first, the symbiont modifies sperm in male hosts which results in the death of fertilized, uninfected embryos. Second, when females are infected with a compatible strain, the symbiont reverses sperm modification in the fertilized egg, allowing offspring of infected females to survive and spread the symbiont to high frequencies in a population. Although CI plays a role in arthropod evolution, the mechanism of CI is unknown for many symbionts. Cardinium hertigii is a common CI-inducing symbiont of arthropods, including parasitoid wasps like Encarsia partenopea. This wasp harbors two Cardinium strains, cEina2 and cEina3, and exhibits strong CI. The strains infect wasps at different densities, with the cEina3 present at a lower density than cEina2, and it was previously not known which strain caused CI. By differentially curing wasps of cEina3, we found that this low-density symbiont is responsible for CI and modifies males during their pupal stage. cEina2 does not modify host reproduction and may spread by 'hitchhiking' with cEina3 CI or by conferring an unknown benefit. The cEina3 strain also shows a unique localization pattern in male reproductive tissues. Instead of infecting sperm like other CI-inducing symbionts, cEina3 cells are found in somatic cells at the testis base and around the seminal vesicle. This may allow the low-density cEina3 to efficiently modify host males and suggests that cEina3 uses a different modification strategy than sperm-infecting CI symbionts.}, } @article {pmid35124068, year = {2022}, author = {Alonso, AC and Stein, M and Matías Hisgen, C and Micieli, MV}, title = {Abiotic factors affecting the prevalence of Wolbachia (Rickettsiaceae) in immature Aedes albopictus (Skuse) (Culicidae).}, journal = {Journal of invertebrate pathology}, volume = {189}, number = {}, pages = {107730}, doi = {10.1016/j.jip.2022.107730}, pmid = {35124068}, issn = {1096-0805}, mesh = {*Aedes/microbiology ; Animals ; Prevalence ; *Rickettsiaceae ; Symbiosis ; *Wolbachia ; }, abstract = {Wolbachia is a genus of gram-negative endosymbiotic bacterium of maternal transmission, located mainly in the gonads of arthropods, including mosquitoes such as Aedes albopictus. The current distribution of Ae. albopictus in Argentina is restricted to the subtropical northeastern region of the country. Here, we studied the seasonal prevalence of Wolbachia detected in Ae. albopictus larvae and the relationship between the abiotic factors of the larval microhabitat and the infection status, in Eldorado city, Misiones province, subtropical region. The prevalence of Wolbachia infection found was 76.89% (n = 312). From the total samples examined, 52.80% (n = 214) showed double infection with the wAlbA/wAlbB strains, 23.84% (n = 97) infection only with wAlbB, and 0.25% (n = 1) only with wAlbA. The prevalence of double infection did not present statistically significant differences between the sites studied. For single infection, the lowest prevalence value of the wAlbB strain (13.33%) was found in the natural park, whereas the highest was found in the family dwellings and cemeteries. Tire repair shops showed an intermediate value. The wAlbA single infection was identified once. Our results also showed an association between temperature and slightly turbid waters with exposure to the sun in the larval habitats and the probability of infection by Wolbachia.}, } @article {pmid35116014, year = {2021}, author = {Gendrin, M and Favia, G and Herren, JK}, title = {Editorial: Microbiota: A Consequential Third Wheel in the Mosquito-Pathogen Relationship.}, journal = {Frontiers in microbiology}, volume = {12}, number = {}, pages = {811880}, doi = {10.3389/fmicb.2021.811880}, pmid = {35116014}, issn = {1664-302X}, } @article {pmid35115867, year = {2022}, author = {Raupach, MJ and Rulik, B and Spelda, J}, title = {Surprisingly high genetic divergence of the mitochondrial DNA barcode fragment (COI) within Central European woodlice species (Crustacea, Isopoda, Oniscidea).}, journal = {ZooKeys}, volume = {1082}, number = {}, pages = {103-125}, pmid = {35115867}, issn = {1313-2989}, abstract = {DNA barcoding has become the most popular approach for species identification in recent years. As part of the German Barcode of Life project, the first DNA barcode library for terrestrial and freshwater isopods from Germany is presented. The analyzed barcode library included 38 terrestrial (78% of the documented species of Germany) and five freshwater (63%) species. A total of 513 new barcodes was generated and 518 DNA barcodes were analyzed. This analysis revealed surprisingly high intraspecific genetic distances for numerous species, with a maximum of 29.4% for Platyarthrushoffmannseggii Brandt, 1833. The number of BINs per species ranged from one (32 species, 68%) to a maximum of six for Trachelipusrathkii (Brandt, 1833). In spite of such high intraspecific variability, interspecific distances with values between 12.6% and 29.8% allowed a valid species assignment of all analyzed isopods. The observed high intraspecific distances presumably result from phylogeographic events, Wolbachia infections, atypical mitochondrial DNAs, heteroplasmy, or various combinations of these factors. Our study represents the first step in generating an extensive reference library of DNA barcodes for terrestrial and freshwater isopods for future molecular biodiversity assessment studies.}, } @article {pmid35115648, year = {2022}, author = {Katlav, A and Nguyen, DT and Morrow, JL and Spooner-Hart, RN and Riegler, M}, title = {Endosymbionts moderate constrained sex allocation in a haplodiploid thrips species in a temperature-sensitive way.}, journal = {Heredity}, volume = {128}, number = {3}, pages = {169-177}, pmid = {35115648}, issn = {1365-2540}, mesh = {Animals ; Bacteroidetes ; Female ; Male ; Sex Ratio ; Symbiosis/genetics ; Temperature ; *Thysanoptera/genetics/microbiology ; *Wolbachia/genetics ; }, abstract = {Maternally inherited bacterial endosymbionts that affect host fitness are common in nature. Some endosymbionts colonise host populations by reproductive manipulations (such as cytoplasmic incompatibility; CI) that increase the reproductive fitness of infected over uninfected females. Theory predicts that CI-inducing endosymbionts in haplodiploid hosts may also influence sex allocation, including in compatible crosses, however, empirical evidence for this is scarce. We examined the role of two common CI-inducing endosymbionts, Cardinium and Wolbachia, in the sex allocation of Pezothrips kellyanus, a haplodiploid thrips species with a split sex ratio. In this species, irrespective of infection status, some mated females are constrained to produce extremely male-biased broods, whereas other females produce extremely female-biased broods. We analysed brood sex ratio of females mated with males of the same infection status at two temperatures. We found that at 20 °C the frequency of constrained sex allocation in coinfected pairs was reduced by 27% when compared to uninfected pairs. However, at 25 °C the constrained sex allocation frequency increased and became similar between coinfected and uninfected pairs, resulting in more male-biased population sex ratios at the higher temperature. This temperature-dependent pattern occurred without changes in endosymbiont densities and compatibility. Our findings indicate that endosymbionts affect sex ratios of haplodiploid hosts beyond the commonly recognised reproductive manipulations by causing female-biased sex allocation in a temperature-dependent fashion. This may contribute to a higher transmission efficiency of CI-inducing endosymbionts and is consistent with previous models that predict that CI by itself is less efficient in driving endosymbiont invasions in haplodiploid hosts.}, } @article {pmid35112745, year = {2022}, author = {Sigle, LT and Jones, M and Novelo, M and Ford, SA and Urakova, N and Lymperopoulos, K and Sayre, RT and Xi, Z and Rasgon, JL and McGraw, EA}, title = {Assessing Aedes aegypti candidate genes during viral infection and Wolbachia-mediated pathogen blocking.}, journal = {Insect molecular biology}, volume = {31}, number = {3}, pages = {356-368}, pmid = {35112745}, issn = {1365-2583}, support = {R01 AI116636/AI/NIAID NIH HHS/United States ; R01 AI143758/AI/NIAID NIH HHS/United States ; R56 AI116636/AI/NIAID NIH HHS/United States ; }, mesh = {*Aedes/genetics ; Animals ; *Chikungunya virus ; *Dengue Virus/genetics ; Mosquito Vectors/genetics ; *Virus Diseases ; *Wolbachia/physiology ; }, abstract = {One approach to control dengue virus transmission is the symbiont Wolbachia, which limits viral infection in mosquitoes. Despite plans for its widespread use in Aedes aegypti, Wolbachia's mode of action remains poorly understood. Many studies suggest that the mechanism is likely multifaceted, involving aspects of immunity, cellular stress and nutritional competition. A previous study from our group used artificial selection to identify a new mosquito candidate gene related to viral blocking; alpha-mannosidase-2a (alpha-Mann-2a) with a predicted role in protein glycosylation. Protein glycosylation pathways tend to be involved in complex host-viral interactions; however, the function of alpha-mannosidases has not been described in mosquito-virus interactions. We examined alpha-Mann-2a expression in response to virus and Wolbachia infections and whether reduced gene expression, caused by RNA interference, affected viral loads. We show that dengue virus (DENV) infection affects the expression of alpha-Mann-2a in a tissue- and time-dependent manner, whereas Wolbachia infection had no effect. In the midgut, DENV prevalence increased following knockdown of alpha-Mann-2a expression in Wolbachia-free mosquitoes, suggesting that alpha-Mann-2a interferes with infection. Expression knockdown had the same effect on the togavirus chikungunya virus, indicating that alpha-Mann-2a may have broad antivirus effects in the midgut. Interestingly, we were unable to knockdown the expression in Wolbachia-infected mosquitoes. We also provide evidence that alpha-Mann-2a may affect the transcriptional level of another gene predicted to be involved in viral blocking and cell adhesion; cadherin87a. These data support the hypothesis that glycosylation and adhesion pathways may broadly be involved in viral infection in Ae. aegypti.}, } @article {pmid35092614, year = {2022}, author = {Rodrigues, LR and Zélé, F and Santos, I and Magalhães, S}, title = {No evidence for the evolution of mating behavior in spider mites due to Wolbachia-induced cytoplasmic incompatibility.}, journal = {Evolution; international journal of organic evolution}, volume = {76}, number = {3}, pages = {623-635}, doi = {10.1111/evo.14429}, pmid = {35092614}, issn = {1558-5646}, support = {COMPCON GA 725419//H2020 European Research Council/ ; SFRH/BD/87628/2012//Fundação para a Ciência e a Tecnologia/ ; EXPL/BIA-EVL/0131/2021//Fundação para a Ciência e a Tecnologia/ ; FCT-ANR//BIA- EVF/0013/2012//FCT-ANR collaboration/ ; }, mesh = {Animals ; *Arthropods ; Cytoplasm ; Female ; Male ; Reproduction ; *Tetranychidae/genetics ; *Wolbachia/genetics ; }, abstract = {Arthropods are often infected with Wolbachia inducing cytoplasmic incompatibility (CI), whereby crosses between uninfected females and infected males yield unviable fertilized offspring. Although uninfected females benefit from avoiding mating with Wolbachia-infected males, this behavior is not always present in host populations and its evolution may hinge upon various factors. Here, we used spider mites to test whether CI could select for mate preference in uninfected females in absence of kin recognition. We found that uninfected females from several field-derived populations showed no preference for infected or uninfected males, nor evolved a preference after being exposed to CI for 12-15 generations by maintaining uninfected females with both infected and uninfected males (i.e., stable "infection polymorphism"). This suggests that Wolbachia-mediated mate choice evolution may require very specific conditions in spider mites. However, after experimental evolution, the copulation duration of Wolbachia-infected control males was significantly higher than that of uninfected control males, but not than that of uninfected males from the "infection polymorphism" regime. This result illustrates how gene flow may oppose Wolbachia-driven divergence between infected and uninfected hosts in natural populations.}, } @article {pmid35073749, year = {2022}, author = {Sun, G and Zhang, M and Chen, H and Hochstrasser, M}, title = {The CinB Nuclease from wNo Wolbachia Is Sufficient for Induction of Cytoplasmic Incompatibility in Drosophila.}, journal = {mBio}, volume = {13}, number = {1}, pages = {e0317721}, pmid = {35073749}, issn = {2150-7511}, support = {R35 GM136325/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Male ; Female ; Humans ; Drosophila/microbiology ; *Wolbachia/genetics ; Saccharomyces cerevisiae ; Animals, Genetically Modified ; Cytoplasm/microbiology ; *Infertility, Male ; }, abstract = {Wolbachia is an obligate intracellular bacterium that can alter reproduction of its arthropod hosts, often through a mechanism called cytoplasmic incompatibility (CI). In CI, uninfected females fertilized by infected males yield few offspring, but if both are similarly infected, normal embryo viability results (called "rescue"). CI factors (Cifs) responsible for CI are pairs of proteins encoded by linked genes. The downstream gene in each pair encodes either a deubiquitylase (CidB) or a nuclease (CinB). The upstream gene products, CidA and CinA, bind their cognate enzymes with high specificity. Expression of CidB or CinB in yeast inhibits growth, but growth is rescued by expression of the cognate CifA protein. By contrast, transgenic Drosophila male germ line expression of both cifA and cifB was reported to be necessary to induce CI-like embryonic arrest; cifA expression alone in females is sufficient for rescue. This pattern, seen with genes from several Wolbachia strains, has been called the "2-by-1" model. Here, we show that male germ line expression of the cinB gene alone, from a distinct clade of cif genes from wNo Wolbachia, is sufficient to induce nearly complete loss of embryo viability. This male sterility is fully rescued by cognate cinA[w][No] expression in the female germ line. The proteins behave similarly in yeast. CinB[w][No] toxicity depends on its nuclease active site. These results demonstrate that highly divergent CinB nucleases can induce CI, that rescue by cognate CifA factors is a general feature of Wolbachia CI systems, and that CifA is not strictly required in males for CI induction. IMPORTANCE Wolbachia bacteria live within the cells of many insects. Like mitochondria, they are only inherited from females. Wolbachia often increases the number of infected females to promote spread of infection using a type of male sterility called cytoplasmic incompatibility (CI): when uninfected females mate with infected males, most embryos die; if both are similarly infected, embryos develop normally, giving infected females an advantage in producing offspring. CI is being used against disease-carrying mosquitoes and agricultural pests. Wolbachia proteins called CifA and CifB, which bind one another, cause CI, but how they work has been unclear. Here, we show that a CifB protein singly produced in fruit fly males causes sterility in crosses to normal females, but this is rescued if the females produce the CifA partner. These findings clarify a broad range of observations on CI and will allow more rational approaches to using it for insect control.}, } @article {pmid35069682, year = {2021}, author = {Rašić, G and Lobo, NF and Jeffrey Gutiérrez, EH and Sánchez C, HM and Marshall, JM}, title = {Monitoring Needs for Gene Drive Mosquito Projects: Lessons From Vector Control Field Trials and Invasive Species.}, journal = {Frontiers in genetics}, volume = {12}, number = {}, pages = {780327}, pmid = {35069682}, issn = {1664-8021}, abstract = {As gene drive mosquito projects advance from contained laboratory testing to semi-field testing and small-scale field trials, there is a need to assess monitoring requirements to: i) assist with the effective introduction of the gene drive system at field sites, and ii) detect unintended spread of gene drive mosquitoes beyond trial sites, or resistance mechanisms and non-functional effector genes that spread within trial and intervention sites. This is of particular importance for non-localized gene drive projects, as the potential scale of intervention means that monitoring is expected to be more costly than research, development and deployment. Regarding monitoring needs for population replacement systems, lessons may be learned from experiences with Wolbachia-infected mosquitoes, and for population suppression systems, from experiences with releases of genetically sterile male mosquitoes. For population suppression systems, assessing monitoring requirements for tracking population size and detecting rare resistant alleles are priorities, while for population replacement systems, allele frequencies must be tracked, and pressing concerns include detection of gene drive alleles with non-functional effector genes, and resistance of pathogens to functional effector genes. For spread to unintended areas, open questions relate to the optimal density and placement of traps and frequency of sampling in order to detect gene drive alleles, drive-resistant alleles or non-functional effector genes while they can still be effectively managed. Invasive species management programs face similar questions, and lessons may be learned from these experiences. We explore these monitoring needs for gene drive mosquito projects progressing through the phases of pre-release, release and post-release.}, } @article {pmid35068955, year = {2021}, author = {Urfer, K and Spasojevic, T and Klopfstein, S and Baur, H and Lasut, L and Kropf, C}, title = {Incongruent molecular and morphological variation in the crab spider Synemaglobosum (Araneae, Thomisidae) in Europe.}, journal = {ZooKeys}, volume = {1078}, number = {}, pages = {107-134}, pmid = {35068955}, issn = {1313-2989}, abstract = {Establishing species boundaries is one of the challenges taxonomists around the world have been tackling for centuries. The relation between intraspecific and interspecific variability is still under discussion and in many taxa it remains understudied. Here the hypothesis of single versus multiple species of the crab spider Synemaglobosum (Fabricius) is tested. The wide distribution range as well as its high morphological variability makes this species an interesting candidate for re-evaluation using an integrative approach. This study combines information from barcoding, phylogenetic reconstruction based on mitochondrial CO1 and ITS2 of more than 60 specimens collected over a wide range of European localities, and morphology. The findings show deep clades with up to 6% mean pairwise distance in the CO1 barcode without any biogeographical pattern. The nuclear ITS2 gene did not support the CO1 clades. Morphological assessment of somatic and genital characters in males and females and a morphometric analysis of the male palp uncovered high intraspecific variation that does not match the CO1 or ITS2 phylogenies or biogeography either. Screening for endosymbiotic Wolbachia bacteria was conducted and only a single infected specimen was found. Several scenarios might explain these inconsistent patterns. While the deep divergences in the barcoding marker might suggest cryptic or ongoing speciation or geographical isolation in the past, the lack of congruent variation in the nuclear ITS2 gene or the studied morphological character systems, especially the male palp, indicates that S.globosum might simply be highly polymorphic both in terms of its mtDNA and morphology. Therefore, more data on ecology and behaviour and full genome sequences are necessary to ultimately resolve this taxonomically intriguing case.}, } @article {pmid35068604, year = {2022}, author = {Sureshan, SC and Mohideen, HS and Nair, TS}, title = {Gut Metagenomic Profiling of Gossypol Induced Oxycarenus laetus (Hemiptera: Lygaeidae) Reveals Gossypol Tolerating Bacterial Species.}, journal = {Indian journal of microbiology}, volume = {62}, number = {1}, pages = {54-60}, pmid = {35068604}, issn = {0046-8991}, abstract = {UNLABELLED: Oxycarenus laetus is a cotton pest that primarily feeds on seeds that are rich in gossypol. Though gossypol is toxic to general herbivores, O. laetus does not show such complications and instead grows and reproduces well on cotton plants compared to its other hosts. In this study, we have fed O. laetus with natural and induced gossypol-based diets to explore the difference in its gut microbiota. We performed NGS 16S rRNA amplicon sequencing on the Illumina MiSeq platform and analyzed the data using the QIIME2 pipeline supplemented with Greengenes and EZBioCloud reference databases. We also used culture-based methods to identify a few abundant gut bacteria present in O. laetus. Enterococcus faecalis, Wolbachia bourtzisii, Wolbachia pipientis, Corynebacterium glyciniphilum, Staphylococcus sciuri, and Kocuria rosea were some of the major species that formed the core gut microbiome of O. laetus. We have also observed that some species were present only in the sample with the highest concentration of gossypol, signifying that they might have the potential to degrade gossypol.

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

RESULTS: The newly designed primer pair (BeeCox1F1/BeeCox1R2) outperformed the standard barcoding primer (LCO1490/HCO2198). By generating barcodes for a small test set of bees we found that the new primer pair produced high-quality sequences in all cases for unambiguous species identification using BOLD. Conversely, the standard barcoding primers often co-amplified the homologous Wolbachia gene and resulted in mixed chromatogram signals. These sequences showed high similarity with the bacterial endosymbiont instead of the host.}, } @article {pmid34820166, year = {2021}, author = {Price, DC and Brennan, JR and Wagner, NE and Egizi, AM}, title = {Comparative hologenomics of two Ixodes scapularis tick populations in New Jersey.}, journal = {PeerJ}, volume = {9}, number = {}, pages = {e12313}, pmid = {34820166}, issn = {2167-8359}, abstract = {Tick-borne diseases, such as those transmitted by the blacklegged tick Ixodes scapularis, are a significant and growing public health problem in the US. There is mounting evidence that co-occurring non-pathogenic microbes can also impact tick-borne disease transmission. Shotgun metagenome sequencing enables sampling of the complete tick hologenome-the collective genomes of the tick and all of the microbial species contained therein, whether pathogenic, commensal or symbiotic. This approach simultaneously uncovers taxonomic composition and allows the detection of intraspecific genetic variation, making it a useful tool to compare spatial differences across tick populations. We evaluated this approach by comparing hologenome data from two tick samples (N = 6 ticks per location) collected at a relatively fine spatial scale, approximately 23 km apart, within a single US county. Several intriguing variants in the data between the two sites were detected, including polymorphisms in both in the tick's own mitochondrial DNA and that of a rickettsial endosymbiont. The two samples were broadly similar in terms of the microbial species present, including multiple known tick-borne pathogens (Borrelia burgdorferi, Babesia microti, and Anaplasma phagocytophilum), filarial nematodes, and Wolbachia and Babesia species. We assembled the complete genome of the rickettsial endosymbiont (most likely Rickettsia buchneri) from both populations. Our results provide further evidence for the use of shotgun metagenome sequencing as a tool to compare tick hologenomes and differentiate tick populations across localized spatial scales.}, } @article {pmid34819638, year = {2021}, author = {Adams, KL and Abernathy, DG and Willett, BC and Selland, EK and Itoe, MA and Catteruccia, F}, title = {Wolbachia cifB induces cytoplasmic incompatibility in the malaria mosquito vector.}, journal = {Nature microbiology}, volume = {6}, number = {12}, pages = {1575-1582}, pmid = {34819638}, issn = {2058-5276}, support = {/HHMI/Howard Hughes Medical Institute/United States ; }, mesh = {Aedes/genetics/microbiology/physiology ; Animals ; Anopheles/genetics/*microbiology/*physiology ; Bacterial Proteins/genetics/*metabolism ; *Extrachromosomal Inheritance ; Female ; Infertility, Male ; Malaria/transmission ; Male ; Mosquito Vectors/genetics/*microbiology/*physiology ; Wolbachia/genetics/*metabolism ; }, abstract = {Wolbachia, a maternally inherited intracellular bacterial species, can manipulate host insect reproduction by cytoplasmic incompatibility (CI), which results in embryo lethality in crosses between infected males and uninfected females. CI is encoded by two prophage genes, cifA and cifB. Wolbachia, coupled with the sterile insect technique, has been used in field trials to control populations of the dengue vector Aedes albopictus, but CI-inducing strains are not known to infect the malaria vector Anopheles gambiae. Here we show that cifA and cifB can induce conditional sterility in the malaria vector An. gambiae. We used transgenic expression of these Wolbachia-derived genes in the An. gambiae germline to show that cifB is sufficient to cause embryonic lethality and that cifB-induced sterility is rescued by cifA expression in females. When we co-expressed cifA and cifB in male mosquitoes, the CI phenotype was attenuated. In female mosquitoes, cifB impaired fertility, which was overcome by co-expression of cifA. Our findings pave the way towards using CI to control malaria mosquito vectors.}, } @article {pmid34792280, year = {2022}, author = {Sanaei, E and Lin, YP and Cook, LG and Engelstädter, J}, title = {Wolbachia in scale insects: a distinct pattern of infection frequencies and potential transfer routes via ant associates.}, journal = {Environmental microbiology}, volume = {24}, number = {3}, pages = {1326-1339}, doi = {10.1111/1462-2920.15833}, pmid = {34792280}, issn = {1462-2920}, mesh = {Animals ; *Ants ; *Arthropods ; Biological Evolution ; *Hemiptera ; Symbiosis ; *Wolbachia/genetics ; }, abstract = {Wolbachia is one of the most successful endosymbiotic bacteria of arthropods. Known as the 'master of manipulation', Wolbachia can induce a wide range of phenotypes in its host that can have far-reaching ecological and evolutionary consequences and may be exploited for disease and pest control. However, our knowledge of Wolbachia's distribution and the infection rate is unevenly distributed across arthropod groups such as scale insects. We fitted a distribution of within-species prevalence of Wolbachia to our data and compared it to distributions fitted to an up-to-date dataset compiled from surveys across all arthropods. The estimated distribution parameters indicate a Wolbachia infection frequency of 43.6% (at a 10% prevalence threshold) in scale insects. Prevalence of Wolbachia in scale insects follows a distribution similar to exponential decline (most species are predicted to have low prevalence infections), in contrast to the U-shaped distribution estimated for other taxa (most species have a very low or very high prevalence). We observed no significant associations between Wolbachia infection and scale insect traits. Finally, we screened for Wolbachia in scale insect's ecological associates. We found a positive correlation between Wolbachia infection in scale insects and their ant associates, pointing to a possible route of horizontal transfer of Wolbachia.}, } @article {pmid34791181, year = {2022}, author = {Mostoufi, SL and Singh, ND}, title = {Diet-induced changes in titer support a discrete response of Wolbachia-associated plastic recombination in Drosophila melanogaster.}, journal = {G3 (Bethesda, Md.)}, volume = {12}, number = {1}, pages = {}, pmid = {34791181}, issn = {2160-1836}, support = {T32 GM007413/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Drosophila melanogaster/physiology ; Phenotype ; Plastics ; Recombination, Genetic ; Symbiosis ; *Wolbachia/genetics ; }, abstract = {Plastic recombination in Drosophila melanogaster has been associated with a variety of extrinsic and intrinsic factors such as temperature, starvation, and parasite infection. The bacterial endosymbiont Wolbachia pipientis has also been associated with plastic recombination in D. melanogaster. Wolbachia infection is pervasive in arthropods and this infection induces a variety of phenotypes in its hosts, the strength of which can depend on bacterial titer. Here, we test the hypothesis that the magnitude of Wolbachia-associated plastic recombination in D. melanogaster depends on titer. To manipulate titer, we raised Wolbachia-infected and uninfected flies on diets that have previously been shown to increase or decrease Wolbachia titer relative to controls. We measured recombination in treated and control individuals using a standard backcrossing scheme with two X-linked visible markers. Our results recapitulate previous findings that Wolbachia infection is associated with increased recombination rate across the yellow-vermillion interval of the X chromosome. Our data show no significant effect of diet or diet by Wolbachia interactions on recombination, suggesting that diet-induced changes in Wolbachia titer have no effect on the magnitude of plastic recombination. These findings represent one of the first steps toward investigating Wolbachia-associated plastic recombination and demonstrate that the phenotype is a discrete response rather than a continuous one.}, } @article {pmid34765121, year = {2021}, author = {Gupta, M and Kaur, R and Gupta, A and Raychoudhury, R}, title = {Are ecological communities the seat of endosymbiont horizontal transfer and diversification? A case study with soil arthropod community.}, journal = {Ecology and evolution}, volume = {11}, number = {21}, pages = {14490-14508}, pmid = {34765121}, issn = {2045-7758}, abstract = {Maternally inherited endosymbionts of arthropods are one of the most abundant and diverse group of bacteria. These bacterial endosymbionts also show extensive horizontal transfer to taxonomically unrelated hosts and widespread recombination in their genomes. Such horizontal transfers can be enhanced when different arthropod hosts come in contact like in an ecological community. Higher rates of horizontal transfer can also increase the probability of recombination between endosymbionts, as they now share the same host cytoplasm. However, reports of community-wide endosymbiont data are rare as most studies choose few host taxa and specific ecological interactions among the hosts. To better understand endosymbiont spread within host populations, we investigated the incidence, diversity, extent of horizontal transfer, and recombination of three endosymbionts (Wolbachia, Cardinium, and Arsenophonus) in a specific soil arthropod community. Wolbachia strains were characterized with MLST genes whereas 16S rRNA gene was used for Cardinium and Arsenophonus. Among 3,509 individual host arthropods, belonging to 390 morphospecies, 12.05% were infected with Wolbachia, 2.82% with Cardinium and 2.05% with Arsenophonus. Phylogenetic incongruence between host and endosymbiont indicated extensive horizontal transfer of endosymbionts within this community. Three cases of recombination between Wolbachia supergroups and eight incidences of within-supergroup recombination were also found. Statistical tests of similarity indicated supergroup A Wolbachia and Cardinium show a pattern consistent with extensive horizontal transfer within the community but not for supergroup B Wolbachia and Arsenophonus. We highlight the importance of extensive community-wide studies for a better understanding of the spread of endosymbionts across global arthropod communities.}, } @article {pmid34749528, year = {2021}, author = {Mancini, MV and Ant, TH and Herd, CS and Martinez, J and Murdochy, SM and Gingell, DD and Mararo, E and Johnson, PCD and Sinkins, SP}, title = {High Temperature Cycles Result in Maternal Transmission and Dengue Infection Differences Between Wolbachia Strains in Aedes aegypti.}, journal = {mBio}, volume = {12}, number = {6}, pages = {e0025021}, pmid = {34749528}, issn = {2150-7511}, support = {/WT_/Wellcome Trust/United Kingdom ; 108508/A/15/Z/WT_/Wellcome Trust/United Kingdom ; 202888/Z/16/Z/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Aedes/growth & development/*microbiology/virology ; Animals ; Dengue/transmission/virology ; Dengue Virus/physiology ; Ecosystem ; Female ; Humans ; Larva/*growth & development/microbiology/virology ; Male ; Mosquito Control ; Mosquito Vectors/growth & development/*microbiology/virology ; Population Dynamics ; Temperature ; Wolbachia/genetics/*physiology ; }, abstract = {Environmental factors play a crucial role in the population dynamics of arthropod endosymbionts, and therefore in the deployment of Wolbachia symbionts for the control of dengue arboviruses. The potential of Wolbachia to invade, persist, and block virus transmission depends in part on its intracellular density. Several recent studies have highlighted the importance of larval rearing temperature in modulating Wolbachia densities in adults, suggesting that elevated temperatures can severely impact some strains, while having little effect on others. The effect of a replicated tropical heat cycle on Wolbachia density and levels of virus blocking was assessed using Aedes aegypti lines carrying strains wMel and wAlbB, two Wolbachia strains currently used for dengue control. Impacts on intracellular density, maternal transmission fidelity, and dengue inhibition capacity were observed for wMel. In contrast, wAlbB-carrying Ae. aegypti maintained a relatively constant intracellular density at high temperatures and conserved its capacity to inhibit dengue. Following larval heat treatment, wMel showed a degree of density recovery in aging adults, although this was compromised by elevated air temperatures. IMPORTANCE In the past decades, dengue incidence has dramatically increased all over the world. An emerging dengue control strategy utilizes Aedes aegypti mosquitoes artificially transinfected with the bacterial symbiont Wolbachia, with the ultimate aim of replacing wild mosquito populations. However, the rearing temperature of mosquito larvae is known to impact on some Wolbachia strains. In this study, we compared the effects of a temperature cycle mimicking natural breeding sites in tropical climates on two Wolbachia strains, currently used for open field trials. When choosing the Wolbachia strain to be used in a dengue control program it is important to consider the effects of environmental temperatures on invasiveness and virus inhibition. These results underline the significance of understanding the impact of environmental factors on released mosquitoes, in order to ensure the most efficient strategy for dengue control.}, } @article {pmid34725401, year = {2021}, author = {Rakotonirina, A and Caruzzo, C and Ballan, V and Kainiu, M and Marin, M and Colot, J and Richard, V and Dupont-Rouzeyrol, M and Selmaoui-Folcher, N and Pocquet, N}, title = {Wolbachia detection in Aedes aegypti using MALDI-TOF MS coupled to artificial intelligence.}, journal = {Scientific reports}, volume = {11}, number = {1}, pages = {21355}, pmid = {34725401}, issn = {2045-2322}, mesh = {Aedes/*microbiology ; Animals ; Artificial Intelligence ; Mosquito Control/methods ; Mosquito Vectors/*microbiology ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Wolbachia/chemistry/*isolation & purification ; }, abstract = {The mosquito Aedes aegypti is the major vector of arboviruses like dengue, Zika and chikungunya viruses. Attempts to reduce arboviruses emergence focusing on Ae. aegypti control has proven challenging due to the increase of insecticide resistances. An emerging strategy which consists of releasing Ae. aegypti artificially infected with Wolbachia in natural mosquito populations is currently being developed. The monitoring of Wolbachia-positive Ae. aegypti in the field is performed in order to ensure the program effectiveness. Here, the reliability of the Matrix‑Assisted Laser Desorption Ionization‑Time Of Flight (MALDI‑TOF) coupled with the machine learning methods like Convolutional Neural Network (CNN) to detect Wolbachia in field Ae. aegypti was assessed for the first time. For this purpose, laboratory reared and field Ae. aegypti were analyzed. The results showed that the CNN recognized Ae. aegypti spectral patterns associated with Wolbachia-infection. The MALDI-TOF coupled with the CNN (sensitivity = 93%, specificity = 99%, accuracy = 97%) was more efficient than the loop-mediated isothermal amplification (LAMP), and as efficient as qPCR for Wolbachia detection. It therefore represents an interesting method to evaluate the prevalence of Wolbachia in field Ae. aegypti mosquitoes.}, } @article {pmid34719095, year = {2022}, author = {Malkeyeva, D and Kiseleva, E and Fedorova, SA}, title = {Loss of Hsp67Bc leads to autolysosome enlargement in the Drosophila brain.}, journal = {Cell biology international}, volume = {46}, number = {2}, pages = {203-212}, doi = {10.1002/cbin.11721}, pmid = {34719095}, issn = {1095-8355}, support = {0259-2021-0011//Ministry of Science and Higher Education of the Russian Federation/ ; }, mesh = {Animals ; Brain/metabolism ; *Drosophila/genetics ; *Drosophila Proteins/genetics/metabolism ; Drosophila melanogaster/metabolism ; Heat-Shock Proteins/metabolism ; Lysosomes/metabolism ; }, abstract = {Hsp67Bc is a small heat shock protein found in Drosophila melanogaster. Apart from performing a function (common for all small heat shock proteins) of preventing aggregation of misfolded proteins, it is involved in macroautophagy regulation alongside the Starvin protein. Overexpression of the D. melanogaster Hsp67Bc gene has been shown to stimulate macroautophagy in S2 cell culture. Nonetheless, it has been unknown how the absence of the Hsp67Bc gene may affect it. Here, we studied the effect of Hsp67Bc gene deletion on the macroautophagy induced by the pathogenic Wolbachia wMelPop strain in D. melanogaster. We detected Wolbachia inside autophagic vacuoles in fly neurons, thereby proving that these endosymbionts were being eliminated via macroautophagy. Nevertheless, we did not register any difference in brain bacterial load between Hsp67Bc-null and control flies at all tested stages of ontogenesis. Moreover, the abundance of autophagic vacuoles was similar between neurons of the mutant and control flies, yet the cross-sectional area of autolysosomes on ultrathin sections was more than 1.5-fold larger in Hsp67Bc-null fly brains than in the control line. Our findings suggest that the product of the Hsp67Bc gene does not participate in the initiation of endosymbiont-induced macroautophagy but may mediate autophagosome maturation: the deletion of the Hsp67Bc gene leads to the increase in autolysosome size.}, } @article {pmid34717929, year = {2021}, author = {Prichard, RK}, title = {Macrocyclic lactone resistance in Dirofilaria immitis: risks for prevention of heartworm disease.}, journal = {International journal for parasitology}, volume = {51}, number = {13-14}, pages = {1121-1132}, doi = {10.1016/j.ijpara.2021.08.006}, pmid = {34717929}, issn = {1879-0135}, mesh = {Animals ; *Cat Diseases/drug therapy ; Cats ; *Dirofilaria immitis/genetics ; *Dirofilariasis/prevention & control ; *Dog Diseases/parasitology/prevention & control ; Dogs ; Genetic Markers ; Lactones/pharmacology/therapeutic use ; Microfilariae ; }, abstract = {Heartworm disease, caused by Dirofilaria immitis, can be lethal in dogs and cats. It is transmitted by mosquitoes, and occurs in many parts of the world. Prevention relies on macrocyclic lactones. Macrocyclic lactones used are ivermectin, selamectin, abamectin, eprinomectin, milbemycin oxime and moxidectin, administered at 30-day intervals during the transmission season. Some moxidectin formulations are long-acting injectables. In the USA, preventives are recommended throughout the year. Loss of efficacy of macrocyclic lactone preventives was reported in 2005 and proof of resistance in the USA was published a decade later. Understanding factors which promote resistance is important to maintain control. Factors important for resistance development are discussed. Better, inexpensive tests to confirm resistance are needed. Infection in animals under chemoprophylaxis per se does not imply resistance because lack of compliance in preventive use could be the reason. In vivo confirmation of resistance is expensive, slow and ethically questionable. A microfilariae suppression test can be a surrogate test, but requires a high dose of a macrocyclic lactone and repeated blood microfilaria counts 2-4 weeks later. DNA single nucleotide polymorphism markers have been successfully used. However, the specific genetic changes which cause resistance are unknown. Surveys to map and follow the extent of resistance are needed. Long acting mosquito repellants and insecticides can play a useful role. High dose rate formulations of moxidectin, coupled with mosquito biting mitigation may reduce transmission of resistant genotypes. Doxycycline, daily for 28 days, as anti-Wolbachia treatment, can reduce transmission and remove adult parasites. However, new classes of heartworm preventives are needed. While any preventive strategy must be highly effective, registration requirements for 100% efficacy may hinder development of useful new classes of preventives. Continued reliance on macrocyclic lactone preventives, when they do not work against resistant genotypes, will spread resistance, and allow for more disease.}, } @article {pmid34710095, year = {2021}, author = {Yang, Y and He, Y and Zhu, G and Zhang, J and Gong, Z and Huang, S and Lu, G and Peng, Y and Meng, Y and Hao, X and Wang, C and Sun, J and Shang, S}, title = {Prevalence and molecular characterization of Wolbachia in field-collected Aedes albopictus, Anopheles sinensis, Armigeres subalbatus, Culex pipiens and Cx. tritaeniorhynchus in China.}, journal = {PLoS neglected tropical diseases}, volume = {15}, number = {10}, pages = {e0009911}, pmid = {34710095}, issn = {1935-2735}, mesh = {Aedes/*microbiology ; Animals ; Anopheles/*microbiology ; China ; Culex/*microbiology ; Female ; Male ; Mosquito Vectors/microbiology ; Multilocus Sequence Typing ; Phylogeny ; Wolbachia/classification/genetics/*isolation & purification ; }, abstract = {Wolbachia are maternally transmitted intracellular bacteria that can naturally and artificially infect arthropods and nematodes. Recently, they were applied to control the spread of mosquito-borne pathogens by causing cytoplasmic incompatibility (CI) between germ cells of females and males. The ability of Wolbachia to induce CI is based on the prevalence and polymorphism of Wolbachia in natural populations of mosquitoes. In this study, we screened the natural infection level and diversity of Wolbachia in field-collected mosquitoes from 25 provinces of China based on partial sequence of Wolbachia surface protein (wsp) gene and multilocus sequence typing (MLST). Among the samples, 2489 mosquitoes were captured from 24 provinces between July and September, 2014 and the remaining 1025 mosquitoes were collected month-by-month in Yangzhou, Jiangsu province between September 2013 and August 2014. Our results showed that the presence of Wolbachia was observed in mosquitoes of Aedes albopictus (97.1%, 331/341), Armigeres subalbatus (95.8%, 481/502), Culex pipiens (87.0%, 1525/1752), Cx. tritaeniorhynchus (17.1%, 14/82), but not Anopheles sinensis (n = 88). Phylogenetic analysis indicated that high polymorphism of wsp and MLST loci was observed in Ae. albopictus mosquitoes, while no or low polymorphisms were in Ar. subalbatus and Cx. pipiens mosquitoes. A total of 12 unique mutations of deduced amino acid were identified in the wsp sequences obtained in this study, including four mutations in Wolbachia supergroup A and eight mutations in supergroup B. This study revealed the prevalence and polymorphism of Wolbachia in mosquitoes in large-scale regions of China and will provide some useful information when performing Wolbachia-based mosquito biocontrol strategies in China.}, } @article {pmid34704919, year = {2021}, author = {Ortiz-Baez, AS and Shi, M and Hoffmann, AA and Holmes, EC}, title = {RNA virome diversity and Wolbachia infection in individual Drosophila simulans flies.}, journal = {The Journal of general virology}, volume = {102}, number = {10}, pages = {}, pmid = {34704919}, issn = {1465-2099}, mesh = {Animals ; Drosophila simulans/*microbiology/virology ; Female ; Phylogeny ; RNA Viruses/classification/genetics/isolation & purification/*physiology ; Symbiosis ; Virome/genetics/*physiology ; Wolbachia/isolation & purification/*physiology ; }, abstract = {The endosymbiont bacteria of the genus Wolbachia are associated with multiple mutualistic effects on insect biology, including nutritional and antiviral properties. Members of the genus Wolbachia naturally occur in fly species of the genus Drosophila, providing an operational model host for studying how virome composition may be affected by its presence. Drosophila simulans populations can carry a variety of strains of members of the genus Wolbachia, with the wAu strain associated with strong antiviral protection under experimental conditions. We used D. simulans sampled from the Perth Hills, Western Australia, to investigate the potential virus protective effect of the wAu strain of Wolbachia on individual wild-caught flies. Our data revealed no appreciable variation in virus composition and abundance between individuals infected or uninfected with Wolbachia associated with the presence or absence of wAu. However, it remains unclear whether wAu might affect viral infection and host survival by increasing tolerance rather than inducing complete resistance. These data also provide new insights into the natural virome diversity of D. simulans. Despite the small number of individuals sampled, we identified a repertoire of RNA viruses, including nora virus, galbut virus, thika virus and La Jolla virus, that have been identified in other species of the genus Drosophila. Chaq virus-like sequences associated with galbut virus were also detected. In addition, we identified five novel viruses from the families Reoviridae, Tombusviridae, Mitoviridae and Bunyaviridae. Overall, this study highlights the complex interaction between Wolbachia and RNA virus infections and provides a baseline description of the natural virome of D. simulans.}, } @article {pmid34680640, year = {2021}, author = {Bell-Sakyi, L and Beliavskaia, A and Hartley, CS and Jones, L and Luu, L and Haines, LR and Hamilton, JGC and Darby, AC and Makepeace, BL}, title = {Isolation in Natural Host Cell Lines of Wolbachia Strains wPip from the Mosquito Culex pipiens and wPap from the Sand Fly Phlebotomus papatasi.}, journal = {Insects}, volume = {12}, number = {10}, pages = {}, pmid = {34680640}, issn = {2075-4450}, support = {BBS/E/I/00007039/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; BBS/E/I/00002118/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; 204806/Z/16/Z and 080961/Z/06/Z/WT_/Wellcome Trust/United Kingdom ; BB/P024270/1, BBS/E/I/00002118 and BBS/E/I/00007039/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; BB/P024270/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; }, abstract = {Endosymbiotic intracellular bacteria of the genus Wolbachia are harboured by many species of invertebrates. They display a wide range of developmental, metabolic and nutritional interactions with their hosts and may impact the transmission of arboviruses and protozoan parasites. Wolbachia have occasionally been isolated during insect cell line generation. Here, we report the isolation of two strains of Wolbachia, wPip and wPap, during cell line generation from their respective hosts, the mosquito Culex pipiens and the sand fly Phlebotomus papatasi. wPip was pathogenic for both new C. pipiens cell lines, CPE/LULS50 and CLP/LULS56, requiring tetracycline treatment to rescue the lines. In contrast, wPap was tolerated by the P. papatasi cell line PPL/LULS49, although tetracycline treatment was applied to generate a Wolbachia-free subline. Both Wolbachia strains were infective for a panel of heterologous insect and tick cell lines, including two novel lines generated from the sand fly Lutzomyia longipalpis, LLE/LULS45 and LLL/LULS52. In all cases, wPip was more pathogenic for the host cells than wPap. These newly isolated Wolbachia strains, and the novel mosquito and sand fly cell lines reported here, will add to the resources available for research on host-endosymbiont relationships, as well as on C. pipiens, P. papatasi, L. longipalpis and the pathogens that they transmit.}, } @article {pmid34680622, year = {2021}, author = {Shapoval, NA and Nokkala, S and Nokkala, C and Kuftina, GN and Kuznetsova, VG}, title = {The Incidence of Wolbachia Bacterial Endosymbiont in Bisexual and Parthenogenetic Populations of the Psyllid Genus Cacopsylla (Hemiptera, Psylloidea).}, journal = {Insects}, volume = {12}, number = {10}, pages = {}, pmid = {34680622}, issn = {2075-4450}, support = {19-14-00202//Russian Science Foundation/ ; }, abstract = {Wolbachia is one of the most common intracellular bacteria; it infects a wide variety of insects, other arthropods, and some nematodes. Wolbachia is ordinarily transmitted vertically from mother to offspring and can manipulate physiology and reproduction of their hosts in different ways, e.g., induce feminization, male killing, and parthenogenesis. Despite the great interest in Wolbachia, many aspects of its biology remain unclear and its incidence across many insect orders, including Hemiptera, is still poorly understood. In this report, we present data on Wolbachia infection in five jumping plant-lice species (Hemiptera, Psylloidea) of the genus Cacopsylla Ossiannilsson, 1970 with different reproductive strategies and test the hypothesis that Wolbachia mediates parthenogenetic and bisexual patterns observed in some Cacopsylla species. We show that the five species studied are infected with a single Wolbachia strain, belonging to the supergroup B. This strain has also been found in different insect orders (Lepidoptera, Hemiptera, Plecoptera, Orthoptera, Hymenoptera, Diptera) and even in acariform mites (Trombidiformes), suggesting extensive horizontal transmission of Wolbachia between representatives of these taxa. Our survey did not reveal significant differences in infection frequency between parthenogenetic and bisexual populations or between males and females within bisexual populations. However, infection rate varied notably in different Cacopsylla species or within distinct populations of the same species. Overall, we demonstrate that Wolbachia infects a high proportion of Cacopsylla individuals and populations, suggesting the essential role of this bacterium in their biology.}, } @article {pmid34677126, year = {2021}, author = {Perlmutter, JI and Meyers, JE and Bordenstein, SR}, title = {A single synonymous nucleotide change impacts the male-killing phenotype of prophage WO gene wmk.}, journal = {eLife}, volume = {10}, number = {}, pages = {}, pmid = {34677126}, issn = {2050-084X}, support = {R21 AI133522/AI/NIAID NIH HHS/United States ; F31 AI143152/AI/NIAID NIH HHS/United States ; P20 GM103418/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Bacterial Proteins/*genetics/metabolism ; Drosophila melanogaster/*microbiology ; Male ; Microorganisms, Genetically-Modified/genetics/physiology ; Prophages/*genetics ; *Symbiosis ; Wolbachia/genetics/*physiology ; }, abstract = {Wolbachia are the most widespread bacterial endosymbionts in animals. Within arthropods, these maternally transmitted bacteria can selfishly hijack host reproductive processes to increase the relative fitness of their transmitting females. One such form of reproductive parasitism called male killing, or the selective killing of infected males, is recapitulated to degrees by transgenic expression of the prophage WO-mediated killing (wmk) gene. Here, we characterize the genotype-phenotype landscape of wmk-induced male killing in D. melanogaster using transgenic expression. While phylogenetically distant wmk homologs induce no sex-ratio bias, closely-related homologs exhibit complex phenotypes spanning no death, male death, or death of all hosts. We demonstrate that alternative start codons, synonymous codons, and notably a single synonymous nucleotide in wmk can ablate killing. These findings reveal previously unrecognized features of transgenic wmk-induced killing and establish new hypotheses for the impacts of post-transcriptional processes in male killing variation. We conclude that synonymous sequence changes are not necessarily silent in nested endosymbiotic interactions with life-or-death consequences.}, } @article {pmid34666103, year = {2021}, author = {Poopandi, S and Sundaraj, R and Rajmichael, R and Thangaraj, S and Dhamodharan, P and Biswal, J and Malaisamy, V and Jeyaraj Pandian, C and Jeyaraman, J}, title = {Computational screening of potential inhibitors targeting MurF of Brugia malayi Wolbachia through multi-scale molecular docking, molecular dynamics and MM-GBSA analysis.}, journal = {Molecular and biochemical parasitology}, volume = {246}, number = {}, pages = {111427}, doi = {10.1016/j.molbiopara.2021.111427}, pmid = {34666103}, issn = {1872-9428}, mesh = {Animals ; *Brugia malayi ; *Elephantiasis, Filarial/parasitology ; Molecular Docking Simulation ; Molecular Dynamics Simulation ; *Wolbachia/metabolism ; }, abstract = {Lymphatic filariasis is a parasitic disease caused by the worms Wuchereria bancrofti, Brugia malayi and Brugia timori. Three anti-filarial drugs namely Diethylcarbamazine, Ivermectin and Albendazole and their combinations are used as the control strategies for filariasis. The disease has received much attention in drug discovery due to the unavailability of vaccines and the toxic pharmaceutical properties of the existing drugs. In Wolbachia endosymbiont Brugia malayi, the UDP-N-acetylmuramoyl-tripeptide-d-alanyl-d-alanine ligase (MurF) plays a key role in peptidoglycan biosynthesis pathway and therefore can be considered as effective drug target against filariasis disease. Therefore, in the present study, MurF was selected as the therapeutic target to identify specific inhibitors against filariasis. Homology modeling was performed to predict the three-dimensional structure of MurF due to the absence of the experimental structure. Further molecular dynamics simulation and structure-based high throughput virtual screening with three different chemical databases (Zinc, Maybridge and Specs) were carried out to identify potent inhibitors and also to check their conformations inside the binding site of MurF, respectively. Top three compounds with high docking score and high relative binding affinity against MurF were selected. Further, validation studies, including predicted ADME (Absorption, Distribution, Metabolism, Excretion) assessment, binding free energy using MM-GBSA (Molecular Mechanics Generalized Born Surface Area) and DFT (Density Functional Theory) calculations were performed for the top three compounds. From the results, it was observed that all the three compounds were predicted to show high reactivity, acceptable range of pharmacokinetic properties and high binding affinity with the drug target MurF. Overall, the results could provide more understanding on the inhibition of MurF enzyme and the screened compounds could lead to the development of new specific anti-filarial drugs.}, } @article {pmid34662426, year = {2022}, author = {Hill, T and Unckless, RL and Perlmutter, JI}, title = {Positive Selection and Horizontal Gene Transfer in the Genome of a Male-Killing Wolbachia.}, journal = {Molecular biology and evolution}, volume = {39}, number = {1}, pages = {}, pmid = {34662426}, issn = {1537-1719}, support = {P20 GM103418/GM/NIGMS NIH HHS/United States ; P20 GM103638/GM/NIGMS NIH HHS/United States ; R00 GM114714/GM/NIGMS NIH HHS/United States ; R01 AI139154/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Drosophila/genetics/microbiology ; Drosophila melanogaster/genetics ; Gene Transfer, Horizontal ; Genome ; Male ; *Wolbachia/genetics ; }, abstract = {Wolbachia are a genus of widespread bacterial endosymbionts in which some strains can hijack or manipulate arthropod host reproduction. Male killing is one such manipulation in which these maternally transmitted bacteria benefit surviving daughters in part by removing competition with the sons for scarce resources. Despite previous findings of interesting genome features of microbial sex ratio distorters, the population genomics of male-killers remain largely uncharacterized. Here, we uncover several unique features of the genome and population genomics of four Arizonan populations of a male-killing Wolbachia strain, wInn, that infects mushroom-feeding Drosophila innubila. We first compared the wInn genome with other closely related Wolbachia genomes of Drosophila hosts in terms of genome content and confirm that the wInn genome is largely similar in overall gene content to the wMel strain infecting D. melanogaster. However, it also contains many unique genes and repetitive genetic elements that indicate lateral gene transfers between wInn and non-Drosophila eukaryotes. We also find that, in line with literature precedent, genes in the Wolbachia prophage and Octomom regions are under positive selection. Of all the genes under positive selection, many also show evidence of recent horizontal transfer among Wolbachia symbiont genomes. These dynamics of selection and horizontal gene transfer across the genomes of several Wolbachia strains and diverse host species may be important underlying factors in Wolbachia's success as a male-killer of divergent host species.}, } @article {pmid34654483, year = {2021}, author = {Gloder, G and Bourne, ME and Verreth, C and Wilberts, L and Bossaert, S and Crauwels, S and Dicke, M and Poelman, EH and Jacquemyn, H and Lievens, B}, title = {Parasitism by endoparasitoid wasps alters the internal but not the external microbiome in host caterpillars.}, journal = {Animal microbiome}, volume = {3}, number = {1}, pages = {73}, pmid = {34654483}, issn = {2524-4671}, support = {FWO//Fonds Wetenschappelijk Onderzoek/ ; G.0961.19N//Fonds Wetenschappelijk Onderzoek/ ; ALWOP.343//Nederlandse Organisatie voor Wetenschappelijk Onderzoek/ ; ALWOP.368//Nederlandse Organisatie voor Wetenschappelijk Onderzoek/ ; }, abstract = {BACKGROUND: The microbiome of many insects consists of a diverse community of microorganisms that can play critical roles in the functioning and overall health of their hosts. Although the microbial communities of insects have been studied thoroughly over the past decade, little is still known about how biotic interactions affect the microbial community structure in and on the bodies of insects. In insects that are attacked by parasites or parasitoids, it can be expected that the microbiome of the host insect is affected by the presence of these parasitic organisms that develop in close association with their host. In this study, we used high-throughput amplicon sequencing targeting both bacteria and fungi to test the hypothesis that parasitism by the endoparasitoid Cotesia glomerata affected the microbiome of its host Pieris brassicae. Healthy and parasitized caterpillars were collected from both natural populations and a laboratory culture.

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

CONCLUSIONS: The results provide a baseline for monitoring and management of resistance as well as knowledge of Vssc genotype frequencies required in Wolbachia release populations to ensure homogeneity with the target field population. Vssc mutation haplotypes observed show some similarity with those from Ae. aegypti in southeast Asia and the Indo-Pacific, but the presence of the triple mutant haplotype in three genotypes indicates that the species in this region may have a unique population history.}, } @article {pmid34233062, year = {2021}, author = {Duplouy, A and Nair, A and Nyman, T and van Nouhuys, S}, title = {Long-term spatiotemporal genetic structure of an accidental parasitoid introduction, and local changes in prevalence of its associated Wolbachia symbiont.}, journal = {Molecular ecology}, volume = {30}, number = {18}, pages = {4368-4380}, doi = {10.1111/mec.16065}, pmid = {34233062}, issn = {1365-294X}, mesh = {Animals ; *Butterflies/genetics ; Humans ; Microsatellite Repeats ; Prevalence ; *Wasps/genetics ; *Wolbachia/genetics ; }, abstract = {Population bottlenecks associated with founder events strongly impact the establishment and genetic makeup of populations. In addition to their genotype, founding individuals also bring along parasites, as well as symbionts that can manipulate the phenotype of their host, affecting the host population establishment, dynamics and evolution. Thus, to understand introduction, invasion, and spread, we should identify the roles played by accompanying symbionts. In 1991, the parasitoid wasp, Hyposoter horticola, and its associated hyperparasitoid were accidentally introduced from the main Åland islands, Finland, to an isolated island in the archipelago, along with their host, the Glanville fritillary butterfly. Though the receiving island was unoccupied, the butterfly was present on some of the small islands in the vicinity. The three introduced species have persisted locally ever since. A strain of the endosymbiotic bacterium Wolbachia has an intermediate prevalence in the parasitoid H. horticola across the main Åland population. The infection increases its susceptibility of to hyperparasitism. We investigated the establishment and spread of the parasitoid, along with patterns of prevalence of its symbiont using 323 specimens collected between 1992 and 2013, from five localities across Åland, including the source and introduced populations. Using 14 microsatellites and one mitochondrial marker, we suggest that the relatively diverse founding population and occasional migration between islands might have facilitated the persistence of all isolated populations, despite multiple local population crashes. We also show that where the hyperparasitoid is absent, and thus selection against infected wasp genotypes is relaxed, there is near-fixation of Wolbachia.}, } @article {pmid34229954, year = {2021}, author = {Johnston, KL and Hong, WD and Turner, JD and O'Neill, PM and Ward, SA and Taylor, MJ}, title = {Anti-Wolbachia drugs for filariasis.}, journal = {Trends in parasitology}, volume = {37}, number = {12}, pages = {1068-1081}, doi = {10.1016/j.pt.2021.06.004}, pmid = {34229954}, issn = {1471-5007}, support = {MR/R025401/1/MRC_/Medical Research Council/United Kingdom ; NC/M00175X/1/NC3RS_/National Centre for the Replacement, Refinement and Reduction of Animals in Research/United Kingdom ; }, mesh = {Anti-Bacterial Agents/pharmacology/therapeutic use ; Drug Discovery ; *Elephantiasis, Filarial/drug therapy ; Humans ; *Nematode Infections/drug therapy ; *Onchocerciasis/drug therapy ; *Wolbachia ; }, abstract = {The mutualistic association between Wolbachia endosymbionts and their filarial nematode hosts has been exploited as a validated drug target delivering macrofilaricidal outcomes. Limitations of existing antibiotics to scale-up have driven the search for new drugs, which are effective in shorter regimens of 7 days or less. Here, we review the last 14 years of anti-Wolbachia drug discovery by the anti-Wolbachia (A·WOL) consortium, which has screened more than two million compounds, delivering thousands of hit compounds. Refined screening models integrated with robust pharmacokinetic/pharmacodynamic (PK/PD) driven optimisation and selection strategies have delivered the first two drug candidates specifically designed to target Wolbachia. AWZ1066S and ABBV-4083 are currently progressing through clinical trials with the aim of delivering safe and effective macrofilaricides to support the elimination of onchocerciasis and lymphatic filariasis.}, } @article {pmid34221442, year = {2021}, author = {Chan, PK and Hawley, JR and Lappin, MR}, title = {Evaluation of the role of Babesia species and Cytauxzoon felis in feline anemia cases in Colorado, USA.}, journal = {JFMS open reports}, volume = {7}, number = {1}, pages = {20551169211024967}, pmid = {34221442}, issn = {2055-1169}, abstract = {OBJECTIVES: The aim of this study was to evaluate the blood of cats in Colorado, USA, with suspected infectious causes of anemia for the presence of Babesia species and Cytauxzoon felis DNA. Results of PCR testing for other common vector-borne diseases potentially associated with anemia are also reported.

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

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

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

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

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

CONCLUSIONS: We identified the I. ricinus microbiota and documented marked shifts in tick microbiota dynamics over time. Statistically, we showed strong relationships between the presence of specific pathogens and the structure of the I. ricinus microbiota. We detected close links between some tick symbionts and the potential presence of either pathogenic Rickettsia or a parasitoid in ticks. These new findings pave the way for the development of new strategies for the control of ticks and tick-borne diseases. Video abstract.}, } @article {pmid34208681, year = {2021}, author = {Ourry, M and Crosland, A and Lopez, V and Derocles, SAP and Mougel, C and Cortesero, AM and Poinsot, D}, title = {Influential Insider: Wolbachia, an Intracellular Symbiont, Manipulates Bacterial Diversity in Its Insect Host.}, journal = {Microorganisms}, volume = {9}, number = {6}, pages = {}, pmid = {34208681}, issn = {2076-2607}, support = {SE 0000282 MP-P10026 EB06//Institut National de la Recherche Agronomique/ ; }, abstract = {Facultative intracellular symbionts like the α-proteobacteria Wolbachia influence their insect host phenotype but little is known about how much they affect their host microbiota. Here, we quantified the impact of Wolbachia infection on the bacterial community of the cabbage root fly Delia radicum by comparing the microbiota of Wolbachia-free and infected adult flies of both sexes. We used high-throughput DNA sequencing (Illumina MiSeq, 16S rRNA, V5-V7 region) and performed a community and a network analysis. In both sexes, Wolbachia infection significantly decreased the diversity of D. radicum bacterial communities and modified their structure and composition by reducing abundance in some taxa but increasing it in others. Infection by Wolbachia was negatively correlated to 8 bacteria genera (Erwinia was the most impacted), and positively correlated to Providencia and Serratia. We suggest that Wolbachia might antagonize Erwinia for being entomopathogenic (and potentially intracellular), but would favor Providencia and Serratia because they might protect the host against chemical plant defenses. Although they might seem prisoners in a cell, endocellular symbionts can impact the whole microbiota of their host, hence its extended phenotype, which provides them with a way to interact with the outside world.}, } @article {pmid34197460, year = {2021}, author = {Kupritz, J and Martin, J and Fischer, K and Curtis, KC and Fauver, JR and Huang, Y and Choi, YJ and Beatty, WL and Mitreva, M and Fischer, PU}, title = {Isolation and characterization of a novel bacteriophage WO from Allonemobius socius crickets in Missouri.}, journal = {PloS one}, volume = {16}, number = {7}, pages = {e0250051}, pmid = {34197460}, issn = {1932-6203}, mesh = {Animals ; Bacteriophages/classification/*genetics/isolation & purification ; Capsid Proteins/genetics ; DNA, Bacterial/chemistry/metabolism ; DNA, Viral/chemistry/metabolism ; Female ; *Genome, Viral ; Gryllidae/*microbiology/virology ; Membrane Proteins/genetics ; Missouri ; Open Reading Frames/genetics ; Phylogeny ; Whole Genome Sequencing ; Wolbachia/genetics/isolation & purification/virology ; }, abstract = {Wolbachia are endosymbionts of numerous arthropod and some nematode species, are important for their development and if present can cause distinct phenotypes of their hosts. Prophage DNA has been frequently detected in Wolbachia, but particles of Wolbachia bacteriophages (phage WO) have been only occasionally isolated. Here, we report the characterization and isolation of a phage WO of the southern ground cricket, Allonemobius socius, and provided the first whole-genome sequence of phage WO from this arthropod family outside of Asia. We screened A. socius abdomen DNA extracts from a cricket population in eastern Missouri by quantitative PCR for Wolbachia surface protein and phage WO capsid protein and found a prevalence of 55% and 50%, respectively, with many crickets positive for both. Immunohistochemistry using antibodies against Wolbachia surface protein showed many Wolbachia clusters in the reproductive system of female crickets. Whole-genome sequencing using Oxford Nanopore MinION and Illumina technology allowed for the assembly of a high-quality, 55 kb phage genome containing 63 open reading frames (ORF) encoding for phage WO structural proteins and host lysis and transcriptional manipulation. Taxonomically important regions of the assembled phage genome were validated by Sanger sequencing of PCR amplicons. Analysis of the nucleotides sequences of the ORFs encoding the large terminase subunit (ORF2) and minor capsid (ORF7) frequently used for phage WO phylogenetics showed highest homology to phage WOAu of Drosophila simulans (94.46% identity) and WOCin2USA1 of the cherry fruit fly, Rhagoletis cingulata (99.33% identity), respectively. Transmission electron microscopy examination of cricket ovaries showed a high density of phage particles within Wolbachia cells. Isolation of phage WO revealed particles characterized by 40-62 nm diameter heads and up to 190 nm long tails. This study provides the first detailed description and genomic characterization of phage WO from North America that is easily accessible in a widely distributed cricket species.}, } @article {pmid34190587, year = {2021}, author = {Leitner, M and Bishop, C and Asgari, S}, title = {Transcriptional Response of Wolbachia to Dengue Virus Infection in Cells of the Mosquito Aedes aegypti.}, journal = {mSphere}, volume = {6}, number = {3}, pages = {e0043321}, pmid = {34190587}, issn = {2379-5042}, abstract = {Aedes aegypti transmits one of the most significant mosquito-borne viruses, dengue virus (DENV). The absence of effective vaccines and clinical treatments and the emergence of insecticide resistance in A. aegypti necessitate novel vector control strategies. A new approach uses the endosymbiotic bacterium Wolbachia pipientis to reduce the spread of arboviruses. However, the Wolbachia-mediated antiviral mechanism is not well understood. To shed light on this mechanism, we investigated an unexplored aspect of Wolbachia-virus-mosquito interaction. We used RNA sequencing to examine the transcriptional response of Wolbachia to DENV infection in A. aegypti Aag2 cells transinfected with the wAlbB strain of Wolbachia. Our results suggest that genes encoding an endoribonuclease (RNase HI), a regulator of sigma 70-dependent gene transcription (6S RNA), essential cellular, transmembrane, and stress response functions and primary type I and IV secretion systems were upregulated, while a number of transport and binding proteins of Wolbachia, ribosome structure, and elongation factor-associated genes were downregulated due to DENV infection. Furthermore, bacterial retrotransposon, transposable, and phage-related elements were found among the up- and downregulated genes. We show that Wolbachia elicits a transcriptional response to virus infection and identify differentially expressed Wolbachia genes mostly at the early stages of virus infection. These findings highlight Wolbachia's ability to alter its gene expression in response to DENV infection of the host cell. IMPORTANCE Aedes aegypti is a vector of several pathogenic viruses, including dengue, Zika, chikungunya, and yellow fever viruses, which are of importance to human health. Wolbachia is an endosymbiotic bacterium currently used in transinfected mosquitoes to suppress replication and transmission of dengue viruses. However, the mechanism of Wolbachia-mediated virus inhibition is not fully understood. While several studies have shown mosquitoes' transcriptional responses to dengue virus infection, none have investigated these responses in Wolbachia, which may provide clues to the inhibition mechanism. Our results suggest changes in the expression of a number of functionally important Wolbachia genes upon dengue virus infection, including those involved in stress responses, providing insights into the endosymbiont's reaction to virus infection.}, } @article {pmid34188180, year = {2021}, author = {Zhang, Y and Cai, T and Ren, Z and Liu, Y and Yuan, M and Cai, Y and Yu, C and Shu, R and He, S and Li, J and Wong, ACN and Wan, H}, title = {Decline in symbiont-dependent host detoxification metabolism contributes to increased insecticide susceptibility of insects under high temperature.}, journal = {The ISME journal}, volume = {15}, number = {12}, pages = {3693-3703}, pmid = {34188180}, issn = {1751-7370}, mesh = {Animals ; Drosophila melanogaster ; *Hemiptera ; Insecta ; Insecticide Resistance ; *Insecticides/toxicity ; Temperature ; }, abstract = {The interactions between insects and their bacterial symbionts are shaped by a variety of abiotic factors, including temperature. As global temperatures continue to break high records, a great deal of uncertainty surrounds how agriculturally important insect pests and their symbionts may be affected by elevated temperatures, and its implications for future pest management. In this study, we examine the role of bacterial symbionts in the brown planthopper Nilaparvata lugens response to insecticide (imidacloprid) under different temperature scenarios. Our results reveal that the bacterial symbionts orchestrate host detoxification metabolism via the CncC pathway to promote host insecticide resistance, whereby the symbiont-inducible CncC pathway acts as a signaling conduit between exogenous abiotic stimuli and host metabolism. However, this insect-bacterial partnership function is vulnerable to high temperature, which causes a significant decline in host-bacterial content. In particular, we have identified the temperature-sensitive Wolbachia as a candidate player in N. lugens detoxification metabolism. Wolbachia-dependent insecticide resistance was confirmed through a series of insecticide assays and experiments comparing Wolbachia-free and Wolbachia-infected N. lugens and also Drosophila melanogaster. Together, our research reveals elevated temperatures negatively impact insect-bacterial symbiosis, triggering adverse consequences on host response to insecticide (imidacloprid) and potentially other xenobiotics.}, } @article {pmid34156702, year = {2021}, author = {Cruz, MA and Magalhães, S and Sucena, É and Zélé, F}, title = {Wolbachia and host intrinsic reproductive barriers contribute additively to postmating isolation in spider mites.}, journal = {Evolution; international journal of organic evolution}, volume = {75}, number = {8}, pages = {2085-2101}, doi = {10.1111/evo.14286}, pmid = {34156702}, issn = {1558-5646}, mesh = {Animals ; Diploidy ; *Mites ; Reproduction ; Reproductive Isolation ; Symbiosis ; *Tetranychidae/genetics ; *Wolbachia ; }, abstract = {Wolbachia are maternally-inherited bacteria that induce cytoplasmic incompatibility in many arthropod species. However, the ubiquity of this isolation mechanism for host speciation processes remains elusive, as only few studies have examined Wolbachia-induced incompatibilities when host populations are not genetically compatible. Here, we used three populations of two genetically differentiated colour forms of the haplodiploid spider mite Tetranychus urticae to dissect the interaction between Wolbachia-induced and host-associated incompatibilities, and their relative contribution to postmating isolation. We found that these two sources of incompatibility act through different mechanisms in an additive fashion. Host-associated incompatibility contributes 1.5 times more than Wolbachia-induced incompatibility in reducing hybrid production, the former through an overproduction of haploid sons at the expense of diploid daughters (ca. 75% decrease) and the latter by increasing the embryonic mortality of daughters (by ca. 49%). Furthermore, regardless of cross direction, we observed near-complete F1 hybrid sterility and complete F2 hybrid breakdown between populations of the two forms, but Wolbachia did not contribute to this outcome. We thus show mechanistic independence and an additive nature of host-intrinsic and Wolbachia-induced sources of isolation. Wolbachia may contribute to reproductive isolation in this system, thereby potentially affecting host differentiation and distribution in the field.}, } @article {pmid34152527, year = {2021}, author = {Dângelo, RAC and Michereff-Filho, M and Inoue-Nagata, AK and da Silva, PS and Chediak, M and Guedes, RNC}, title = {Area-wide insecticide resistance and endosymbiont incidence in the whitefly Bemisia tabaci MEAM1 (B biotype): A Neotropical context.}, journal = {Ecotoxicology (London, England)}, volume = {30}, number = {6}, pages = {1056-1070}, pmid = {34152527}, issn = {1573-3017}, mesh = {Animals ; *Hemiptera ; Humans ; Incidence ; Insecticide Resistance ; *Insecticides/toxicity ; Symbiosis ; }, abstract = {Agriculture insecticides are used against insect pest species, but are able to change community structure in contaminated habitats, and also the genetic pool of exposed individuals. In fact, the latter effect is a relevant tool to in situ biomonitoring of pollutant contamination and impact, besides its practical economic and management concerns. This takes place because the emergence of individuals with resistance to insecticides is particularly frequent among insect pest species and usually enhances insecticide overuse and crop losses. Pest insects of global prominence such as whiteflies are a focus of attention due to problems with insecticide resistance and association with endosymbionts, as the case of the invasive putative species Bemisia tabaci MEAM1. The scenario is particularly complex in the Neotropics, where insecticide use is ubiquitous, but whose spatial scale of occurrence is usually neglected. Here we explored the spatial-dependence of both phenomena in MEAM1 whiteflies recording resistance to two widely used insecticides, lambda-cyhalothrin and spiromesifen, and endosymbiont co-occurrence. Resistance to both insecticides was frequent exhibiting low to moderate frequency of lambda-cyhalothrin resistance and moderate to high frequency of spiromesifen resistance. Among the prevailing whitefly endosymbionts, Wolbachia, Cardinium and Arsenophonus were markedly absent. In contrast, Hamiltonella and Rickettsia prevailed and their incidence was correlated. Furthermore, Rickettsia endosymbionts were particularly associated with lambda-cyhalothrin susceptibility. These traits were spatially dependent with significant variation taking place within an area of about 700 Km[2]. Such findings reinforce the notion of endosymbiont-associated resistance to insecticides, and also of their local incidence allowing spatial mapping and locally-targeted mitigation.}, } @article {pmid34149963, year = {2021}, author = {Ponce, GE and Fuse, M and Chan, A and Connor, EF}, title = {The Localization of Phytohormones within the Gall-inducing Insect Eurosta solidaginis (Diptera: Tephritidae).}, journal = {Arthropod-plant interactions}, volume = {15}, number = {3}, pages = {375-385}, pmid = {34149963}, issn = {1872-8855}, support = {R25 GM048972/GM/NIGMS NIH HHS/United States ; }, abstract = {The phytohormone production hypothesis suggests that organisms, including insects, induce galls by producing and secreting plant growth hormones. Auxins and cytokinins are classes of phytohormones that induce cell growth and cell division, which could contribute to the plant tissue proliferation which constitutes the covering gall. Bacteria, symbiotic with insects, may also play a part in gall induction by insects through the synthesis of phytohormones or other effectors. Past studies have shown that concentrations of cytokinins and auxins in gall-inducing insects are higher than in their host plants. However, these analyses have involved whole-body extractions. Using immunolocalization of cytokinin and auxin, in the gall inducing stage of Eurosta solidaginis, we found both phytohormones to localize almost exclusively to the salivary glands. Co-localization of phytohormone label with a nucleic acid stain in the salivary glands revealed the absence of Wolbachia sp., the bacterial symbiont of E. solidaginis, which suggests that phytohormone production is symbiont independent. Our findings are consistent with the hypothesis that phytohormones are synthesized in and secreted from the salivary glands of E. solidaginis into host-plant tissues for the purpose of manipulating the host plant.}, } @article {pmid34145422, year = {2021}, author = {York, A}, title = {Defeating dengue with Wolbachia.}, journal = {Nature reviews. Microbiology}, volume = {19}, number = {8}, pages = {482}, pmid = {34145422}, issn = {1740-1534}, } @article {pmid34143770, year = {2021}, author = {Duarte, EH and Carvalho, A and López-Madrigal, S and Costa, J and Teixeira, L}, title = {Forward genetics in Wolbachia: Regulation of Wolbachia proliferation by the amplification and deletion of an addictive genomic island.}, journal = {PLoS genetics}, volume = {17}, number = {6}, pages = {e1009612}, pmid = {34143770}, issn = {1553-7404}, mesh = {Animals ; Bacterial Load ; Dicistroviridae/growth & development/pathogenicity ; Drosophila melanogaster/immunology/*microbiology/virology ; Female ; Gene Editing/methods ; *Genome, Bacterial ; Genomic Islands ; Longevity/*immunology ; Male ; Phenotype ; Symbiosis/*genetics ; Wolbachia/*genetics/growth & development/metabolism ; }, abstract = {Wolbachia is one of the most prevalent bacterial endosymbionts, infecting approximately 40% of terrestrial arthropod species. Wolbachia is often a reproductive parasite but can also provide fitness benefits to its host, as, for example, protection against viral pathogens. This protective effect is currently being applied to fight arboviruses transmission by releasing Wolbachia-transinfected mosquitoes. Titre regulation is a crucial aspect of Wolbachia biology. Higher titres can lead to stronger phenotypes and fidelity of transmission but can have a higher cost to the host. Since Wolbachia is maternally transmitted, its fitness depends on host fitness, and, therefore, its cost to the host may be under selection. Understanding how Wolbachia titres are regulated and other aspects of Wolbachia biology has been hampered by the lack of genetic tools. Here we developed a forward genetic screen to identify new Wolbachia over-proliferative mutant variants. We characterized in detail two new mutants, wMelPop2 and wMelOctoless, and show that the amplification or loss of the Octomom genomic region lead to over-proliferation. These results confirm previous data and expand on the complex role of this genomic region in the control of Wolbachia proliferation. Both new mutants shorten the host lifespan and increase antiviral protection. Moreover, we show that Wolbachia proliferation rate in Drosophila melanogaster depends on the interaction between Octomom copy number, the host developmental stage, and temperature. Our analysis also suggests that the life shortening and antiviral protection phenotypes of Wolbachia are dependent on different, but related, properties of the endosymbiont; the rate of proliferation and the titres near the time of infection, respectively. We also demonstrate the feasibility of a novel and unbiased experimental approach to study Wolbachia biology, which could be further adapted to characterize other genetically intractable bacterial endosymbionts.}, } @article {pmid34125999, year = {2022}, author = {Norte, AC and Harris, DJ and Silveira, D and Nunes, CS and Núncio, MS and Martínez, EG and Giménez, A and de Sousa, R and Lopes de Carvalho, I and Perera, A}, title = {Diversity of microorganisms in Hyalomma aegyptium collected from spur-thighed tortoise (Testudo graeca) in North Africa and Anatolia.}, journal = {Transboundary and emerging diseases}, volume = {69}, number = {4}, pages = {1951-1962}, doi = {10.1111/tbed.14188}, pmid = {34125999}, issn = {1865-1682}, mesh = {Animals ; *Borrelia ; Ehrlichia ; Humans ; *Ixodidae/microbiology ; *Rickettsia/genetics ; *Ticks/microbiology ; Tunisia/epidemiology ; Turkey/epidemiology ; *Turtles/parasitology ; }, abstract = {Ticks carry a diverse community of microorganisms including non-pathogenic symbionts, commensals, and pathogens, such as viruses, bacteria, protozoans, and fungi. The assessment of tick-borne microorganisms (TBM) in tortoises and their ticks is essential to understand their eco-epidemiology, and to map and monitor potential pathogens to humans and other animals. The aim of this study was to characterize the diversity of microorganisms found in ticks collected from the spur-thighed tortoise (Testudo graeca) in North Africa and Anatolia. Ticks feeding on wild T. graeca were collected, and pathogens were screened by polymerase chain reaction using group-specific primers. In total, 131 adult Hyalomma aegyptium ticks were collected from 92 T. graeca in Morocco (n = 48), Tunisia (n = 2), Algeria (n = 70), and Turkey (n = 11). Bacteria and protozoa detected included Hemolivia mauritanica (22.9%), Midichloria mitochondrii (11.4%), relapsing-fever borreliae (8.4%), Ehrlichia spp. (7.6%), Rickettsia spp. (3.4%), Borrelia burgdorferi s.l. (0.9%), Francisella spp. (0.9%), and Wolbachia spp. (0.8%). The characterization of Rickettsia included R. sibirica mongolitimonae (Algeria), R. aeschlimannii (Turkey), and R.africae (Morocco). Hemolivia mauritanica and Ehrlichia spp. prevalence varied significantly with the sampling region/country. We did not detect significant associations in microorganism presence within ticks, nor between microorganism presence and tick mitochondrial DNA haplogroups. This is the first report of Francisella persica-like, relapsing fever borreliae, M. mitochondrii, and Wolbachia spp. in H. aegyptium ticks collected from wild hosts from the South and Eastern Mediterranean region, and of R. sibirica mongolitimonae and R. africae in H. aegyptium from Algeria and Morocco, respectively. Given that T. graeca is a common species in commercial and non-commercial pet trade, the evaluation of the role of this species and its ticks as hosts for TBM is particularly relevant for public health.}, } @article {pmid34122367, year = {2021}, author = {Demirbas-Uzel, G and Augustinos, AA and Doudoumis, V and Parker, AG and Tsiamis, G and Bourtzis, K and Abd-Alla, AMM}, title = {Interactions Between Tsetse Endosymbionts and Glossina pallidipes Salivary Gland Hypertrophy Virus in Glossina Hosts.}, journal = {Frontiers in microbiology}, volume = {12}, number = {}, pages = {653880}, pmid = {34122367}, issn = {1664-302X}, abstract = {Tsetse flies are the sole cyclic vector for trypanosomosis, the causative agent for human African trypanosomosis or sleeping sickness and African animal trypanosomosis or nagana. Tsetse population control is the most efficient strategy for animal trypanosomosis control. Among all tsetse control methods, the Sterile Insect Technique (SIT) is one of the most powerful control tactics to suppress or eradicate tsetse flies. However, one of the challenges for the implementation of SIT is the mass production of target species. Tsetse flies have a highly regulated and defined microbial fauna composed of three bacterial symbionts (Wigglesworthia, Sodalis and Wolbachia) and a pathogenic Glossina pallidipes Salivary Gland Hypertrophy Virus (GpSGHV) which causes reproduction alterations such as testicular degeneration and ovarian abnormalities with reduced fertility and fecundity. Interactions between symbionts and GpSGHV might affect the performance of the insect host. In the present study, we assessed the possible impact of GpSGHV on the prevalence of tsetse endosymbionts under laboratory conditions to decipher the bidirectional interactions on six Glossina laboratory species. The results indicate that tsetse symbiont densities increased over time in tsetse colonies with no clear impact of the GpSGHV infection on symbionts density. However, a positive correlation between the GpSGHV and Sodalis density was observed in Glossina fuscipes species. In contrast, a negative correlation between the GpSGHV density and symbionts density was observed in the other taxa. It is worth noting that the lowest Wigglesworthia density was observed in G. pallidipes, the species which suffers most from GpSGHV infection. In conclusion, the interactions between GpSGHV infection and tsetse symbiont infections seems complicated and affected by the host and the infection density of the GpSGHV and tsetse symbionts.}, } @article {pmid34110943, year = {2021}, author = {Christensen, D and Khoshmanesh, A and Perez-Guaita, D and Iturbe-Ormaetxe, I and O'Neill, S and Wood, BR}, title = {Detection and Identification of Wolbachia pipientis Strains in Mosquito Eggs Using Attenuated Total Reflection Fourier Transform Infrared (ATR FT-IR) Spectroscopy.}, journal = {Applied spectroscopy}, volume = {75}, number = {8}, pages = {1003-1011}, doi = {10.1177/00037028211027140}, pmid = {34110943}, issn = {1943-3530}, mesh = {*Aedes ; Animals ; Fourier Analysis ; Mosquito Vectors ; Spectroscopy, Fourier Transform Infrared ; *Wolbachia ; }, abstract = {The global fight against mosquito-borne viral diseases has in recent years been bolstered by the introduction of the endosymbiotic bacteria Wolbachia to vector populations, which in host mosquitoes suppresses the transmissibility of several viruses. Researchers engaged on this front of the battle often need to know the Wolbachia infection status of individual mosquitoes, as the intervention progresses and the mosquitoes become established in the target population. Previously, we successfully applied attenuated total reflection Fourier transform infrared spectroscopy to the detection of Wolbachia in adult Aedes aegypti mosquitoes; here we apply the same principles to Aedes eggs, with sensitivity and selectivity > 0.95. Further, we successfully distinguish between infections in eggs of the wMel and wMelPop strains of Wolbachia pipientis, with a classification error of 3%. The disruption of host lipid profile by Wolbachia is found to be a key driver in spectral differences between these sample classes.}, } @article {pmid34108021, year = {2021}, author = {Novelo, M and Audsley, MD and McGraw, EA}, title = {The effects of DENV serotype competition and co-infection on viral kinetics in Wolbachia-infected and uninfected Aedes aegypti mosquitoes.}, journal = {Parasites & vectors}, volume = {14}, number = {1}, pages = {314}, pmid = {34108021}, issn = {1756-3305}, support = {R01 AI143758/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/*microbiology/physiology/*virology ; Animals ; Dengue Virus/chemistry/classification/genetics/*physiology ; Female ; Kinetics ; Mosquito Vectors/*microbiology/physiology/*virology ; Viral Load ; Virus Replication ; Wolbachia/genetics/*physiology ; }, abstract = {BACKGROUND: The Aedes aegypti mosquito is responsible for the transmission of several medically important arthropod-borne viruses, including multiple serotypes of dengue virus (DENV-1, -2, -3, and -4). Competition within the mosquito between DENV serotypes can affect viral infection dynamics, modulating the transmission potential of the pathogen. Vector control remains the main method for limiting dengue fever. The insect endosymbiont Wolbachia pipientis is currently being trialed in field releases globally as a means of biological control because it reduces virus replication inside the mosquito. It is not clear how co-infection between DENV serotypes in the same mosquito might alter the pathogen-blocking phenotype elicited by Wolbachia in Ae. aegypti.

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

CONCLUSIONS: This study supports a previous hypothesis that suggests that Torix Rickettsia are overrepresented in aquatic insects. In addition, multiple methods reveal further putative hot spots of Torix Rickettsia infection, including in phloem-feeding bugs, parasitoid wasps, spiders, and vectors of disease. The unknown host effects and transmission strategies of these endosymbionts make these newly discovered associations important to inform future directions of investigation involving the understudied Torix Rickettsia.}, } @article {pmid33760987, year = {2021}, author = {Abrun, P and Ashouri, A and Duplouy, A and Farahani, HK}, title = {Wolbachia impairs post-eclosion host preference in a parasitoid wasp.}, journal = {Die Naturwissenschaften}, volume = {108}, number = {2}, pages = {13}, pmid = {33760987}, issn = {1432-1904}, support = {120586//Marie-Curie Sklodowska Individual fellowship/ ; }, mesh = {Animals ; Female ; Host-Parasite Interactions/*physiology ; Moths/*parasitology ; Wasps/*microbiology ; Wolbachia/*physiology ; }, abstract = {Host preference behavior can result in adaptive advantages with important consequences for the fitness of individuals. Hopkin's host-selection principle (HHSP) suggests that organisms at higher trophic levels demonstrate a preference for the host species on which they developed during their own larval stage. Although investigated in many herbivorous and predatory insects, the HHSP has, to our knowledge, never been tested in the context of insects hosting selfish endosymbiotic passengers. Here, we investigated the effect of infection with the facultative bacterial symbiont Wolbachia on post-eclosion host preference in the parasitoid wasp Trichogramma brassicae (Hymenoptera: Trichogrammatidae). We compared host preference in Wolbachia-infected individuals and uninfected adult female parasitoids after rearing them on two different Lepidopteran hosts, namely the flour moth Ephestia kuehniella Zeller (Lepidoptera: Pyralidae) or the grain moth Sitotroga cerealella (Lepidoptera: Gelechiidae) in choice and no choice experimental design (n = 120 wasps per each choice/no choice experiments). We showed that in T. brassicae, Wolbachia affects the post-eclosion host preference of female wasps. Wolbachia-infected wasps did not show any host preference and more frequently switched hosts in the laboratory, while uninfected wasps significantly preferred to lay eggs on the host species they developed on. Additionally, Wolbachia significantly improved the emergence rate of infected wasps when reared on new hosts. Altogether, our results revealed that the wasp's infection with Wolbachia may lead to impairment of post-eclosion host preference and facilitates growing up on different host species. The impairment of host preference by Wolbachia may allow T. brassicae to shift between hosts, a behavior that might have important evolutionary consequences for the wasp and its symbiont.}, } @article {pmid33717014, year = {2021}, author = {Wang, YT and Shen, RX and Xing, D and Zhao, CP and Gao, HT and Wu, JH and Zhang, N and Zhang, HD and Chen, Y and Zhao, TY and Li, CX}, title = {Metagenome Sequencing Reveals the Midgut Microbiota Makeup of Culex pipiens quinquefasciatus and Its Possible Relationship With Insecticide Resistance.}, journal = {Frontiers in microbiology}, volume = {12}, number = {}, pages = {625539}, pmid = {33717014}, issn = {1664-302X}, abstract = {Midgut microbiota can participate in the detoxification and metabolism processes in insects, but there are few reports on the relationship between midgut microbiota and insecticide resistance in mosquitoes. In this study, we performed metagenomic sequencing on a susceptible strain (SS), a field-collected Hainan strain (HN), and a deltamethrin-resistant strain (RR) of Culex pipiens quinquefasciatus to understand the diversity and functions of their midgut microbiota. The results revealed differences in midgut microbiota among the three strains of Cx. pipiens quinquefasciatus. At the phylum level, Proteobacteria was the most prominent, accounting for nearly 70% of their midgut microbes. At the genus level, Aeromonas made up the highest proportion. In addition, Aeromonas, Morganella, Elizabethkingia, Enterobacter, Cedecea, and Thorsellia showed significant differences between strains. At the species level, Bacillus cereus, Enterobacter cloacae complex sp. 4DZ3-17B2, Streptomyces sp. CNQ329, and some species of Pseudomonas and Wolbachia were more abundant in the two resistant strains. Principal component analysis (PCA) showed that the SS strain had significantly different metagenomic functions than the two deltamethrin-resistant strains (HN and RR strain). The HN and RR strains differed from the SS strain in more than 10 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. The analysis of species abundance and functional diversity can provide directions for future studies.}, } @article {pmid35526996, year = {2021}, author = {Fujita, R}, title = {[Osugoroshi virus, a male-killer virus].}, journal = {Uirusu}, volume = {71}, number = {1}, pages = {63-70}, doi = {10.2222/jsv.71.63}, pmid = {35526996}, issn = {0042-6857}, mesh = {Animals ; DNA Viruses ; Male ; *Moths/microbiology ; *RNA Viruses ; Sex Ratio ; *Spiroplasma ; Symbiosis ; *Viruses, Unclassified ; *Wolbachia ; }, abstract = {In insects, sex ratio bias is sometimes introduced by feminization, parthenogenesis, cytoplasmic incompatibility, or male-killing. Some intracellular bacteria such as Wolbachia or Spiroplasma has been known as male-killing agents. Here I introduce an example of non-bacterial male-killing agent, Osugoroshi virus found in oriental tea tortrix.}, } @article {pmid34392954, year = {2020}, author = {Ciuca, L and Vismarra, A and Lebon, W and Beugnet, F and Morchon, R and Rinaldi, L and Cringoli, G and Kramer, L and Genchi, M}, title = {New insights into the biology, diagnosis and immune response to Dirofilaria repens in the canine host.}, journal = {Veterinary parasitology}, volume = {277S}, number = {}, pages = {100029}, doi = {10.1016/j.vpoa.2020.100029}, pmid = {34392954}, issn = {1873-2550}, abstract = {Dogs are the primary host for Dirofilaria repens, therefore it is mandatory to accurately diagnose the canine infection and to expand our current knowledge on parasite biology and the immune response of the infected host for a better prevention.Thus, the aim of the present study was to provide new insights from experimental infections of dogs with D. repens, focusing on the evaluation of: 1) the pre-patent period and 2) the antibody response against D. repens somatic antigens and against the Wolbachia endosymbiont. Briefly, on Day 0, twenty purpose-bred Beagle dogs were experimentally infected with 50 infective larvae (L3) of D. repens. Starting from Day 58 until the last day of the study (Day 281), blood samples were collected on a monthly basis for detection of antibodies against D. repens (Dr) and recombinant Wolbachia surface protein (rWSP) by non-commercial IgG-ELISAs. Additional samples were collected on Days 220, 245 and 281 for the detection of microfilariae (mff) using the modified Knott's test and biomolecular analysis, following two PCR protocols: Gioia et al. (2010; protocol A) and Rishniw et al. (2006- protocol B). The results were analysed by univariate statistical analyses using 2×2 contingency tables and K Cohen was calculated to assess the agreement among all the diagnostic techniques. Overall, the outcome of the study revealed that out of the 20 dogs experimentally infected with D. repens, 16 (80 %) were microfilaraemic, 17 (85 %) were positive at DNA detection in the blood, 18 (90 %) had D. repens antibodies and 16 (80 %) had Wolbachia antibodies on the last day of the study. The overall k agreement between Knott's and PCR protocol B was 0.442 (P=0.0001) and increased throughout the study, reaching 0.828 (P=0.0001) on Day 281. To the authors knowledge, this is only the second study reporting antibody response to D. repens somatic antigen in experimentally infected dogs. ELISA results showed that an antibody response develops before the onset of patency, and steadily increases with time. Results would suggest that the development of an immunological response to infection could lead to application in epidemiological studies, risk assessment and as an aid in the diagnostic approach in dogs, in particular for early infections without mff.}, } @article {pmid34900237, year = {2019}, author = {Velez, ID and Santacruz, E and Kutcher, SC and Duque, SL and Uribe, A and Barajas, J and Gonzalez, S and Patino, AC and Zuluaga, L and Martínez, L and Muñoz, E and Mejia, MC and Arbelaez, MP and Pulido, H and Jewell, NP and Dufault, SM and O'Neill, SL and Simmons, CP and Anders, KL and Tanamas, SK}, title = {The impact of city-wide deployment of Wolbachia-carrying mosquitoes on arboviral disease incidence in Medellín and Bello, Colombia: study protocol for an interrupted time-series analysis and a test-negative design study.}, journal = {F1000Research}, volume = {8}, number = {}, pages = {1327}, pmid = {34900237}, issn = {2046-1402}, support = {/WT_/Wellcome Trust/United Kingdom ; }, abstract = {Background: Dengue, chikungunya and Zika are viral infections transmitted by Aedes aegypti mosquitoes, and present major public health challenges in tropical regions. Traditional vector control methods have been ineffective at halting disease transmission. The World Mosquito Program has developed a novel approach to arbovirus control using Ae. aegypti stably transfected with the Wolbachia bacterium, which have significantly reduced ability to transmit dengue, Zika and chikungunya in laboratory experiments. Field releases in eight countries have demonstrated Wolbachia establishment in local Ae. aegypti populations. Methods: We describe a pragmatic approach to measuring the epidemiological impact of city-wide Wolbachia deployments in Bello and Medellín, Colombia. First, an interrupted time-series analysis will compare the incidence of dengue, chikungunya and Zika case notifications before and after Wolbachia releases, across the two municipalities. Second, a prospective case-control study using a test-negative design will be conducted in one quadrant of Medellín. Three of the six contiguous release zones in the case-control area were allocated to receive the first Wolbachia deployments in the city and three to be treated last, approximating a parallel two-arm trial for the >12-month period during which Wolbachia exposure remains discordant. Allocation, although non-random, aimed to maximise balance between arms in historical dengue incidence and demographics. Arboviral disease cases and arbovirus-negative controls will be enrolled concurrently from febrile patients presenting to primary care, with case/control status classified retrospectively following laboratory diagnostic testing. Intervention effect is estimated from an aggregate odds ratio comparing Wolbachia-exposure odds among test-positive cases versus test-negative controls. Discussion: The study findings will add to an accumulating body of evidence from global field sites on the efficacy of the Wolbachia method in reducing arboviral disease incidence, and can inform decisions on wider public health implementation of this intervention in the Americas and beyond. Trial registration: ClinicalTrials.gov: NCT03631719. Registered on 15 August 2018.}, } @article {pmid34541196, year = {2017}, author = {Fink, C and Von Frieling, J and Knop, M and Roeder, T}, title = {Drosophila Fecal Sampling.}, journal = {Bio-protocol}, volume = {7}, number = {18}, pages = {e2547}, pmid = {34541196}, issn = {2331-8325}, abstract = {Fecal sampling is a non-invasive method which raises the possibility to study the development and the changes in the microbial community throughout different time points of a fly population or throughout different treatments. This method allows precise manipulation to trigger the fly's physiology by nutritional interventions, bacterial infections or other stressors. As in most other animals, the intestinal microbiota is essential for a healthy fly-life. Because Drosophila only harbors a relative simple bacterial community with a small variety of round about 8 to 10 different species, it is rather easy to build up the microbial community and to investigate microbial changes after treatment. Another positive effect using the fly's feces is that bacteria that are not part of the intestinal microbiome, for example Wolbachia, can be excluded directly from the analysis because they are not excreted. Using this method, the generated datasets may reflect a good paradigm to study microbiome associated diseases in a simple fly model or furthermore, to test drugs in a high-throughput approach.}, } @article {pmid33716790, year = {2021}, author = {Martins, M and Ramos, LFC and Murillo, JR and Torres, A and de Carvalho, SS and Domont, GB and de Oliveira, DMP and Mesquita, RD and Nogueira, FCS and Maciel-de-Freitas, R and Junqueira, M}, title = {Comprehensive Quantitative Proteome Analysis of Aedes aegypti Identifies Proteins and Pathways Involved in Wolbachia pipientis and Zika Virus Interference Phenomenon.}, journal = {Frontiers in physiology}, volume = {12}, number = {}, pages = {642237}, pmid = {33716790}, issn = {1664-042X}, abstract = {Zika virus (ZIKV) is a global public health emergency due to its association with microcephaly, Guillain-Barré syndrome, neuropathy, and myelitis in children and adults. A total of 87 countries have had evidence of autochthonous mosquito-borne transmission of ZIKV, distributed across four continents, and no antivirus therapy or vaccines are available. Therefore, several strategies have been developed to target the main mosquito vector, Aedes aegypti, to reduce the burden of different arboviruses. Among such strategies, the use of the maternally-inherited endosymbiont Wolbachia pipientis has been applied successfully to reduce virus susceptibility and decrease transmission. However, the mechanisms by which Wolbachia orchestrate resistance to ZIKV infection remain to be elucidated. In this study, we apply isobaric labeling quantitative mass spectrometry (MS)-based proteomics to quantify proteins and identify pathways altered during ZIKV infection; Wolbachia infection; co-infection with Wolbachia/ZIKV in the A. aegypti heads and salivary glands. We show that Wolbachia regulates proteins involved in reactive oxygen species production, regulates humoral immune response, and antioxidant production. The reduction of ZIKV polyprotein in the presence of Wolbachia in mosquitoes was determined by MS and corroborates the idea that Wolbachia helps to block ZIKV infections in A. aegypti. The present study offers a rich resource of data that may help to elucidate mechanisms by which Wolbachia orchestrate resistance to ZIKV infection in A. aegypti, and represents a step further on the development of new targeted methods to detect and quantify ZIKV and Wolbachia directly in complex tissues.}, } @article {pmid33704487, year = {2021}, author = {Dobson, SL}, title = {When More is Less: Mosquito Population Suppression Using Sterile, Incompatible and Genetically Modified Male Mosquitoes.}, journal = {Journal of medical entomology}, volume = {58}, number = {5}, pages = {1980-1986}, doi = {10.1093/jme/tjab025}, pmid = {33704487}, issn = {1938-2928}, mesh = {Animals ; *Culicidae/microbiology/radiation effects ; Male ; Mosquito Control/*methods ; *Mosquito Vectors/microbiology/radiation effects ; Wolbachia/*physiology ; }, abstract = {The current review of the Sterile Insect Technique (SIT) is motivated by new technologies and the recent renaissance of male release field trials, which is driving an evolution in mosquito control and regulation. Practitioners that are releasing male mosquitoes would do well to learn from past successes and failures, including political and public engagement complications. With examples that include nuanced integrations of the different technologies, e.g., combinations of Wolbachia and irradiation, it is critical that scientists understand and communicate accurately about the technologies, including their evolving management by different regulatory agencies in the USA. Some male release approaches are considered 'pesticides' and regulated by federal and state agencies, while other male release approaches are unregulated. It is important to consider how the new technologies fit with the more 'traditional' chemical applications of adulticides and larvicides. The economics of male release programs are substantially different from traditional control costs, which can be a challenge to their adoption by abatement districts. However, there is substantial need to overcome these complications and challenges, because the problem with invasive mosquitoes grows ever worse with factors that include insecticide resistance, globalization and climate change.}, } @article {pmid33683351, year = {2021}, author = {Shropshire, JD and Rosenberg, R and Bordenstein, SR}, title = {The impacts of cytoplasmic incompatibility factor (cifA and cifB) genetic variation on phenotypes.}, journal = {Genetics}, volume = {217}, number = {1}, pages = {1-13}, pmid = {33683351}, issn = {1943-2631}, support = {R01 AI132581/AI/NIAID NIH HHS/United States ; R01 AI143725/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Drosophila melanogaster ; Female ; Genetic Fitness ; *Genetic Variation ; Infertility/microbiology ; Male ; *Phenotype ; Prophages/*genetics ; Spermatozoa/microbiology ; Viral Proteins/*genetics ; Wolbachia/pathogenicity/virology ; }, abstract = {Wolbachia are maternally transmitted, intracellular bacteria that can often selfishly spread through arthropod populations via cytoplasmic incompatibility (CI). CI manifests as embryonic death when males expressing prophage WO genes cifA and cifB mate with uninfected females or females harboring an incompatible Wolbachia strain. Females with a compatible cifA-expressing strain rescue CI. Thus, cif-mediated CI confers a relative fitness advantage to females transmitting Wolbachia. However, whether cif sequence variation underpins incompatibilities between Wolbachia strains and variation in CI penetrance remains unknown. Here, we engineer Drosophila melanogaster to transgenically express cognate and non-cognate cif homologs and assess their CI and rescue capability. Cognate expression revealed that cifA;B native to D. melanogaster causes strong CI, and cognate cifA;B homologs from two other Drosophila-associated Wolbachia cause weak transgenic CI, including the first demonstration of phylogenetic type 2 cifA;B CI. Intriguingly, non-cognate expression of cifA and cifB alleles from different strains revealed that cifA homologs generally contribute to strong transgenic CI and interchangeable rescue despite their evolutionary divergence, and cifB genetic divergence contributes to weak or no transgenic CI. Finally, we find that a type 1 cifA can rescue CI caused by a genetically divergent type 2 cifA;B in a manner consistent with unidirectional incompatibility. By genetically dissecting individual CI functions for type 1 and 2 cifA and cifB, this work illuminates new relationships between cif genotype and CI phenotype. We discuss the relevance of these findings to CI's genetic basis, phenotypic variation patterns, and mechanism.}, } @article {pmid33671250, year = {2021}, author = {Zheng, Z and Hu, X and Xu, Y and Wei, C and He, H}, title = {Bacterial Composition and Diversity of the Digestive Tract of Odontomachus monticola Emery and Ectomomyrmex javanus Mayr.}, journal = {Insects}, volume = {12}, number = {2}, pages = {}, pmid = {33671250}, issn = {2075-4450}, support = {Grant Number 31570388//the Natural Science Foundation of China/ ; Grant Number 2452019174.//the Fundamental Research Funds for the Central Universities/ ; }, abstract = {Ponerine ants are generalist predators feeding on a variety of small arthropods, annelids, and isopods; however, knowledge of their bacterial communities is rather limited. This study investigated the bacterial composition and diversity in the digestive tract (different gut sections and the infrabuccal pockets (IBPs)) of two ponerine ant species (Odontomachus monticola Emery and Ectomomyrmex javanus Mayr) distributed in northwestern China using high-throughput sequencing. We found that several dominant bacteria that exist in other predatory ants were also detected in these two ponerine ant species, including Wolbachia, Mesoplasma, and Spiroplasma. Bacterial communities of these two ant species were differed significantly from each other, and significant differences were also observed across their colonies, showing distinctive inter-colony characteristics. Moreover, bacterial communities between the gut sections (crops, midguts, and hindguts) of workers were highly similar within colony, but they were clearly different from those in IBPs. Further, bacterial communities in the larvae of O. monticola were similar to those in the IBPs of workers, but significantly different from those in gut sections. We presume that the bacterial composition and diversity in ponerine ants are related to their social behavior and feeding habits, and bacterial communities in the IBPs may play a potential role in their social life.}, } @article {pmid33664278, year = {2021}, author = {Ma, YJ and He, HP and Zhao, HM and Xian, YD and Guo, H and Liu, B and Xue, K}, title = {Microbiome diversity of cotton aphids (Aphis gossypii) is associated with host alternation.}, journal = {Scientific reports}, volume = {11}, number = {1}, pages = {5260}, pmid = {33664278}, issn = {2045-2322}, mesh = {Animals ; Aphids/genetics/*microbiology ; Genetic Variation/*genetics ; Gossypium/genetics/microbiology ; High-Throughput Nucleotide Sequencing ; Host-Pathogen Interactions/*genetics ; Microbiota/*genetics ; Symbiosis/genetics ; }, abstract = {Aphids are infected by a series of bacteria that can help them survive on specific host plants. However, the associations between aphids and these bacteria are not clear, and the bacterial communities in many aphid species are poorly characterized. Here, we investigated the bacterial communities of cotton aphids (Aphis gossypii) on 2 representative winter host plants and transferred to 3 summer host plants by 16S rDNA sequencing using the Illumina MiSeq platform. Our results revealed that the bacterial communities varied among cotton aphids on hibiscus, cotton aphids on pomegranate, cotton aphids on cotton transferred from hibiscus, cotton aphids on muskmelon transferred from hibiscus, cotton aphids on cucumber transferred from hibiscus,. The diversity and richness of the bacterial communities were significantly higher in aphids on muskmelon and aphids on cucumber than in the other treatments. There were two main factors influencing the distribution of internal bacterial OTUs revealed by principal component analysis, including the differences among Punicaceae, Malvaceae and Cucurbitaceae. There were 28 bacterial communities with significant differences between two arbitrary treatments, which could be grouped into 6 main clusters depending on relative abundance. Moreover, our results indicated that in addition to the obligate endosymbiont Buchnera, with a dominant position (> 52%), A. gossypii also harbored 3 facultative endosymbiotic bacteria (Serratia, Arsenophonus, and Wolbachia) and 3 possibly symbiotic bacteria (Acinetobacter, Pantoea, and Flavobacterium). There were several correspondences between the symbiotic bacteria in cotton aphids and the specific host plants of the aphids. This study provides a better understanding of the interactions among symbiotic bacteria, aphids and host plants, suggesting that the selection pressure on aphid bacterial communities is likely to be exerted by the species of host plants.}, } @article {pmid33659853, year = {2020}, author = {Romanov, DA and Zakharov, IA and Shaikevich, EV}, title = {Wolbachia, Spiroplasma, and Rickettsia symbiotic bacteria in aphids (Aphidoidea).}, journal = {Vavilovskii zhurnal genetiki i selektsii}, volume = {24}, number = {6}, pages = {673-682}, doi = {10.18699/VJ20.661}, pmid = {33659853}, issn = {2500-0462}, abstract = {Aphids are a diverse family of crop pests. Aphids formed a complex relationship with intracellular bacteria. Depending on the region of study, the species composition of both aphids and their facultative endosymbionts varies. The aim of the work was to determine the occurrence and genetic diversity of Wolbachia, Spiroplasma and Rickettsia symbionts in aphids collected in 2018-2019 in Moscow. For these purposes, 578 aphids from 32 collection sites were tested by PCR using specific primers. At least 21 species of aphids from 14 genera and four families were identified by barcoding method, of which 11 species were infected with endosymbionts. Rickettsia was found in six species, Wolbachia in two species, Spiroplasma in one species. The presence of Rickettsia in Impatientinum asiaticum, Myzus cerasi, Hyalopterus pruni, Eucallipterus tiliae, Chaitophorus tremulae and Wolbachia in Aphis pomi and C. tremulae has been described for the first time. A double infection with Rickettsia and Spiroplasma was detected in a half of pea aphid (Acyrthosiphon pisum) individuals. For the first time was found that six species of aphids are infected with Rickettsia that are genetically different from previously known. It was first discovered that A. pomi is infected with two Wolbachia strains, one of which belongs to supergroup B and is genetically close to Wolbachia from C. tremulae. The second Wolbachia strain from A. pomi belongs to the supergroup M, recently described in aphid species. Spiroplasma, which we observed in A. pisum, is genetically close to male killing Spiroplasma from aphids, ladybirds and moths. Both maternal inheritance and horizontal transmission are the pathways for the distribution of facultative endosymbiotic bacteria in aphids.}, } @article {pmid33646378, year = {2021}, author = {Wangkeeree, J and Sanit, P and Roddee, J and Hanboonsong, Y}, title = {Phylogeny and Strain Typing of Wolbachia from Yamatotettix flavovittatus Matsumura Leafhoppers.}, journal = {Current microbiology}, volume = {78}, number = {4}, pages = {1367-1376}, pmid = {33646378}, issn = {1432-0991}, support = {226/2563//National Research Council of Thailand/ ; }, mesh = {Animals ; Female ; *Hemiptera ; Multilocus Sequence Typing ; Phylogeny ; *Saccharum ; *Wolbachia/genetics ; }, abstract = {Wolbachia is a maternally inherited bacterium of insects that can alter the reproduction, biology, and fitness of the hosts. It was detected in natural populations of the Yamatotettix flavovittatus Matsumura leafhoppers, the vector of phytoplasma, which is responsible for sugarcane white leaf disease. Wolbachia infection prolongs the longevity of female leafhoppers and promotes a strong reproductive incompatibility; importantly, highly maternal transmission rate was observed. However, limited data on the diversity or strain typing of Wolbachia in Y. flavovittatus are available. We aimed here to detect the presence of Wolbachia in different populations by amplification of the wsp gene, which was then sequenced. Multilocus sequence typing (MLST) was also performed to explore the diversity of the Wolbachia strains. Based on the wsp sequences, Wolbachia in the Y. flavovittatus leafhoppers belonged to supergroup B, and formed a distinct evolutionary lineage; therefore, we designated this new specific strain as wYfla. The MLST profiles revealed ten potential new sequence types (STs) in different leafhopper populations. Multiple STs were detected in individual leafhoppers, among which the ST-wYfla1 strain was predominant. Furthermore, we obtained congruent results for the phylogenetic analyses using the wsp gene and MLST loci. To the best of our knowledge, this is the first study characterizing Wolbachia strains in Y. flavovittatus. Our results reveal a novel strain and multiple STs of Wolbachia, and these data may prove useful in the exploitation of Wolbachia as a biological Y. flavovittatus control agent.}, } @article {pmid33644932, year = {2021}, author = {Burdina, EV and Bykov, RA and Menshanov, PN and Ilinsky, YY and Gruntenko, NЕ}, title = {Unique Wolbachia strain wMelPlus increases heat stress resistance in Drosophila melanogaster.}, journal = {Archives of insect biochemistry and physiology}, volume = {106}, number = {4}, pages = {e21776}, doi = {10.1002/arch.21776}, pmid = {33644932}, issn = {1520-6327}, support = {0259-2021-0016//Ministry of Science and Higher Education of the Russian Federation/ ; 20-04-00579//Russian Foundation for Basic Research/ ; }, mesh = {Alkaline Phosphatase/metabolism ; Animals ; Dopamine/metabolism ; Drosophila melanogaster/metabolism/*microbiology ; Genotype ; Heat-Shock Response ; Symbiosis/physiology ; *Thermotolerance/genetics/physiology ; Wolbachia/*genetics ; }, abstract = {Maternally inherited endosymbiotic bacterium Wolbachia infects Drosophila melanogaster populations worldwide. Its genetic diversity includes several closely related genotypes, which can be attributed to two main genotype groups: wMel and wMelCS. Here, we studied eight D. melanogaster lines carrying the nuclear background of wild type interbred Bi90 line and cytoplasmic backgrounds with or without Wolbachia of different origin, each of which belongs to wMelCS genotype group. We analyzed the effect these seven Wolbachia strains had on the heat stress resistance and dopamine metabolism in D. melanogaster females. Survival under heat stress (38°C, 3 h 30 min) was increased only in the line infected with bacteria of the wMelPlus strain. At the same time, the activity of alkaline phosphatase (an enzyme regulating the pool of dopamine precursor tyrosine) was increased under normal conditions in females infected with all strains under study and retained the response to heat stress typical for the uninfected line. Thus, we found the unique Wolbachia strain that provides an increase of the host stress resistance, and demonstrated that the mechanism of this resistance is not dopamine-mediated.}, } @article {pmid33629719, year = {2021}, author = {Benedict, MQ}, title = {Sterile Insect Technique: Lessons From the Past.}, journal = {Journal of medical entomology}, volume = {58}, number = {5}, pages = {1974-1979}, doi = {10.1093/jme/tjab024}, pmid = {33629719}, issn = {1938-2928}, mesh = {Mosquito Control/instrumentation/*methods/statistics & numerical data ; *Mosquito Vectors ; }, abstract = {When E.F. Knipling conceived of the release of sexually sterile insects to suppress wild populations, he laid down several fundamental qualities that characterized suitable target species-some of which mosquitoes generally violate-including high reproductive rates and large population numbers. Regardless of this, their global importance in public health has led numerous research teams to attempt to use the mosquito sterile insect technique against several species. Because of the degree of financial commitment required for suppression programs, most releases have consisted of preliminary investigations of male performance, population characteristics, and production methods. Those that have accomplished suppression provide important insights regarding the challenges of production, dispersal, and immigration. Insights gained from these studies remain relevant today, regardless of the genetic control technology being applied. In this article, I highlight studies that were notable for the insights that were gained, the intrinsic difficulties that mosquitoes present, and synthesize these into recommendations for successful applications of the sterile insect technique and newer technologies to mosquitoes.}, } @article {pmid33627171, year = {2021}, author = {Bulman, CA and Chappell, L and Gunderson, E and Vogel, I and Beerntsen, B and Slatko, BE and Sullivan, W and Sakanari, JA}, title = {The Eagle effect in the Wolbachia-worm symbiosis.}, journal = {Parasites & vectors}, volume = {14}, number = {1}, pages = {118}, pmid = {33627171}, issn = {1756-3305}, support = {OPP1017584//Bill and Melinda Gates Foundation/ ; }, mesh = {Animals ; Anti-Bacterial Agents/pharmacology ; Brugia malayi/drug effects/microbiology/*physiology ; Doxycycline/pharmacology ; Female ; Male ; Microfilariae/drug effects/*microbiology/physiology ; Onchocerca/drug effects/microbiology/*physiology ; *Symbiosis/drug effects ; Wolbachia/drug effects/*physiology ; }, abstract = {BACKGROUND: Onchocerciasis (river blindness) and lymphatic filariasis (elephantiasis) are two human neglected tropical diseases that cause major disabilities. Mass administration of drugs targeting the microfilarial stage has reduced transmission and eliminated these diseases in several countries but a macrofilaricidal drug that kills or sterilizes the adult worms is critically needed to eradicate the diseases. The causative agents of onchocerciasis and lymphatic filariasis are filarial worms that harbor the endosymbiotic bacterium Wolbachia. Because filarial worms depend on Wolbachia for reproduction and survival, drugs targeting Wolbachia hold great promise as a means to eliminate these diseases.

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

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

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

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

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

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

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

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

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

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

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

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

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

CONCLUSIONS: The HIFI-SE pipeline represents an efficient way to produce standard full-length barcodes, while the reasonable cost and high sensitivity of our method can contribute considerably more DNA barcodes under the same budget. Our method thereby advances DNA-based species identification from diverse ecosystems and increases the number of relevant applications.}, } @article {pmid33263885, year = {2020}, author = {Attardo, GM and Scolari, F and Malacrida, A}, title = {Bacterial Symbionts of Tsetse Flies: Relationships and Functional Interactions Between Tsetse Flies and Their Symbionts.}, journal = {Results and problems in cell differentiation}, volume = {69}, number = {}, pages = {497-536}, pmid = {33263885}, issn = {0080-1844}, mesh = {Animals ; Enterobacteriaceae/*physiology ; Female ; *Host Microbial Interactions ; Male ; Microbiota ; *Symbiosis ; Trypanosoma ; Tsetse Flies/*microbiology/parasitology ; Wolbachia/*physiology ; }, abstract = {Tsetse flies (Glossina spp.) act as the sole vectors of the African trypanosome species that cause Human African Trypanosomiasis (HAT or African Sleeping Sickness) and Nagana in animals. These flies have undergone a variety of specializations during their evolution including an exclusive diet consisting solely of vertebrate blood for both sexes as well as an obligate viviparous reproductive biology. Alongside these adaptations, Glossina species have developed intricate relationships with specific microbes ranging from mutualistic to parasitic. These relationships provide fundamental support required to sustain the specializations associated with tsetse's biology. This chapter provides an overview on the knowledge to date regarding the biology behind these relationships and focuses primarily on four bacterial species that are consistently associated with Glossina species. Here their interactions with the host are reviewed at the morphological, biochemical and genetic levels. This includes: the obligate symbiont Wigglesworthia, which is found in all tsetse species and is essential for nutritional supplementation to the blood-specific diet, immune system maturation and facilitation of viviparous reproduction; the commensal symbiont Sodalis, which is a frequently associated symbiont optimized for survival within the fly via nutritional adaptation, vertical transmission through mating and may alter vectorial capacity of Glossina for trypanosomes; the parasitic symbiont Wolbachia, which can manipulate Glossina via cytoplasmic incompatibility and shows unique interactions at the genetic level via horizontal transmission of its genetic material into the genome in two Glossina species; finally, knowledge on recently observed relations between Spiroplasma and Glossina is explored and potential interactions are discussed based on knowledge of interactions between this bacterial Genera and other insect species. These flies have a simple microbiome relative to that of other insects. However, these relationships are deep, well-studied and provide a window into the complexity and function of host/symbiont interactions in an important disease vector.}, } @article {pmid33263882, year = {2020}, author = {Lefoulon, E and Foster, JM and Truchon, A and Carlow, CKS and Slatko, BE}, title = {The Wolbachia Symbiont: Here, There and Everywhere.}, journal = {Results and problems in cell differentiation}, volume = {69}, number = {}, pages = {423-451}, pmid = {33263882}, issn = {0080-1844}, mesh = {Animals ; Arthropods/*microbiology ; Filarioidea/*microbiology ; Host Microbial Interactions ; *Symbiosis ; *Wolbachia ; }, abstract = {Wolbachia symbionts, first observed in the 1920s, are now known to be present in about 30-70% of tested arthropod species, in about half of tested filarial nematodes (including the majority of human filarial nematodes), and some plant-parasitic nematodes. In arthropods, they are generally viewed as parasites while in nematodes they appear to be mutualists although this demarcation is not absolute. Their presence in arthropods generally leads to reproductive anomalies, while in nematodes, they are generally required for worm development and reproduction. In mosquitos, Wolbachia inhibit RNA viral infections, leading to populational reductions in human RNA virus pathogens, whereas in filarial nematodes, their requirement for worm fertility and survival has been channeled into their use as drug targets for filariasis control. While much more research on these ubiquitous symbionts is needed, they are viewed as playing significant roles in biological processes, ranging from arthropod speciation to human health.}, } @article {pmid33263871, year = {2020}, author = {Russell, SL and Castillo, JR}, title = {Trends in Symbiont-Induced Host Cellular Differentiation.}, journal = {Results and problems in cell differentiation}, volume = {69}, number = {}, pages = {137-176}, pmid = {33263871}, issn = {0080-1844}, support = {K99 GM135583/GM/NIGMS NIH HHS/United States ; R00 GM135583/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Arthropods/*microbiology ; *Cell Differentiation ; Genome, Bacterial ; Nematoda/*microbiology ; Reproduction ; *Symbiosis ; *Wolbachia/genetics ; }, abstract = {Bacteria participate in a wide diversity of symbiotic associations with eukaryotic hosts that require precise interactions for bacterial recognition and persistence. Most commonly, host-associated bacteria interfere with host gene expression to modulate the immune response to the infection. However, many of these bacteria also interfere with host cellular differentiation pathways to create a hospitable niche, resulting in the formation of novel cell types, tissues, and organs. In both of these situations, bacterial symbionts must interact with eukaryotic regulatory pathways. Here, we detail what is known about how bacterial symbionts, from pathogens to mutualists, control host cellular differentiation across the central dogma, from epigenetic chromatin modifications, to transcription and mRNA processing, to translation and protein modifications. We identify four main trends from this survey. First, mechanisms for controlling host gene expression appear to evolve from symbionts co-opting cross-talk between host signaling pathways. Second, symbiont regulatory capacity is constrained by the processes that drive reductive genome evolution in host-associated bacteria. Third, the regulatory mechanisms symbionts exhibit correlate with the cost/benefit nature of the association. And, fourth, symbiont mechanisms for interacting with host genetic regulatory elements are not bound by native bacterial capabilities. Using this knowledge, we explore how the ubiquitous intracellular Wolbachia symbiont of arthropods and nematodes may modulate host cellular differentiation to manipulate host reproduction. Our survey of the literature on how infection alters gene expression in Wolbachia and its hosts revealed that, despite their intermediate-sized genomes, different strains appear capable of a wide diversity of regulatory manipulations. Given this and Wolbachia's diversity of phenotypes and eukaryotic-like proteins, we expect that many symbiont-induced host differentiation mechanisms will be discovered in this system.}, } @article {pmid33257667, year = {2020}, author = {Souto-Maior, C and King, JG and Sartori, LM and Maciel-de-Freitas, R and Gomes, MGM}, title = {Reply to: "Enhancement of Aedes aegypti susceptibility to dengue by Wolbachia is not supported".}, journal = {Nature communications}, volume = {11}, number = {1}, pages = {6113}, pmid = {33257667}, issn = {2041-1723}, mesh = {*Aedes ; Animals ; *Dengue ; *Wolbachia ; }, } @article {pmid33257651, year = {2020}, author = {Ant, TH and Mancini, MV and Martinez, J and Sinkins, SP}, title = {Enhancement of Aedes aegypti susceptibility to dengue by Wolbachia is not supported.}, journal = {Nature communications}, volume = {11}, number = {1}, pages = {6111}, pmid = {33257651}, issn = {2041-1723}, support = {202888/Z/16/Z/WT_/Wellcome Trust/United Kingdom ; }, mesh = {*Aedes ; Animals ; *Dengue ; *Wolbachia ; }, } @article {pmid33248417, year = {2020}, author = {Madhav, M and Baker, D and Morgan, JAT and Asgari, S and James, P}, title = {Wolbachia: A tool for livestock ectoparasite control.}, journal = {Veterinary parasitology}, volume = {288}, number = {}, pages = {109297}, doi = {10.1016/j.vetpar.2020.109297}, pmid = {33248417}, issn = {1873-2550}, mesh = {Animals ; Camelus/parasitology ; Cattle/parasitology ; Cimicidae/microbiology ; Diptera/microbiology ; Ectoparasitic Infestations/parasitology/prevention & control/*veterinary ; Goats/parasitology ; Horses/parasitology ; *Livestock ; Mites/microbiology ; Phthiraptera/microbiology ; Sheep, Domestic/parasitology ; Siphonaptera/microbiology ; Sus scrofa/parasitology ; Ticks/microbiology ; Wolbachia/*physiology ; }, abstract = {Ectoparasites and livestock-associated insects are a major concern throughout the world because of their economic and welfare impacts. Effective control is challenging and relies mainly on the use of chemical insecticides and acaricides. Wolbachia, an arthropod and nematode-infecting, maternally-transmitted endosymbiont is currently of widespread interest for use in novel strategies for the control of a range of arthropod-vectored human diseases and plant pests but to date has received only limited consideration for use in the control of diseases of veterinary concern. Here, we review the currently available information on Wolbachia in veterinary ectoparasites and disease vectors, consider the feasibility for use of Wolbachia in the control of livestock pests and diseases and highlight critical issues which need further investigation.}, } @article {pmid33240234, year = {2020}, author = {Momtaz, AZ and Ahumada Sabagh, AD and Gonzalez Amortegui, JG and Salazar, SA and Finessi, A and Hernandez, J and Christensen, S and Serbus, LR}, title = {A Role for Maternal Factors in Suppressing Cytoplasmic Incompatibility.}, journal = {Frontiers in microbiology}, volume = {11}, number = {}, pages = {576844}, pmid = {33240234}, issn = {1664-302X}, abstract = {Wolbachia are maternally transmitted bacterial endosymbionts, carried by approximately half of all insect species. Wolbachia prevalence in nature stems from manipulation of host reproduction to favor the success of infected females. The best known reproductive modification induced by Wolbachia is referred to as sperm-egg Cytoplasmic Incompatibility (CI). In CI, the sperm of Wolbachia-infected males cause embryonic lethality, attributed to paternal chromatin segregation defects during early mitotic divisions. Remarkably, the embryos of Wolbachia-infected females "rescue" CI lethality, yielding egg hatch rates equivalent to uninfected female crosses. Several models have been discussed as the basis for Rescue, and functional evidence indicates a major contribution by Wolbachia CI factors. A role for host contributions to Rescue remains largely untested. In this study, we used a chemical feeding approach to test for CI suppression capabilities by Drosophila simulans. We found that uninfected females exhibited significantly higher CI egg hatch rates in response to seven chemical treatments that affect DNA integrity, cell cycle control, and protein turnover. Three of these treatments suppressed CI induced by endogenous wRi Wolbachia, as well as an ectopic wMel Wolbachia infection. The results implicate DNA integrity as a focal aspect of CI suppression for different Wolbachia strains. The framework presented here, applied to diverse CI models, will further enrich our understanding of host reproductive manipulation by insect endosymbionts.}, } @article {pmid33237151, year = {2020}, author = {Pimentel, AC and Beraldo, CS and Cogni, R}, title = {Host-shift as the cause of emerging infectious diseases: Experimental approaches using Drosophila-virus interactions.}, journal = {Genetics and molecular biology}, volume = {44}, number = {1 Suppl 1}, pages = {e20200197}, pmid = {33237151}, issn = {1415-4757}, abstract = {Host shifts, when a cross-species transmission of a pathogen can lead to successful infections, are the main cause of emerging infectious diseases, such as COVID-19. A complex challenge faced by the scientific community is to address the factors that determine whether the cross-species transmissions will result in spillover or sustained onwards infections. Here we review recent literature and present a perspective on current approaches we are using to understand the mechanisms underlying host shifts. We highlight the usefulness of the interactions between Drosophila species and viruses as an ideal study model. Additionally, we discuss how cross-infection experiments - when pathogens from a natural reservoir are intentionally injected in novel host species- can test the effect cross-species transmissions may have on the fitness of virus and host, and how the host phylogeny may influence this response. We also discuss experiments evaluating how cooccurrence with other viruses or the presence of the endosymbiont bacteria Wolbachia may affect the performance of new viruses in a novel host. Finally, we discuss the need of surveys of virus diversity in natural populations using next-generation sequencing technologies. In the long term, these approaches can contribute to a better understanding of the basic biology of host shifts.}, } @article {pmid33222623, year = {2022}, author = {Amala, M and Richard, M and Saritha, P and Prabhu, D and Veerapandiyan, M and Surekha, K and Jeyakanthan, J}, title = {Molecular evolution, binding site interpretation and functional divergence of aspartate semialdehyde dehydrogenase.}, journal = {Journal of biomolecular structure & dynamics}, volume = {40}, number = {7}, pages = {3223-3241}, doi = {10.1080/07391102.2020.1846619}, pmid = {33222623}, issn = {1538-0254}, mesh = {Amino Acid Sequence ; *Aspartate-Semialdehyde Dehydrogenase/chemistry/genetics ; Binding Sites ; *Evolution, Molecular ; Phylogeny ; }, abstract = {Aspartate Semialdehyde Dehydrogenase (ASDH) is an important enzyme essential for the viability of pathogenic microorganisms. ASDH is mainly involved in amino acid and cell wall biosynthesis of microorganisms, hence it is considered to be a promising target for drug design. This enzyme depicts similar mechanistic function in all microorganisms; although, the kinetic efficiency of an enzyme differs according to their active site residual composition. Therefore, understanding the residual variation and kinetic efficiency of the enzyme would pave new insights in structure-based drug discovery and a novel drug molecule against ASDH. Here, ASDH from Wolbachia endosymbiont of Brugia malayi is used as a prime enzyme to execute evolutionary studies. The phylogenetic analysis was opted to classify 400 sequences of ASDH enzymes based on their structure and electrostatic surfaces. Analysis resulted in 37 monophyletic clades of diverse pathogenic and non-pathogenic organisms. The representative structures of 37 ASDHs from different clades were further deciphered to structural homologues. These enzymes exhibited presence of more positively charged surfaces than negatively charged surfaces in the active site pocket which restrains evolutionary significance. Docking studies of NADP[+] with 37 enzymes reveals that site-specific residual variation in the active site pocket modulates the binding affinity (ranges of -13 to -9 kcal/mol). Type-I and Type-II divergence studies show, no significant functional divergence among ASDH, but residual changes were found among the enzyme that modulates the biochemical characteristics and catalytic efficiency. The present study not only explores residual alteration and catalytic variability, it also aids in the design of species-specific inhibitors.Communicated by Ramaswamy H. Sarma.}, } @article {pmid33216144, year = {2021}, author = {Cormier, A and Chebbi, MA and Giraud, I and Wattier, R and Teixeira, M and Gilbert, C and Rigaud, T and Cordaux, R}, title = {Comparative Genomics of Strictly Vertically Transmitted, Feminizing Microsporidia Endosymbionts of Amphipod Crustaceans.}, journal = {Genome biology and evolution}, volume = {13}, number = {1}, pages = {}, pmid = {33216144}, issn = {1759-6653}, mesh = {Amphipoda/*genetics ; Animals ; Female ; Feminization/*genetics/parasitology ; *Genomics ; Host-Parasite Interactions ; Male ; Microsporidia/*genetics ; Nosema ; Phylogeny ; Wolbachia/genetics ; }, abstract = {Microsporidia are obligate intracellular eukaryotic parasites of vertebrates and invertebrates. Microsporidia are usually pathogenic and undergo horizontal transmission or a mix of horizontal and vertical transmission. However, cases of nonpathogenic microsporidia, strictly vertically transmitted from mother to offspring, have been reported in amphipod crustaceans. Some of them further evolved the ability to feminize their nontransmitting male hosts into transmitting females. However, our understanding of the evolution of feminization in microsporidia is hindered by a lack of genomic resources. We report the sequencing and analysis of three strictly vertically transmitted microsporidia species for which feminization induction has been demonstrated (Nosema granulosis) or is strongly suspected (Dictyocoela muelleri and Dictyocoela roeselum), along with a draft genome assembly of their host Gammarus roeselii. Contrary to horizontally transmitted microsporidia that form environmental spores that can be purified, feminizing microsporidia cannot be easily isolated from their host cells. Therefore, we cosequenced symbiont and host genomic DNA and devised a computational strategy to obtain genome assemblies for the different partners. Genomic comparison with feminizing Wolbachia bacterial endosymbionts of isopod crustaceans indicated independent evolution of feminization in microsporidia and Wolbachia at the molecular genetic level. Feminization thus represents a remarkable evolutionary convergence of eukaryotic and prokaryotic microorganisms. Furthermore, a comparative genomics analysis of microsporidia allowed us to identify several candidate genes for feminization, involving functions such as DNA binding and membrane fusion. The genomic resources we generated contribute to establish Gammarus roeselii and its microsporidia symbionts as a new model to study the evolution of symbiont-mediated feminization.}, } @article {pmid33198811, year = {2020}, author = {Tongkrajang, N and Ruenchit, P and Tananchai, C and Chareonviriyaphap, T and Kulkeaw, K}, title = {Molecular identification of native Wolbachia pipientis in Anopheles minimus in a low-malaria transmission area of Umphang Valley along the Thailand-Myanmar border.}, journal = {Parasites & vectors}, volume = {13}, number = {1}, pages = {579}, pmid = {33198811}, issn = {1756-3305}, support = {RSA6280102//Research Career Development Grant from Thailand Science Research and Innovation/ ; R016233004//Faculty of Medicine Siriraj Hospital, Mahidol University (TH)/ ; }, mesh = {Animals ; Anopheles/*microbiology ; Female ; Malaria/prevention & control ; Mosquito Vectors/*microbiology ; Myanmar/epidemiology ; Phylogeny ; Thailand/epidemiology ; *Wolbachia/classification/isolation & purification ; }, abstract = {BACKGROUND: Wolbachia, obligate intracellular bacteria, infect the majority of arthropods, including many mosquito species of medical importance. Some Wolbachia strains interfere with the development of Plasmodium parasites in female Anopheles, a major vector of malaria. The use of Wolbachia as a means to block malaria transmission is an emerging vector control strategy in highly endemic areas. Hence, identification of native Wolbachia strains in areas where malaria transmission is low may uncover a particular Wolbachia strain capable of Plasmodium interference. This study aims to identify native Wolbachia strains in female Anopheles spp. that are predominant in a low-malaria transmission area in mainland Southeast Asia.

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

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

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

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

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

CONCLUSIONS: Different Wolbachia numbers are detected in natural populations of C. pipiens pallens collected from Nanjing and Wuxi of Jiangsu Province and Tangkou of Shandong Province, and WD0513 gene may be involved in the Wolbachia-induced cytoplasmic incompatibility among three natural populations of C. pipiens pallens.}, } @article {pmid33180772, year = {2020}, author = {Arias-Giraldo, LM and Muñoz, M and Hernández, C and Herrera, G and Velásquez-Ortiz, N and Cantillo-Barraza, O and Urbano, P and Ramírez, JD}, title = {Species-dependent variation of the gut bacterial communities across Trypanosoma cruzi insect vectors.}, journal = {PloS one}, volume = {15}, number = {11}, pages = {e0240916}, pmid = {33180772}, issn = {1932-6203}, mesh = {Animals ; Biodiversity ; Chagas Disease/transmission ; Colombia ; DNA, Bacterial/genetics ; DNA, Protozoan/genetics ; *Gastrointestinal Microbiome/genetics ; Genotype ; Host Specificity ; Humans ; Insect Vectors/classification/*microbiology/*parasitology ; RNA, Ribosomal, 16S/genetics ; Triatominae/classification/*microbiology/*parasitology ; Trypanosoma cruzi/classification/genetics/*pathogenicity ; }, abstract = {Triatomines (Hemiptera: Reduviidae) are the insect vectors of Trypanosoma cruzi, the causative agent of Chagas disease. The gut bacterial communities affect the development of T. cruzi inside the vector, making the characterization of its composition important in the understanding of infection development. We collected 54 triatomine bugs corresponding to four genera in different departments of Colombia. DNA extraction and PCR were performed to evaluate T. cruzi presence and to determine the discrete typing unit (DTU) of the parasite. PCR products of the bacterial 16S rRNA gene were pooled and sequenced. Resulting reads were denoised and QIIME 2 was used for the identification of amplicon sequence variants (ASVs). Diversity (alpha and beta diversity) and richness analyses, Circos plots, and principal component analysis (PCA) were also performed. The overall T. cruzi infection frequency was 75.9%, with TcI being the predominant DTU. Approximately 500,000 sequences were analyzed and 27 bacterial phyla were identified. The most abundant phyla were Proteobacteria (33.9%), Actinobacteria (32.4%), Firmicutes (19.6%), and Bacteroidetes (7.6%), which together accounted for over 90% of the gut communities identified in this study. Genera were identified for these main bacterial phyla, revealing the presence of important bacteria such as Rhodococcus, Serratia, and Wolbachia. The composition of bacterial phyla in the gut of the insects was significantly different between triatomine species, whereas no significant difference was seen between the state of T. cruzi infection. We suggest further investigation with the evaluation of additional variables and a larger sample size. To our knowledge, this study is the first characterization of the gut bacterial structure of the main triatomine genera in Colombia.}, } @article {pmid33170730, year = {2021}, author = {Onah, IE and Sumner, S}, title = {DNA barcodes and new primers for nature's pest controllers: the social wasps.}, journal = {Genome}, volume = {64}, number = {5}, pages = {581-590}, doi = {10.1139/gen-2019-0193}, pmid = {33170730}, issn = {1480-3321}, mesh = {Animals ; Biodiversity ; DNA/analysis ; DNA Barcoding, Taxonomic/*methods ; *DNA Primers ; Insecta/genetics ; *Pest Control ; Phylogeny ; Polymerase Chain Reaction ; Wasps/classification/*genetics ; }, abstract = {Globally, biodiversity is declining because of anthropogenic pressures, and this could lead to extinction of some species before they are discovered. The loss of insect taxa is of prime concern, given recent reports of significant declines in the populations of many taxa across the globe. Efforts to document biodiversity have met with several challenges, amongst which are the difficulties in using morphological features to discriminate species, especially in insects. DNA barcoding is a rapid and reliable method for species identification and discovery but choosing appropriate primers to amplify the barcode region without co-amplifying contaminants remains a key challenge. We developed and tested a set of primers for PCR amplification of the DNA barcode region of the COI gene in polistine wasps. We tested their efficacy in 36 species of vespid wasps, and the solitary wasp Zethus miniatus Saussure. Samples were obtained from Africa, Americas, Asia, and Europe. The polistine-specific primers successfully amplified the barcode region for all polistines tested, without amplifying any Wolbachia present; they also worked with many species from the other Vespidae wasp subfamilies. The new primers are valuable for the discovery and accurate documentation of polistine wasps in the four continents.}, } @article {pmid33168082, year = {2020}, author = {Torres, R and Hernandez, E and Flores, V and Ramirez, JL and Joyce, AL}, title = {Wolbachia in mosquitoes from the Central Valley of California, USA.}, journal = {Parasites & vectors}, volume = {13}, number = {1}, pages = {558}, pmid = {33168082}, issn = {1756-3305}, support = {U01CK000516/ACL/ACL HHS/United States ; }, mesh = {Aedes/microbiology ; Animals ; Anopheles/microbiology ; Bacterial Typing Techniques ; California ; Culex/microbiology ; Culicidae/*microbiology ; DNA, Bacterial/genetics ; Female ; Mosquito Vectors/microbiology ; Multilocus Sequence Typing ; *Phylogeny ; RNA, Ribosomal, 16S ; Wolbachia/*classification/isolation & purification ; }, abstract = {BACKGROUND: Wolbachia bacteria are widely distributed throughout terrestrial arthropod species. These bacteria can manipulate reproduction and influence the vector competence of their hosts. Recently, Wolbachia have been integrated into vector control programmes for mosquito management. A number of supergroups and strains exist for Wolbachia, and they have yet to be characterized for many mosquito species. In this study, we examined Wolbachia prevalence and their phylogenetic relationship to other Wolbachia, using mosquitoes collected in Merced County in the Central Valley of California.

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

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

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

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

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

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

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

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

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

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

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

CONCLUSIONS: Our data showed that the apparently healthy buffalo were naturally infected by bacteria in the family Anaplasmataceae at a relatively high prevalence. We also report the finding of A. platys-like infections in water buffalo in Thailand for the first time. Water buffalo serving as the reservoir host of anaplasmosis is of concern for managing the disease control and prevention in ruminants.}, } @article {pmid33027888, year = {2020}, author = {Bakovic, V and Schebeck, M and Stauffer, C and Schuler, H}, title = {Wolbachia-Mitochondrial DNA Associations in Transitional Populations of Rhagoletis cerasi.}, journal = {Insects}, volume = {11}, number = {10}, pages = {}, pmid = {33027888}, issn = {2075-4450}, support = {I 2604/FWF_/Austrian Science Fund FWF/Austria ; J 3527/FWF_/Austrian Science Fund FWF/Austria ; P 31441/FWF_/Austrian Science Fund FWF/Austria ; }, abstract = {The endosymbiont Wolbachia can manipulate arthropod host reproduction by inducing cytoplasmic incompatibility (CI), which results in embryonic mortality when infected males mate with uninfected females. A CI-driven invasion of Wolbachia can result in a selective sweep of associated mitochondrial haplotype. The co-inheritance of Wolbachia and host mitochondrial DNA can therefore provide significant information on the dynamics of an ongoing Wolbachia invasion. Therefore, transition zones (i.e., regions where a Wolbachia strain is currently spreading from infected to uninfected populations) represent an ideal area to investigate the relationship between Wolbachia and host mitochondrial haplotype. Here, we studied Wolbachia-mitochondrial haplotype associations in the European cherry fruit fly, Rhagoletis cerasi, in two transition zones in the Czech Republic and Hungary, where the CI-inducing strain wCer2 is currently spreading. The wCer2-infection status of 881 individuals was compared with the two known R. cerasi mitochondrial haplotypes, HT1 and HT2. In accordance with previous studies, wCer2-uninfected individuals were associated with HT1, and wCer2-infected individuals were mainly associated with HT2. We found misassociations only within the transition zones, where HT2 flies were wCer2-uninfected, suggesting the occurrence of imperfect maternal transmission. We did not find any HT1 flies that were wCer2-infected, suggesting that Wolbachia was not acquired horizontally. Our study provides new insights into the dynamics of the early phase of a Wolbachia invasion.}, } @article {pmid33024036, year = {2020}, author = {Hague, MTJ and Caldwell, CN and Cooper, BS}, title = {Pervasive Effects of Wolbachia on Host Temperature Preference.}, journal = {mBio}, volume = {11}, number = {5}, pages = {}, pmid = {33024036}, issn = {2150-7511}, support = {R35 GM124701/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Behavior, Animal ; *Body Temperature ; Body Temperature Regulation ; Cytoplasm/microbiology ; Drosophila melanogaster/microbiology/*physiology ; Female ; Genotype ; Host Microbial Interactions/*physiology ; Male ; Phenotype ; Phylogeny ; Symbiosis ; Wolbachia/*physiology ; }, abstract = {Heritable symbionts can modify a range of ecologically important host traits, including behavior. About half of all insect species are infected with maternally transmitted Wolbachia, a bacterial endosymbiont known to alter host reproduction, nutrient acquisition, and virus susceptibility. Here, we broadly test the hypothesis that Wolbachia modifies host behavior by assessing the effects of eight different Wolbachia strains on the temperature preference of six Drosophila melanogaster subgroup species. Four of the seven host genotypes infected with A-group Wolbachia strains (wRi in Drosophila simulans, wHa in D. simulans, wSh in Drosophila sechellia, and wTei in Drosophila teissieri) prefer significantly cooler temperatures relative to uninfected genotypes. Contrastingly, when infected with divergent B-group wMau, Drosophila mauritiana prefers a warmer temperature. For most strains, changes to host temperature preference do not alter Wolbachia titer. However, males infected with wSh and wTei tend to experience an increase in titer when shifted to a cooler temperature for 24 h, suggesting that Wolbachia-induced changes to host behavior may promote bacterial replication. Our results indicate that Wolbachia modifications to host temperature preference are likely widespread, which has important implications for insect thermoregulation and physiology. Understanding the fitness consequences of these Wolbachia effects is crucial for predicting evolutionary outcomes of host-symbiont interactions, including how Wolbachia spreads to become common.IMPORTANCE Microbes infect a diversity of species, influencing the performance and fitness of their hosts. Maternally transmitted Wolbachia bacteria infect most insects and other arthropods, making these bacteria some of the most common endosymbionts in nature. Despite their global prevalence, it remains mostly unknown how Wolbachia influence host physiology and behavior to proliferate. We demonstrate pervasive effects of Wolbachia on Drosophila temperature preference. Most hosts infected with A-group Wolbachia prefer cooler temperatures, whereas the one host species infected with divergent B-group Wolbachia prefers warmer temperatures, relative to uninfected genotypes. Changes to host temperature preference generally do not alter Wolbachia abundance in host tissues, but for some A-group strains, adult males have increased Wolbachia titer when shifted to a cooler temperature. This suggests that Wolbachia-induced changes to host behavior may promote bacterial replication. Our results help elucidate the impact of endosymbionts on their hosts amid the global Wolbachia pandemic.}, } @article {pmid33023928, year = {2020}, author = {Lemon, A and Bordenstein, SR and Bordenstein, SR}, title = {Discover the Microbes Within! The Wolbachia Project: Citizen Science and Student-Based Discoveries for 15 Years and Counting.}, journal = {Genetics}, volume = {216}, number = {2}, pages = {263-268}, pmid = {33023928}, issn = {1943-2631}, mesh = {Animals ; Awards and Prizes ; Citizen Science/*methods/organization & administration ; Genetic Techniques ; Genetics/*education/organization & administration ; Humans ; *Microbiota ; Wolbachia/*genetics/pathogenicity ; }, abstract = {The Elizabeth W. Jones Award for Excellence in Education recognizes an individual who has had a significant impact on genetics education at any education level. Seth R. Bordenstein, Ph.D., Centennial Professor of Biological Sciences at Vanderbilt University and Founding Director of the Vanderbilt Microbiome Initiative, is the 2020 recipient in recognition of his cofounding, developing, and expanding Discover the Microbes Within! The Wolbachia Project.}, } @article {pmid33022249, year = {2020}, author = {Hoffmann, A}, title = {Wolbachia.}, journal = {Current biology : CB}, volume = {30}, number = {19}, pages = {R1113-R1114}, doi = {10.1016/j.cub.2020.08.039}, pmid = {33022249}, issn = {1879-0445}, mesh = {Animals ; *Bacterial Load ; *Host-Pathogen Interactions ; Invertebrates/*parasitology ; *Reproduction ; Wolbachia/*physiology ; }, abstract = {Ary Hoffman introduces Wolbachia.}, } @article {pmid33016318, year = {2020}, author = {Moussa, A and Passera, A and Sanna, F and Faccincani, M and Casati, P and Bianco, PA and Mori, N and Quaglino, F}, title = {Bacterial microbiota associated with insect vectors of grapevine Bois noir disease in relation to phytoplasma infection.}, journal = {FEMS microbiology ecology}, volume = {96}, number = {11}, pages = {}, doi = {10.1093/femsec/fiaa203}, pmid = {33016318}, issn = {1574-6941}, mesh = {Insect Vectors ; *Microbiota ; Phylogeny ; Phytoplasma Disease ; Plant Diseases ; *Vitis ; }, abstract = {Bois noir is a grapevine disease causing severe yield loss in vineyards worldwide. It is associated with 'Candidatus Phytoplasma solani', a phloem-limited bacterium transmitted by polyphagous insects. Due to its complex epidemiology, it is difficult to organize effective containment measures. This study aimed to describe the bacterial microbiota associated with 'Candidatus Phytoplasma solani' infected and non-infected insect hosts and vectors to investigate if phytoplasma presence can shape the microbiota. Alpha-diversity analysis showed a low microbiota diversity in these insects, in which few genera were highly abundant. Beta-diversity analysis revealed that the xylem- and phloem-feeding behavior influences the microbiota structure. Moreover, it highlighted that phytoplasma infection is associated with a restructuring of microbiota exclusively in Deltocephalinae insect vectors. Obtained data showed that 'Candidatus Phytoplasma solani' may have adverse effects on the endosymbionts Sulcia and Wolbachia, suggesting a possible fitness modification in the insects. The phytoplasma-antagonistic Dyella was not found in any of the examined insect species. The results indicate an interesting perspective regarding the microbial signatures associated with xylem- and phloem-feeding insects, and determinants that could be relevant to establish whether an insect species can be a vector or not, opening up new avenues for developing microbial resource management-based approaches.}, } @article {pmid33004433, year = {2020}, author = {Mathers, TC and Mugford, ST and Hogenhout, SA and Tripathi, L}, title = {Genome Sequence of the Banana Aphid, Pentalonia nigronervosa Coquerel (Hemiptera: Aphididae) and Its Symbionts.}, journal = {G3 (Bethesda, Md.)}, volume = {10}, number = {12}, pages = {4315-4321}, pmid = {33004433}, issn = {2160-1836}, support = {BB/R01227X/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; BB/P012574/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; }, mesh = {Animals ; *Aphids/genetics ; *Babuvirus ; *Buchnera ; *Musa ; *Wolbachia ; }, abstract = {The banana aphid, Pentalonia nigronervosa Coquerel (Hemiptera: Aphididae), is a major pest of cultivated bananas (Musa spp., order Zingiberales), primarily due to its role as a vector of Banana bunchy top virus (BBTV), the most severe viral disease of banana worldwide. Here, we generated a highly complete genome assembly of P. nigronervosa using a single PCR-free Illumina sequencing library. Using the same sequence data, we also generated complete genome assemblies of the P. nigronervosa symbiotic bacteria Buchnera aphidicola and Wolbachia To improve our initial assembly of P. nigronervosa we developed a k-mer based deduplication pipeline to remove genomic scaffolds derived from the assembly of haplotigs (allelic variants assembled as separate scaffolds). To demonstrate the usefulness of this pipeline, we applied it to the recently generated assembly of the aphid Myzus cerasi, reducing the duplication of conserved BUSCO genes by 25%. Phylogenomic analysis of P. nigronervosa, our improved M. cerasi assembly, and seven previously published aphid genomes, spanning three aphid tribes and two subfamilies, reveals that P. nigronervosa falls within the tribe Macrosiphini, but is an outgroup to other Macrosiphini sequenced so far. As such, the genomic resources reported here will be useful for understanding both the evolution of Macrosphini and for the study of P. nigronervosa. Furthermore, our approach using low cost, high-quality, Illumina short-reads to generate complete genome assemblies of understudied aphid species will help to fill in genomic black spots in the diverse aphid tree of life.}, } @article {pmid33002324, year = {2021}, author = {Izraeli, Y and Lalzar, M and Netanel, N and Mozes-Daube, N and Steinberg, S and Chiel, E and Zchori-Fein, E}, title = {Wolbachia influence on the fitness of Anagyrus vladimiri (Hymenoptera: Encyrtidae), a bio-control agent of mealybugs.}, journal = {Pest management science}, volume = {77}, number = {2}, pages = {1023-1034}, doi = {10.1002/ps.6117}, pmid = {33002324}, issn = {1526-4998}, support = {20-02-0092//Chief Scientist of Agriculture, Israel/ ; }, mesh = {Animals ; Female ; *Hymenoptera ; Multilocus Sequence Typing ; Reproduction ; Symbiosis ; *Wasps ; *Wolbachia/genetics ; }, abstract = {BACKGROUND: Like numerous other animals, biocontrol agents (BCAs) of arthropod pests carry various microorganisms that may have diverse effects on the biology of their eukaryote hosts. We postulated that it is possible to improve the efficacy of BCAs by manipulating the composition of their associated microbiota. The parasitoid wasp Anagyrus vladimiri (Hymenoptera: Encyrtidae) from a mass-rearing facility was chosen for testing this hypothesis.

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

CONCLUSIONS: The findings suggest that W[+] A. vladimiri have no clear fitness benefit under mass-rearing conditions and may be disadvantageous under lab-controlled conditions. In a broader view, the results suggest that augmentative biological control can benefit from manipulation of the microbiome of natural enemies.}, } @article {pmid32975515, year = {2020}, author = {Shropshire, JD and Leigh, B and Bordenstein, SR}, title = {Symbiont-mediated cytoplasmic incompatibility: what have we learned in 50 years?.}, journal = {eLife}, volume = {9}, number = {}, pages = {}, pmid = {32975515}, issn = {2050-084X}, support = {R01 AI143725/AI/NIAID NIH HHS/United States ; R01 AI132581/AI/NIAID NIH HHS/United States ; F32 AI140694/AI/NIAID NIH HHS/United States ; IOS1456778//National Science Foundation/International ; DGE144519//National Science Foundation/International ; }, mesh = {Animals ; Arthropods/genetics/*physiology ; *Cytoplasm/genetics/metabolism/microbiology/physiology ; Female ; Male ; *Reproduction/genetics/physiology ; Species Specificity ; Symbiosis/*physiology ; Ubiquitins/genetics/metabolism ; }, abstract = {Cytoplasmic incompatibility (CI) is the most common symbiont-induced reproductive manipulation. Specifically, symbiont-induced sperm modifications cause catastrophic mitotic defects in the fertilized embryo and ensuing lethality in crosses between symbiotic males and either aposymbiotic females or females harboring a different symbiont strain. However, if the female carries the same symbiont strain, then embryos develop properly, thereby imparting a relative fitness benefit to symbiont-transmitting mothers. Thus, CI drives maternally-transmitted bacteria to high frequencies in arthropods worldwide. In the past two decades, CI experienced a boom in interest due to its (i) deployment in worldwide efforts to curb mosquito-borne diseases, (ii) causation by bacteriophage genes, cifA and cifB, that modify sexual reproduction, and (iii) important impacts on arthropod speciation. This review serves as a gateway to experimental, conceptual, and quantitative themes of CI and outlines significant gaps in understanding CI's mechanism that are ripe for investigation from diverse subdisciplines in the life sciences.}, } @article {pmid32969551, year = {2020}, author = {Price, TAR and Windbichler, N and Unckless, RL and Sutter, A and Runge, JN and Ross, PA and Pomiankowski, A and Nuckolls, NL and Montchamp-Moreau, C and Mideo, N and Martin, OY and Manser, A and Legros, M and Larracuente, AM and Holman, L and Godwin, J and Gemmell, N and Courret, C and Buchman, A and Barrett, LG and Lindholm, AK}, title = {Resistance to natural and synthetic gene drive systems.}, journal = {Journal of evolutionary biology}, volume = {33}, number = {10}, pages = {1345-1360}, pmid = {32969551}, issn = {1420-9101}, support = {NIH R35GM119515/NH/NIH HHS/United States ; P300PA_177906/SNSF_/Swiss National Science Foundation/Switzerland ; P300PA_177830/SNSF_/Swiss National Science Foundation/Switzerland ; ZSEZ0_180460/SNSF_/Swiss National Science Foundation/Switzerland ; 31003A_160328//Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung/ ; R35 GM119515/GM/NIGMS NIH HHS/United States ; }, mesh = {*Biological Evolution ; *Gene Drive Technology ; *Selection, Genetic ; }, abstract = {Scientists are rapidly developing synthetic gene drive elements intended for release into natural populations. These are intended to control or eradicate disease vectors and pests, or to spread useful traits through wild populations for disease control or conservation purposes. However, a crucial problem for gene drives is the evolution of resistance against them, preventing their spread. Understanding the mechanisms by which populations might evolve resistance is essential for engineering effective gene drive systems. This review summarizes our current knowledge of drive resistance in both natural and synthetic gene drives. We explore how insights from naturally occurring and synthetic drive systems can be integrated to improve the design of gene drives, better predict the outcome of releases and understand genomic conflict in general.}, } @article {pmid32966874, year = {2020}, author = {Guo, Y and Khan, J and Zheng, XY and Wu, Y}, title = {Wolbachia increase germ cell mitosis to enhance the fecundity of Laodelphax striatellus.}, journal = {Insect biochemistry and molecular biology}, volume = {127}, number = {}, pages = {103471}, doi = {10.1016/j.ibmb.2020.103471}, pmid = {32966874}, issn = {1879-0240}, mesh = {Animals ; Female ; Fertility ; Hemiptera/microbiology/*physiology ; *Mitosis ; Ovum/*physiology ; Wolbachia/*physiology ; }, abstract = {Wolbachia are intracellular bacteria that infect a wide range of invertebrates and have evolved various strategies to alter host reproduction for their own survival and dissemination. In small brown planthopper Laodelphax striatellus, Wolbachia-infected females lay more eggs than uninfected females. Our previous study has shown that Wolbachia are abundant in ovarian cells of L. striatellus and change the number of apoptotic nurse cells in a caspase-dependent manner to provide nutrition for oogenesis. The cellular and molecular bases of the Wolbachia-mediated alterations in L. striatellus oogenesis remain largely unknown. Here, we investigated whether germ cell mitosis, which has been implicated in determination of egg production rates, influences the interaction between fecundity and Wolbachia in L.striatellus. We used an anti-phospho-histone 3 (pH3) antibody to label and visualize mitotic cells. Microscopic observations indicated that the Wolbachia strain wStri increased the number of ovarioles that contained mitotic germ cells. The increased fecundity of Wolbachia-infected females was a result of mitosis of germ cells; the frequency of germ cell mitosis was much higher in infected females than in uninfected females. In addition, mitosis inhibition by Cdc20, CDK1, and CycB messenger RNA interference in Wolbachia-infected L. striatellus markedly decreased egg numbers. Live Wolbachia recolonization enhanced the egg production of uninfected L. striatellus by directly affecting mitosis regulators. Together, these data suggest that wStri might increase germ cell mitosis to enhance the fecundity of L. striatellus in a mitosis-regulating manner. Our findings establish a link between Wolbachia-induced mitosis and Wolbachia-mediated egg production effects.}, } @article {pmid32948809, year = {2020}, author = {Koh, C and Islam, MN and Ye, YH and Chotiwan, N and Graham, B and Belisle, JT and Kouremenos, KA and Dayalan, S and Tull, DL and Klatt, S and Perera, R and McGraw, EA}, title = {Dengue virus dominates lipid metabolism modulations in Wolbachia-coinfected Aedes aegypti.}, journal = {Communications biology}, volume = {3}, number = {1}, pages = {518}, pmid = {32948809}, issn = {2399-3642}, support = {R01 AI151166/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/*metabolism/microbiology/pathogenicity/virology ; Animals ; Dengue/genetics/metabolism/microbiology/virology ; Dengue Virus/*genetics/metabolism/pathogenicity ; Humans ; Insect Vectors/genetics/microbiology/virology ; Lipid Metabolism/*genetics ; Pest Control, Biological ; Virus Replication/genetics ; Wolbachia/*genetics/metabolism/pathogenicity ; }, abstract = {Competition between viruses and Wolbachia for host lipids is a proposed mechanism of Wolbachia-mediated virus blocking in insects. Yet, the metabolomic interaction between virus and symbiont within the mosquito has not been clearly defined. We compare the lipid profiles of Aedes aegypti mosquitoes bearing mono- or dual-infections of the Wolbachia wMel strain and dengue virus serotype 3 (DENV3). We found metabolic signatures of infection-induced intracellular events but little evidence to support direct competition between Wolbachia and virus for host lipids. Lipid profiles of dual-infected mosquitoes resemble those of DENV3 mono-infected mosquitoes, suggesting virus-driven modulation dominates over that of Wolbachia. Interestingly, knockdown of key metabolic enzymes suggests cardiolipins are host factors for DENV3 and Wolbachia replication. These findings define the Wolbachia-DENV3 metabolic interaction as indirectly antagonistic, rather than directly competitive, and reveal new research avenues with respect to mosquito × virus interactions at the molecular level.}, } @article {pmid32945029, year = {2021}, author = {Xia, X and Peng, CW and Cui, JR and Jin, PY and Yang, K and Hong, XY}, title = {Wolbachia affects reproduction in the spider mite Tetranychus truncatus (Acari: Tetranychidae) by regulating chorion protein S38-like and Rop.}, journal = {Insect molecular biology}, volume = {30}, number = {1}, pages = {18-29}, doi = {10.1111/imb.12669}, pmid = {32945029}, issn = {1365-2583}, mesh = {Animals ; Egg Proteins/metabolism ; Female ; Fertility ; Gene Expression ; *Host-Parasite Interactions ; Oviposition ; RNA Interference ; RNA-Seq ; Reproduction ; Symbiosis ; Tetranychidae/genetics/*microbiology/physiology ; Wolbachia/*physiology ; }, abstract = {Wolbachia-induced reproductive regulation in hosts has been used to control pest populations, but little is known about the molecular mechanism underlying Wolbachia regulation of host genes. Here, reproductive regulation by Wolbachia in the spider mite Tetranychus truncatus was studied at the molecular level. Infection with Wolbachia resulted in decreasing oviposition and cytoplasmic incompatibility in T. truncatus. Further RNA-seq revealed genes regulated by Wolbachia in T. truncatus. Real-time quantitative polymerase chain reaction (qPCR) showed that genes, including chorion protein S38-like and Rop were down-regulated by Wolbachia. RNA interference (RNAi) of chorion protein S38-like and Rop in Wolbachia-uninfected T. truncatus decreased oviposition, which was consistent with Wolbachia-induced oviposition decrease. Interestingly, suppressing Rop in Wolbachia-infected T. truncatus led to increased Wolbachia titres in eggs; however, this did not occur after RNAi of chorion protein S38-like. This is the first study to show that chorion protein S38-like and Rop facilitate Wolbachia-mediated changes in T. truncatus fertility. In addition, RNAi of Rop turned the body colour of Wolbachia-uninfected T. truncatus black, which indicates that the role of Rop is not limited to the reproductive regulation of T. truncatus.}, } @article {pmid32944607, year = {2020}, author = {Arham, AF and Amin, L and Mustapa, MAC and Mahadi, Z and Arham, AF and Yaacob, M and Ibrahim, M and Norizan, NS}, title = {Perceived benefits and risks: A survey data set towards Wolbachia-infected Aedes Mosquitoes in Klang Valley.}, journal = {Data in brief}, volume = {32}, number = {}, pages = {106262}, pmid = {32944607}, issn = {2352-3409}, abstract = {Perceived Benefits and Risks: A survey data set towards Wolbachia-infected Aedes Mosquitoes in the Klang Valley, Malaysia. Introduction: The paper presents data collected using measures of perceived benefits, perceived risks, trust in key players, attitude towards nature versus material, attitude towards technology, religiosity, and attitude towards the Wolbachia-infected Aedes mosquitoes (WiAM) technique. The validated questionnaires were used to randomly survey targeted stakeholders in the Klang Valley, Malaysia, who had been asked to voluntarily participate in face-to-face interviews. Completed questionnaires were received from 399 respondents (adults above 18 years old) and comprised two stakeholder groups: scientists (n = 202), and the public (n = 197). The detailed findings serve numerous opportunities to examine the social acceptance of Wolbachia-infected Aedes mosquitoes, to ensure the development of policy and action plans, and to encourage further study by other researchers interested in the measures and data presented.}, } @article {pmid32937243, year = {2020}, author = {Shin, PT and Baptista, RP and O'Neill, CM and Wallis, C and Reaves, BJ and Wolstenholme, AJ}, title = {Comparative sequences of the Wolbachia genomes of drug-sensitive and resistant isolates of Dirofilaria immitis.}, journal = {Veterinary parasitology}, volume = {286}, number = {}, pages = {109225}, pmid = {32937243}, issn = {1873-2550}, support = {R01 AI103140/AI/NIAID NIH HHS/United States ; R21 AI125899/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Dirofilaria immitis/*drug effects/microbiology ; *Drug Resistance ; *Genetic Variation ; *Genome, Bacterial ; Lactones/*pharmacology ; Macrocyclic Compounds/pharmacology ; Wolbachia/*genetics ; }, abstract = {The recent identification of isolates of D. immitis with confirmed resistance to the macrocyclic lactone preventatives presents an opportunity for comparative genomic studies using these isolates, and examining the genetic diversity within and between them. We studied the genomes of Wolbachia endosymbionts of five isolates of D. immitis maintained at the University of Georgia. Missouri and Georgia-2 are maintained as drug susceptible isolates, and JYD-27, Yazoo-2013 and Metairie-2014 are resistant to the macrocyclic lactone preventatives. We used whole genome amplification followed by Illumina-based sequencing from 8 to 12 individual microfilariae from each of the five isolates, obtaining a depth of coverage of approximately 40-75 fold for each. The Illumina sequences were used to create new genome assemblies for all the Wolbachia isolates studied. Comparisons of the Wolbachia sequences revealed more than 3000 sequence variations in each isolate. We identified 67 loci specific in resistant isolates but not in susceptible isolates, including 18 genes affected.Phylogenetic analysis suggested that the endosymbionts of the drug-susceptible isolates are more closely related to each other than to those from any of the resistant parasites. This level of variation in the Wolbachia endosymbionts of D. immitis isolates suggests a potential for selection for resistance against drugs targeting them.}, } @article {pmid32932887, year = {2020}, author = {Ge, C and Hu, J and Zhao, Z and Hoffmann, AA and Ma, S and Shen, L and Fang, J and Zhu, J and Yu, W and Jiang, W}, title = {Phylogeny and Density Dynamics of Wolbachia Infection of the Health Pest Paederus fuscipes Curtis (Coleoptera: Staphylinidae).}, journal = {Insects}, volume = {11}, number = {9}, pages = {}, pmid = {32932887}, issn = {2075-4450}, support = {31401997//National Natural Science Foundation of China/ ; 2019112//Shanghai Municipal Human Resources and Social Security Bureau/ ; }, abstract = {The maternally inherited obligate intracellular bacteria Wolbachia infects the reproductive tissues of a wide range of arthropods and affects host reproduction. Wolbachia is a credible biocontrol agent for reducing the impact of diseases associated with arthropod vectors. Paederus fuscipes is a small staphylinid beetle that causes dermatitis linearis and conjunctivitis in humans when they come into contact with skin. Wolbachia occur in this beetle, but their relatedness to other Wolbachia, their infection dynamics, and their potential host effects remain unknown. In this study, we report the phylogenetic position and density dynamics of Wolbachia in P. fuscipes. The phylogeny of Wolbachia based on an analysis of MLST genotyping showed that the bacteria from P. fuscipes belong to supergroup B. Quantitative PCR indicated that the infection density in adults was higher than in any other life stage (egg, larva or pupa), and that reproductive tissue in adults had the highest infection densities, with similar densities in the sexes. These findings provide a starting point for understanding the Wolbachia infection dynamics in P. fuscipes, and interactions with other components of the microbiota.}, } @article {pmid32927527, year = {2020}, author = {Kakumanu, ML and DeVries, ZC and Barbarin, AM and Santangelo, RG and Schal, C}, title = {Bed bugs shape the indoor microbial community composition of infested homes.}, journal = {The Science of the total environment}, volume = {743}, number = {}, pages = {140704}, doi = {10.1016/j.scitotenv.2020.140704}, pmid = {32927527}, issn = {1879-1026}, mesh = {Aged ; Animals ; *Bedbugs ; Hot Temperature ; Housing ; Humans ; *Microbiota ; Poverty ; }, abstract = {Indoor pests, and the allergens they produce, adversely affect human health. Surprisingly, however, their effects on indoor microbial communities have not been assessed. Bed bug (Cimex lectularius) infestations pose severe challenges in elderly and low-income housing. They void large amounts of liquid feces into the home environment, which might alter the indoor microbial community composition. In this study, using bed bug-infested and uninfested homes, we showed a strong impact of bed bug infestations on the indoor microbial diversity. Floor dust samples were collected from uninfested and bed bug-infested homes and their microbiomes were analyzed before and after heat interventions that eliminated bed bugs. The microbial communities of bed bug-infested homes were radically different from those of uninfested homes, and the bed bug endosymbiont Wolbachia was the major driver of this difference. After bed bugs were eliminated, the microbial community gradually shifted toward the community composition of uninfested homes, strongly implicating bed bugs in shaping the dust-associated environmental microbiome. Further studies are needed to understand the viability of these microbial communities and the potential risks that bed bug-associated microbes and their metabolites pose to human health.}, } @article {pmid32913052, year = {2020}, author = {Manokaran, G and Flores, HA and Dickson, CT and Narayana, VK and Kanojia, K and Dayalan, S and Tull, D and McConville, MJ and Mackenzie, JM and Simmons, CP}, title = {Modulation of acyl-carnitines, the broad mechanism behind Wolbachia-mediated inhibition of medically important flaviviruses in Aedes aegypti.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {117}, number = {39}, pages = {24475-24483}, pmid = {32913052}, issn = {1091-6490}, support = {/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Aedes/chemistry/metabolism/*microbiology/*virology ; Animals ; Carnitine/chemistry/*metabolism ; Female ; Mosquito Vectors/chemistry/metabolism/microbiology/virology ; Wolbachia/*physiology ; Zika Virus/*physiology ; }, abstract = {Wolbachia-infected mosquitoes are refractory to flavivirus infections, but the role of lipids in Wolbachia-mediated virus blocking remains to be elucidated. Here, we use liquid chromatography mass spectrometry to provide a comprehensive picture of the lipidome of Aedes aegypti (Aag2) cells infected with Wolbachia only, either dengue or Zika virus only, and Wolbachia-infected Aag2 cells superinfected with either dengue or Zika virus. This approach identifies a class of lipids, acyl-carnitines, as being down-regulated during Wolbachia infection. Furthermore, treatment with an acyl-carnitine inhibitor assigns a crucial role for acyl-carnitines in the replication of dengue and Zika viruses. In contrast, depletion of acyl-carnitines increases Wolbachia density while addition of commercially available acyl-carnitines impairs Wolbachia production. Finally, we show an increase in flavivirus infection of Wolbachia-infected cells with the addition of acyl-carnitines. This study uncovers a previously unknown role for acyl-carnitines in this tripartite interaction that suggests an important and broad mechanism that underpins Wolbachia-mediated pathogen blocking.}, } @article {pmid32905752, year = {2020}, author = {Caragata, EP and Dong, S and Dong, Y and Simões, ML and Tikhe, CV and Dimopoulos, G}, title = {Prospects and Pitfalls: Next-Generation Tools to Control Mosquito-Transmitted Disease.}, journal = {Annual review of microbiology}, volume = {74}, number = {}, pages = {455-475}, doi = {10.1146/annurev-micro-011320-025557}, pmid = {32905752}, issn = {1545-3251}, mesh = {Animals ; *Biological Control Agents ; *Communicable Disease Control ; Communicable Diseases/*parasitology/transmission/*virology ; Culicidae/*genetics/parasitology/physiology/virology ; Humans ; Infertility ; Malaria ; Mosquito Control/*methods ; Wolbachia/genetics ; }, abstract = {Mosquito-transmitted diseases, including malaria and dengue, are a major threat to human health around the globe, affecting millions each year. A diverse array of next-generation tools has been designed to eliminate mosquito populations or to replace them with mosquitoes that are less capable of transmitting key pathogens. Many of these new approaches have been built on recent advances in CRISPR/Cas9-based genome editing. These initiatives have driven the development of pathogen-resistant lines, new genetics-based sexing methods, and new methods of driving desirable genetic traits into mosquito populations. Many other emerging tools involve microorganisms, including two strategies involving Wolbachia that are achieving great success in the field. At the same time, other mosquito-associated bacteria, fungi, and even viruses represent untapped sources of new mosquitocidal or antipathogen compounds. Although there are still hurdles to be overcome, the prospect that such approaches will reduce the impact of these diseases is highly encouraging.}, } @article {pmid32904796, year = {2020}, author = {Ciuca, L and Vismarra, A and Lebon, W and Beugnet, F and Morchon, R and Rinaldi, L and Cringoli, G and Kramer, L and Genchi, M}, title = {New insights into the biology, diagnosis and immune response to Dirofilaria repens in the canine host.}, journal = {Veterinary parasitology: X}, volume = {4}, number = {}, pages = {100029}, pmid = {32904796}, issn = {2590-1389}, abstract = {Dogs are the primary host for Dirofilaria repens, therefore it is mandatory to accurately diagnose the canine infection and to expand our current knowledge on parasite biology and the immune response of the infected host for a better prevention.Thus, the aim of the present study was to provide new insights from experimental infections of dogs with D. repens, focusing on the evaluation of: 1) the pre-patent period and 2) the antibody response against D. repens somatic antigens and against the Wolbachia endosymbiont. Briefly, on Day 0, twenty purpose-bred Beagle dogs were experimentally infected with 50 infective larvae (L3) of D. repens. Starting from Day 58 until the last day of the study (Day 281), blood samples were collected on a monthly basis for detection of antibodies against D. repens (Dr) and recombinant Wolbachia surface protein (rWSP) by non-commercial IgG-ELISAs. Additional samples were collected on Days 220, 245 and 281 for the detection of microfilariae (mff) using the modified Knott's test and biomolecular analysis, following two PCR protocols: Gioia et al. (2010; protocol A) and Rishniw et al. (2006- protocol B). The results were analysed by univariate statistical analyses using 2 × 2 contingency tables and K Cohen was calculated to assess the agreement among all the diagnostic techniques. Overall, the outcome of the study revealed that out of the 20 dogs experimentally infected with D. repens, 16 (80 %) were microfilaraemic, 17 (85 %) were positive at DNA detection in the blood, 18 (90 %) had D. repens antibodies and 16 (80 %) had Wolbachia antibodies on the last day of the study. The overall k agreement between Knott's and PCR protocol B was 0.442 (P = 0.0001) and increased throughout the study, reaching 0.828 (P = 0.0001) on Day 281. To the authors knowledge, this is only the second study reporting antibody response to D. repens somatic antigen in experimentally infected dogs. ELISA results showed that an antibody response develops before the onset of patency, and steadily increases with time. Results would suggest that the development of an immunological response to infection could lead to application in epidemiological studies, risk assessment and as an aid in the diagnostic approach in dogs, in particular for early infections without mff.}, } @article {pmid32888935, year = {2020}, author = {Lee, JM and Yek, SH and Wilson, RF and Rahman, S}, title = {Characterization of the Aedes albopictus (Diptera: Culicidae) holobiome: bacterial composition across land use type and mosquito sex in Malaysia.}, journal = {Acta tropica}, volume = {212}, number = {}, pages = {105683}, doi = {10.1016/j.actatropica.2020.105683}, pmid = {32888935}, issn = {1873-6254}, mesh = {Aedes/*microbiology ; Animals ; Bacteria/*isolation & purification ; Female ; Malaysia ; Male ; Microbial Interactions ; *Microbiota ; Wolbachia/isolation & purification ; }, abstract = {Understanding the diversity and dynamics of the microbiota within the mosquito holobiome is of great importance to apprehend how the microbiota modulates various complex processes and interactions. This study examined the bacterial composition of Aedes albopictus across land use type and mosquito sex in the state of Selangor, Malaysia using 16S rRNA sequencing. The bacterial community structure in mosquitoes was found to be influenced by land use type and mosquito sex, with the environment and mosquito diet respectively identified to be the most likely sources of microbes. We found that approximately 70% of the microbiota samples were dominated by Wolbachia and removing Wolbachia from analyses revealed the relatively even composition of the remaining bacterial microbiota. Furthermore, microbial interaction network analysis highlighted the prevalence of co-exclusionary patterns in all networks regardless of land use and mosquito sex, with Wolbachia exhibiting co-exclusionary interactions with other residential bacteria such as Xanthomonas, Xenophilus and Zymobacter.}, } @article {pmid32884660, year = {2020}, author = {Noh, P and Oh, SY and Park, S and Kwon, T and Kim, Y and Choe, JC and Jeong, G}, title = {Association between host wing morphology polymorphism and Wolbachia infection in Vollenhovia emeryi (Hymenoptera: Myrmicinae).}, journal = {Ecology and evolution}, volume = {10}, number = {16}, pages = {8827-8837}, pmid = {32884660}, issn = {2045-7758}, abstract = {Many eusocial insects, including ants, show complex colony structures, distributions, and reproductive strategies. In the ant Vollenhovia emeryi Wheeler (Hymenoptera: Myrmicinae), queens and males are produced clonally, while sterile workers arise sexually, unlike other ant species and Hymenopteran insects in general. Furthermore, there is a wing length polymorphism in the queen caste. Despite its evolutionary remarkable traits, little is known about the population structure of this ant species, which may provide insight into its unique reproductive mode and polymorphic traits. We performed in-depth analyses of ant populations from Korea, Japan, and North America using three mitochondrial genes (COI, COII, and Cytb). The long-winged (L) morph is predominant in Korean populations, and the short-winged (S) morph is very rare. Interestingly, all L morphs were infected with Wolbachia, while all Korean S morphs lacked Wolbachia, demonstrating a association between a symbiont and a phenotypic trait. A phylogenetic analysis revealed that the S morph is derived from the L morph. We propose that the S morph is associated with potential resistance to Wolbachia infection and that Wolbachia infection does not influence clonal reproduction (as is the case in other ant species).}, } @article {pmid32881871, year = {2020}, author = {Zhang, D and Xi, Z and Li, Y and Wang, X and Yamada, H and Qiu, J and Liang, Y and Zhang, M and Wu, Y and Zheng, X}, title = {Toward implementation of combined incompatible and sterile insect techniques for mosquito control: Optimized chilling conditions for handling Aedes albopictus male adults prior to release.}, journal = {PLoS neglected tropical diseases}, volume = {14}, number = {9}, pages = {e0008561}, pmid = {32881871}, issn = {1935-2735}, mesh = {Aedes/*microbiology/*physiology ; Animals ; Cold Temperature ; Female ; Male ; Mosquito Control/*methods ; Reproduction ; Wolbachia/*physiology ; }, abstract = {Combined incompatible and sterile insect technique (IIT-SIT) has been considered to be an effective and safe approach to control mosquito populations. Immobilization of male adults by chilling is a crucial process required for the packing, transportation and release of the mosquitoes during the implementation of IIT-SIT for mosquito control. In this study, effects of chilling on the Aedes albopictus males with triple Wolbachia infections (HC line), a powerful weapon to fight against the wild type Ae. albopictus population via IIT-SIT, were evaluated under both laboratory and field conditions. Irradiated HC (IHC) males were exposed to 1, 5 and 10°C for 1, 2, 3, 6 and 24 h. The survival rate of the post-chilled IHC males was then monitored. Longevity of post-chilled IHC males was compared to non-chilled males under laboratory and semi-field conditions. Mating competitiveness of IHC/HC males after exposure to 5 or 10°C for 0, 3 and 24 h was then evaluated. Effects of compaction and transportation under chilled conditions on the survival rate of IHC males were also monitored. The optimal chilling conditions for handling IHC males were temperatures between 5 and 10°C for a duration of less than 3 h with no negative impacts on survival rate, longevity and mating competitiveness when compared to non-chilled males. However, the overall quality of post-chilled IHC/HC males decreased when exposed to low temperatures for 24 h. Reduced survival was observed when IHC males were stored at 5°C under a compaction height of 8 cm. Transportation with chilling temperatures fluctuating from 8 to 12°C has no negative impact on the survival of IHC males. This study identified the optimal chilling temperature and duration for the handling and transportation of Ae. albopictus IHC male adults without any detrimental effect on their survival, longevity and mating competitiveness. Further studies are required to develop drone release systems specific for chilled mosquitoes to improve release efficiency, as well as to compare the population suppression efficiency between release of post-chilled and non-chilled males in the field.}, } @article {pmid32878094, year = {2020}, author = {Castillo, AM and Saltonstall, K and Arias, CF and Chavarria, KA and Ramírez-Camejo, LA and Mejía, LC and De León, LF}, title = {The Microbiome of Neotropical Water Striders and Its Potential Role in Codiversification.}, journal = {Insects}, volume = {11}, number = {9}, pages = {}, pmid = {32878094}, issn = {2075-4450}, support = {No. 270-2013-284//Secretaría Nacional de Ciencia, Tecnología e Innovación/ ; No. FID16-116//Secretaría Nacional de Ciencia, Tecnología e Innovación/ ; 152-2018//Sistema Nacional de Investigación, SNI/ ; }, abstract = {Insects host a highly diverse microbiome, which plays a crucial role in insect life. However, the composition and diversity of microbiomes associated with Neotropical freshwater insects is virtually unknown. In addition, the extent to which diversification of this microbiome is associated with host phylogenetic divergence remains to be determined. Here, we present the first comprehensive analysis of bacterial communities associated with six closely related species of Neotropical water striders in Panama. We used comparative phylogenetic analyses to assess associations between dominant bacterial linages and phylogenetic divergence among species of water striders. We found a total of 806 16S rRNA amplicon sequence variants (ASVs), with dominant bacterial taxa belonging to the phyla Proteobacteria (76.87%) and Tenericutes (19.51%). Members of the α- (e.g., Wolbachia) and γ- (e.g., Acinetobacter, Serratia) Proteobacteria, and Mollicutes (e.g., Spiroplasma) were predominantly shared across species, suggesting the presence of a core microbiome in water striders. However, some bacterial lineages (e.g., Fructobacillus, Fluviicola and Chryseobacterium) were uniquely associated with different water strider species, likely representing a distinctive feature of each species' microbiome. These findings indicate that both host identity and environmental context are important drivers of microbiome diversity in water striders. In addition, they suggest that diversification of the microbiome is associated with diversification in water striders. Although more research is needed to establish the evolutionary consequences of host-microbiome interaction in water striders, our findings support recent work highlighting the role of bacterial community host-microbiome codiversification.}, } @article {pmid32857984, year = {2021}, author = {Brattig, NW and Cheke, RA and Garms, R}, title = {Onchocerciasis (river blindness) - more than a century of research and control.}, journal = {Acta tropica}, volume = {218}, number = {}, pages = {105677}, doi = {10.1016/j.actatropica.2020.105677}, pmid = {32857984}, issn = {1873-6254}, mesh = {Animals ; Humans ; Insect Vectors/parasitology ; Onchocerciasis, Ocular/epidemiology/*prevention & control/transmission ; Prevalence ; }, abstract = {This review summarises more than a century of research on onchocerciasis, also known as river blindness, and its control. River blindness is an infection caused by the tissue filaria Onchocerca volvulus affecting the skin, subcutaneous tissue and eyes and leading to blindness in a minority of infected persons. The parasite is transmitted by its intermediate hosts Simulium spp. which breed in rivers. Featured are history and milestones in onchocerciasis research and control, state-of-the-art data on the parasite, its endobacteria Wolbachia, on the vectors, previous and current prevalence of the infection, its diagnostics, the interaction between the parasite and its host, immune responses and the pathology of onchocerciasis. Detailed information is documented on the time course of control programmes in the afflicted countries in Africa and the Americas, a long road from previous programmes to current successes in control of the transmission of this infectious disease. By development, adjustment and optimization of the control measures, transmission by the vector has been interrupted in foci of countries in the Americas, in Uganda, in Sudan and elsewhere, followed by onchocerciasis eliminations. The current state and future perspectives for control, elimination and eradication within the next 20-30 years are described and discussed. This review contributes to a deeper comprehension of this disease by a tissue-dwelling filaria and it will be helpful in efforts to control and eliminate other filarial infections.}, } @article {pmid32855244, year = {2020}, author = {Neupane, S and Bonilla, SI and Manalo, AM and Pelz-Stelinski, KS}, title = {Near-Complete Genome Sequences of a Wolbachia Strain Isolated from Diaphorina citri Kuwayama (Hemiptera: Liviidae).}, journal = {Microbiology resource announcements}, volume = {9}, number = {35}, pages = {}, pmid = {32855244}, issn = {2576-098X}, abstract = {Wolbachia strains are one of three endosymbionts associated with the insect vector of "Candidatus Liberibacter asiaticus," Diaphorina citri Kuwayama (Hemiptera: Liviidae). We report three near-complete genome sequences of samples of Wolbachia from D. citri (wDi), with sizes of 1,518,595, 1,542,468, and 1,538,523 bp.}, } @article {pmid32854447, year = {2020}, author = {Adegoke, A and Kumar, D and Bobo, C and Rashid, MI and Durrani, AZ and Sajid, MS and Karim, S}, title = {Tick-Borne Pathogens Shape the Native Microbiome Within Tick Vectors.}, journal = {Microorganisms}, volume = {8}, number = {9}, pages = {}, pmid = {32854447}, issn = {2076-2607}, support = {P20 GM103476/GM/NIGMS NIH HHS/United States ; }, abstract = {Ticks are blood-feeding arthropods and transmit a variety of medically important viral, bacterial, protozoan pathogens to animals and humans. Ticks also harbor a diverse community of microbes linked to their biological processes, such as hematophagy, and hence affect vector competence. The interactions between bacterial and/or protozoan pathogens and the tick microbiome is a black-box, and therefore we tested the hypothesis that the presence of a protozoan or bacterial pathogen will alter the microbial composition within a tick. Hence, this study was designed to define the microbial composition of two tick species, Hyalomma (H.) anatolicum and Rhipicephalus (R.) microplus. We used a combination of PCR based pathogen (Anaplasma marginale and Theileria species) and symbiont (Wolbachia species) identification followed by metagenomic sequencing and comparison of the microbial communities in PCR positive and negative ticks. A total of 1786 operational taxonomic units was identified representing 25 phyla, 50 classes, and 342 genera. The phylum Proteobacteria, Firmicutes, Actinobacteriota, and Bacteroidota were the most represented bacteria group. Alpha and beta diversity were not significantly affected in the presence or absence of Theileria sp. and A. marginale as see with H. anatolicum ticks. Interestingly, bacterial communities were significantly reduced in Theileria sp. infected R. microplus ticks, while also exhibiting a significant reduction in microbial richness and evenness. Putting these observations together, we referred to the effect the presence of Theileria sp. has on R. microplus a "pathogen-induced dysbiosis". We also identify the presence of Plasmodium falciparum, the causative agent of human malaria from the microbiome of both H. anatolicum and R. microplus ticks. These findings support the presence of a "pathogen-induced dysbiosis" within the tick and further validation experiments are required to investigate how they are important in the vector competence of ticks. Understanding the mechanism of "pathogen-induced dysbiosis" on tick microbial composition may aid the discovery of intervention strategies for the control of emerging tick-borne infections.}, } @article {pmid32854141, year = {2020}, author = {Altinli, M and Lequime, S and Atyame, C and Justy, F and Weill, M and Sicard, M}, title = {Wolbachia modulates prevalence and viral load of Culex pipiens densoviruses in natural populations.}, journal = {Molecular ecology}, volume = {29}, number = {20}, pages = {4000-4013}, doi = {10.1111/mec.15609}, pmid = {32854141}, issn = {1365-294X}, mesh = {Animals ; *Culex/genetics ; *Densovirus/genetics ; Phylogeny ; Prevalence ; Viral Load ; *Wolbachia/genetics ; }, abstract = {The inadequacy of standard mosquito control strategies calls for ecologically safe novel approaches, for example the use of biological agents such as the endosymbiotic α-proteobacteria Wolbachia or insect-specific viruses (ISVs). Understanding the ecological interactions between these "biocontrol endosymbionts" is thus a fundamental step. Wolbachia are transmitted vertically from mother to offspring and modify their hosts' phenotypes, including reproduction (e.g., cytoplasmic incompatibility) and survival (e.g., viral interference). In nature, Culex pipiens (sensu lato) mosquitoes are always found infected with genetically diverse Wolbachia called wPip that belong to five phylogenetic groups. In recent years, ISVs have also been discovered in these mosquito species, although their interactions with Wolbachia in nature are unknown. Here, we studied the interactions between a widely prevalent ISV, the Culex pipiens densovirus (CpDV, Densovirinae), and Wolbachia in northern Tunisian C. pipiens populations. We showed an influence of different Wolbachia groups on CpDV prevalence and a general positive correlation between Wolbachia and CpDV loads. By investigating the putative relationship between CpDV diversification and wPip groups in the different sites, we detected a signal linked to wPip groups in CpDV phylogeny in sites where all larvae were infected by the same wPip group. However, no such signal was detected where the wPip groups coexisted, suggesting CpDV horizontal transfer between hosts. Overall, our results provide good evidence for an ecological influence of Wolbachia on an ISV, CpDV, in natural populations and highlight the importance of integrating Wolbachia in our understanding of ISV ecology in nature.}, } @article {pmid32853197, year = {2020}, author = {Sim, S and Ng, LC and Lindsay, SW and Wilson, AL}, title = {A greener vision for vector control: The example of the Singapore dengue control programme.}, journal = {PLoS neglected tropical diseases}, volume = {14}, number = {8}, pages = {e0008428}, pmid = {32853197}, issn = {1935-2735}, support = {/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; MR/P027873/1/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Aedes/virology ; Animals ; Community Participation ; Dengue/epidemiology/*prevention & control ; Humans ; Mosquito Control/*methods/*organization & administration ; Mosquito Vectors/virology ; Singapore/epidemiology ; }, abstract = {Vector-borne diseases are a major cause of morbidity and mortality worldwide. Aedes-borne diseases, in particular, including dengue, chikungunya, yellow fever, and Zika, are increasing at an alarming rate due to urbanisation, population movement, weak vector control programmes, and climate change. The World Health Organization calls for strengthening of vector control programmes in line with the Global Vector Control Response (GVCR) strategy, and many vector control programmes are transitioning to this new approach. The Singapore dengue control programme, situated within the country's larger vision of a clean, green, and sustainable environment for the health and well-being of its citizens, provides an excellent example of the GVCR approach in action. Since establishing vector control operations in the 1960s, the Singapore dengue control programme succeeded in reducing the dengue force of infection 10-fold by the 1990s and has maintained it at low levels ever since. Key to this success is consideration of dengue as an environmental disease, with a strong focus on source reduction and other environmental management methods as the dominant vector control strategy. The programme collaborates closely with other government ministries, as well as town councils, communities, the private sector, and academic and research institutions. Community engagement programmes encourage source reduction, and house-to-house inspections accompanied by a strong legislative framework with monetary penalties help to support compliance. Strong vector and epidemiological surveillance means that routine control activities can be heightened to specifically target dengue clusters. Despite its success, the programme continues to innovate to tackle challenges such as climate change, low herd immunity, and manpower constraints. Initiatives include development of novel vector controls such as Wolbachia-infected males and spatiotemporal models for dengue risk assessment. Lessons learnt from the Singapore programme can be applied to other settings, even those less well-resourced than Singapore, for more effective vector control.}, } @article {pmid32852571, year = {2021}, author = {Hubert, J and Nesvorna, M and Green, SJ and Klimov, PB}, title = {Microbial Communities of Stored Product Mites: Variation by Species and Population.}, journal = {Microbial ecology}, volume = {81}, number = {2}, pages = {506-522}, pmid = {32852571}, issn = {1432-184X}, support = {GA19-09998S//Grantová Agentura České Republiky/ ; 19-14-00004//Russian Science Foundation/ ; }, mesh = {Acaridae/classification/growth & development/*microbiology ; Animals ; Bacteria/classification/genetics/isolation & purification ; Diet ; Feces/microbiology ; Host Microbial Interactions ; *Microbiota ; Ovum/microbiology ; Phylogeny ; }, abstract = {Arthropod-associated microorganisms are important because they affect host fitness, protect hosts from pathogens, and influence the host's ability to vector pathogens. Stored product mites (Astigmata) often establish large populations in various types of food items, damaging the food by direct feeding and introducing contaminants, including their own bodies, allergen-containing feces, and associated microorganisms. Here we access the microbial structure and abundance in rearing diets, eggs, feces fraction, and mite bodies of 16 mite populations belonging to three species (Carpoglyphus lactis, Acarus siro, and Tyrophagus putrescentiae) using quantitative PCR and 16S ribosomal RNA (rRNA) gene amplicon sequencing. The mite microbiomes had a complex structure dominated by the following bacterial taxa (OTUs): (a) intracellular symbionts of the genera Cardinium and Wolbachia in the mite bodies and eggs; (b) putative gut symbionts of the genera Solitalea, Bartonella, and Sodalis abundant in mite bodies and also present in mite feces; (c) feces-associated or environmental bacteria of the genera Bacillus, Staphylococcus, and Kocuria in the diet, mite bodies, and feces. Interestingly and counterintuitively, the differences between microbial communities in various conspecific mite populations were higher than those between different mite species. To explain some of these differences, we hypothesize that the intracellular bacterial symbionts can affect microbiome composition in mite bodies, causing differences between microbial profiles. Microbial profiles differed between various sample types, such as mite eggs, bodies, and the environment (spent growth medium-SPGM). Low bacterial abundances in eggs may result in stochastic effects in parent-offspring microbial transmission, except for the intracellular symbionts. Bacteria in the rearing diet had little effect on the microbial community structure in SPGM and mite bodies. Mite fitness was positively correlated with bacterial abundance in SPGM and negatively correlated with bacterial abundances in mite bodies. Our study demonstrates critical host-microbe interactions, affecting all stages of mite growth and leading to alteration of the environmental microbiome. Correlational evidence based on absolute quantitation of bacterial 16S rRNA gene copies suggests that mite-associated microorganisms are critical for modulating important pest properties of mites by altering population growth.}, } @article {pmid32851048, year = {2020}, author = {Esteban-Mendoza, MV and Arcila-Quiceno, V and Albarracín-Navas, J and Hernández, I and Flechas-Alarcón, MC and Morchón, R}, title = {Current Situation of the Presence of Dirofilaria immitis in Dogs and Humans in Bucaramanga, Colombia.}, journal = {Frontiers in veterinary science}, volume = {7}, number = {}, pages = {488}, pmid = {32851048}, issn = {2297-1769}, abstract = {The cardiopulmonary dirofilariosis caused by Dirofilaria immitis, is a vector-borne infection, which can be transmitted to humans. The main hosts are both domestic and wild canids. This species mainly occurs in tropical and subtropical climates, and temperature and humidity are the main factors that favor the presence and proliferation of culicid mosquitoes as vectors of the disease. There are few reports of cardiopulmonary dirofilariosis in dogs and humans in Colombia, a region with favorable climatic conditions which favors the presence of mosquitoes that act as vectors of the disease. Therefore, this study aimed to examine its current prevalence in dogs and the risk of human exposure to the disease in Bucaramanga, one of the most populated areas in Colombia located at the center of the country. Furthermore, its demographic and environmental characteristics could be useful as a study model for other similar locations and neighboring countries. Serum samples from 351 dogs and 506 humans from the Bucaramanga Metropolitan area were analyzed. All dog samples were analyzed by Knott's technique and tested with a commercial immunochromatographic to detect the presence of circulating antigens of D. immitis. Human samples were analyzed using a non-commercial ELISA test kit to detect IgG against the somatic antigens of adult D. immitis and Wolbachia. Positive results were further confirmed using western blot analysis. Thirty-eight dogs tested positive with a overall prevalence of 10.82%. Of these dogs, 18 showed D. immitis microfilariae, being 5.12% of the total population. The overall seroprevalence in humans was 6.71%; seroprevalence was significantly higher in individuals aged 16-34 years-old and in women than in men. To our knowledge, this study describes seropositivity to D. immitis for the first time in a Colombian human population located in the same area as that of dogs infected with D. immitis, which represents a potential threat to public health. In humans, age and gender can be considered risk factors for exposure to D. immitis.}, } @article {pmid32849379, year = {2020}, author = {Lu, P and Sun, Q and Fu, P and Li, K and Liang, X and Xi, Z}, title = {Wolbachia Inhibits Binding of Dengue and Zika Viruses to Mosquito Cells.}, journal = {Frontiers in microbiology}, volume = {11}, number = {}, pages = {1750}, pmid = {32849379}, issn = {1664-302X}, support = {R01 AI080597/AI/NIAID NIH HHS/United States ; }, abstract = {As traditional approaches to the control of dengue and Zika are insufficient, significant efforts have been made to develop utilization of the endosymbiotic bacterium Wolbachia to reduce the ability of mosquitoes to transmit pathogens. Although Wolbachia is known to inhibit flaviviruses in mosquitoes, including dengue virus (DENV) and Zika virus (ZIKV), it remains unclear how the endosymbiont interferes with viral replication cycle. In this study, we have carried out viral binding assays to investigate the impact of the Wolbachia strain wAlbB on the attachment of DENV serotype 2 (DENV-2) and ZIKV to Aedes aegypti Aag-2 cells. RNA interference (RNAi) was used to silence a variety of putative mosquito receptors of DENV that were differentially regulated by wAlbB in Aag-2 cells, in order to identify host factors involved in the inhibition of viral binding. Our results showed that, in addition to suppression of viral replication, Wolbachia strongly inhibited binding of both DENV-2 and ZIKV to Aag-2 cells. Moreover, the expression of two putative mosquito DENV receptors - dystroglycan and tubulin - was downregulated by wAlbB, and their knock-down resulted in the inhibition of DENV-2 binding to Aag-2 cells. These results will aid in understanding the Wolbachia-DENV interactions in mosquito and the development of novel control strategies for mosquito-borne diseases.}, } @article {pmid32831122, year = {2020}, author = {Wuliandari, JR and Hoffmann, AA and Tantowijoyo, W and Endersby-Harshman, NM}, title = {Frequency of kdr mutations in the voltage-sensitive sodium channel (VSSC) gene in Aedes aegypti from Yogyakarta and implications for Wolbachia-infected mosquito trials.}, journal = {Parasites & vectors}, volume = {13}, number = {1}, pages = {429}, pmid = {32831122}, issn = {1756-3305}, support = {1037003//National Health and Medical Research Council/ ; 1132412//National Health and Medical Research Council/ ; 1118640//NHMRC Fellowship/ ; }, mesh = {*Aedes/drug effects/genetics/microbiology ; Animals ; Biological Control Agents ; Dengue/transmission ; Genotype ; Genotyping Techniques ; Indonesia ; Insect Proteins/genetics ; Insecticide Resistance ; Insecticides/pharmacology ; *Mosquito Control ; Mosquito Vectors/drug effects/genetics/microbiology ; Mutation ; Mutation Rate ; Pest Control, Biological ; Pyrethrins/pharmacology ; Voltage-Gated Sodium Channels/*genetics ; Wolbachia/pathogenicity ; }, abstract = {BACKGROUND: In the inner city of Yogyakarta, Indonesia, insecticide resistance is expected in the main dengue vector, Aedes aegypti, because of the intensive local application of pyrethroid insecticides. However, detailed information about the nature of resistance in this species is required to assist the release of Wolbachia mosquitoes in a dengue control program, so that we can ensure that insecticide resistance in the strain of Ae. aegypti being released matches that of the background population.

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

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

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

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

CONCLUSIONS: We demonstrate that spatial genetics can provide a robust characterization of mosquito dispersal. With the decreasing cost of next-generation sequencing, the production of spatial genetic data is increasingly accessible. Given the challenges of conventional MRR methods, and the importance of quantified dispersal in operational vector control decisions, we recommend genetic-based dispersal characterization as the more desirable means of parameterization.}, } @article {pmid32813725, year = {2020}, author = {Shropshire, JD and Kalra, M and Bordenstein, SR}, title = {Evolution-guided mutagenesis of the cytoplasmic incompatibility proteins: Identifying CifA's complex functional repertoire and new essential regions in CifB.}, journal = {PLoS pathogens}, volume = {16}, number = {8}, pages = {e1008794}, pmid = {32813725}, issn = {1553-7374}, support = {R01 AI132581/AI/NIAID NIH HHS/United States ; R01 AI143725/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Animals, Genetically Modified/microbiology/physiology ; Bacterial Proteins/genetics/*metabolism ; *Biological Evolution ; Cytoplasm/*metabolism/microbiology ; Drosophila melanogaster/*microbiology/physiology ; Female ; Gram-Negative Bacterial Infections/*metabolism/microbiology/pathology ; Male ; *Mutation ; Wolbachia/*physiology ; }, abstract = {Wolbachia are the world's most common, maternally-inherited, arthropod endosymbionts. Their worldwide distribution is due, in part, to a selfish drive system termed cytoplasmic incompatibility (CI) that confers a relative fitness advantage to females that transmit Wolbachia to their offspring. CI results in embryonic death when infected males mate with uninfected females but not infected females. Under the Two-by-One genetic model of CI, males expressing the two phage WO proteins CifA and CifB cause CI, and females expressing CifA rescue CI. While each protein is predicted to harbor three functional domains, there is no knowledge on how sites across these Cif domains, rather than in any one particular domain, contribute to CI and rescue. Here, we use evolution-guided, substitution mutagenesis of conserved amino acids across the Cif proteins, coupled with transgenic expression in uninfected Drosophila melanogaster, to determine the functional impacts of conserved residues evolving mostly under purifying selection. We report that amino acids in CifA's N-terminal unannotated region and annotated catalase-related domain are important for both complete CI and rescue, whereas C-terminal residues in CifA's putative domain of unknown function are solely important for CI. Moreover, conserved CifB amino acids in the predicted nucleases, peptidase, and unannotated regions are essential for CI. Taken together, these findings indicate that (i) all CifA amino acids determined to be crucial in rescue are correspondingly crucial in CI, (ii) an additional set of CifA amino acids are uniquely important in CI, and (iii) CifB amino acids across the protein, rather than in one particular domain, are all crucial for CI. We discuss how these findings advance an expanded view of Cif protein evolution and function, inform the mechanistic and biochemical bases of Cif-induced CI/rescue, and continue to substantiate the Two-by-One genetic model of CI.}, } @article {pmid32813707, year = {2020}, author = {Seabourn, P and Spafford, H and Yoneishi, N and Medeiros, M}, title = {The Aedes albopictus (Diptera: Culicidae) microbiome varies spatially and with Ascogregarine infection.}, journal = {PLoS neglected tropical diseases}, volume = {14}, number = {8}, pages = {e0008615}, pmid = {32813707}, issn = {1935-2735}, support = {P20 GM125508/GM/NIGMS NIH HHS/United States ; }, mesh = {Aedes/*microbiology/*parasitology ; Animal Diseases ; Animals ; Apicomplexa ; Bacteria/*classification/genetics ; Host-Parasite Interactions ; Larva/microbiology/parasitology ; Microbiota/*physiology ; Protozoan Infections, Animal/*microbiology ; Symbiosis ; }, abstract = {The mosquito microbiome alters the physiological traits of medically important mosquitoes, which can scale to impact how mosquito populations sustain disease transmission. The mosquito microbiome varies significantly within individual mosquitoes and among populations, however the ecological and environmental factors that contribute to this variation are poorly understood. To further understand the factors that influence variation and diversity of the mosquito microbiome, we conducted a survey of the bacterial microbiome in the medically important mosquito, Aedes albopictus, on the high Pacific island of Maui, Hawai'i. We detected three bacterial Phyla and twelve bacterial families: Proteobacteria, Acitinobacteria, and Firmicutes; and Anaplasmataceae, Acetobacteraceae, Enterobacteriaceae, Burkholderiaceae, Xanthobacteraceae, Pseudomonadaceae, Streptomycetaceae, Staphylococcaceae, Xanthomonadaceae, Beijerinckiaceae, Rhizobiaceae, and Sphingomonadaceae. The Ae. albopictus bacterial microbiota varied among geographic locations, but temperature and rainfall were uncorrelated with this spatial variation. Infection status with an ampicomplexan pathosymbiont Ascogregarina taiwanensis was significantly associated with the composition of the Ae. albopictus bacteriome. The bacteriomes of mosquitoes with an A. taiwanensis infection were more likely to include several bacterial symbionts, including the most abundant lineage of Wolbachia sp. Other symbionts like Asaia sp. and several Enterobacteriaceae lineages were less prevalent in A. taiwanensis-infected mosquitoes. This highlights the possibility that inter- and intra-domain interactions may structure the Ae. albopictus microbiome.}, } @article {pmid32803130, year = {2020}, author = {Indriani, C and Tantowijoyo, W and Rancès, E and Andari, B and Prabowo, E and Yusdi, D and Ansari, MR and Wardana, DS and Supriyati, E and Nurhayati, I and Ernesia, I and Setyawan, S and Fitriana, I and Arguni, E and Amelia, Y and Ahmad, RA and Jewell, NP and Dufault, SM and Ryan, PA and Green, BR and McAdam, TF and O'Neill, SL and Tanamas, SK and Simmons, CP and Anders, KL and Utarini, A}, title = {Reduced dengue incidence following deployments of Wolbachia-infected Aedes aegypti in Yogyakarta, Indonesia: a quasi-experimental trial using controlled interrupted time series analysis.}, journal = {Gates open research}, volume = {4}, number = {}, pages = {50}, pmid = {32803130}, issn = {2572-4754}, support = {R56 AI134724/AI/NIAID NIH HHS/United States ; }, abstract = {Background: Ae. aegypti mosquitoes stably transfected with the intracellular bacterium Wolbachia pipientis (wMel strain) have been deployed for biocontrol of dengue and related arboviral diseases in multiple countries. Field releases in northern Australia have previously demonstrated near elimination of local dengue transmission from Wolbachia-treated communities, and pilot studies in Indonesia have demonstrated the feasibility and acceptability of the method. We conducted a quasi-experimental trial to evaluate the impact of scaled Wolbachia releases on dengue incidence in an endemic setting in Indonesia. Methods: In Yogyakarta City, Indonesia, following extensive community engagement, wMel Wolbachia-carrying mosquitoes were released every two weeks for 13-15 rounds over seven months in 2016-17, in a contiguous 5 km [2] area (population 65,000). A 3 km [2] area (population 34,000) on the opposite side of the city was selected a priori as an untreated control area. Passive surveillance data on notified hospitalised dengue patients was used to evaluate the epidemiological impact of Wolbachia deployments, using controlled interrupted time-series analysis. Results: Rapid and sustained introgression of wMel Wolbachia into local Ae. aegypti populations was achieved. Thirty-four dengue cases were notified from the intervention area and 53 from the control area (incidence 26 vs 79 per 100,000 person-years) during 24 months following Wolbachia deployment. This corresponded in the regression model to a 73% reduction in dengue incidence (95% confidence interval 49%,86%) associated with the Wolbachia intervention. Exploratory analysis including 6 months additional post-intervention observations showed a small strengthening of this effect (30 vs 115 per 100,000 person-years; 76% reduction in incidence, 95%CI 60%,86%). Conclusions: We demonstrate a significant reduction in dengue incidence following successful introgression of Wolbachia into local Ae. aegypti populations in an endemic setting in Indonesia. These findings are consistent with previous field trials in northern Australia, and support the effectiveness of this novel approach for dengue control.}, } @article {pmid32797213, year = {2021}, author = {Martinez, J and Klasson, L and Welch, JJ and Jiggins, FM}, title = {Life and Death of Selfish Genes: Comparative Genomics Reveals the Dynamic Evolution of Cytoplasmic Incompatibility.}, journal = {Molecular biology and evolution}, volume = {38}, number = {1}, pages = {2-15}, pmid = {32797213}, issn = {1537-1719}, support = {/WT_/Wellcome Trust/United Kingdom ; 281668/ERC_/European Research Council/International ; WT202888/Z/16/Z/WT_/Wellcome Trust/United Kingdom ; WT094664MA/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Animals ; Drosophila/microbiology/physiology ; *Evolution, Molecular ; Female ; *Genome, Bacterial ; Host-Pathogen Interactions/*genetics ; Male ; *Repetitive Sequences, Nucleic Acid ; Spermatozoa/physiology ; Wolbachia/*genetics ; }, abstract = {Cytoplasmic incompatibility is a selfish reproductive manipulation induced by the endosymbiont Wolbachia in arthropods. In males Wolbachia modifies sperm, leading to embryonic mortality in crosses with Wolbachia-free females. In females, Wolbachia rescues the cross and allows development to proceed normally. This provides a reproductive advantage to infected females, allowing the maternally transmitted symbiont to spread rapidly through host populations. We identified homologs of the genes underlying this phenotype, cifA and cifB, in 52 of 71 new and published Wolbachia genome sequences. They are strongly associated with cytoplasmic incompatibility. There are up to seven copies of the genes in each genome, and phylogenetic analysis shows that Wolbachia frequently acquires new copies due to pervasive horizontal transfer between strains. In many cases, the genes have subsequently acquired loss-of-function mutations to become pseudogenes. As predicted by theory, this tends to occur first in cifB, whose sole function is to modify sperm, and then in cifA, which is required to rescue the cross in females. Although cif genes recombine, recombination is largely restricted to closely related homologs. This is predicted under a model of coevolution between sperm modification and embryonic rescue, where recombination between distantly related pairs of genes would create a self-incompatible strain. Together, these patterns of gene gain, loss, and recombination support evolutionary models of cytoplasmic incompatibility.}, } @article {pmid32796890, year = {2020}, author = {Straub, TJ and Shaw, WR and Marcenac, P and Sawadogo, SP and Dabiré, RK and Diabaté, A and Catteruccia, F and Neafsey, DE}, title = {The Anopheles coluzzii microbiome and its interaction with the intracellular parasite Wolbachia.}, journal = {Scientific reports}, volume = {10}, number = {1}, pages = {13847}, pmid = {32796890}, issn = {2045-2322}, support = {R21 AI117313/AI/NIAID NIH HHS/United States ; U19 AI110818/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Anopheles/*microbiology/*physiology ; Burkina Faso ; Disease Vectors ; Female ; Host Microbial Interactions/*physiology ; *Insect Vectors ; Malaria/transmission ; Mosquito Control/methods ; Oviposition ; Vector Borne Diseases/transmission ; *Wolbachia ; }, abstract = {Wolbachia, an endosymbiotic alpha-proteobacterium commonly found in insects, can inhibit the transmission of human pathogens by mosquitoes. Biocontrol programs are underway using Aedes aegypti mosquitoes trans-infected with a non-natural Wolbachia strain to reduce dengue virus transmission. Less is known about the impact of Wolbachia on the biology and vectorial capacity of Anopheles mosquitoes, the vectors of malaria parasites. A naturally occurring strain of Wolbachia, wAnga, infects populations of the major malaria vectors Anopheles gambiae and Anopheles coluzzii in Burkina Faso. Previous studies found wAnga infection was negatively correlated with Plasmodium infection in the mosquito and wAnga influenced mosquito egg-laying behavior. Here, we investigate wAnga in natural populations of An. coluzzii and its interactions with other resident microbiota using targeted 16S sequencing. Though we find no major differences in microbiota composition associated with wAnga infection, we do find several taxa that correlate with the presence or absence of wAnga in female mosquitoes following oviposition, with the caveat that we could not rule out batch effects due to the unanticipated impact of wAnga on oviposition timing. These data suggest wAnga may influence or interact with the Anopheles microbiota, which may contribute to the impact of wAnga on Anopheles biology and vectorial capacity.}, } @article {pmid32792343, year = {2020}, author = {Ferguson, KB and Kursch-Metz, T and Verhulst, EC and Pannebakker, BA}, title = {Hybrid Genome Assembly and Evidence-Based Annotation of the Egg Parasitoid and Biological Control Agent Trichogramma brassicae.}, journal = {G3 (Bethesda, Md.)}, volume = {10}, number = {10}, pages = {3533-3540}, pmid = {32792343}, issn = {2160-1836}, mesh = {Animals ; Biological Control Agents ; Germany ; *Hymenoptera/genetics ; *Moths ; Random Amplified Polymorphic DNA Technique ; *Wasps/genetics ; }, abstract = {Trichogramma brassicae (Bezdenko) are egg parasitoids that are used throughout the world as biological control agents and in laboratories as model species. Despite this ubiquity, few genetic resources exist beyond COI, ITS2, and RAPD markers. Aided by a Wolbachia infection, a wild-caught strain from Germany was reared for low heterozygosity and sequenced in a hybrid de novo strategy, after which several assembling strategies were evaluated. The best assembly, derived from a DBG2OLC-based pipeline, yielded a genome of 235 Mbp made up of 1,572 contigs with an N50 of 556,663 bp. Following a rigorous ab initio-, homology-, and evidence-based annotation, 16,905 genes were annotated and functionally described. As an example of the utility of the genome, a simple ortholog cluster analysis was performed with sister species T. pretiosum, revealing over 6000 shared clusters and under 400 clusters unique to each species. The genome and transcriptome presented here provides an essential resource for comparative genomics of the commercially relevant genus Trichogramma, but also for research into molecular evolution, ecology, and breeding of T. brassicae.}, } @article {pmid32788982, year = {2020}, author = {N Miyata, M and Nomura, M and Kageyama, D}, title = {Wolbachia have made it twice: Hybrid introgression between two sister species of Eurema butterflies.}, journal = {Ecology and evolution}, volume = {10}, number = {15}, pages = {8323-8330}, pmid = {32788982}, issn = {2045-7758}, abstract = {Wolbachia, cytoplasmically inherited endosymbionts of arthropods, are known to hijack their host reproduction in various ways to increase their own vertical transmission. This may lead to the selective sweep of associated mitochondria, which can have a large impact on the evolution of mitochondrial lineages. In Japan, two different Wolbacahia strains (wCI and wFem) are found in two sister species of pierid butterflies, Eurema mandarina and Eurema hecabe. In both species, females infected with wCI (C females) produce offspring with a nearly 1:1 sex ratio, while females infected with both wCI and wFem (CF females) produce all-female offspring. Previous studies have suggested the historical occurrence of hybrid introgression in C individuals between the two species. Furthermore, hybrid introgression in CF individuals is suggested by the distinct mitochondrial lineages between C females and CF females of E. mandarina. In this study, we performed phylogenetic analyses based on nuclear DNA and mitochondrial DNA markers of E. hecabe with previously published data on E. mandarina. We found that the nuclear DNA of this species significantly diverged from that of E. mandarina. By contrast, mitochondrial DNA haplotypes comprised two clades, mostly reflecting Wolbachia infection status rather than the individual species. Collectively, our results support the previously suggested occurrence of two independent historical events wherein the cytoplasms of CF females and C females moved between E. hecabe and E. mandarina through hybrid introgression.}, } @article {pmid32787974, year = {2020}, author = {Wong, ML and Liew, JWK and Wong, WK and Pramasivan, S and Mohamed Hassan, N and Wan Sulaiman, WY and Jeyaprakasam, NK and Leong, CS and Low, VL and Vythilingam, I}, title = {Natural Wolbachia infection in field-collected Anopheles and other mosquito species from Malaysia.}, journal = {Parasites & vectors}, volume = {13}, number = {1}, pages = {414}, pmid = {32787974}, issn = {1756-3305}, support = {LR002C-2018//Ministry of Education Malaysia/ ; }, mesh = {Aedes/microbiology ; Animals ; Anopheles/microbiology ; Bacterial Outer Membrane Proteins/genetics ; Culex/microbiology ; Culicidae/*microbiology ; Genes, Bacterial ; Insect Control ; Malaysia/epidemiology ; Malvaceae/microbiology ; Mosquito Vectors/microbiology ; Pathology, Molecular ; Phylogeny ; Prevalence ; RNA, Ribosomal, 16S/genetics ; Vector Borne Diseases/prevention & control ; *Wolbachia/genetics/isolation & purification ; }, abstract = {BACKGROUND: The endosymbiont bacterium Wolbachia is maternally inherited and naturally infects some filarial nematodes and a diverse range of arthropods, including mosquito vectors responsible for disease transmission in humans. Previously, it has been found infecting most mosquito species but absent in Anopheles and Aedes aegypti. However, recently these two mosquito species were found to be naturally infected with Wolbachia. We report here the extent of Wolbachia infections in field-collected mosquitoes from Malaysia based on PCR amplification of the Wolbachia wsp and 16S rRNA genes.

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

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

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

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

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

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

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

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

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

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

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

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

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

Potentials of LAB to prevent and/or alleviate the onset and pathogenesis of neurodegenerative diseases such as AD, supporting brain health strategies along the gut-brain axis.}, } @article {pmid32639986, year = {2020}, author = {Gunderson, EL and Vogel, I and Chappell, L and Bulman, CA and Lim, KC and Luo, M and Whitman, JD and Franklin, C and Choi, YJ and Lefoulon, E and Clark, T and Beerntsen, B and Slatko, B and Mitreva, M and Sullivan, W and Sakanari, JA}, title = {The endosymbiont Wolbachia rebounds following antibiotic treatment.}, journal = {PLoS pathogens}, volume = {16}, number = {7}, pages = {e1008623}, pmid = {32639986}, issn = {1553-7374}, support = {R01 AI081803/AI/NIAID NIH HHS/United States ; R01 GM097435/GM/NIGMS NIH HHS/United States ; R56 AI081803/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Brugia pahangi/*microbiology ; Female ; Filariasis/*microbiology ; Filaricides/*pharmacology ; Gerbillinae ; Rifampin/*pharmacology ; Wolbachia/*drug effects ; }, abstract = {Antibiotic treatment has emerged as a promising strategy to sterilize and kill filarial nematodes due to their dependence on their endosymbiotic bacteria, Wolbachia. Several studies have shown that novel and FDA-approved antibiotics are efficacious at depleting the filarial nematodes of their endosymbiont, thus reducing female fecundity. However, it remains unclear if antibiotics can permanently deplete Wolbachia and cause sterility for the lifespan of the adult worms. Concerns about resistance arising from mass drug administration necessitate a careful exploration of potential Wolbachia recrudescence. In the present study, we investigated the long-term effects of the FDA-approved antibiotic, rifampicin, in the Brugia pahangi jird model of infection. Initially, rifampicin treatment depleted Wolbachia in adult worms and simultaneously impaired female worm fecundity. However, during an 8-month washout period, Wolbachia titers rebounded and embryogenesis returned to normal. Genome sequence analyses of Wolbachia revealed that despite the population bottleneck and recovery, no genetic changes occurred that could account for the rebound. Clusters of densely packed Wolbachia within the worm's ovarian tissues were observed by confocal microscopy and remained in worms treated with rifampicin, suggesting that they may serve as privileged sites that allow Wolbachia to persist in worms while treated with antibiotic. To our knowledge, these clusters have not been previously described and may be the source of the Wolbachia rebound.}, } @article {pmid32635864, year = {2020}, author = {Rosenwald, LC and Sitvarin, MI and White, JA}, title = {Endosymbiotic Rickettsiella causes cytoplasmic incompatibility in a spider host.}, journal = {Proceedings. Biological sciences}, volume = {287}, number = {1930}, pages = {20201107}, pmid = {32635864}, issn = {1471-2954}, mesh = {Animals ; Coxiellaceae/*physiology ; Cytoplasm/*microbiology ; Female ; Host-Pathogen Interactions ; Male ; Spiders/*microbiology ; Symbiosis ; }, abstract = {Many arthropod hosts are infected with bacterial endosymbionts that manipulate host reproduction, but few bacterial taxa have been shown to cause such manipulations. Here, we show that a bacterial strain in the genus Rickettsiella causes cytoplasmic incompatibility (CI) between infected and uninfected hosts. We first surveyed the bacterial community of the agricultural spider Mermessus fradeorum (Linyphiidae) using high throughput sequencing and found that individual spiders can be infected with up to five different strains of maternally inherited symbiont from the genera Wolbachia, Rickettsia, and Rickettsiella. The Rickettsiella strain was pervasive, found in all 23 tested spider matrilines. We used antibiotic curing to generate uninfected matrilines that we reciprocally crossed with individuals infected only with Rickettsiella. We found that only 13% of eggs hatched when uninfected females were mated with Rickettsiella-infected males; in contrast, at least 83% of eggs hatched in the other cross types. This is the first documentation of Rickettsiella, or any Gammaproteobacteria, causing CI. We speculate that induction of CI may be much more widespread among maternally inherited bacteria than previously appreciated. Further, our results reinforce the importance of thoroughly characterizing and assessing the inherited microbiome before attributing observed host phenotypes to well-characterized symbionts such as Wolbachia.}, } @article {pmid32631404, year = {2020}, author = {Faria Waziry, PA and Raja, A and Salmon, C and Aldana, N and Damodar, S and Fukushima, AR and Mayi, BS}, title = {Impact of pyriproxyfen on virus behavior: implications for pesticide-induced virulence and mechanism of transmission.}, journal = {Virology journal}, volume = {17}, number = {1}, pages = {93}, pmid = {32631404}, issn = {1743-422X}, mesh = {Aedes/virology ; Animals ; Brazil ; Cell Survival/drug effects ; Dengue Virus/drug effects ; Extracellular Vesicles/drug effects/virology ; Flavivirus Infections/*transmission ; Green Fluorescent Proteins/genetics ; HeLa Cells ; Humans ; Insecticides/*pharmacology ; Jurkat Cells ; Pyridines/*pharmacology ; Trophoblasts/drug effects/virology ; Vesiculovirus/*drug effects ; Virulence ; Virus Replication/*drug effects ; Zika Virus/drug effects ; }, abstract = {BACKGROUND: More than 3 years since the last Zika virus (ZIKV) outbreak in Brazil, researchers are still deciphering the molecular mechanisms of neurovirulence and vertical transmission, as well as the best way to control spread of ZIKV, a flavivirus. The use of pesticides was the main strategy of mosquito control during the last ZIKV outbreak.

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

CONCLUSION: Rampant CNV in C. felis has dire implications for gene-targeting pest control measures and stands to complicate standard normalization procedures utilized in comparative transcriptomics analysis. Coupled with co-infection by novel Wolbachia endosymbionts-potential tools for blocking pathogen transmission-these oddities highlight a unique and underappreciated disease vector.}, } @article {pmid32559297, year = {2020}, author = {Wangkeeree, J and Tewaruxsa, P and Roddee, J and Hanboonsong, Y}, title = {Wolbachia (Rickettsiales: Alphaproteobacteria) Infection in the Leafhopper Vector of Sugarcane White Leaf Disease.}, journal = {Journal of insect science (Online)}, volume = {20}, number = {3}, pages = {}, pmid = {32559297}, issn = {1536-2442}, mesh = {Animals ; Female ; Hemiptera/growth & development/*microbiology ; Insect Vectors/growth & development/microbiology ; Male ; Nymph/growth & development/microbiology ; Ovum/growth & development/microbiology ; Plant Diseases/microbiology ; Saccharum/microbiology ; *Symbiosis ; Wolbachia/*physiology ; }, abstract = {Wolbachia is a maternally inherited bacterium ubiquitous in insects that has attracted interest as a prospective insect pest-control agent. Here, we detected and characterized Wolbachia in the leafhoppers Matsumuratettix hiroglyphicus (Matsumura) (Cicadellidae: Hemiptera) and Yamatotettix flavovittatus Matsumura (Cicadellidae: Hemiptera), insect vectors of the phytoplasma that cause white leaf disease in sugarcane. The 16S rRNA and wsp gene markers revealed that Wolbachia was not present in the M. hiroglyphicus but naturally occurs in Y. flavovittatus. Additionally, the infection rates in adult leafhoppers ranged from 0 to 100% depending on geographic location. Moreover, Wolbachia was detected in the eggs and first- to fifth-instar nymphs of Y. flavovittatus. A phylogenic tree of Wolbachia indicated that it resided in the monophyletic supergroup B clade and clustered in the Ori subgroup. Furthermore, fluorescence in situ hybridization revealed that Wolbachia localized to the egg apices, randomly distributed in the egg cytoplasm, and was concentrated in the nymph and adult bacteriomes, as well as occasional detection in the thorax and abdomen. To the best of our knowledge, the present study is the first to demonstrate the prevalence of Wolbachia in the leafhopper Y. flavovittatus. The obtained results would provide useful information for the future development of Wolbachia as a biological control agent for the leafhopper vectors.}, } @article {pmid32555677, year = {2020}, author = {Bhattacharya, T and Newton, ILG and Hardy, RW}, title = {Viral RNA is a target for Wolbachia-mediated pathogen blocking.}, journal = {PLoS pathogens}, volume = {16}, number = {6}, pages = {e1008513}, pmid = {32555677}, issn = {1553-7374}, support = {R01 AI144430/AI/NIAID NIH HHS/United States ; R21 AI121849/AI/NIAID NIH HHS/United States ; R21 AI153785/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes ; Animals ; Cell Line ; Chlorocebus aethiops ; Cricetinae ; Drosophila melanogaster ; Flavivirus/genetics/*metabolism ; RNA, Viral/genetics/*metabolism ; Togaviridae/genetics/*metabolism ; Vero Cells ; Wolbachia/genetics/*metabolism ; }, abstract = {The ability of the endosymbiont Wolbachia pipientis to restrict RNA viruses is presently being leveraged to curb global transmission of arbovirus-induced diseases. Past studies have shown that virus replication is limited early in arthropod cells colonized by the bacterium, although it is unclear if this phenomenon is replicated in mosquito cells that first encounter viruses obtained through a vertebrate blood meal. Furthermore, these cellular events neither explain how Wolbachia limits dissemination of viruses between mosquito tissues, nor how it prevents transmission of infectious viruses from mosquitoes to vertebrate host. In this study, we try to address these issues using an array of mosquito cell culture models, with an additional goal being to identify a common viral target for pathogen blocking. Our results establish the viral RNA as a cellular target for Wolbachia-mediated inhibition, with the incoming viral RNA experiencing rapid turnover following internalization in cells. This early block in replication in mosquito cells initially infected by the virus thus consequently reduces the production of progeny viruses from these same cells. However, this is not the only contributor to pathogen blocking. We show that the presence of Wolbachia reduces the per-particle infectivity of progeny viruses on naïve mosquito and vertebrate cells, consequently limiting virus dissemination and transmission, respectively. Importantly, we demonstrate that this aspect of pathogen blocking is independent of any particular Wolbachia-host association and affects viruses belonging to Togaviridae and Flaviviridae families of RNA viruses. Finally, consistent with the idea of the viral RNA as a target, we find that the encapsidated virion RNA is less infectious for viruses produced from Wolbachia-colonized cells. Collectively, our findings present a common mechanism of pathogen blocking in mosquitoes that establish a link between virus inhibition in the cell to virus dissemination and transmission.}, } @article {pmid32546497, year = {2020}, author = {Hague, MTJ and Mavengere, H and Matute, DR and Cooper, BS}, title = {Environmental and Genetic Contributions to Imperfect wMel-Like Wolbachia Transmission and Frequency Variation.}, journal = {Genetics}, volume = {215}, number = {4}, pages = {1117-1132}, pmid = {32546497}, issn = {1943-2631}, support = {R01 GM121750/GM/NIGMS NIH HHS/United States ; R35 GM124701/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Drosophila/classification/genetics/growth & development/*microbiology ; Environmental Exposure/*analysis ; Female ; *Gene-Environment Interaction ; Gram-Negative Bacterial Infections/microbiology/*transmission ; *Host-Pathogen Interactions ; Male ; Wolbachia/*pathogenicity ; }, abstract = {Maternally transmitted Wolbachia bacteria infect about half of all insect species. They usually show imperfect maternal transmission and often produce cytoplasmic incompatibility (CI). Irrespective of CI, Wolbachia frequencies tend to increase when rare only if they benefit host fitness. Several Wolbachia, including wMel that infects Drosophila melanogaster, cause weak or no CI and persist at intermediate frequencies. On the island of São Tomé off West Africa, the frequencies of wMel-like Wolbachia infecting Drosophila yakuba (wYak) and Drosophila santomea (wSan) fluctuate, and the contributions of imperfect maternal transmission, fitness effects, and CI to these fluctuations are unknown. We demonstrate spatial variation in wYak frequency and transmission on São Tomé. Concurrent field estimates of imperfect maternal transmission do not predict spatial variation in wYak frequencies, which are highest at high altitudes where maternal transmission is the most imperfect. Genomic and genetic analyses provide little support for D. yakuba effects on wYak transmission. Instead, rearing at cool temperatures reduces wYak titer and increases imperfect transmission to levels observed on São Tomé. Using mathematical models of Wolbachia frequency dynamics and equilibria, we infer that temporally variable imperfect transmission or spatially variable effects on host fitness and reproduction are required to explain wYak frequencies. In contrast, spatially stable wSan frequencies are plausibly explained by imperfect transmission, modest fitness effects, and weak CI. Our results provide insight into causes of wMel-like frequency variation in divergent hosts. Understanding this variation is crucial to explain Wolbachia spread and to improve wMel biocontrol of human disease in transinfected mosquito systems.}, } @article {pmid32545688, year = {2020}, author = {Suwantika, AA and Kautsar, AP and Supadmi, W and Zakiyah, N and Abdulah, R and Ali, M and Postma, MJ}, title = {Cost-Effectiveness of Dengue Vaccination in Indonesia: Considering Integrated Programs with Wolbachia-Infected Mosquitos and Health Education.}, journal = {International journal of environmental research and public health}, volume = {17}, number = {12}, pages = {}, pmid = {32545688}, issn = {1660-4601}, mesh = {Animals ; Child ; Cost-Benefit Analysis ; *Culicidae ; *Dengue ; Health Education ; Humans ; Indonesia ; Vaccination ; *Wolbachia ; }, abstract = {Despite the fact that morbidity and mortality rates due to dengue infection in Indonesia are relatively high, a dengue vaccination has not yet been introduced. Next to vaccination, Wolbachia-infected mosquitoes and health education have been considered to be potential interventions to prevent dengue infection in Indonesia. This study was aimed to analyse the cost-effectiveness of dengue vaccination in Indonesia whilst taking Wolbachia and health education programs into account. An age-structured decision tree model was developed to assess the cost-effectiveness. Approximately 4,701,100 children were followed-up in a 10-year time horizon within a 1-year analytical cycle. We compared three vaccination strategies: one focussing on vaccination only, another combining vaccination and a Wolbachia program, and a third scenario combining vaccination and health education. All scenarios were compared with a no-intervention strategy. The result showed that only vaccination would reduce dengue fever (DF), dengue haemorrhagic fever (DHF), and dengue shock syndrome (DSS) by 123,203; 97,140 and 283 cases, respectively. It would save treatment cost at $10.3 million and $6.2 million from the healthcare and payer perspectives, respectively. The combination of vaccination and a Wolbachia program would reduce DF, DHF and DSS by 292,488; 230,541; and 672 cases, respectively. It would also save treatment cost at $24.3 million and $14.6 million from the healthcare and payer perspectives, respectively. The combination of vaccination and health education would reduce DF, DHF, and DSS by 187,986; 148,220; and 432 cases, respectively. It would save treatment cost at $15.6 million and $9.4 million from the healthcare and payer perspectives, respectively. The incremental cost-effectiveness ratios (ICERs) from the healthcare perspective were estimated to be $9,995, $4,460, and $6,399 per quality-adjusted life year (QALY) gained for the respective scenarios. ICERs from the payer perspective were slightly higher. It can be concluded that vaccination combined with a Wolbachia program was confirmed to be the most cost-effective intervention. Dengue infection rate, vaccine efficacy, cost of Wolbachia program, underreporting factor for hospitalization, vaccine price and mortality rate were considered to be the most influential parameters affecting the ICERs.}, } @article {pmid32530921, year = {2020}, author = {Staunton, KM and Crawford, JE and Cornel, D and Yeeles, P and Desnoyer, M and Livni, J and Holeman, J and Mulligan, FS and Snoad, N and Ritchie, SA}, title = {Environmental influences on Aedes aegypti catches in Biogents Sentinel traps during a Californian "rear and release" program: Implications for designing surveillance programs.}, journal = {PLoS neglected tropical diseases}, volume = {14}, number = {6}, pages = {e0008367}, pmid = {32530921}, issn = {1935-2735}, mesh = {Aedes/*physiology ; Animals ; California ; Ecosystem ; *Environmental Monitoring ; Female ; Housing ; Male ; Mosquito Control ; Mosquito Vectors/*physiology ; Wolbachia ; }, abstract = {As Aedes aegypti continues to expand its global distribution, the diseases it vectors (dengue, Zika, chikungunya and yellow fever) are of increasing concern. Modern efforts to control this species include "rear and release" strategies where lab-reared mosquitoes are distributed throughout the landscape to replace or suppress invasive populations. These programs require intensive surveillance efforts to monitor their success, and the Biogents Sentinel (BGS) trap is one of the most effective tools for sampling adult Ae. aegypti. BGS trap catches can be highly variable throughout landscapes, so we investigated the potential impacts of environmental factors on adult Ae. aegypti capture rates during a "rear and release" program in California to better understand the relative contributions of true variability in population density across a landscape and trap context. We recorded male and female Ae. aegypti catches from BGS traps, with and without CO2, throughout control sites where no mosquitoes were released and in treatment sites where males infected with Wolbachia were released. BGS trap catches were positively influenced by higher proportions of shade or bushes in the front yard of the premises as well as the presence of potential larval habitats such as subterranean vaults. In contrast, an increase in residential habitat within a 100 m radius of trap locations negatively influenced BGS trap catches. For male Ae. aegypti, increased visual complexity of the trap location positively influenced capture rates, and the presence of yard drains negatively affected catch rates in control sites. Lastly, for BGS traps using CO2, higher catch rates were noted from traps placed greater than one meter from walls or fences for both male and female mosquitoes. These results have important implications for surveillance programs of Ae. aegypti throughout the Californian urban environment including adult monitoring during "rear and release" programs.}, } @article {pmid32529355, year = {2020}, author = {Pina, T and Sabater-Muñoz, B and Cabedo-López, M and Cruz-Miralles, J and Jaques, JA and Hurtado-Ruiz, MA}, title = {Molecular characterization of Cardinium, Rickettsia, Spiroplasma and Wolbachia in mite species from citrus orchards.}, journal = {Experimental & applied acarology}, volume = {81}, number = {3}, pages = {335-355}, doi = {10.1007/s10493-020-00508-z}, pmid = {32529355}, issn = {1572-9702}, support = {AGL2014-55616-C3-3-R//Ministerio de Economía y Competitividad/ ; PICD//Universitat Jaume I/ ; FPI BES-2015-074570//Ministerio de Economía y Competitividad/ ; UJI-B2017-24//Universitat Jaume I/ ; }, mesh = {Animals ; *Citrus ; *Mites/microbiology ; Phylogeny ; *Rickettsia/genetics ; *Spiroplasma ; *Tetranychidae ; *Wolbachia/genetics ; }, abstract = {Tetranychidae spider mites are considered key citrus pests in some production areas, especially Tetranychus urticae Koch. Over the past decades, pesticide overuse seems to have promoted T. urticae population selection in citrus orchards. However, the microbiota has also been pointed out as a plausible explanation for population structure or plant host specialisation observed in several arthropod species. In this work, we have determined the incidence of Cardinium, Rickettsia, Spiroplasma and Wolbachia as representatives of major distorter bacteria genera in Aplonobia histricina (Berlese), Eutetranychus banksi (McGregor), Eutetranychus orientalis (Klein), Panonychus citri (McGregor), Tetranychus evansi Baker and Pritchard, Tetranychus turkestani Ugarov and Nikolskii, and T. urticae populations from Spanish citrus orchards. Only Wolbachia was detected by PCR. The multilocus alignment approach and phylogenetic inference indicated that all detected Wolbachia belong to supergroup B. The deep analysis of each 16S rDNA, ftsZ and wsp gene sequences allowed identifying several phylogenetically different Wolbachia sequences. It probably indicates the presence of several different races or strains, all of them belonging to supergroup B. The wsp sequence typing analysis unveiled the presence of the two already identified alleles (61 and 370) and allowed to contribute with five new alleles, supporting the presence of different but related B-races in the studied mite populations. The results are discussed and related to T. urticae population structure, previously observed in Spanish citrus orchards.}, } @article {pmid32522243, year = {2020}, author = {Madhav, M and Brown, G and Morgan, JAT and Asgari, S and McGraw, EA and James, P}, title = {Transinfection of buffalo flies (Haematobia irritans exigua) with Wolbachia and effect on host biology.}, journal = {Parasites & vectors}, volume = {13}, number = {1}, pages = {296}, pmid = {32522243}, issn = {1756-3305}, mesh = {Animals ; Female ; Fertility ; Host Microbial Interactions ; Life Cycle Stages ; Longevity ; Male ; Microinjections ; Muscidae/*microbiology ; Pest Control, Biological/*methods ; Wolbachia/genetics/*physiology ; }, abstract = {BACKGROUND: Buffalo flies (Haematobia irritans exigua) (BF) and closely related horn flies (Haematobia irritans irritans) (HF) are invasive haematophagous parasites with significant economic and welfare impacts on cattle production. Wolbachia are intracellular bacteria found widely in insects and currently of much interest for use in novel strategies for the area wide control of insect pests and insect-vectored diseases. In this paper, we report the transinfection of BF towards the development of area-wide controls.

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

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

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

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

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

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

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

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

CONCLUSION: The SEM technique is an excellent tool for differential recognition of the parasite surface morphology and molecular techniques could differentiate and identify Dirofilaria spp.}, } @article {pmid32488484, year = {2020}, author = {Semiatizki, A and Weiss, B and Bagim, S and Rohkin-Shalom, S and Kaltenpoth, M and Chiel, E}, title = {Effects, interactions, and localization of Rickettsia and Wolbachia in the house fly parasitoid, Spalangia endius.}, journal = {Microbial ecology}, volume = {80}, number = {3}, pages = {718-728}, doi = {10.1007/s00248-020-01520-x}, pmid = {32488484}, issn = {1432-184X}, support = {435/18//Planning and Budgeting Committee of the Council for Higher Education of Israel/ ; CoG 819585/ERC_/European Research Council/International ; }, mesh = {Animals ; Female ; *Host-Parasite Interactions ; Houseflies/parasitology ; Male ; Reproduction ; Rickettsia/*physiology ; *Symbiosis ; Wasps/*microbiology/physiology ; Wolbachia/*physiology ; }, abstract = {Many insect species harbor facultative microbial symbionts that affect their biology in diverse ways. Here, we studied the effects, interactions, and localization of two bacterial symbionts-Wolbachia and Rickettsia-in the parasitoid Spalangia endius. We crossed between four S. endius colonies-Wolbachia only (W), Rickettsia only (R), both (WR), and none (aposymbiotic, APS) (16 possible crosses) and found that Wolbachia induces incomplete cytoplasmic incompatibility (CI), both when the males are W or WR. Rickettsia did not cause reproductive manipulations and did not rescue the Wolbachia-induced CI. However, when R females were crossed with W or WR males, significantly less offspring were produced compared with that of control crosses. In non-CI crosses, the presence of Wolbachia in males caused a significant reduction in offspring numbers. Females' developmental time was significantly prolonged in the R colony, with adults starting to emerge one day later than the other colonies. Other fitness parameters did not differ significantly between the colonies. Using fluorescence in situ hybridization microscopy in females, we found that Wolbachia is localized alongside Rickettsia inside oocytes, follicle cells, and nurse cells in the ovaries. However, Rickettsia is distributed also in muscle cells all over the body, in ganglia, and even in the brain.}, } @article {pmid32485988, year = {2020}, author = {Papadopoulos, C and Karas, PA and Vasileiadis, S and Ligda, P and Saratsis, A and Sotiraki, S and Karpouzas, DG}, title = {Host Species Determines the Composition of the Prokaryotic Microbiota in Phlebotomus Sandflies.}, journal = {Pathogens (Basel, Switzerland)}, volume = {9}, number = {6}, pages = {}, pmid = {32485988}, issn = {2076-0817}, abstract = {Phlebotomine sandflies are vectors of the humans' and mammals' parasite Leishmania spp. Although the role of gut microbiome in the biological cycle of insects is acknowledged, we still know little about the factors modulating the composition of the gut microbiota of sandflies. We tested whether host species impose a strong structural effect on the gut microbiota of Phlebotomus spp. Sandflies were collected from the island of Leros, Greece, and classified to P. papatasi, P. neglectus, P. tobbi, and P. similis, all being negative to Leishmania spp. The prokaryotic gut microbiota was determined via 16S rRNA gene amplicon sequencing. Phlebotomus species supported distinct microbial communities (p < 0.001). P. papatasi microbiota was the most distinct over-dominated by three Spiroplasma, Wolbachia and Paenibacillus operational taxonomic units (OTUs), while another Wolbachia OTU prevailed in P. neglectus. Conversely, the microbiota of P. tobbi and P. similis was composed of several less dominant OTUs. Archaea showed low presence with the dominant OTUs belonging to methanogenic Euryarcheota, ammonia-oxidizing Thaumarcheota, and Nanoarchaeota. We provide first insights into the composition of the bacterial and archaeal community of Phlebotomus sandflies and showed that, in the absence of Leishmania, host genotype is the major modulator of Phlebotomus sandfly gut microbiota.}, } @article {pmid32477411, year = {2020}, author = {Mazzucco, R and Nolte, V and Vijayan, T and Schlötterer, C}, title = {Long-Term Dynamics Among Wolbachia Strains During Thermal Adaptation of Their Drosophila melanogaster Hosts.}, journal = {Frontiers in genetics}, volume = {11}, number = {}, pages = {482}, pmid = {32477411}, issn = {1664-8021}, support = {P 27630/FWF_/Austrian Science Fund FWF/Austria ; W 1225/FWF_/Austrian Science Fund FWF/Austria ; }, abstract = {Climate change is a major evolutionary force triggering thermal adaptation in a broad range of species. While the consequences of global warming are being studied for an increasing number of species, limited attention has been given to the evolutionary dynamics of endosymbionts in response to climate change. Here, we address this question by studying the dynamics of Wolbachia, a well-studied endosymbiont of Drosophila melanogaster. D. melanogaster populations infected with 13 different Wolbachia strains were exposed to novel hot and cold laboratory environments for up to 180 generations. The short-term dynamics suggested a temperature-related fitness difference resulting in the increase of clade V strains in the cold environment only. Our long-term analysis now uncovers that clade V dominates in all replicates after generation 60 irrespective of temperature treatment. We propose that adaptation of the Drosophila host to either temperature or Drosophila C virus (DCV) infection are the cause of the replicated, temporally non-concordant Wolbachia dynamics. Our study provides an interesting case demonstrating that even simple, well-controlled experiments can result in complex, but repeatable evolutionary dynamics, thus providing a cautionary note on too simple interpretations on the impact of climate change.}, } @article {pmid32477320, year = {2020}, author = {Sharma, P and Rani, J and Chauhan, C and Kumari, S and Tevatiya, S and Das De, T and Savargaonkar, D and Pandey, KC and Dixit, R}, title = {Altered Gut Microbiota and Immunity Defines Plasmodium vivax Survival in Anopheles stephensi.}, journal = {Frontiers in immunology}, volume = {11}, number = {}, pages = {609}, pmid = {32477320}, issn = {1664-3224}, mesh = {Animals ; Anopheles/immunology/microbiology/*parasitology ; Gastrointestinal Microbiome/*physiology ; Plasmodium vivax/*growth & development ; RNA-Seq ; Symbiosis ; }, abstract = {Blood-feeding enriched gut-microbiota boosts mosquitoes' anti-Plasmodium immunity. Here, we ask how Plasmodium vivax alters gut-microbiota, anti-Plasmodial immunity, and impacts tripartite Plasmodium-mosquito-microbiota interactions in the gut lumen. We used a metagenomics and RNAseq strategy to address these questions. In naïve mosquitoes, Elizabethkingia meningitis and Pseudomonas spp. are the dominant bacteria and blood-feeding leads to a heightened detection of Elizabethkingia, Pseudomonas and Serratia 16S rRNA. A parallel RNAseq analysis of blood-fed midguts also shows the presence of Elizabethkingia-related transcripts. After, P. vivax infected blood-meal, however, we do not detect bacterial 16S rRNA until circa 36 h. Intriguingly, the transcriptional expression of a selected array of antimicrobial arsenal cecropins 1-2, defensin-1, and gambicin remained low during the first 36 h-a time frame when ookinetes/early oocysts invaded the gut. We conclude during the preinvasive phase, P. vivax outcompetes midgut-microbiota. This microbial suppression likely negates the impact of mosquito immunity which in turn may enhance the survival of P. vivax. Detection of sequences matching to mosquito-associated Wolbachia opens a new inquiry for its exploration as an agent for "paratransgenesis-based" mosquito control.}, } @article {pmid32473140, year = {2020}, author = {Burki, T}, title = {Wolbachia, a bacterium fighting on our side.}, journal = {The Lancet. Infectious diseases}, volume = {20}, number = {6}, pages = {662-663}, doi = {10.1016/S1473-3099(20)30384-4}, pmid = {32473140}, issn = {1474-4457}, mesh = {Aedes/*microbiology ; Animals ; Host-Pathogen Interactions ; Humans ; Malaria/*prevention & control/*transmission ; Pest Control, Biological/*methods ; Wolbachia/*physiology ; }, } @article {pmid32471038, year = {2020}, author = {Tseng, SP and Hsu, PW and Lee, CC and Wetterer, JK and Hugel, S and Wu, LH and Lee, CY and Yoshimura, T and Yang, CS}, title = {Evidence for Common Horizontal Transmission of Wolbachia among Ants and Ant Crickets: Kleptoparasitism Added to the List.}, journal = {Microorganisms}, volume = {8}, number = {6}, pages = {}, pmid = {32471038}, issn = {2076-2607}, support = {N/A//Kyoto University/ ; }, abstract = {While Wolbachia, an intracellular bacterial symbiont, is primarily transmitted maternally in arthropods, horizontal transmission between species has been commonly documented. We examined kleptoparasitism as a potential mechanism for Wolbachia horizontal transmission, using ant crickets and their host ants as the model system. We compared prevalence and diversity of Wolbachia across multiple ant cricket species with different degrees of host specificity/integration level. Our analyses revealed at least three cases of inter-ordinal Wolbachia transfer among ant and ant crickets, and also showed that ant cricket species with high host-integration and host-specificity tend to harbor a higher Wolbachia prevalence and diversity than other types of ant crickets. This study provides empirical evidence that distribution of Wolbachia across ant crickets is largely attributable to horizontal transmission, but also elucidates the role of intimate ecological association in successful Wolbachia horizontal transmission.}, } @article {pmid32458175, year = {2020}, author = {Zhang, X and Liu, Q and Zhu, H}, title = {Modeling and dynamics of Wolbachia-infected male releases and mating competition on mosquito control.}, journal = {Journal of mathematical biology}, volume = {81}, number = {1}, pages = {243-276}, doi = {10.1007/s00285-020-01509-7}, pmid = {32458175}, issn = {1432-1416}, mesh = {*Aedes/microbiology ; Animals ; Humans ; Male ; *Models, Biological ; *Mosquito Control/methods/statistics & numerical data ; Mosquito Vectors/microbiology ; *Vector Borne Diseases/prevention & control ; *Wolbachia/physiology ; }, abstract = {Despite centuries of continuous efforts, mosquito-borne diseases (MBDs) remain enormous health threat of human life worldwide. Lately, the USA government has approved an innovative technology of releasing Wolbachia-infected male mosquitoes to suppress the wild mosquito population. In this paper we first introduce a stage-structured model for natural mosquitos, then we establish a new model considering the releasing of Wolbachia-infected male mosquitoes and the mating competition between the natural male mosquitoes and infected males on the suppression of natural mosquitoes. Dynamical analysis of the two models, including the existence and local stability of the equilibria and bifurcation analysis, reveals the existence of a forward bifurcation or a backward bifurcation with multiple attractors. Moreover, globally dynamical properties are further explored by using Lyapunov function and theory of monotone operators, respectively. Our findings suggest that infected male augmentation itself cannot always guarantee the success of population eradication, but leads to three possible levels of population suppression, so we define the corresponding suppression rate and estimate the minimum release ratio for population eradication. Furthermore, we study how the release ratio of infected males and natural ones, mating competition, the rate of cytoplasmic incompatibility and the basic offspring number affect the suppression rate of natural mosquitoes. Our results show that the successful eradication relies on assessing the reproductive capacity of natural mosquitoes, a selection of suitable Wolbachia strains and an appropriate release amount of infected males. This study will be helpful for public health authorities in designing proper strategies to control vector mosquitoes and prevent the epidemics of MBDs.}, } @article {pmid32456036, year = {2020}, author = {Yen, PS and Failloux, AB}, title = {A Review: Wolbachia-Based Population Replacement for Mosquito Control Shares Common Points with Genetically Modified Control Approaches.}, journal = {Pathogens (Basel, Switzerland)}, volume = {9}, number = {5}, pages = {}, pmid = {32456036}, issn = {2076-0817}, support = {n_ANR-10-LABX-62-IBEID//French Government's Investissement d'Avenir program, Laboratoire d'Excellence "Integrative Biology of Emerging Infectious Diseases, IBEID"/ ; NHRI-PP-108-0324-01-17-07//National Health Research Institutes/ ; }, abstract = {The growing expansion of mosquito vectors has made mosquito-borne arboviral diseases a global threat to public health, and the lack of licensed vaccines and treatments highlight the urgent need for efficient mosquito vector control. Compared to genetically modified control strategies, the intracellular bacterium Wolbachia, endowing a pathogen-blocking phenotype, is considered an environmentally friendly strategy to replace the target population for controlling arboviral diseases. However, the incomplete knowledge regarding the pathogen-blocking mechanism weakens the reliability of a Wolbachia-based population replacement strategy. Wolbachia infections are also vulnerable to environmental factors, temperature, and host diet, affecting their densities in mosquitoes and thus the virus-blocking phenotype. Here, we review the properties of the Wolbachia strategy as an approach to control mosquito populations in comparison with genetically modified control methods. Both strategies tend to limit arbovirus infections but increase the risk of selecting arbovirus escape mutants, rendering these strategies less reliable.}, } @article {pmid32455576, year = {2020}, author = {Laidoudi, Y and Marie, JL and Tahir, D and Watier-Grillot, S and Mediannikov, O and Davoust, B}, title = {Detection of Canine Vector-Borne Filariasis and Their Wolbachia Endosymbionts in French Guiana.}, journal = {Microorganisms}, volume = {8}, number = {5}, pages = {}, pmid = {32455576}, issn = {2076-2607}, abstract = {In French Guiana, canine heartworm disease is well known, but the diversity of filarial parasites of dogs remains largely unknown. A total of 98 canine blood samples from Cayenne and Kourou were assessed by a blood wet mount preparation, heartworm antigen test and molecular exploration of filarioid and Wolbachia DNAs, followed by a multiplex species-specific qPCR's identification and a subsequent sequencing analysis. Thereafter, a phylogeny based on maximum likelihood was carried out to facilitate specific identification. Five dogs were microfilaremic. Heartworm antigens were detected in 15 (15.3%) dogs. Of these, six (6.1%) were considered as occult infections as neither microfilariae nor Dirofilaria immitis DNA were detected. The 11 (11.2%) D. immitis isolates corresponded to a low virulent strain. Six of the D. immitis isolates were positive for Wolbachia endosymbionts of D. immitis belonging to the clade C DNA. Acanthocheilonema reconditum DNA was detected in 3 (3.1%) samples. Of these latter, one was found co-infected with the Brugia sp. genotype and the DNA of the clade D of the Wolbachia endosymbiont of Brugia species. This latter was also detected in two filarioid DNA-free samples. Finally, two samples were positive for Cercopithifilaria bainae genotype, which is distinct from those identified in Europe. The present study highlights the urgent need to implement chemoprophylaxis associated with anti-Wolbachia drugs to control these potential zoonoses.}, } @article {pmid32450914, year = {2020}, author = {Anders, KL and Indriani, C and Ahmad, RA and Tantowijoyo, W and Arguni, E and Andari, B and Jewell, NP and Dufault, SM and Ryan, PA and Tanamas, SK and Rancès, E and O'Neill, SL and Simmons, CP and Utarini, A}, title = {Update to the AWED (Applying Wolbachia to Eliminate Dengue) trial study protocol: a cluster randomised controlled trial in Yogyakarta, Indonesia.}, journal = {Trials}, volume = {21}, number = {1}, pages = {429}, pmid = {32450914}, issn = {1745-6215}, support = {R56 AI070043/AI/NIAID NIH HHS/United States ; R56 AI134724/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/*microbiology ; Animals ; Dengue/epidemiology/*prevention & control/transmission/virology ; Dengue Virus/*pathogenicity ; Humans ; Incidence ; Indonesia/epidemiology ; Mosquito Vectors/*microbiology ; Pest Control, Biological/methods ; Randomized Controlled Trials as Topic ; Time Factors ; Wolbachia/*physiology ; }, abstract = {BACKGROUND: The AWED (Applying Wolbachia to Eliminate Dengue) trial is a parallel, two-arm, non-blinded cluster randomised controlled trial that is under way in Yogyakarta, Indonesia, with the aim of measuring the efficacy of Wolbachia-infected Aedes aegypti deployments in reducing dengue incidence in an endemic setting. Enrolment began in January 2018 and is ongoing. The original study protocol was published in April 2018. Here, we describe amendments that have been made to the study protocol since commencement of the trial.

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

CONCLUSIONS: The Cardinium phylogeny shows evidence of diversification within particular host lineages, and also of host shifts among trophic levels within parasitoid-host communities. Like Wolbachia, the relatedness of Cardinium strains does not necessarily predict their reproductive phenotypes. Lastly, the genetic tools proposed in this study may help future authors to characterize new strains and add to our understanding of Cardinium evolution.}, } @article {pmid31774393, year = {2019}, author = {Beckmann, JF and Sharma, GD and Mendez, L and Chen, H and Hochstrasser, M}, title = {The Wolbachia cytoplasmic incompatibility enzyme CidB targets nuclear import and protamine-histone exchange factors.}, journal = {eLife}, volume = {8}, number = {}, pages = {}, pmid = {31774393}, issn = {2050-084X}, support = {R01 GM053756/GM/NIGMS NIH HHS/United States ; R37 GM046904/GM/NIGMS NIH HHS/United States ; R01 GM046904/GM/NIGMS NIH HHS/United States ; GM053756/GF/NIH HHS/United States ; S10 OD018034/OD/NIH HHS/United States ; GM046904/GF/NIH HHS/United States ; }, mesh = {Active Transport, Cell Nucleus ; Animals ; Animals, Genetically Modified ; Centromere Protein A/genetics/*metabolism ; Crosses, Genetic ; Cytoplasm/*metabolism ; Drosophila Proteins/genetics/*metabolism ; Drosophila melanogaster/genetics/metabolism/microbiology ; Escherichia coli/metabolism ; Female ; Gene Expression Regulation ; Histones/*metabolism ; Karyopherins/genetics/metabolism ; Male ; Protamines/*metabolism ; Recombinant Proteins ; Reproduction ; Saccharomyces cerevisiae/metabolism ; Saccharomyces cerevisiae Proteins/genetics/metabolism ; Wolbachia/*metabolism ; alpha Karyopherins/isolation & purification/metabolism ; }, abstract = {Intracellular Wolbachia bacteria manipulate arthropod reproduction to promote their own inheritance. The most prevalent mechanism, cytoplasmic incompatibility (CI), traces to a Wolbachia deubiquitylase, CidB, and CidA. CidB has properties of a toxin, while CidA binds CidB and rescues embryonic viability. CidB is also toxic to yeast where we identified both host effects and high-copy suppressors of toxicity. The strongest suppressor was karyopherin-α, a nuclear-import receptor; this required nuclear localization-signal binding. A protein-interaction screen of Drosophila extracts using a substrate-trapping catalytic mutant, CidB*, also identified karyopherin-α; the P32 protamine-histone exchange factor bound as well. When CidB* bound CidA, these host protein interactions disappeared. These associations would place CidB at the zygotic male pronucleus where CI defects first manifest. Overexpression of karyopherin-α, P32, or CidA in female flies suppressed CI. We propose that CidB targets nuclear-protein import and protamine-histone exchange and that CidA rescues embryos by restricting CidB access to its targets.}, } @article {pmid31769394, year = {2020}, author = {Lau, MJ and Endersby-Harshman, NM and Axford, JK and Ritchie, SA and Hoffmann, AA and Ross, PA}, title = {Measuring the Host-Seeking Ability of Aedes aegypti Destined for Field Release.}, journal = {The American journal of tropical medicine and hygiene}, volume = {102}, number = {1}, pages = {223-231}, pmid = {31769394}, issn = {1476-1645}, support = {108508/A/15/Z/WT_/Wellcome Trust/United Kingdom ; 108508/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Aedes/*microbiology/*physiology ; Animals ; Feeding Behavior/*physiology ; Female ; Mosquito Control/methods ; Pest Control, Biological/methods ; Wolbachia ; }, abstract = {Host seeking is an essential process in mosquito reproduction. Field releases of modified mosquitoes for population replacement rely on successful host seeking by female mosquitoes, but host-seeking ability is rarely tested in a realistic context. We tested the host-seeking ability of female Aedes aegypti mosquitoes using a semi-field system. Females with different Wolbachia infection types (wMel-, wAlbB-infected, and uninfected) or from different origins (laboratory and field) were released at one end of a semi-field cage and recaptured as they landed on human experimenters 15 m away. Mosquitoes from each population were then identified with molecular tools or through minimal dusting with fluorescent powder. Wolbachia-infected and uninfected populations had similar average durations to landing and overall recapture proportions, as did laboratory and field-sourced Ae. aegypti. These results indicate that the host-seeking ability of mosquitoes is not negatively affected by Wolbachia infection or long-term laboratory maintenance. This method provides an approach to study the host-seeking ability of mosquitoes in a realistic setting, which will be useful when evaluating strains of mosquitoes that are planned for releases into the field to suppress arbovirus transmission.}, } @article {pmid31767943, year = {2020}, author = {Ju, JF and Bing, XL and Zhao, DS and Guo, Y and Xi, Z and Hoffmann, AA and Zhang, KJ and Huang, HJ and Gong, JT and Zhang, X and Hong, XY}, title = {Wolbachia supplement biotin and riboflavin to enhance reproduction in planthoppers.}, journal = {The ISME journal}, volume = {14}, number = {3}, pages = {676-687}, pmid = {31767943}, issn = {1751-7370}, mesh = {Animals ; Biotin/*metabolism ; Fertility ; Genomics ; Hemiptera/*microbiology/*physiology ; Reproduction ; Riboflavin/*metabolism ; Symbiosis ; Vitamin B Complex/metabolism ; Wolbachia/genetics/*metabolism ; }, abstract = {Symbiont-mediated nutritional mutualisms can contribute to the host fitness of insects, especially for those that feed exclusively on nutritionally unbalanced diets. Here, we elucidate the importance of B group vitamins in the association of endosymbiotic bacteria Wolbachia with two plant-sap feeding insects, the small brown planthopper, Laodelphax striatellus (Fallén), and the brown planthopper, Nilaparvata lugens (Stål). Infected planthoppers of both species laid more eggs than uninfected planthoppers, while the experimental transfer of Wolbachia into uninfected lines of one planthopper species rescued this fecundity deficit. The genomic analysis showed that Wolbachia strains from the two planthopper species encoded complete biosynthesis operons for biotin and riboflavin, while a metabolic analysis revealed that Wolbachia-infected planthoppers of both species had higher titers of biotin and riboflavin. Furthermore, experimental supplementation of food with a mixture of biotin and riboflavin recovered the fecundity deficit of Wolbachia-uninfected planthoppers. In addition, comparative genomic analysis suggested that the riboflavin synthesis genes are conserved among Wolbachia supergroups. Biotin operons are rare in Wolbachia, and those described share a recent ancestor that may have been horizontally transferred from Cardinium bacteria. Our research demonstrates a type of mutualism that involves a facultative interaction between Wolbachia and plant-sap feeding insects involving vitamin Bs.}, } @article {pmid31767914, year = {2019}, author = {Liu, QQ and Zhou, JC and Zhang, C and Ning, SF and Duan, LJ and Dong, H}, title = {Co-occurrence of thelytokous and bisexual Trichogramma dendrolimi Matsumura (Hymenoptera: Trichogrammatidae) in a natural population.}, journal = {Scientific reports}, volume = {9}, number = {1}, pages = {17480}, pmid = {31767914}, issn = {2045-2322}, mesh = {Animals ; Behavior, Animal ; Bisexuality ; DNA, Bacterial/genetics ; DNA, Intergenic/genetics ; Electron Transport Complex IV/genetics ; Female ; Genetic Introgression ; Insect Proteins/genetics ; Male ; Phylogeny ; Wasps/*classification/microbiology/*physiology ; Wolbachia/*genetics ; }, abstract = {Trichogramma dendrolimi is one of the most successful biocontrol agents in China. However, an inundative condition is necessary to obtain acceptable parasitism effect. A good solution to this is the application of its thelytokous counterparts which unfortunately are scarce in field. We here report the first case of a natural T. dendrolimi population in China comprising both bisexual wasps and an extremely low proportion of thelytokous wasps. These two forms of T. dendrolimi are phylogenetically related based on the reconstructions of ITS-2 and COI genes. Also, the phylogenetic results suggested a potentially Wolbachia-drived ITS-2 variation. The expression of thelytoky was hardly affected by temperature, which might help control Asian corn borer and Dendrolimus punctatus. Wolbachia are responsible for current thelytoky according to phylogenetic analyses, antibiotic treatment and introgression experiment. We also present the third case of paternal sex ratio chromosome that restrains the expansion of Wolbachia. Moreover, the low frequency of thelytoky may be common in natural populations. Consequently if for biological control it is determined that a thelytokous strain is to be preferred, then large number of field collected females should be set up as isofemale lines, to detect the rare thelytoky.}, } @article {pmid31763192, year = {2019}, author = {Lucchetti, C and Genchi, M and Venco, L and Bazzocchi, C and Kramer, LH and Vismarra, A}, title = {Optimized protocol for DNA/RNA co-extraction from adults of Dirofilaria immitis.}, journal = {MethodsX}, volume = {6}, number = {}, pages = {2601-2605}, pmid = {31763192}, issn = {2215-0161}, abstract = {Dirofilaria immitis, the etiologic agent of canine heartworm disease, like several other filarial nematodes, harbors the bacterial endosymbiont Wolbachia. To investigate metabolic and functional pathways of D. immitis and Wolbachia individually, along with their interactions, the use of both transcriptomic and genome analysis has becoming increasingly popular. Although several commercial kits are available for the single extraction of either DNA or RNA, no specific protocol has been described for simultaneous extraction of DNA and RNA from such a large organism like an adult D. immitis, where female worms generally reach ∼25 cm in length. More importantly, adult worms of D. immitis can only be obtained either through necropsy of experimentally infected dogs or by minimally-invasive surgical heartworm removal of naturally infected dogs. This makes each individual worm sample extremely important. Thus, in the context of a project aimed at the evaluation of both gene expression analysis and Wolbachia population assessment following different treatments, an optimized protocol for co-extraction of DNA and RNA from a single sample of adult D. immitis has been developed. •An optimized method for DNA/RNA co-extraction from large size nematodes using TRIzol® reagent.•Allows maximum exploitation of unique samples as adults of D. immitis.}, } @article {pmid31761702, year = {2019}, author = {Nazni, WA and Hoffmann, AA and NoorAfizah, A and Cheong, YL and Mancini, MV and Golding, N and Kamarul, GMR and Arif, MAK and Thohir, H and NurSyamimi, H and ZatilAqmar, MZ and NurRuqqayah, M and NorSyazwani, A and Faiz, A and Irfan, FMN and Rubaaini, S and Nuradila, N and Nizam, NMN and Irwan, SM and Endersby-Harshman, NM and White, VL and Ant, TH and Herd, CS and Hasnor, AH and AbuBakar, R and Hapsah, DM and Khadijah, K and Kamilan, D and Lee, SC and Paid, YM and Fadzilah, K and Topek, O and Gill, BS and Lee, HL and Sinkins, SP}, title = {Establishment of Wolbachia Strain wAlbB in Malaysian Populations of Aedes aegypti for Dengue Control.}, journal = {Current biology : CB}, volume = {29}, number = {24}, pages = {4241-4248.e5}, pmid = {31761702}, issn = {1879-0445}, support = {108508/A/15/Z/WT_/Wellcome Trust/United Kingdom ; 202888/Z/16/Z/WT_/Wellcome Trust/United Kingdom ; 202888/WT_/Wellcome Trust/United Kingdom ; MC_UU_12014/8/MRC_/Medical Research Council/United Kingdom ; /WT_/Wellcome Trust/United Kingdom ; 108508/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Aedes/genetics/metabolism/*microbiology ; Animals ; Dengue/*prevention & control ; Dengue Virus/metabolism/pathogenicity ; Female ; Humans ; Insect Vectors ; Malaysia ; Male ; Mosquito Vectors ; Pest Control, Biological/*methods ; Wolbachia/genetics/*metabolism ; }, abstract = {Dengue has enormous health impacts globally. A novel approach to decrease dengue incidence involves the introduction of Wolbachia endosymbionts that block dengue virus transmission into populations of the primary vector mosquito, Aedes aegypti. The wMel Wolbachia strain has previously been trialed in open releases of Ae. aegypti; however, the wAlbB strain has been shown to maintain higher density than wMel at high larval rearing temperatures. Releases of Ae. aegypti mosquitoes carrying wAlbB were carried out in 6 diverse sites in greater Kuala Lumpur, Malaysia, with high endemic dengue transmission. The strain was successfully established and maintained at very high population frequency at some sites or persisted with additional releases following fluctuations at other sites. Based on passive case monitoring, reduced human dengue incidence was observed in the release sites when compared to control sites. The wAlbB strain of Wolbachia provides a promising option as a tool for dengue control, particularly in very hot climates.}, } @article {pmid31750894, year = {2019}, author = {López-Madrigal, S and Duarte, EH}, title = {Titer regulation in arthropod-Wolbachia symbioses.}, journal = {FEMS microbiology letters}, volume = {366}, number = {23}, pages = {}, doi = {10.1093/femsle/fnz232}, pmid = {31750894}, issn = {1574-6968}, mesh = {Animals ; Arthropods/*microbiology ; Bacterial Load ; Environment ; Symbiosis/*physiology ; Wolbachia/*physiology ; }, abstract = {Symbiosis between intracellular bacteria (endosymbionts) and animals are widespread. The alphaproteobacterium Wolbachia pipientis is known to maintain a variety of symbiotic associations, ranging from mutualism to parasitism, with a wide range of invertebrates. Wolbachia infection might deeply affect host fitness (e.g. reproductive manipulation and antiviral protection), which is thought to explain its high prevalence in nature. Bacterial loads significantly influence both the infection dynamics and the extent of bacteria-induced host phenotypes. Hence, fine regulation of bacterial titers is considered as a milestone in host-endosymbiont interplay. Here, we review both environmental and biological factors modulating Wolbachia titers in arthropods.}, } @article {pmid31747432, year = {2019}, author = {Jasper, ME and Yang, Q and Ross, PA and Endersby-Harshman, N and Bell, N and Hoffmann, AA}, title = {A LAMP assay for the rapid and robust assessment of Wolbachia infection in Aedes aegypti under field and laboratory conditions.}, journal = {PloS one}, volume = {14}, number = {11}, pages = {e0225321}, pmid = {31747432}, issn = {1932-6203}, support = {/WT_/Wellcome Trust/United Kingdom ; 108508/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Aedes/*microbiology ; Animals ; DNA, Bacterial/chemistry/genetics ; Nucleic Acid Amplification Techniques/*methods/standards ; Sensitivity and Specificity ; Wolbachia/*genetics/pathogenicity ; }, abstract = {With Wolbachia-based arbovirus control programs being scaled and operationalised around the world, cost effective and reliable detection of Wolbachia in field samples and laboratory stocks is essential for quality control. Here we validate a modified loop-mediated isothermal amplification (LAMP) assay for routine scoring of Wolbachia in mosquitoes from laboratory cultures and the field, applicable to any setting. We show that this assay is a rapid and robust method for highly sensitive and specific detection of wAlbB Wolbachia infection within Aedes aegypti under a variety of conditions. We test the quantitative nature of the assay by evaluating pooled mixtures of Wolbachia-infected and uninfected mosquitoes and show that it is capable of estimating infection frequencies, potentially circumventing the need to perform large-scale individual analysis for wAlbB infection status in the course of field monitoring. These results indicate that LAMP assays are useful for routine screening particularly under field conditions away from laboratory facilities.}, } @article {pmid31736900, year = {2019}, author = {Zhao, C and Zhao, H and Zhang, S and Luo, J and Zhu, X and Wang, L and Zhao, P and Hua, H and Cui, J}, title = {The Developmental Stage Symbionts of the Pea Aphid-Feeding Chrysoperla sinica (Tjeder).}, journal = {Frontiers in microbiology}, volume = {10}, number = {}, pages = {2454}, pmid = {31736900}, issn = {1664-302X}, abstract = {Chrysoperla sinica (Tjeder) is widely recognized as an important holometabolous natural enemy of various insect pests in different cropping systems and as a non-target surrogate in environmental risk assessment of Bt rice (i.e., genetically modified rice to express a toxin gene from Bacillus thuringiensis). Like other complex organisms, abundant microbes live inside C. sinica; however, to date, microbiome composition and diversity of the whole life cycle in C. sinica has not yet been well characterized. In the current study, we analyze the composition and biodiversity of microbiota across the whole life cycle of C. sinica by using high-throughput Illumina sequencing of the 16S ribosomal RNA gene. Collectively, Proteobacteria and Firmicutes dominated the microenvironment at all stages, but their relative abundances fluctuated by host developmental stage. Interestingly, eggs, neonates, and adults shared similar microbes, including an abundance of Rickettsia and Wolbachia. After larva feeding, Staphylococcus, Enterobacteriaceae, and Serratia were enriched in larvae and pupa, suggesting that food may serve as a major factor contributing to altered microbial community divergence at different developmental stages. Our findings demonstrated that C. sinica harbor a variety of bacteria, and that dynamic changes in community composition and relative abundances of members of its microbiome occur during different life cycle stages. Evaluating the role of these bacterial symbionts in this natural enemy may assist in developing environmental risk assessments and novel biological control strategies.}, } @article {pmid31726321, year = {2020}, author = {Ote, M and Yamamoto, D}, title = {Impact of Wolbachia infection on Drosophila female germline stem cells.}, journal = {Current opinion in insect science}, volume = {37}, number = {}, pages = {8-15}, doi = {10.1016/j.cois.2019.10.001}, pmid = {31726321}, issn = {2214-5753}, mesh = {Animals ; Bacterial Proteins/metabolism ; Drosophila Proteins/metabolism ; Drosophila melanogaster/genetics/*microbiology ; Female ; Oogonial Stem Cells/metabolism/*microbiology ; RNA/metabolism ; Symbiosis ; Wolbachia/*physiology ; }, abstract = {Wolbachia pipientis, one of the most dominant insect-symbiotic bacteria, highjacks the female germline of insects for its own propagation across host generations. Such strict dependence on female gametes in trans-generational propagation has driven Wolbachia to devise ingenious strategies to enhance female fertility. In Drosophila melanogaster females with female-sterile mutant alleles of the master sex-determining gene Sex-lethal (Sxl), Wolbachia colonizing female germline stem cells (GSCs) support the maintenance of GSCs, thereby rescuing the defective ovarian development. In the germ cell cytoplasm, Wolbachia are often found in proximity to ribonucleoprotein-complex processing bodies (P bodies), where the Wolbachia-derived protein TomO interacts with RNAs encoding Nanos and Orb proteins, which support the GSC maintenance and oocyte polarization, respectively. Thus, manipulation of host RNA is the key to successful vertical transmission of Wolbachia.}, } @article {pmid31723729, year = {2019}, author = {, }, title = {The World Health Organization 2030 goals for onchocerciasis: Insights and perspectives from mathematical modelling: NTD Modelling Consortium Onchocerciasis Group.}, journal = {Gates open research}, volume = {3}, number = {}, pages = {1545}, pmid = {31723729}, issn = {2572-4754}, abstract = {The World Health Organization (WHO) has embarked on a consultation process to refine the 2030 goals for priority neglected tropical diseases (NTDs), onchocerciasis among them. Current goals include elimination of transmission (EOT) by 2020 in Latin America, Yemen and selected African countries. The new goals propose that, by 2030, EOT be verified in 10 countries; mass drug administration (MDA) with ivermectin be stopped in at least one focus in 34 countries; and that the proportion of the population no longer in need of MDA be equal or greater than 25%, 50%, 75% and 100% in at least 16, 14, 12, and 10 countries, respectively. The NTD Modelling Consortium onchocerciasis teams have used EPIONCHO and ONCHOSIM to provide modelling insights into these goals. EOT appears feasible in low-moderate endemic areas with long-term MDA at high coverage (≥75%), but uncertain in areas of higher endemicity, poor coverage and adherence, and where MDA has not yet, or only recently, started. Countries will have different proportions of their endemic areas classified according to these categories, and this distribution of pre-intervention prevalence and MDA duration and programmatic success will determine the feasibility of achieving the proposed MDA cessation goals. Highly endemic areas would benefit from switching to biannual or quarterly MDA and implementing vector control where possible (determining optimal frequency and duration of anti-vectorial interventions requires more research). Areas without loiasis that have not yet initiated MDA should implement biannual (preferably with moxidectin) or quarterly MDA from the start. Areas with loiasis not previously treated would benefit from implementing test-and(not)-treat-based interventions, vector control, and anti- Wolbachia therapies, but their success will depend on the levels of screening and coverage achieved and sustained. The diagnostic performance of IgG4 Ov16 serology for assessing EOT is currently uncertain. Verification of EOT requires novel diagnostics at the individual- and population-levels.}, } @article {pmid31715700, year = {2019}, author = {Yin, X and Zhao, S and Yan, B and Tian, Y and Ba, T and Zhang, J and Wang, Y}, title = {Bartonella rochalimae, B. grahamii, B. elizabethae, and Wolbachia spp. in Fleas from Wild Rodents near the China-Kazakhstan Border.}, journal = {The Korean journal of parasitology}, volume = {57}, number = {5}, pages = {553-559}, pmid = {31715700}, issn = {1738-0006}, support = {2016IK264//General Administration of Quality Supervision, Inspection and Quarantine/ ; 81560338//National Natural Science Foundation of China/ ; 2018ZX10101002-007//National Key Research and Development Program of China/ ; }, mesh = {Animals ; Animals, Wild/microbiology ; Bartonella/classification/genetics/*isolation & purification ; Bartonella Infections/microbiology/*veterinary ; China ; Genotype ; Kazakhstan ; Phylogeny ; Rodent Diseases/*microbiology ; Rodentia/classification/*microbiology ; Siphonaptera/classification/*microbiology ; Wolbachia/classification/genetics/*isolation & purification ; }, abstract = {The Alataw Pass, near the Ebinur Lake Wetland (northwest of China) and Taldykorgan (east of Kazakhstan), is a natural habitat for wild rodents. To date, little has been done on the surveillance of Bartonella spp. and Wolbachia spp. from fleas in the region. Here we molecularly detected Bartonella spp. and Wolbachia spp. in wild rodent fleas during January and October of 2016 along the Alataw Pass-Kazakhstan border. A total of 1,706 fleas belonging to 10 species were collected from 6 rodent species. Among the 10 flea species, 4 were found to be positive for Wolbachia, and 5 flea species were positive for Bartonella. Molecular analysis indicated that i) B. rochalimae was firstly identified in Xenopsylla gerbilli minax and X. conforms conforms, ii) B. grahamii was firstly identified in X. gerbilli minax, and iii) B. elizabethae was firstly detected in Coptopsylla lamellifer ardua, Paradoxopsyllus repandus, and Nosopsyllus laeviceps laeviceps. Additionally, 3 Wolbachia endosymbionts were firstly found in X. gerbilli minax, X. conforms conforms, P. repandus, and N. laeviceps laeviceps. BLASTn analysis indicated 3 Bartonella species showed genotypic variation. Phylogenetic analysis revealed 3 Wolbachia endosymbionts were clustered into the non-Siphonaptera Wolbachia group. These findings extend our knowledge of the geographical distribution and carriers of B. rochalimae, B. grahamii, B. elizabethae, and Wolbachia spp. In the future, there is a need for China-Kazakhstan cooperation to strengthen the surveillance of flea-borne pathogens in wildlife.}, } @article {pmid31709323, year = {2019}, author = {Adekunle, AI and Meehan, MT and McBryde, ES}, title = {Mathematical analysis of a Wolbachia invasive model with imperfect maternal transmission and loss of Wolbachia infection.}, journal = {Infectious Disease Modelling}, volume = {4}, number = {}, pages = {265-285}, pmid = {31709323}, issn = {2468-0427}, abstract = {Arboviral infections, especially dengue, continue to cause significant health burden in their endemic regions. One of the strategies to tackle these infections is to replace the main vector agent, Ae. aegypti, with the ones incapable of transmitting the virus. Wolbachia, an intracellular bacterium, has shown promise in achieving this goal. However, key factors such as imperfect maternal transmission, loss of Wolbachia infection, reduced reproductive capacity and shortened life-span affect the dynamics of Wolbachia in different forms in the Ae. aegypti population. In this study, we developed a Wolbachia transmission dynamic model adjusting for imperfect maternal transmission and loss of Wolbachia infection. The invasive reproductive number that determines the likelihood of replacement of the Wolbachia-uninfected (WU) population is derived and with it, we established the local and global stability of the equilibrium points. This analysis clearly shows that cytoplasmic incompatibility (CI) does not guarantee establishment of the Wolbachia-infected (WI) mosquitoes as imperfect maternal transmission and loss of Wolbachia infection could outweigh the gains from CI. Optimal release programs depending on the level of imperfect maternal transmission and loss of Wolbachia infection are shown. Hence, it is left to decision makers to either aim for replacement or co-existence of both populations.}, } @article {pmid31699757, year = {2019}, author = {Basting, PJ and Bergman, CM}, title = {Complete Genome Assemblies for Three Variants of the Wolbachia Endosymbiont of Drosophila melanogaster.}, journal = {Microbiology resource announcements}, volume = {8}, number = {45}, pages = {}, pmid = {31699757}, issn = {2576-098X}, abstract = {Here, we report genome assemblies for three strains of Wolbachia pipientis, assembled from unenriched, unfiltered long-read shotgun sequencing data of geographically distinct strains of Drosophila melanogaster Our simple methodology can be applied to long-read data sets of other Wolbachia-infected species with limited Wolbachia-host lateral gene transfers to produce complete assemblies for this important model symbiont.}, } @article {pmid31698562, year = {2019}, author = {Almeida, L and Duprez, MG and Privat, Y and Vauchelet, N}, title = {Mosquito population control strategies for fighting against arboviruses.}, journal = {Mathematical biosciences and engineering : MBE}, volume = {16}, number = {6}, pages = {6274-6297}, doi = {10.3934/mbe.2019313}, pmid = {31698562}, issn = {1551-0018}, mesh = {Aedes ; Algorithms ; Animals ; Arbovirus Infections/*prevention & control/therapy ; Arboviruses ; Computer Simulation ; Culicidae/microbiology/virology ; Female ; Life Cycle Stages ; Male ; Models, Statistical ; Mosquito Control/*methods ; Mosquito Vectors/microbiology/*virology ; Pest Control, Biological/*methods ; Population Dynamics ; Probability ; Wolbachia ; }, abstract = {In the fight against vector-borne arboviruses, an important strategy of control of epidemic consists in controlling the population of the vector, Aedes mosquitoes in this case. Among possible actions, two techniques consist either in releasing sterile mosquitoes to reduce the size of the population (Sterile Insect Technique) or in replacing the wild population by one carrying a bacteria, called Wolbachia, blocking the transmission of viruses from insects to humans. This article addresses the issue of optimizing the dissemination protocol for each of these strategies, in order to get as close as possible to these objectives. Starting from a mathematical model describing population dynamics, we study the control problem and introduce the cost function standing for population replacement and sterile insect technique. Then, we establish some properties of the optimal control and illustrate them with numerical simulations.}, } @article {pmid31698114, year = {2020}, author = {Karut, K and Castle, SJ and Karut, ŞT and Karaca, MM}, title = {Secondary endosymbiont diversity of Bemisia tabaci and its parasitoids.}, journal = {Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases}, volume = {78}, number = {}, pages = {104104}, doi = {10.1016/j.meegid.2019.104104}, pmid = {31698114}, issn = {1567-7257}, mesh = {Animals ; Arizona ; Bacteria/*classification/genetics/isolation & purification ; Gammaproteobacteria/genetics/isolation & purification ; Genotyping Techniques/*methods ; Hemiptera/*classification/genetics/*microbiology ; Mediterranean Region ; Phylogeography ; Rickettsia/genetics/isolation & purification ; Species Specificity ; Symbiosis ; Turkey ; Wolbachia/genetics/isolation & purification ; }, abstract = {Cotton whitefly, Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) is one of the most important insect pests worldwide. It is known as a species complex consisting of at least 40 cryptic species. Although there are substantial data regarding species composition, parasitoids and endosymbionts of B. tabaci, data on relationship between the pest, parasitoids and endosymbionts are very restricted. Therefore, in this study, secondary endosymbionts in populations of B. tabaci and their parasitoids collected from Turkey and the USA were determined by PCR-based DNA analysis. Whitefly populations in Turkey represented both Mediterranean (MED) and Middle East-Asia Minor1 (MEAM1) genotypes from single or mixed populations of both genotypes. Arsenophonus, Rickettsia and Wolbachia were found in MED, while Hamiltonella and Rickettsia in MEAM1. Whitefly populations collected from Arizona were all MEAM1 and dually infected with Hamiltonella and Rickettsia. The aphelinid parasitoids Encarsia lutea and Eretmocerus mundus predominated in all Turkish populations. While almost all En. lutea populations were infected with Wolbachia, no endosymbionts were detected in any Er. mundus. Parasitoid species and the pattern of secondary endosymbiont infection in Arizona populations were different with Rickettsia detected only from Encarsia sophia while both Rickettsia and Wolbachia were found in Eretmocerus species. As a result, four secondary endosymbionts, namely, Rickettsia, Hamiltonella, Arsenophonus and Wolbachia, were detected from B.tabaci and its parasitoids. Among them only Wolbachia and Rickettsia were found in both the pest and parasitoids. It is conclude that further studies should be pursued to determine effect of these endosymbionts on biology of the parasitoids and success in biological control of B. tabaci.}, } @article {pmid31690673, year = {2019}, author = {Layton, EM and On, J and Perlmutter, JI and Bordenstein, SR and Shropshire, JD}, title = {Paternal Grandmother Age Affects the Strength of Wolbachia-Induced Cytoplasmic Incompatibility in Drosophila melanogaster.}, journal = {mBio}, volume = {10}, number = {6}, pages = {}, pmid = {31690673}, issn = {2150-7511}, support = {R21 HD086833/HD/NICHD NIH HHS/United States ; P30 DK058404/DK/NIDDK NIH HHS/United States ; R21 AI133522/AI/NIAID NIH HHS/United States ; F31 AI143152/AI/NIAID NIH HHS/United States ; R01 AI132581/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Cytoplasm/*genetics/*microbiology ; Drosophila melanogaster/*genetics/*microbiology ; Female ; Grandparents ; Male ; Reproduction/genetics ; Wolbachia/*genetics ; }, abstract = {Wolbachia are obligate intracellular bacteria that are globally distributed in half of all arthropod species. As the most abundant maternally inherited microbe in animals, Wolbachia manipulate host reproduction via reproductive parasitism strategies, including cytoplasmic incompatibility (CI). CI manifests as embryonic death when Wolbachia-modified sperm fertilize uninfected eggs but not maternally infected eggs. Thus, CI can provide a relative fitness advantage to Wolbachia-infected females and drive the infection through a population. In the genetic model Drosophila melanogaster, the Wolbachia strain wMel induces variable CI, making mechanistic studies in D. melanogaster cumbersome. Here, we demonstrate that sons of older paternal D. melanogaster grandmothers induce stronger CI than sons of younger paternal grandmothers, and we term this relationship the "paternal grandmother age effect" (PGAE). Moreover, the embryos and adult sons of older D. melanogaster grandmothers have higher Wolbachia densities, correlating with their ability to induce stronger CI. In addition, we report that Wolbachia density positively correlates with female age and decreases after mating, suggesting that females transmit Wolbachia loads that are proportional to their own titers. These findings reveal a transgenerational impact of age on wMel-induced CI, elucidate Wolbachia density dynamics in D. melanogaster, and provide a methodological advance to studies aimed at understanding wMel-induced CI in the D. melanogaster model.IMPORTANCE Unidirectional cytoplasmic incompatibility (CI) results in a postfertilization incompatibility between Wolbachia-infected males and uninfected females. CI contributes to reproductive isolation between closely related species and is used in worldwide vector control programs to drastically lower arboviral vector population sizes or to replace populations that transmit arboviruses with those resistant to transmission. Despite decades of research on the factors that influence CI, penetrance is often variable under controlled laboratory conditions in various arthropods, suggesting that additional variables influence CI strength. Here, we demonstrate that paternal D. melanogaster grandmother age influences the strength of CI induced by their sons. Older D. melanogaster females have higher Wolbachia densities and produce offspring with higher Wolbachia densities that associate with stronger CI. This work reveals a multigenerational impact of age on CI and expands our understanding of host-Wolbachia interactions and the biology of CI induced by the Wolbachia strain infecting the most widely used arthropod model, D. melanogaster.}, } @article {pmid31683719, year = {2019}, author = {Liang, Y and Hong, Y and Mai, Z and Zhu, Q and Guo, L}, title = {Internal and External Microbial Community of the Thitarodes Moth, the Host of Ophiocordyceps sinensis.}, journal = {Microorganisms}, volume = {7}, number = {11}, pages = {}, pmid = {31683719}, issn = {2076-2607}, support = {No. 2018A030313094//Natural Science Foundation of Guangdong Province/ ; No. 20185071521641//Social Science and Technology Development Project of Dongguan/ ; No. 81303155//National Natural Science Foundation of China/ ; No. 2018GkQNCX050//Project for Young Innovative Talents in Ordinary Higher University of Guangdong Province/ ; }, abstract = {Ophiocordyceps sinensis is a widely known medicinal entomogenous fungus, which parasitizes the soil-borne larva of Thitarodes (Hepialidae, Lepidoptera) distributed in the Qinghai-Tibetan Plateau and its adjacent areas. Previous research has involved artificial cultivation of Chinese cordyceps (the fungus-caterpillar complex), but it is difficult to achieve large-scale cultivation because the coupling relation between the crucial microbes and their hosts is not quite clear. To clarify the influence of the internal microbial community on the occurrence of Chinese cordyceps, in this study, the unfertilized eggs of Thitarodes of different sampling sites were chosen to analyze the bacterial and fungal communities via 16S rRNA and ITS sequencing for the first time. The results showed that for bacteria, 348 genera (dominant genera include Wolbachia, Spiroplasma, Carnobacterium, Sphingobium, and Acinetobacter) belonging to 26 phyla (dominant phyla include Proteobacteria, Firmicutes, Tenericutes, Actinobacteria, Acidobacteria, and Bacteroidetes), 58 classes, 84 orders, and 120 families were identified from 1294 operational taxonomic units (OTUs). The dominant bacterial genus (Spiroplasma) may be an important bacterial factor promoting the occurrence of Chinese cordyceps. For fungi, 289 genera, mainly including Aureobasidium, Candida, and Cryptococcus, were identified, and they belonged to 5 phyla (Ascomycota, Basidiomycota, Chytridiomycota, Glomeromycota, and Zygomycota), 26 classes, 82 orders, and 165 families. Eight bacterial OTUs and 12 fungal OTUs were shared among all of the detected samples and were considered as core species. Among them, Wolbachia, Spiroplasma, Carnobacterium, Aureobasidium, and Phoma may play important roles in helping the host larva to digest foods, adapt to extreme environments, or resist pathogens. On the other hand, the external (soil) microbial community was synchronously and comparatively analyzed. Comparative analysis revealed that external microbial factors might play a more significant role in the occurrence of Chinese cordyceps, owing to the significant differences revealed by α-diversity and β-diversity analyses among different groups. In summary, the results of this study may contribute to the large-scale cultivation of Chinese cordyceps.}, } @article {pmid31665056, year = {2019}, author = {Kieran, TJ and Arnold, KMH and Thomas, JC and Varian, CP and Saldaña, A and Calzada, JE and Glenn, TC and Gottdenker, NL}, title = {Regional biogeography of microbiota composition in the Chagas disease vector Rhodnius pallescens.}, journal = {Parasites & vectors}, volume = {12}, number = {1}, pages = {504}, pmid = {31665056}, issn = {1756-3305}, support = {DGE-1545433//National Science Foundation/ ; COL11-043//Secretaría Nacional de Ciencia, Tecnología e Innovación/ ; }, mesh = {Actinobacteria/classification/genetics ; Analysis of Variance ; Animals ; Bacteria/*classification/genetics ; Bacteroidetes/classification/genetics ; Biodiversity ; Chagas Disease/*transmission ; DNA Barcoding, Taxonomic ; Ecosystem ; Firmicutes/classification/genetics ; Gene Library ; Humans ; Insect Vectors/*microbiology/physiology ; *Microbiota ; Panama ; Phylogeography ; Polymerase Chain Reaction ; Proteobacteria/classification/genetics ; RNA, Ribosomal, 16S/chemistry ; Rhodnius/*microbiology/physiology ; }, abstract = {BACKGROUND: Triatomine bugs are vectors of the protozoan parasite Trypanosoma cruzi, which causes Chagas disease. Rhodnius pallescens is a major vector of Chagas disease in Panama. Understanding the microbial ecology of disease vectors is important in the development of vector management strategies that target vector survival and fitness. In this study we examined the whole-body microbial composition of R. pallescens from three locations in Panama.

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

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

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

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

CONCLUSIONS/SIGNIFICANCE: Our study represents the first open field release of sterile Ae. aegypti males developed from a combined SIT/IIT approach. Entomological assessment using ovitraps, adult sticky traps, and portable vacuum aspirators confirmed the success in reducing natural populations of Ae. aegypti females in treated areas. Public awareness through media resulted in positive support for practical use of this strategy in wider areas. Further study using a systematic randomized trial is needed to determine whether this approach could have a significant impact on the diseases transmitted by Ae. aegypti vector.}, } @article {pmid31649084, year = {2019}, author = {Gasser, MT and Chung, M and Bromley, RE and Nadendla, S and Dunning Hotopp, JC}, title = {Complete Genome Sequence of wAna, the Wolbachia Endosymbiont of Drosophila ananassae.}, journal = {Microbiology resource announcements}, volume = {8}, number = {43}, pages = {}, pmid = {31649084}, issn = {2576-098X}, support = {R01 CA206188/CA/NCI NIH HHS/United States ; U19 AI110820/AI/NIAID NIH HHS/United States ; }, abstract = {Here, we present the complete genome sequence of the Wolbachia endosymbiont wAna, isolated from Drosophila ananassae and derived from Oxford Nanopore and Illumina sequencing. We anticipate that this will aid in Wolbachia comparative genomics and the assembly of D. ananassae specifically in regions containing extensive lateral gene transfer events.}, } @article {pmid31647706, year = {2019}, author = {Staunton, KM and Usher, L and Prachar, T and Ritchie, SA and Snoad, N and Johnson, BJ}, title = {A Novel Methodology For Recording Wing Beat Frequencies of Untethered Male and Female Aedes aegypti.}, journal = {Journal of the American Mosquito Control Association}, volume = {35}, number = {3}, pages = {169-177}, doi = {10.2987/18-6799.1}, pmid = {31647706}, issn = {1943-6270}, mesh = {Aedes/*physiology ; Animals ; Entomology/methods ; Female ; Male ; Mosquito Control/*methods ; Mosquito Vectors/*physiology ; Wings, Animal/*physiology ; }, abstract = {Aedes aegypti is a vector of many significant arboviruses worldwide, including dengue, Zika, chikungunya, and yellow fever viruses. With vector control methodology pivoting toward rearing and releasing large numbers of insects for either population suppression or virus-blocking, economical remote (sentinel) surveillance methods for release tracking become increasingly necessary. Recent steps in this direction include advances in optical sensors that identify and classify insects based on their wing beat frequency (WBF). As these traps are being developed, there is a strong need to better understand the environmental and biological factors influencing mosquito WBFs. Here, we developed new untethered-subject methodology to detect changes in WBFs of male and female Ae. aegypti. This new methodology involves directing an ultrasonic transducer at a free-flying subject and measuring the Doppler shift of the reflected ultrasonic continuous wave signal. This system's utility was assessed by determining its ability to confirm previous reports on the effect of temperature, body size, and age on the WBFs generated from acoustic or optical-based experiments. The presented ultrasonic method successfully detected expected trends for each factor for both male and female Ae. aegypti without the need for subject manipulation and potential impediment of natural flight dynamics due to tethering. As a result, this ultrasonic methodology provides a new method for understanding the environmental and physiological determinants of male and female WBFs that can inform the design of remote mosquito surveillance systems.}, } @article {pmid31644794, year = {2019}, author = {Chen, R and Su, X and Chen, J and Jiang, L and Qiao, GX}, title = {Wolbachia Infection in Two Species: Novel Views on the Colonization Ability of Wolbachia in Aphids.}, journal = {Environmental entomology}, volume = {48}, number = {6}, pages = {1388-1393}, doi = {10.1093/ee/nvz122}, pmid = {31644794}, issn = {1938-2936}, mesh = {Animals ; *Aphids ; China ; Geography ; *Infections ; *Wolbachia ; }, abstract = {Wolbachia pipientis (Rickettsiales: Anaplasmataceae) is an intracellular symbiont residing in arthropods and filarial nematodes. Sixteen supergroups have been described from different host taxa. Four supergroups A, B, M, and N were found in aphids according to prior studies. The cotton aphid, Aphis gossypii, and the green peach aphid, Myzus persicae, are typical polyphagous species with global distributions. We conducted an extensive and systematic survey of Wolbachia infections in these aphids from China. High incidences of Wolbachia infection were detected. The total infection incidence was 60% in A. gossypii and 88% in M. persicae. Both aphid species were infected with supergroups A, B and M. Different incidences of infection were observed among the seven geographical regions in China, which suggested a positive relationship between Wolbachia infections and the geographical distribution of aphid species. Furthermore, multiple infection patterns (M, B, A&M, B&M, and A&B&M) were observed. Infection patterns M and B&M were detected in almost all populations. Patterns A&B&M and B showed geographical restriction in North China. Three factors can possibly influence the Wolbachia infection incidences and patterns: the geographical distribution, aphid species, and different supergroup types.}, } @article {pmid31641465, year = {2019}, author = {Gómez-Zurita, J}, title = {Assessment of the role of Wolbachia in mtDNA paraphyly and the evolution of unisexuality in Calligrapha (Coleoptera: Chrysomelidae).}, journal = {Ecology and evolution}, volume = {9}, number = {19}, pages = {11198-11214}, pmid = {31641465}, issn = {2045-7758}, abstract = {Calligrapha is a New World leaf beetle genus that includes several unisexual species in northeastern North America. Each unisexual species had an independent hybrid origin involving different combinations of bisexual species. However, surprisingly, they all cluster in a single mtDNA clade and with some individuals of their parental species, which are in turn deeply polyphyletic for mtDNA. This pattern is suggestive of a selective sweep which, together with mtDNA taxonomic incongruence and occurrence of unisexuality in Calligrapha, led to hypothesize that Wolbachia might be responsible. I tested this hypothesis studying the correlation between diversity of Wolbachia and well-established mtDNA lineages in >500 specimens of two bisexual species of Calligrapha and their derived unisexual species. Wolbachia appears highly prevalent (83.4%), and fifteen new supergroup-A strains of the bacteria are characterized, belonging to three main classes: wCallA, occupying the whole species ranges, and wCallB and wCallC, narrowly parapatric, infecting beetles with highly divergent mtDNAs where they coexist. Most beetles (71.6%) carried double infections of wCallA with another sequence class. Bayesian inference of ancestral character states and association tests between bacterial diversity and the mtDNA genealogy show that each mtDNA lineage of Calligrapha has specific types of infection. Moreover, shifts can be explained by horizontal or vertical transfer from local populations to an expanding lineage and cytoplasmic incompatibility between wCallB and wCallC types, suggesting that the symbionts hitchhike with the host and are not responsible for selective mtDNA sweeps. Lack of evidence for sweeps and the fact that individuals in the unisexual clade are uninfected or infected by the widespread wCallA type indicate that Wolbachia does not induce unisexuality in Calligrapha, although they may manipulate host reproduction through cytoplasmic incompatibility.}, } @article {pmid31632650, year = {2019}, author = {König, K and Zundel, P and Krimmer, E and König, C and Pollmann, M and Gottlieb, Y and Steidle, JLM}, title = {Reproductive isolation due to prezygotic isolation and postzygotic cytoplasmic incompatibility in parasitoid wasps.}, journal = {Ecology and evolution}, volume = {9}, number = {18}, pages = {10694-10706}, pmid = {31632650}, issn = {2045-7758}, abstract = {The reproductive barriers that prevent gene flow between closely related species are a major topic in evolutionary research. Insect clades with parasitoid lifestyle are among the most species-rich insects and new species are constantly described, indicating that speciation occurs frequently in this group. However, there are only very few studies on speciation in parasitoids. We studied reproductive barriers in two lineages of Lariophagus distinguendus (Chalcidoidea: Hymenoptera), a parasitoid wasp of pest beetle larvae that occur in human environments. One of the two lineages occurs in households preferably attacking larvae of the drugstore beetle Stegobium paniceum ("DB-lineage"), the other in grain stores with larvae of the granary weevil Sitophilus granarius as main host ("GW-lineage"). Between two populations of the DB-lineage, we identified slight sexual isolation as intraspecific barrier. Between populations from both lineages, we found almost complete sexual isolation caused by female mate choice, and postzygotic isolation, which is partially caused by cytoplasmic incompatibility induced by so far undescribed endosymbionts which are not Wolbachia or Cardinium. Because separation between the two lineages is almost complete, they should be considered as separate species according to the biological species concept. This demonstrates that cryptic species within parasitoid Hymenoptera also occur in Central Europe in close contact to humans.}, } @article {pmid31631529, year = {2020}, author = {Bi, J and Wang, YF}, title = {The effect of the endosymbiont Wolbachia on the behavior of insect hosts.}, journal = {Insect science}, volume = {27}, number = {5}, pages = {846-858}, pmid = {31631529}, issn = {1744-7917}, support = {31672352//National Natural Science Foundation of China/ ; 2017AHB050//International Cooperation Projects of Science and Technology of Hubei Province, China/ ; 2017AHB050//the International Cooperation Projects of Science and Technology of Hubei Province, China/ ; }, mesh = {Aggression ; Animals ; *Behavior, Animal ; Feeding Behavior ; Insecta/*microbiology/*physiology ; Learning ; Memory ; Sexual Behavior, Animal ; Sleep ; *Symbiosis ; Wolbachia/*physiology ; }, abstract = {As one of the most successful intracellular symbiotic bacteria, Wolbachia can infect many arthropods and nematodes. Wolbachia infection usually affects the reproduction of their hosts to promote their own proliferation and transmission. Currently, most of the studies focus on the mechanisms of Wolbachia interactions with host reproduction. However, in addition to distribution in the reproductive tissues, Wolbachia also infect various somatic tissues of their hosts, including the brain. This raises the potential that Wolbachia may influence some somatic processes, such as behaviors in their hosts. So far, information about the effects of Wolbachia infection on host behavior is still very limited. The present review presents the current literature on different aspects of the influence of Wolbachia on various behaviors, including sleep, learning and memory, mating, feeding and aggression in their insect hosts. We then highlight ongoing scientific efforts in the field that need addressing to advance this field, which can have significant implications for further developing Wolbachia as environmentally friendly biocontrol agents to control insect-borne diseases and agricultural pests.}, } @article {pmid31622546, year = {2020}, author = {Galis, F and van Alphen, JJM}, title = {Parthenogenesis and developmental constraints.}, journal = {Evolution & development}, volume = {22}, number = {1-2}, pages = {205-217}, doi = {10.1111/ede.12324}, pmid = {31622546}, issn = {1525-142X}, mesh = {Animals ; *Biological Evolution ; Invertebrates/*growth & development ; *Parthenogenesis ; Vertebrates/*growth & development ; }, abstract = {The absence of a paternal contribution in an unfertilized ovum presents two developmental constraints against the evolution of parthenogenesis. We discuss the constraint caused by the absence of a centrosome and the one caused by the missing set of chromosomes and how they have been broken in specific taxa. They are examples of only a few well-underpinned examples of developmental constraints acting at macro-evolutionary scales in animals. Breaking of the constraint of the missing chromosomes is the best understood and generally involves rare occasions of drastic changes of meiosis. These drastic changes can be best explained by having been induced, or at least facilitated, by sudden cytological events (e.g., repeated rounds of hybridization, endosymbiont infections, and contagious infections). Once the genetic and developmental machinery is in place for regular or obligate parthenogenesis, shifts to other types of parthenogenesis can apparently rather easily evolve, for example, from facultative to obligate parthenogenesis, or from pseudoarrhenotoky to haplodiploidy. We argue that the combination of the two developmental constraints forms a near-absolute barrier against the gradual evolution from sporadic to obligate or regular facultative parthenogenesis, which can probably explain why the occurrence of the highly advantageous mode of regular facultative parthenogenesis is so rare and entirely absent in vertebrates.}, } @article {pmid31620116, year = {2019}, author = {Li, F and Hua, H and Ali, A and Hou, M}, title = {Characterization of a Bacterial Symbiont Asaia sp. in the White-Backed Planthopper, Sogatella furcifera, and Its Effects on Host Fitness.}, journal = {Frontiers in microbiology}, volume = {10}, number = {}, pages = {2179}, pmid = {31620116}, issn = {1664-302X}, abstract = {The white-backed planthopper (WBPH), Sogatella furcifera Horváth (Hemiptera: Delphacidae), is an economically significant rice insect pest that harbors a primary fungal yeast-like symbiont (YLS), and some secondary bacterial symbionts like Wolbachia and Cardinium. In the present study, an additional bacterial symbiont in WBPH was characterized. Phylogenetic analysis employing the 16S rRNA gene showed a bacterium closely related to Asaia of Nilaparvata lugens and Nysius expressus, and Asaia krungthepensis. TEM observation of the bacterium showed the typical morphology of Asaia sp. with signature filamentous structures in the nucleoid region. These results indicate that the bacterium belongs to Asaia. The Asaia bacterium was detected in all the tested individual adults and tissues of the laboratory WBPH population but showed varying infection rates (ca 45%) in the field collected WBPH populations. Quantitative PCR analysis revealed that Asaia sp. were significantly more abundant in WBPH females than males, and mainly distributed in the guts, fatty bodies, and salivary glands. Asaia-infected WBPH were of shorter nymphal duration and heavier adult weight than Asaia-free WBPH, while Asaia-free WBPH comparatively fed more, indicating that Asaia plays a role in improving WBPH fitness through involvement in host's nutrient supply.}, } @article {pmid31617302, year = {2020}, author = {Biwot, JC and Zhang, HB and Liu, C and Qiao, JX and Yu, XQ and Wang, YF}, title = {Wolbachia-induced expression of kenny gene in testes affects male fertility in Drosophila melanogaster.}, journal = {Insect science}, volume = {27}, number = {5}, pages = {869-882}, doi = {10.1111/1744-7917.12730}, pmid = {31617302}, issn = {1744-7917}, support = {31672352//National Natural Science Foundation of China/ ; }, mesh = {Animals ; Drosophila Proteins/*genetics/metabolism ; Drosophila melanogaster/genetics/microbiology/*physiology ; Fertility/*genetics ; Gene Expression ; I-kappa B Kinase/*genetics/metabolism ; Male ; Testis/metabolism ; *Up-Regulation ; Wolbachia/*physiology ; }, abstract = {Wolbachia are Gram-negative endosymbionts that are known to cause embryonic lethality when infected male insects mate with uninfected females or with females carrying a different strain of Wolbachia, a situation characterized as cytoplasmic incompatibility (CI). However, the mechanism of CI is not yet fully understood, although recent studies on Drosophila melanogaster have achieved great progress. Here, we found that Wolbachia infection caused changes in the expressions of several immunity-related genes, including significant upregulation of kenny (key), in the testes of D. melanogaster. Overexpression of key in fly testes led to a significant decrease in egg hatch rates when these flies mate with wild-type females. Wolbachia-infected females could rescue this embryonic lethality. Furthermore, in key overexpressing testes terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labeling signal was significantly stronger than in the control testes, and the level of reactive oxygen species was significantly increased. Overexpression of key also resulted in alterations of some other immunity-related gene expressions, including the downregulation of Zn72D. Knockdown of Zn72D in fly testes also led to a significant decrease in egg hatch rates. These results suggest that Wolbachia might induce the defect in male host fertility by immunity-related pathways and thus cause an oxidative damage and cell death in male testes.}, } @article {pmid31615889, year = {2019}, author = {Chen, H and Ronau, JA and Beckmann, JF and Hochstrasser, M}, title = {A Wolbachia nuclease and its binding partner provide a distinct mechanism for cytoplasmic incompatibility.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {116}, number = {44}, pages = {22314-22321}, pmid = {31615889}, issn = {1091-6490}, support = {R01 GM046904/GM/NIGMS NIH HHS/United States ; R01 GM053756/GM/NIGMS NIH HHS/United States ; R37 GM046904/GM/NIGMS NIH HHS/United States ; T32 GM007223/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Bacterial Proteins/genetics/*metabolism ; Deoxyribonucleases/genetics/*metabolism ; Drosophila melanogaster/*microbiology/physiology ; *Host-Pathogen Interactions ; Infertility, Male/*microbiology ; Male ; Operon ; Pest Control, Biological ; Protein Binding ; Wolbachia/enzymology/genetics/*pathogenicity ; }, abstract = {Wolbachia are endosymbiotic bacteria that infect nearly half of all arthropod species. This pandemic is due in part to their ability to increase their transmission through the female germline, most commonly by a mechanism called cytoplasmic incompatibility (CI). The Wolbachia cid operon, encoding 2 proteins, CidA and CidB, the latter a deubiquitylating enzyme (DUB), recapitulates CI in transgenic Drosophila melanogaster However, some CI-inducing Wolbachia strains lack a DUB-encoding cid operon; it was therefore proposed that the related cin operon codes for an alternative CI system. Here we show that the Wolbachia cin operon encodes a nuclease, CinB, and a second protein, CinA, that tightly binds CinB. Recombinant CinB has nuclease activity against both single-stranded and double-stranded DNA but not RNA under the conditions tested. Expression of the cin operon in transgenic male flies induces male sterility and embryonic defects typical of CI. Importantly, transgenic CinA can rescue defects in egg-hatch rates when expressed in females. Expression of CinA also rescues CinB-induced growth defects in yeast. CinB has 2 PD-(D/E)xK nuclease domains, and both are required for nuclease activity and for toxicity in yeast and flies. Our data suggest a distinct mechanism for CI involving a nuclease toxin and highlight the central role of toxin-antidote operons in Wolbachia-induced cytoplasmic incompatibility.}, } @article {pmid31615544, year = {2019}, author = {Ghosh, A and Jasperson, D and Cohnstaedt, LW and Brelsfoard, CL}, title = {Transfection of Culicoides sonorensis biting midge cell lines with Wolbachia pipientis.}, journal = {Parasites & vectors}, volume = {12}, number = {1}, pages = {483}, pmid = {31615544}, issn = {1756-3305}, support = {Start up funds//Texas Tech University/ ; }, mesh = {Aedes/cytology ; Animals ; Biological Control Agents ; Cell Line/microbiology ; Ceratopogonidae/immunology/*microbiology ; Immunity/genetics ; In Situ Hybridization, Fluorescence ; Insect Vectors/immunology/*microbiology ; Pest Control, Biological/methods ; Phenotype ; Polymerase Chain Reaction ; Real-Time Polymerase Chain Reaction ; Reproduction ; Transfection/*methods ; Wolbachia/genetics/immunology/*pathogenicity ; }, abstract = {BACKGROUND: Biting midges of the genus Culicoides vector multiple veterinary pathogens and are difficult to control. Endosymbionts particularly Wolbachia pipientis may offer an alternative to control populations of Culicoides and/or impact disease transmission in the form of population suppression or replacement strategies.

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

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

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

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

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

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

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

DISCUSSION: The evolutionary lag observed in our system is not caused by a need for changes at multiple genomic locations. The findings favour hypotheses in which either multiple mutations are required within a single genomic region, or the suppressor mutation is a singularly rare event.}, } @article {pmid31568486, year = {2019}, author = {Voronin, D and Schnall, E and Grote, A and Jawahar, S and Ali, W and Unnasch, TR and Ghedin, E and Lustigman, S}, title = {Pyruvate produced by Brugia spp. via glycolysis is essential for maintaining the mutualistic association between the parasite and its endosymbiont, Wolbachia.}, journal = {PLoS pathogens}, volume = {15}, number = {9}, pages = {e1008085}, pmid = {31568486}, issn = {1553-7374}, support = {R56 AI118936/AI/NIAID NIH HHS/United States ; T32 AI007180/AI/NIAID NIH HHS/United States ; F31 AI131527/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Brugia/genetics/*metabolism/*microbiology ; Brugia malayi/genetics/metabolism/microbiology ; Brugia pahangi/genetics/metabolism/microbiology ; Female ; Filariasis/metabolism/microbiology/parasitology ; Genes, Helminth ; Glycolysis ; Host Microbial Interactions ; Host-Parasite Interactions ; Humans ; Male ; Pyruvic Acid/*metabolism ; Symbiosis ; Wolbachia/genetics/*metabolism ; }, abstract = {Human parasitic nematodes are the causative agents of lymphatic filariasis (elephantiasis) and onchocerciasis (river blindness), diseases that are endemic to more than 80 countries and that consistently rank in the top ten for the highest number of years lived with disability. These filarial nematodes have evolved an obligate mutualistic association with an intracellular bacterium, Wolbachia, a symbiont that is essential for the successful development, reproduction, and survival of adult filarial worms. Elimination of the bacteria causes adult worms to die, making Wolbachia a primary target for developing new interventional tools to combat filariases. To further explore Wolbachia as a promising indirect macrofilaricidal drug target, the essential cellular processes that define the symbiotic Wolbachia-host interactions need to be identified. Genomic analyses revealed that while filarial nematodes encode all the enzymes necessary for glycolysis, Wolbachia does not encode the genes for three glycolytic enzymes: hexokinase, 6-phosphofructokinase, and pyruvate kinase. These enzymes are necessary for converting glucose into pyruvate. Wolbachia, however, has the full complement of genes required for gluconeogenesis starting with pyruvate, and for energy metabolism via the tricarboxylic acid cycle. Therefore, we hypothesized that Wolbachia might depend on host glycolysis to maintain a mutualistic association with their parasitic host. We did conditional experiments in vitro that confirmed that glycolysis and its end-product, pyruvate, sustain this symbiotic relationship. Analysis of alternative sources of pyruvate within the worm indicated that the filarial lactate dehydrogenase could also regulate the local intracellular concentration of pyruvate in proximity to Wolbachia and thus help control bacterial growth via molecular interactions with the bacteria. Lastly, we have shown that the parasite's pyruvate kinase, the enzyme that performs the last step in glycolysis, could be a potential novel anti-filarial drug target. Establishing that glycolysis is an essential component of symbiosis in filarial worms could have a broader impact on research focused on other intracellular bacteria-host interactions where the role of glycolysis in supporting intracellular survival of bacteria has been reported.}, } @article {pmid31568480, year = {2019}, author = {Gawande, SJ and Anandhan, S and Ingle, A and Roylawar, P and Khandagale, K and Gawai, T and Jacobson, A and Asokan, R and Singh, M}, title = {Microbiome profiling of the onion thrips, Thrips tabaci Lindeman (Thysanoptera: Thripidae).}, journal = {PloS one}, volume = {14}, number = {9}, pages = {e0223281}, pmid = {31568480}, issn = {1932-6203}, mesh = {Actinobacteria/classification/*genetics/isolation & purification ; Animals ; Bacterial Typing Techniques ; Bacteroidetes/classification/*genetics/isolation & purification ; Cyanobacteria/classification/*genetics/isolation & purification ; Firmicutes/classification/*genetics/isolation & purification ; Gastrointestinal Microbiome/*genetics ; Genetic Variation ; India ; Phylogeny ; Proteobacteria/classification/*genetics/isolation & purification ; RNA, Ribosomal, 16S/genetics ; Symbiosis/genetics ; Thysanoptera/*microbiology ; Nicotiana/parasitology ; Wolbachia/classification/genetics/isolation & purification ; }, abstract = {The gut microbial community structure of adult Thrips tabaci collected from 10 different agro-climatically diverse locations of India was characterized by using the Illumina MiSeq platform to amplify the V3 region of the 16S rRNA gene of bacteria present in the sampled insects. Analyses were performed to study the bacterial communities associated with Thrips tabaci in India. The complete bacterial metagenome of T. tabaci was comprised of 1662 OTUs of which 62.25% belong to known and 37.7% of unidentified/unknown bacteria. These OTUs constituted 21 bacterial phyla of 276 identified genera. Phylum Proteobacteria was predominant, followed by Actinobacteria, Firmicutes, Bacteroidetes and Cyanobacteria. Additionally, the occurrence of the reproductive endosymbiont, Wolbachia was detected at two locations (0.56%) of the total known OTUs. There is high variation in diversity and species richness among the different locations. Alpha-diversity metrics indicated the higher gut bacterial diversity at Bangalore and lowest at Rahuri whereas higher bacterial species richness at T. tabaci samples from Imphal and lowest at Jhalawar. Beta diversity analyses comparing bacterial communities between the samples showed distinct differences in bacterial community composition of T. tabaci samples from different locations. This paper also constitutes the first record of detailed bacterial communities associated with T. tabaci. The location-wise variation in microbial metagenome profile of T. tabaci suggests that bacterial diversity might be governed by its population genetic structure, environment and habitat.}, } @article {pmid31566662, year = {2019}, author = {Detcharoen, M and Arthofer, W and Schlick-Steiner, BC and Steiner, FM}, title = {Wolbachia megadiversity: 99% of these microorganismic manipulators unknown.}, journal = {FEMS microbiology ecology}, volume = {95}, number = {11}, pages = {}, doi = {10.1093/femsec/fiz151}, pmid = {31566662}, issn = {1574-6941}, mesh = {Animals ; Arthropods/microbiology ; *Biodiversity ; Ecosystem ; Symbiosis ; *Wolbachia/genetics ; }, abstract = {Wolbachia (Alphaproteobacteria) are the most widespread endosymbionts of arthropods, manipulating their hosts by various means to maximize the number of host individuals infected. Based on quantitative analyzes of the published literature from Web of Science® and of DNA sequences of arthropod-hosted Wolbachia from GenBank, we made plausible that less than 1% of the expected 100 000 strains of Wolbachia in arthropods is known. Our findings suggest that more and globally better coordinated efforts in screening arthropods are needed to explore the true Wolbachia diversity and to help us understand the ecology and evolution of these host-endosymbiont interactions.}, } @article {pmid31565105, year = {2019}, author = {Huang, K and Kelly, PJ and Zhang, J and Yang, Y and Liu, W and Kalalah, A and Wang, C}, title = {Molecular Detection of Bartonella spp. in China and St. Kitts.}, journal = {The Canadian journal of infectious diseases & medical microbiology = Journal canadien des maladies infectieuses et de la microbiologie medicale}, volume = {2019}, number = {}, pages = {3209013}, pmid = {31565105}, issn = {1712-9532}, abstract = {Bartonella are vector-borne hemotropic bacteria that infect a wide variety of hosts, including people. While there are PCR assays that can identify individual or groups of Bartonella, there is no reliable molecular method to simultaneously detect all species while maintaining genus specificity and sensitivity. By comparing highly conserved 16S rRNA sequences of the better-recognized Bartonella spp. on GenBank, we selected primers and probes for a genus-specific pan-Bartonella FRET-qPCR. Then, a gltA-based Bartonella PCR was established by selecting primers for a highly variable region of gltA, of which the sequenced amplicons could identify individual Bartonella spp. The pan-Bartonella FRET-qPCR did not detect negative controls (Brucella spp., Anaplasma spp., Rickettsia spp., Coxiella burnetii, and Wolbachia) but reliably detected as few as two copies of the positive control (Bartonella henselae) per reaction. There was complete agreement between the pan-Bartonella FRET-qPCR and the gltA-based Bartonella PCR in detecting Bartonella in convenience test samples from China and St. Kitts: cats (26%; 81/310), Ctenocephalides felis (20%; 12/60), cattle (24%; 23/98), and donkeys (4%; 1/20). Sequencing of the gltA-based Bartonella PCR products revealed B. henselae (70%; 57/81) and B. clarridgeiae (30%; 24/81) in cats and C. felis (67%; 8/12, and 33%; 4/12, respectively) and B. bovis in cattle (23.5%; 23/98) and donkeys (4.0%; 1/24). The pan-Bartonella FRET-qPCR and gltA-based Bartonella PCR we developed are highly sensitive and specific in detecting recognized Bartonella spp. in a single reaction. The pan-Bartonella FRET-qPCR is convenient requiring no gel electrophoresis and providing copy numbers, while the gltA-based Bartonella PCR reliably differentiates individual Bartonella species. The use of these PCRs should greatly facilitate large-scale surveillance studies and the diagnosis of infections in clinical samples.}, } @article {pmid31557154, year = {2019}, author = {Andersen, BJ and Rosa, BA and Kupritz, J and Meite, A and Serge, T and Hertz, MI and Curtis, K and King, CL and Mitreva, M and Fischer, PU and Weil, GJ}, title = {Systems analysis-based assessment of post-treatment adverse events in lymphatic filariasis.}, journal = {PLoS neglected tropical diseases}, volume = {13}, number = {9}, pages = {e0007697}, pmid = {31557154}, issn = {1935-2735}, mesh = {Adolescent ; Adult ; Aged ; Albendazole/administration & dosage/adverse effects ; Antigens, Helminth/blood ; Cytokines/blood/immunology ; Diethylcarbamazine/adverse effects/therapeutic use ; Elephantiasis, Filarial/*drug therapy/genetics/immunology ; Female ; Filaricides/adverse effects/*therapeutic use ; Humans ; Ivermectin/administration & dosage/adverse effects ; Male ; Middle Aged ; Young Adult ; }, abstract = {BACKGROUND: Lymphatic filariasis (LF) is a neglected tropical disease, and the Global Program to Eliminate LF delivers mass drug administration (MDA) to 500 million people every year. Adverse events (AEs) are common after LF treatment.

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

This study provides a pilot data on the high presence of Wolbachia in Cx. quinque-fasciatus and Ae. albopictus mosquitoes prevalent in the study region. Whether the high prevalence of Wolbachia and its genetic diversity in mosquitoes affects the mosquitoes' susceptibility to R. felis infection in Georgia will need further evaluation.}, } @article {pmid31396100, year = {2019}, author = {Liu, L and Zhang, KJ and Rong, X and Li, YY and Liu, H}, title = {Identification of Wolbachia-Responsive miRNAs in the Small Brown Planthopper, Laodelphax striatellus.}, journal = {Frontiers in physiology}, volume = {10}, number = {}, pages = {928}, pmid = {31396100}, issn = {1664-042X}, abstract = {Laodelphax striatellus is naturally infected with the Wolbachia strain wStri, which induces strong cytoplasmic incompatibility of its host. MicroRNAs (miRNAs) are a class of endogenous non-coding small RNAs that play a critical role in the regulation of gene expression at post-transcriptional level in various biological processes. Despite various studies reporting that Wolbachia affects the miRNA expression of their hosts, the molecular mechanism underlying interactions between Wolbachia and their host miRNAs has not been well understood. In order to better understand the impact of Wolbachia infection on its host, we investigated the differentially expressed miRNAs between Wolbachia-infected and Wolbachia-uninfected strains of L. striatellus. Compared with uninfected strains, Wolbachia infection resulted in up-regulation of 18 miRNAs and down-regulation of 6 miRNAs in male, while 25 miRNAs were up-regulated and 15 miRNAs were down-regulated in female. The target genes of these differentially expressed miRNAs involved in immune response regulation, reproduction, redox homeostasis and ecdysteroidogenesis were also annotated in both sexes. We further verified the expression of several significantly differentially expressed miRNAs and their predicted target genes by qRT-PCR method. The results suggested that Wolbachia appears to reduce the expression of genes related to fertility in males and increase the expression of genes related to fecundity in females. At the same time, Wolbachia may enhance the expression of immune-related genes in both sexes. All of the results in this study may be helpful in further exploration of the molecular mechanisms by which Wolbachia affects on its hosts.}, } @article {pmid31384012, year = {2019}, author = {Klinges, JG and Rosales, SM and McMinds, R and Shaver, EC and Shantz, AA and Peters, EC and Eitel, M and Wörheide, G and Sharp, KH and Burkepile, DE and Silliman, BR and Vega Thurber, RL}, title = {Phylogenetic, genomic, and biogeographic characterization of a novel and ubiquitous marine invertebrate-associated Rickettsiales parasite, Candidatus Aquarickettsia rohweri, gen. nov., sp. nov.}, journal = {The ISME journal}, volume = {13}, number = {12}, pages = {2938-2953}, pmid = {31384012}, issn = {1751-7370}, mesh = {Animals ; Aquatic Organisms/*microbiology ; Genome, Bacterial ; Genomics ; Gram-Negative Bacterial Infections/microbiology/*veterinary ; Invertebrates/*microbiology ; Parasites/classification/genetics/isolation & purification ; *Phylogeny ; Rickettsiales/classification/genetics/*isolation & purification ; }, abstract = {Bacterial symbionts are integral to the health and homeostasis of invertebrate hosts. Notably, members of the Rickettsiales genus Wolbachia influence several aspects of the fitness and evolution of their terrestrial hosts, but few analogous partnerships have been found in marine systems. We report here the genome, phylogenetics, and biogeography of a ubiquitous and novel Rickettsiales species that primarily associates with marine organisms. We previously showed that this bacterium was found in scleractinian corals, responds to nutrient exposure, and is associated with reduced host growth and increased mortality. This bacterium, like other Rickettsiales, has a reduced genome indicative of a parasitic lifestyle. Phylogenetic analysis places this Rickettsiales within a new genus we define as "Candidatus Aquarickettsia." Using data from the Earth Microbiome Project and SRA databases, we also demonstrate that members of "Ca. Aquarickettsia" are found globally in dozens of invertebrate lineages. The coral-associated "Candidatus A. rohweri" is the first finished genome in this new clade. "Ca. A. rohweri" lacks genes to synthesize most sugars and amino acids but possesses several genes linked to pathogenicity including Tlc, an antiporter that exchanges host ATP for ADP, and a complete Type IV secretion system. Despite its inability to metabolize nitrogen, "Ca. A. rohweri" possesses the NtrY-NtrX two-component system involved in sensing and responding to extracellular nitrogen. Given these data, along with visualization of the parasite in host tissues, we hypothesize that "Ca. A. rohweri" reduces coral health by consuming host nutrients and energy, thus weakening and eventually killing host cells. Last, we hypothesize that nutrient enrichment, which is increasingly common on coral reefs, encourages unrestricted growth of "Ca. A. rohweri" in its host by providing abundant N-rich metabolites to be scavenged.}, } @article {pmid31381563, year = {2019}, author = {Hübner, MP and Koschel, M and Struever, D and Nikolov, V and Frohberger, SJ and Ehrens, A and Fendler, M and Johannes, I and von Geldern, TW and Marsh, K and Turner, JD and Taylor, MJ and Ward, SA and Pfarr, K and Kempf, DJ and Hoerauf, A}, title = {In vivo kinetics of Wolbachia depletion by ABBV-4083 in L. sigmodontis adult worms and microfilariae.}, journal = {PLoS neglected tropical diseases}, volume = {13}, number = {8}, pages = {e0007636}, pmid = {31381563}, issn = {1935-2735}, mesh = {Animals ; Anti-Bacterial Agents/*pharmacology ; Doxycycline/pharmacology ; Female ; Filariasis ; Filarioidea/drug effects/*microbiology ; Gerbillinae ; Kinetics ; Mice ; Mice, Inbred BALB C ; Microfilariae/drug effects/embryology/*microbiology ; Models, Animal ; Wolbachia/*drug effects/*physiology ; }, abstract = {Depletion of Wolbachia endosymbionts of human pathogenic filariae using 4-6 weeks of doxycycline treatment can lead to permanent sterilization and adult filarial death. We investigated the anti-Wolbachia drug candidate ABBV-4083 in the Litomosoides sigmodontis rodent model to determine Wolbachia depletion kinetics with different regimens. Wolbachia reduction occurred in mice as early as 3 days after the initiation of ABBV-4083 treatment and continued throughout a 10-day treatment period. Importantly, Wolbachia levels continued to decline after a 5-day-treatment from 91.5% to 99.9% during a 3-week washout period. In jirds, two weeks of ABBV-4083 treatment (100mg/kg once-per-day) caused a >99.9% Wolbachia depletion in female adult worms, and the kinetics of Wolbachia depletion were recapitulated in peripheral blood microfilariae. Similar to Wolbachia depletion, inhibition of embryogenesis was time-dependent in ABBV-4083-treated jirds, leading to a complete lack of late embryonic stages (stretched microfilariae) and lack of peripheral microfilariae in 5/6 ABBV-4083-treated jirds by 14 weeks after treatment. Twice daily treatment in comparison to once daily treatment with ABBV-4083 did not significantly improve Wolbachia depletion. Moreover, up to 4 nonconsecutive daily treatments within a 14-dose regimen did not significantly erode Wolbachia depletion. Within the limitations of an animal model that does not fully recapitulate human filarial disease, our studies suggest that Wolbachia depletion should be assessed clinically no earlier than 3-4 weeks after the end of treatment, and that Wolbachia depletion in microfilariae may be a viable surrogate marker for the depletion within adult worms. Furthermore, strict daily adherence to the dosing regimen with anti-Wolbachia candidates may not be required, provided that the full regimen is subsequently completed.}, } @article {pmid31380084, year = {2019}, author = {Yoshida, K and Sanada-Morimura, S and Huang, SH and Tokuda, M}, title = {Influences of two coexisting endosymbionts, CI-inducing Wolbachia and male-killing Spiroplasma, on the performance of their host Laodelphax striatellus (Hemiptera: Delphacidae).}, journal = {Ecology and evolution}, volume = {9}, number = {14}, pages = {8214-8224}, pmid = {31380084}, issn = {2045-7758}, abstract = {The small brown planthopper Laodelphax striatellus (Hemiptera: Delphacidae) is reported to have the endosymbiont Wolbachia, which shows a strong cytoplasmic incompatibility (CI) between infected males and uninfected females. In the 2000s, female-biased L. striatellus populations were found in Taiwan, and this sex ratio distortion was the result of male-killing induced by the infection of another endosymbiont, Spiroplasma. Spiroplasma infection is considered to negatively affect both L. striatellus and Wolbachia because the male-killing halves the offspring of L. striatellus and hinders the spread of Wolbachia infection via CI. Spiroplasma could have traits that increase the fitness of infected L. striatellus and/or coexisting organisms because the coinfection rates of Wolbachia and Spiroplasma were rather high in some areas. In this study, we investigated the influences of the infection of these two endosymbionts on the development, reproduction, and insecticide resistance of L. striatellus in the laboratory. Our results show that the single-infection state of Spiroplasma had a negative influence on the fertility of L. striatellus, while the double-infection state had no significant influence. At late nymphal and adult stages, the abundance of Spiroplasma was lower in the double-infection state than in the single-infection state. In the double-infection state, the reduction of Spiroplasma density may be caused by competition between the two endosymbionts, and the negative influence of Spiroplasma on the fertility of host may be relieved. The resistance of L. striatellus to four insecticides was compared among different infection states of endosymbionts, but Spiroplasma infection did not contribute to increase insecticide resistance. Because positive influences of Spiroplasma infection were not found in terms of the development, reproduction, and insecticide resistance of L. striatellus, other factors improving the fitness of Spiroplasma-infected L. striatellus may be related to the high frequency of double infection in some L. striatellus populations.}, } @article {pmid31380078, year = {2019}, author = {Hu, G and Zhang, L and Yun, Y and Peng, Y}, title = {Taking insight into the gut microbiota of three spider species: No characteristic symbiont was found corresponding to the special feeding style of spiders.}, journal = {Ecology and evolution}, volume = {9}, number = {14}, pages = {8146-8156}, pmid = {31380078}, issn = {2045-7758}, abstract = {Microorganisms in insect guts have been recognized as having a great impact on their hosts' nutrition, health, and behavior. Spiders are important natural enemies of pests, and the composition of the gut microbiota of spiders remains unclear. Will the bacterial taxa in spiders be same as the bacterial taxa in insects, and what are the potential functions of the gut bacteria in spiders? To gain insight into the composition of the gut bacteria in spiders and their potential function, we collected three spider species, Pardosa laura, Pardosa astrigera, and Nurscia albofasciata, in the field, and high-throughput sequencing of the 16S rRNA V3 and V4 regions was used to investigate the diversity of gut microbiota across the three spider species. A total of 23 phyla and 150 families were identified in these three spider species. The dominant bacterial phylum across all samples was Proteobacteria. Burkholderia, Ralstonia, Ochrobactrum, Providencia, Acinetobacter, Proteus, and Rhodoplanes were the dominant genera in the guts of the three spider species. The relative abundances of Wolbachia and Rickettsiella detected in N. albofasciata were significantly higher than those in the other two spider species. The relative abundance of Thermus, Amycolatopsis, Lactococcus, Acinetobacter Microbacterium, and Koribacter detected in spider gut was different among the three spider species. Biomolecular interaction networks indicated that the microbiota in the guts had complex interactions. The results of this study also suggested that at the genus level, some of the gut bacteria taxa in the three spider species were the same as the bacteria in insect guts.}, } @article {pmid31380036, year = {2019}, author = {Su, Q and Wang, X and Ilyas, N and Zhang, F and Yun, Y and Jian, C and Peng, Y}, title = {Combined effects of elevated CO2 concentration and Wolbachia on Hylyphantes graminicola (Araneae: Linyphiidae).}, journal = {Ecology and evolution}, volume = {9}, number = {12}, pages = {7112-7121}, pmid = {31380036}, issn = {2045-7758}, abstract = {The increasing concentration of carbon dioxide in atmosphere is not only a major cause of global warming, but it also adversely affects the ecological diversity of invertebrates. This study was conducted to evaluate the effect of elevated CO2 concentration (ambient, 400 ppm and high, 800 ppm) and Wolbachia (Wolbachia-infected, W[+] and Wolbachia-uninfected, W[-]) on Hylyphantes graminicola. The total survival rate, developmental duration, carapace width and length, body weight, sex ratio, net reproductive rate, nutrition content, and enzyme activity in H. graminicola were examined under four treatments: W[-] 400 ppm, W[-] 800 ppm, W[+] 400 ppm, and W[+] 800 ppm. Results showed that Wolbachia-infected spiders had significantly decreased the total developmental duration. Different instars showed variations up to some extent, but no obvious effect was found under elevated CO2 concentration. Total survival rate, sex ratio, and net reproductive rate were not affected by elevated CO2 concentration or Wolbachia infection. The carapace width of Wolbachia-uninfected spiders decreased significantly under elevated CO2 concentration, while the width, length and weight were not significantly affected in Wolbachia-infected spiders reared at ambient CO2 concentration. The levels of protein, specific activities of peroxidase, and amylase were significantly increased under elevated CO2 concentration or Wolbachia-infected spiders, while the total amino content was only increased in Wolbachia-infected spiders. Thus, our current finding suggested that elevated CO2 concentration and Wolbachia enhance nutrient contents and enzyme activity of H. graminicola and decrease development duration hence explore the interactive effects of factors which were responsible for reproduction regulation, but it also gives a theoretical direction for spider's protection in such a dynamic environment. Increased activities of enzymes and nutrients caused by Wolbachia infection aids for better survival of H. graminicola under stress.}, } @article {pmid31375435, year = {2019}, author = {Dimopoulos, G}, title = {Combining Sterile and Incompatible Insect Techniques for Aedes albopictus Suppression.}, journal = {Trends in parasitology}, volume = {35}, number = {9}, pages = {671-673}, doi = {10.1016/j.pt.2019.07.006}, pmid = {31375435}, issn = {1471-5007}, support = {R01 AI141532/AI/NIAID NIH HHS/United States ; R01 AI122743/AI/NIAID NIH HHS/United States ; R21 AI131574/AI/NIAID NIH HHS/United States ; }, mesh = {*Aedes ; Animals ; *Dengue ; *Infertility ; Male ; Mosquito Vectors ; *Wolbachia ; }, abstract = {Traditional control strategies are failing to contain Aedes albopictus as an emerging major vector for dengue. A combination of approaches (Zheng et al., Nature, 2019) involving an artificial triple Wolbachia superinfection and low-dose irradiation enabled mass production of adult sterile males for release. The resulting suppression of field populations suggests feasibility for area-wide vector control.}, } @article {pmid31369548, year = {2019}, author = {Schneider, DI and Saarman, N and Onyango, MG and Hyseni, C and Opiro, R and Echodu, R and O'Neill, M and Bloch, D and Vigneron, A and Johnson, TJ and Dion, K and Weiss, BL and Opiyo, E and Caccone, A and Aksoy, S}, title = {Spatio-temporal distribution of Spiroplasma infections in the tsetse fly (Glossina fuscipes fuscipes) in northern Uganda.}, journal = {PLoS neglected tropical diseases}, volume = {13}, number = {8}, pages = {e0007340}, pmid = {31369548}, issn = {1935-2735}, support = {D43 TW007391/TW/FIC NIH HHS/United States ; R01 AI068932/AI/NIAID NIH HHS/United States ; R01 AI139525/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Coinfection ; DNA, Ribosomal/genetics ; Female ; Gram-Negative Bacterial Infections/*microbiology/*veterinary ; Insect Vectors/*microbiology/parasitology ; Male ; Prevalence ; Spiroplasma/genetics/*pathogenicity/physiology ; Symbiosis ; Trypanosoma ; Tsetse Flies/*microbiology/parasitology ; Uganda ; Wolbachia ; }, abstract = {Tsetse flies (Glossina spp.) are vectors of parasitic trypanosomes, which cause human (HAT) and animal African trypanosomiasis (AAT) in sub-Saharan Africa. In Uganda, Glossina fuscipes fuscipes (Gff) is the main vector of HAT, where it transmits Gambiense disease in the northwest and Rhodesiense disease in central, southeast and western regions. Endosymbionts can influence transmission efficiency of parasites through their insect vectors via conferring a protective effect against the parasite. It is known that the bacterium Spiroplasma is capable of protecting its Drosophila host from infection with a parasitic nematode. This endosymbiont can also impact its host's population structure via altering host reproductive traits. Here, we used field collections across 26 different Gff sampling sites in northern and western Uganda to investigate the association of Spiroplasma with geographic origin, seasonal conditions, Gff genetic background and sex, and trypanosome infection status. We also investigated the influence of Spiroplasma on Gff vector competence to trypanosome infections under laboratory conditions. Generalized linear models (GLM) showed that Spiroplasma probability was correlated with the geographic origin of Gff host and with the season of collection, with higher prevalence found in flies within the Albert Nile (0.42 vs 0.16) and Achwa River (0.36 vs 0.08) watersheds and with higher prevalence detected in flies collected in the intermediate than wet season. In contrast, there was no significant correlation of Spiroplasma prevalence with Gff host genetic background or sex once geographic origin was accounted for in generalized linear models. Additionally, we found a potential negative correlation of Spiroplasma with trypanosome infection, with only 2% of Spiroplasma infected flies harboring trypanosome co-infections. We also found that in a laboratory line of Gff, parasitic trypanosomes are less likely to colonize the midgut in individuals that harbor Spiroplasma infection. These results indicate that Spiroplasma infections in tsetse may be maintained by not only maternal but also via horizontal transmission routes, and Spiroplasma infections may also have important effects on trypanosome transmission efficiency of the host tsetse. Potential functional effects of Spiroplasma infection in Gff could have impacts on vector control approaches to reduce trypanosome infections.}, } @article {pmid33447371, year = {2019}, author = {Durovni, B and Saraceni, V and Eppinghaus, A and Riback, TIS and Moreira, LA and Jewell, NP and Dufault, SM and O'Neill, SL and Simmons, CP and Tanamas, SK and Anders, KL}, title = {The impact of large-scale deployment of Wolbachia mosquitoes on dengue and other Aedes-borne diseases in Rio de Janeiro and Niterói, Brazil: study protocol for a controlled interrupted time series analysis using routine disease surveillance data.}, journal = {F1000Research}, volume = {8}, number = {}, pages = {1328}, pmid = {33447371}, issn = {2046-1402}, mesh = {Aedes/*virology ; Animals ; Biological Control Agents ; Brazil/epidemiology ; *Chikungunya Fever/epidemiology/prevention & control ; Cities ; *Dengue/epidemiology/prevention & control ; Dengue Virus ; Humans ; Incidence ; Interrupted Time Series Analysis ; *Mosquito Control ; Mosquito Vectors/virology ; *Wolbachia ; Zika Virus ; *Zika Virus Infection/epidemiology/prevention & control ; }, abstract = {Background: Rio de Janeiro and Niterói are neighbouring cities in southeastern Brazil which experience large dengue epidemics every 2 to 5 years, with >100,000 cases notified in epidemic years. Costs of vector control and direct and indirect costs due to the Aedes-borne diseases dengue, chikungunya and Zika were estimated to total $650 million USD in 2016, but traditional vector control strategies have not been effective in preventing mosquito-borne disease outbreaks. The Wolbachia method is a novel and self-sustaining approach for the biological control of Aedes-borne diseases, in which the transmission potential of Aedes aegypti mosquitoes is reduced by stably transfecting them with the Wolbachia bacterium (wMel strain). This paper describes a study protocol for evaluating the effect of large-scale non-randomised releases of Wolbachia--infected mosquitoes on the incidence of dengue, Zika and chikungunya in the two cities of Niterói and Rio de Janeiro. This follows a lead-in period since 2014 involving intensive community engagement, regulatory and public approval, entomological surveys, and small-scale pilot releases. Method: The Wolbachia releases during 2017-2019 covered a combined area of 170 km [2] with a resident population of 1.2 million, across Niterói and Rio de Janeiro. Untreated areas with comparable historical dengue profiles and demographic characteristics have been identified a priori as comparative control areas in each city. The proposed pragmatic epidemiological approach combines a controlled interrupted time series analysis of routinely notified suspected and laboratory-confirmed dengue and chikungunya cases, together with monitoring of Aedes-borne disease activity utilising outbreak signals routinely used in public health disease surveillance. Discussion: If the current project is successful, this model for control of mosquito-borne disease through Wolbachia releases can be expanded nationally and regionally.}, } @article {pmid31362350, year = {2019}, author = {Satjawongvanit, H and Phumee, A and Tiawsirisup, S and Sungpradit, S and Brownell, N and Siriyasatien, P and Preativatanyou, K}, title = {Molecular Analysis of Canine Filaria and Its Wolbachia Endosymbionts in Domestic Dogs Collected from Two Animal University Hospitals in Bangkok Metropolitan Region, Thailand.}, journal = {Pathogens (Basel, Switzerland)}, volume = {8}, number = {3}, pages = {}, pmid = {31362350}, issn = {2076-0817}, abstract = {Canine filariasis is caused by several nematode species, such as Dirofilaria immitis, Dirofilaria repens, Brugia pahangi, Brugia malayi, and Acanthocheilonema reconditum. Zoonotic filariasis is one of the world's neglected tropical diseases. Since 2000, the World Health Organization (WHO) has promoted a global filarial eradication program to eliminate filariasis by 2020. Apart from vector control strategies, the infection control of reservoir hosts is necessary for more effective filariasis control. In addition, many studies have reported that Wolbachia is necessary for the development, reproduction, and survival of the filarial nematode. Consequently, the use of antibiotics to kill Wolbachia in nematodes has now become an alternative strategy to control filariasis. Previously, a case of subconjunctival dirofilariasis caused by Dirofilaria spp. has been reported in a woman who resides in the center of Bangkok, Thailand. Therefore, our study aimed to principally demonstrate the presence of filarial nematodes and Wolbachia bacteria in blood collected from domestic dogs from the Bangkok Metropolitan Region, Thailand. A total of 57 blood samples from dogs with suspected dirofilariasis who had visited veterinary clinics in Bangkok were collected. The investigations for the presence of microfilaria were carried out by using both microscopic and molecular examinations. PCR was used as the molecular detection method for the filarial nematodes based on the COI and ITS1 regions. The demonstration of Wolbachia was performed using PCR to amplify the FtsZ gene. All positive samples by PCR were then cloned and sequenced. The results showed that the filarial nematodes were detected in 16 samples (28.07%) using microscopic examinations. The molecular detection of filarial species using COI-PCR revealed that 50 samples (87.72%) were positive; these consisted of 33 (57.89%), 13 (22.81%), and 4 (7.02%) samples for D. immitis, B. pahangi, and B. malayi, respectively. While the ITS1-PCR showed that 41 samples (71.93%) were positive-30 samples (52.63%) were identified as containing D. immitis and 11 samples (19.30%) were identified to have B. pahangi, whereas B. malayi was not detected. Forty-seven samples (82.45%) were positive for Wolbachia DNA and the phylogenetic tree of all positive Wolbachia was classified into the supergroup C clade. This study has established fundamental data on filariasis associated with Wolbachia infection in domestic dogs in the Bangkok Metropolitan Region. An extensive survey of dog blood samples would provide valuable epidemiologic data on potential zoonotic filariasis in Thailand. In addition, this information could be used for the future development of more effective prevention and control strategies for canine filariasis in Thailand.}, } @article {pmid31358201, year = {2019}, author = {Liu, XD and Guo, HF}, title = {Importance of endosymbionts Wolbachia and Rickettsia in insect resistance development.}, journal = {Current opinion in insect science}, volume = {33}, number = {}, pages = {84-90}, doi = {10.1016/j.cois.2019.05.003}, pmid = {31358201}, issn = {2214-5753}, mesh = {Animals ; Inactivation, Metabolic ; Insecta/*microbiology/physiology ; *Insecticide Resistance ; Insecticides/toxicity ; Rickettsia/*physiology ; Symbiosis ; Wolbachia/*physiology ; }, abstract = {Endosymbionts play important roles in protecting hosts from environmental stress, such as natural enemies, heat, and toxins. Many insects are infected with the facultative nonessential endosymbionts Wolbachia and Rickettsia, which are the crux in this review, although other relevant symbiont genera will also be treated. Insecticide resistance of hosts can be related to infections with Wolbachia and Rickettsia. These endosymbionts commonly increase host susceptibility to chemical insecticides, but cases of increased resistance also exist. The symbiont-mediated insecticide resistance/susceptibility varies with species of insect, species of symbiont, and chemical compound. Changes in insecticide resistance levels of insects can be associated with fluctuations in population density of endosymbionts. Effects of endosymbionts on host fitness, metabolism, immune system, and gene expression may determine how endosymbionts influence insecticide resistance. A clearer understanding of these interactions can improve our knowledge about drivers of decreasing insecticide resistance.}, } @article {pmid31351474, year = {2019}, author = {Singh, JA}, title = {Informed consent and community engagement in open field research: lessons for gene drive science.}, journal = {BMC medical ethics}, volume = {20}, number = {1}, pages = {54}, pmid = {31351474}, issn = {1472-6939}, mesh = {Animals ; *Communicable Disease Control/methods ; *Community Participation ; Community-Based Participatory Research/*ethics/methods/standards ; Culicidae/genetics ; Dengue/prevention & control ; Disease Transmission, Infectious/prevention & control ; Gene Drive Technology/*ethics/methods ; Humans ; *Informed Consent/ethics/standards ; Mosquito Control/ethics/methods ; Mosquito Vectors/genetics ; Randomized Controlled Trials as Topic/ethics ; Trust ; Wolbachia ; }, abstract = {BACKGROUND: The development of the CRISPR/Cas9 gene editing system has generated new possibilities for the use of gene drive constructs to reduce or suppress mosquito populations to levels that do not support disease transmission. Despite this prospect, social resistance to genetically modified organisms remains high. Gene drive open field research thus raises important questions regarding what is owed to those who may not consent to such research, or those could be affected by the proposed research, but whose consent is not solicited. The precise circumstances under which informed consent must be obtained, and from whom, requires careful consideration. Furthermore, appropriate engagement processes should be central to any introduction of genetically modified mosquitos in proposed target settings.

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

CONCLUSIONS: Our results indicate that the bacterial microbiomes of I. scapularis ticks vary by sex and geography, with significantly more diversity in male microbiomes compared to females. We found that sex plays a larger role than geography in shaping the composition/diversity of the I. scapularis microbiome, but that geography affects what additional taxa are represented (beyond Rickettsia) and whether pathogens are found. Furthermore, recent feeding may have a role in shaping the tick microbiome, as evident from a more complex bacterial community in female ticks from dogs compared to the wild-caught questing females. These findings may provide further insight into the differences in the ability of the ticks to acquire, maintain and transmit pathogens. Future studies on possible causes and consequences of these differences will shed additional light on tick microbiome biology and vector competence.}, } @article {pmid31233646, year = {2019}, author = {Hinojosa, JC and Koubínová, D and Szenteczki, MA and Pitteloud, C and Dincă, V and Alvarez, N and Vila, R}, title = {A mirage of cryptic species: Genomics uncover striking mitonuclear discordance in the butterfly Thymelicus sylvestris.}, journal = {Molecular ecology}, volume = {28}, number = {17}, pages = {3857-3868}, doi = {10.1111/mec.15153}, pmid = {31233646}, issn = {1365-294X}, mesh = {Animals ; Bayes Theorem ; Butterflies/*genetics ; Cell Nucleus/*genetics ; Electron Transport Complex IV/genetics ; Genetic Loci ; *Genomics ; Likelihood Functions ; Mitochondria/*genetics ; Phylogeny ; Polymorphism, Single Nucleotide/genetics ; Species Specificity ; }, abstract = {Mitochondrial DNA (mtDNA) sequencing has led to an unprecedented rise in the identification of cryptic species. However, it is widely acknowledged that nuclear DNA (nuDNA) sequence data are also necessary to properly define species boundaries. Next generation sequencing techniques provide a wealth of nuclear genomic data, which can be used to ascertain both the evolutionary history and taxonomic status of putative cryptic species. Here, we focus on the intriguing case of the butterfly Thymelicus sylvestris (Lepidoptera: Hesperiidae). We identified six deeply diverged mitochondrial lineages; three distributed all across Europe and found in sympatry, suggesting a potential case of cryptic species. We then sequenced these six lineages using double-digest restriction-site associated DNA sequencing (ddRADseq). Nuclear genomic loci contradicted mtDNA patterns and genotypes generally clustered according to geography, i.e., a pattern expected under the assumption of postglacial recolonization from different refugia. Further analyses indicated that this strong mtDNA/nuDNA discrepancy cannot be explained by incomplete lineage sorting, sex-biased asymmetries, NUMTs, natural selection, introgression or Wolbachia-mediated genetic sweeps. We suggest that this mitonuclear discordance was caused by long periods of geographic isolation followed by range expansions, homogenizing the nuclear but not the mitochondrial genome. These results highlight T. sylvestris as a potential case of multiple despeciation and/or lineage fusion events. We finally argue, since mtDNA and nuDNA do not necessarily follow the same mechanisms of evolution, their respective evolutionary history reflects complementary aspects of past demographic and biogeographic events.}, } @article {pmid31229556, year = {2020}, author = {Parvathi, A and Jasna, V and Aswathy, VK and Aparna, S and Nathan, VK and Jyothibabu, R}, title = {Dominance of Wolbachia sp. in the deep-sea sediment bacterial metataxonomic sequencing analysis in the Bay of Bengal, Indian Ocean.}, journal = {Genomics}, volume = {112}, number = {1}, pages = {1030-1041}, doi = {10.1016/j.ygeno.2019.06.019}, pmid = {31229556}, issn = {1089-8646}, mesh = {Alphaproteobacteria/isolation & purification ; Bacteria/classification/genetics/isolation & purification ; Biodiversity ; Firmicutes/isolation & purification ; Geologic Sediments/chemistry/*microbiology ; High-Throughput Nucleotide Sequencing ; Indian Ocean ; Metagenomics ; Oceans and Seas ; Proteobacteria/isolation & purification ; Sequence Analysis, DNA ; Wolbachia/genetics/*isolation & purification ; }, abstract = {The Bay of Bengal, located in the north-eastern part of the Indian Ocean is world's largest bay occupying an area of ~8,39,000 mile[2]. The variability in bacterial community structure and function in sediment ecosystems of the Bay of Bengal is examined by Illumina high-throughput metagenomic sequencing. Of five metataxonomics data sets presented, two (SD1 and SD2) were from stations close to the shore and three (SD4, SD5, and SD6) were from the deep-sea (~3000 m depth). Phylum Proteobacteria (90.27 to 92.52%) dominated the deep-sea samples, whereas phylum Firmicutes (65.35 to 90.98%) dominated the coastal samples. Comparative analysis showed that coastal and deep-sea sediments showed distinct microbial communities. Wolbachia species, belonging to class Alphaproteobacteria was the most dominant species in the deep-sea sediments. The gene functions of bacterial communities were predicted for deep-sea and coastal sediment ecosystems. The results indicated that deep-sea sediment bacterial communities were involved in metabolic activities like dehalogenation and sulphide oxidation.}, } @article {pmid31229468, year = {2019}, author = {Bock, W and Jayathunga, Y}, title = {Optimal control of a multi-patch Dengue model under the influence of Wolbachia bacterium.}, journal = {Mathematical biosciences}, volume = {315}, number = {}, pages = {108219}, doi = {10.1016/j.mbs.2019.108219}, pmid = {31229468}, issn = {1879-3134}, mesh = {Aedes/*microbiology ; Animals ; *Basic Reproduction Number ; Dengue/*prevention & control/*transmission ; Humans ; *Models, Biological ; *Mosquito Vectors ; *Pest Control, Biological ; *Wolbachia/pathogenicity ; }, abstract = {In this work, a multi-patch model for dengue transmission dynamics including the bacterium Wolbachia is studied and by that the control efforts to minimize the disease spread by host and vector control are investigated. The multi-patch system models the host movement within the patches which coupled via a residence-time budgeting matrix P. Numerical results confirm that the control mechanism embedded in incidence rates of the disease transmission, effectively reduce the spread of the disease.}, } @article {pmid31227544, year = {2019}, author = {Cooper, BS and Vanderpool, D and Conner, WR and Matute, DR and Turelli, M}, title = {Wolbachia Acquisition by Drosophila yakuba-Clade Hosts and Transfer of Incompatibility Loci Between Distantly Related Wolbachia.}, journal = {Genetics}, volume = {212}, number = {4}, pages = {1399-1419}, pmid = {31227544}, issn = {1943-2631}, support = {R01 GM104325/GM/NIGMS NIH HHS/United States ; R01 GM121750/GM/NIGMS NIH HHS/United States ; R35 GM124701/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Drosophila/*genetics/microbiology/physiology ; Female ; *Gene Transfer, Horizontal ; Genome, Bacterial ; Genome, Insect ; Genome, Mitochondrial ; Host-Pathogen Interactions ; Infertility/*genetics ; Male ; Phylogeny ; Wolbachia/*genetics/pathogenicity ; }, abstract = {Maternally transmitted Wolbachia infect about half of insect species, yet the predominant mode(s) of Wolbachia acquisition remains uncertain. Species-specific associations could be old, with Wolbachia and hosts codiversifying (i.e., cladogenic acquisition), or relatively young and acquired by horizontal transfer or introgression. The three Drosophila yakuba-clade hosts [(D. santomea, D. yakuba) D. teissieri] diverged ∼3 MYA and currently hybridize on the West African islands Bioko and São Tomé. Each species is polymorphic for nearly identical Wolbachia that cause weak cytoplasmic incompatibility (CI)-reduced egg hatch when uninfected females mate with infected males. D. yakuba-clade Wolbachia are closely related to wMel, globally polymorphic in D. melanogaster We use draft Wolbachia and mitochondrial genomes to demonstrate that D. yakuba-clade phylogenies for Wolbachia and mitochondria tend to follow host nuclear phylogenies. However, roughly half of D. santomea individuals, sampled both inside and outside of the São Tomé hybrid zone, have introgressed D. yakuba mitochondria. Both mitochondria and Wolbachia possess far more recent common ancestors than the bulk of the host nuclear genomes, precluding cladogenic Wolbachia acquisition. General concordance of Wolbachia and mitochondrial phylogenies suggests that horizontal transmission is rare, but varying relative rates of molecular divergence complicate chronogram-based statistical tests. Loci that cause CI in wMel are disrupted in D. yakuba-clade Wolbachia; but a second set of loci predicted to cause CI are located in the same WO prophage region. These alternative CI loci seem to have been acquired horizontally from distantly related Wolbachia, with transfer mediated by flanking Wolbachia-specific ISWpi1 transposons.}, } @article {pmid31222094, year = {2019}, author = {Mateos, M and Silva, NO and Ramirez, P and Higareda-Alvear, VM and Aramayo, R and Erickson, JW}, title = {Effect of heritable symbionts on maternally-derived embryo transcripts.}, journal = {Scientific reports}, volume = {9}, number = {1}, pages = {8847}, pmid = {31222094}, issn = {2045-2322}, support = {R03 AI078348/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Drosophila melanogaster/*embryology/genetics/*microbiology ; Embryo, Nonmammalian/*microbiology ; Female ; Genes, Insect/genetics ; Host-Pathogen Interactions/genetics ; Male ; Phenotype ; RNA, Ribosomal ; Reproduction/genetics ; Ribosome Inactivating Proteins/genetics/physiology ; Sequence Analysis, RNA ; Spiroplasma/enzymology ; *Symbiosis ; Transcriptome/*genetics ; Wolbachia ; }, abstract = {Maternally-transmitted endosymbiotic bacteria are ubiquitous in insects. Among other influential phenotypes, many heritable symbionts of arthropods are notorious for manipulating host reproduction through one of four reproductive syndromes, which are generally exerted during early developmental stages of the host: male feminization; parthenogenesis induction; male killing; and cytoplasmic incompatibility (CI). Major advances have been achieved in understanding mechanisms and identifying symbiont factors involved in reproductive manipulation, particularly male killing and cytoplasmic incompatibility. Nonetheless, whether cytoplasmically-transmitted bacteria influence the maternally-loaded components of the egg or early embryo has not been examined. In the present study, we investigated whether heritable endosymbionts that cause different reproductive phenotypes in Drosophila melanogaster influence the mRNA transcriptome of early embryos. We used mRNA-seq to evaluate differential expression in Drosophila embryos lacking endosymbionts (control) to those harbouring the male-killing Spiroplasma poulsonii strain MSRO-Br, the CI-inducing Wolbachia strain wMel, or Spiroplasma poulsonii strain Hyd1; a strain that lacks a reproductive phenotype and is naturally associated with Drosophila hydei. We found no consistent evidence of influence of symbiont on mRNA composition of early embryos, suggesting that the reproductive manipulation mechanism does not involve alteration of maternally-loaded transcripts. In addition, we capitalized on several available mRNA-seq datasets derived from Spiroplasma-infected Drosophila melanogaster embryos, to search for signals of depurination of rRNA, consistent with the activity of Ribosome Inactivating Proteins (RIPs) encoded by Spiroplasma poulsonii. We found small but statistically significant signals of depurination of Drosophila rRNA in the Spiroplasma treatments (both strains), but not in the symbiont-free control or Wolbachia treatment, consistent with the action of RIPs. The depurination signal was slightly stronger in the treatment with the male-killing strain. This result supports a recent report that RIP-induced damage contributes to male embryo death.}, } @article {pmid31215133, year = {2020}, author = {Su, QC and Wang, X and Deng, C and Yun, YL and Zhao, Y and Peng, Y}, title = {Transcriptome responses to elevated CO2 level and Wolbachia-infection stress in Hylyphantes graminicola (Araneae: Linyphiidae).}, journal = {Insect science}, volume = {27}, number = {5}, pages = {908-920}, doi = {10.1111/1744-7917.12701}, pmid = {31215133}, issn = {1744-7917}, support = {2016YFD0200900//State's Key Project of Research and Development Plan/ ; 31672317//National Natural Science Foundation of China/ ; 2016jzxjh012//Competitive Planning Projects of Hubei Academy of Agricultural Sciences/ ; }, mesh = {Animals ; Carbon Dioxide/*physiology ; Female ; Spiders/drug effects/*genetics/microbiology ; Stress, Physiological ; Transcriptome/drug effects/*genetics ; Wolbachia/*physiology ; }, abstract = {Hylyphantes graminicola is a resident spider species found in maize and cotton fields and is an important biological control agent of various pests. Previous studies have demonstrated that stress from elevated CO2 and Wolbachia infection can strongly affect spider species. Thus, based on CO2 levels (400 ppm, current atmospheric CO2 concentration and 800 ppm, high CO2 concentration) and Wolbachia status (Wolbachia-infected, W[+] and Wolbachia-uninfected, W[-]), we divided H. graminicola individuals into four treatment groups: W[-] 400 ppm, W[-] 800 ppm, W[+] 400 ppm, and W[+] 800 ppm. To investigate the effects of elevated CO2 levels (W[-] 400 vs W[-] 800), Wolbachia infection (W[-] 400 vs W[+] 400), and the interactions between these two factors (W[-] 400 vs W[+] 800), high-throughput transcriptome sequencing was employed to characterize the de novo transcriptome of the spiders and identify stress-related differentially expressed genes (DEGs). De novo assembly of complementary DNA sequences generated 86 688 unigenes, 23 938 of which were annotated in public databases. A total of 84, 21, and 157 DEGs were found among W[-] 400 vs W[-] 800, W[-] 400 vs W[+] 400, and W[-] 400 vs W[+] 800, respectively. Functional enrichment analysis revealed that metabolic processes, signaling, and catalytic activity were significantly affected by elevated CO2 levels and Wolbachia infection. Our findings suggest that the impact of elevated CO2 levels and Wolbachia infection on the H. graminicola transcriptome was, to a large extent, on genes involved in metabolic processes. This study is the first description of transcriptome changes in response to elevated CO2 levels and Wolbachia infection in spiders.}, } @article {pmid31214820, year = {2019}, author = {Konecka, E and Olszanowski, Z}, title = {First Evidence of Intracellular Bacteria Cardinium in Thermophilic Mite Microzetorchestes emeryi (Acari: Oribatida): Molecular Screening of Bacterial Endosymbiont Species.}, journal = {Current microbiology}, volume = {76}, number = {9}, pages = {1038-1044}, pmid = {31214820}, issn = {1432-0991}, mesh = {Animals ; Bacteroidetes/classification/*genetics/*isolation & purification/physiology ; DNA, Bacterial/genetics ; DNA, Ribosomal/genetics ; Mites/*microbiology/physiology ; Phylogeny ; Poland ; Symbiosis ; }, abstract = {We undertook the issue of the distribution of intracellular bacteria among Oribatida (Acari). Six genera of bacteria were detected by PCR and Sanger DNA sequencing: Wolbachia, Cardinium, Rickettsia, Spiroplasma, Arsenophonus, and Hamiltonella. Our research, for the first time, revealed the presence of Cardinium in Microzetorchestes emeryi in two subpopulations separated from each other by 300 m. The percentages of infected animals were the same in both subpopulations-ca. 20%. The identity of 16S rDNA sequences of Cardinium between these two subpopulations of M. emeryi was 97%. Phylogenetic analysis showed that the Cardinium in M. emeryi was clustered into the group A. The occurrence of M. emeryi in Poland has not been reported before and our report is the first one. Cardinium maybe help the thermophilic M. emeryi to adapt to low temperatures in the Central Europe.}, } @article {pmid31208002, year = {2019}, author = {Schebeck, M and Feldkirchner, L and Stauffer, C and Schuler, H}, title = {Dynamics of an Ongoing Wolbachia Spread in the European Cherry Fruit Fly, Rhagoletis cerasi (Diptera: Tephritidae).}, journal = {Insects}, volume = {10}, number = {6}, pages = {}, pmid = {31208002}, issn = {2075-4450}, support = {J 3527/FWF_/Austrian Science Fund FWF/Austria ; P 26749/FWF_/Austrian Science Fund FWF/Austria ; I 2604/FWF_/Austrian Science Fund FWF/Austria ; P 31441/FWF_/Austrian Science Fund FWF/Austria ; }, abstract = {Numerous terrestrial arthropods are infected with the alphaproteobacterium Wolbachia. This endosymbiont is usually transmitted vertically from infected females to their offspring and can alter the reproduction of hosts through various manipulations, like cytoplasmic incompatibility (CI), enhancing its spread in new host populations. Studies on the spatial and temporal dynamics of Wolbachia under natural conditions are scarce. Here, we analyzed Wolbachia infection frequencies in populations of the European cherry fruit fly, Rhagoletis cerasi (L.), in central Germany-an area of an ongoing spread of the CI-inducing strain wCer2. In total, 295 individuals from 19 populations were PCR-screened for the presence of wCer2 and their mitochondrial haplotype. Results were compared with historic data to understand the infection dynamics of the ongoing wCer2 invasion. An overall wCer2 infection frequency of about 30% was found, ranging from 0% to 100% per population. In contrast to an expected smooth transition from wCer2-infected to completely wCer2-uninfected populations, a relatively scattered infection pattern across geography was observed. Moreover, a strong Wolbachia-haplotype association was detected, with only a few rare misassociations. Our results show a complex dynamic of an ongoing Wolbachia spread in natural field populations of R. cerasi.}, } @article {pmid31204356, year = {2019}, author = {Watanabe, A and Takaku, S and Yokota, K and Hayashi, S and Tamaki, N and Kokeguchi, S}, title = {A survey of Lasioderma serricorne (Fabricius) in Japanese Dental Clinics.}, journal = {Biocontrol science}, volume = {24}, number = {2}, pages = {117-121}, doi = {10.4265/bio.24.117}, pmid = {31204356}, issn = {1884-0205}, mesh = {Animals ; Bacteria/*classification/*isolation & purification ; Cluster Analysis ; Coleoptera/*microbiology ; DNA, Bacterial/chemistry/genetics ; DNA, Ribosomal/chemistry/genetics ; *Dental Clinics ; Japan ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA ; }, abstract = {This study was to survey the capturing rate in Japanese dental clinics of the Lasioderma serricorne (cigarette beetles) , and to evaluate the beetle's potential as a carrier for transmission of nosocomial pathogens. L. serricorne imagoes were captured in pheromone traps in 14 Japanese dental clinics in August and September 2012 and 2013, and their numbers recorded. Polymerase chain reaction (PCR) for the bacterial antibiotic-resistant genes mecA, vanA, vanB, blaIMP, and blaVIM was performed on the captured L. serricorne imagoes. Bacterial species in the captured specimens were identified by 16S rRNA PCR and sequencing analysis. The L. serricorne imagoes were captured from 10 dental clinics (71.4%) . We failed to detect the presence of nosocomial antibiotic-resistant pathogens in L. serricorne imagoes. The bacterial species detected most commonly in the imagoes was Wolbachia sp., an intracellular proteobacterium infecting certain insect species. Monitoring of insects including L. serricorne should be incorporated into regiment of the infection control.}, } @article {pmid31194893, year = {2020}, author = {Ant, TH and Herd, C and Louis, F and Failloux, AB and Sinkins, SP}, title = {Wolbachia transinfections in Culex quinquefasciatus generate cytoplasmic incompatibility.}, journal = {Insect molecular biology}, volume = {29}, number = {1}, pages = {1-8}, pmid = {31194893}, issn = {1365-2583}, support = {202888/WT_/Wellcome Trust/United Kingdom ; 108508/A/15/Z/WT_/Wellcome Trust/United Kingdom ; 202888/Z/16/Z/WT_/Wellcome Trust/United Kingdom ; MC_UU_12014/8/MRC_/Medical Research Council/United Kingdom ; BB/K004506/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom ; /WT_/Wellcome Trust/United Kingdom ; 108508/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Aedes/microbiology ; Animals ; Culex/*microbiology ; Female ; Male ; Mosquito Control/methods ; Mosquito Vectors/microbiology ; Pest Control, Biological/methods ; Salivary Glands/microbiology ; Symbiosis ; Wolbachia/classification/*physiology ; }, abstract = {Culex quinquefasciatus is an important mosquito vector of a number of viral and protozoan pathogens of humans and animals, and naturally carries the endosymbiont Wolbachia pipientis, strain wPip. Wolbachia are used in two distinct vector control strategies: firstly, population suppression caused by mating incompatibilities between mass-released transinfected males and wild females; and secondly, the spread of pathogen transmission-blocking strains through populations. Using embryonic microinjection, two novel Wolbachia transinfections were generated in C. quinquefasciatus using strains native to the mosquito Aedes albopictus: a wAlbB single infection, and a wPip plus wAlbA superinfection. The wAlbB infection showed full bidirectional cytoplasmic incompatibility (CI) with wild-type C. quinquefasciatus in reciprocal crosses. The wPipwAlbA superinfection showed complete unidirectional CI, and therefore population invasion potential. Whereas the wAlbB strain showed comparatively low overall densities, similar to the native wPip, the wPipwAlbA superinfection reached over 400-fold higher densities in the salivary glands compared to the native wPip, suggesting it may be a candidate for pathogen transmission blocking.}, } @article {pmid31191980, year = {2019}, author = {Koh, C and Audsley, MD and Di Giallonardo, F and Kerton, EJ and Young, PR and Holmes, EC and McGraw, EA}, title = {Sustained Wolbachia-mediated blocking of dengue virus isolates following serial passage in Aedes aegypti cell culture.}, journal = {Virus evolution}, volume = {5}, number = {1}, pages = {vez012}, pmid = {31191980}, issn = {2057-1577}, abstract = {Wolbachia is an intracellular endosymbiont of insects that inhibits the replication of a range of pathogens in its arthropod hosts. The release of Wolbachia into wild populations of mosquitoes is an innovative biocontrol effort to suppress the transmission of arthropod-borne viruses (arboviruses) to humans, most notably dengue virus. The success of the Wolbachia-based approach hinges upon the stable persistence of the 'pathogen blocking' effect, whose mechanistic basis is poorly understood. Evidence suggests that Wolbachia may affect viral replication via a combination of competition for host resources and activation of host immunity. The evolution of resistance against Wolbachia and pathogen blocking in the mosquito or the virus could reduce the public health impact of the symbiont releases. Here, we investigate if dengue 3 virus (DENV-3) is capable of accumulating adaptive mutations that improve its replicative capacity during serial passage in Wolbachia wMel-infected cells. During the passaging regime, viral isolates in Wolbachia-infected cells exhibited greater variation in viral loads compared to controls. The viral loads of these isolates declined rapidly during passaging due to the blocking effects of Wolbachia carriage, with several being lost all together and the remainder recovering to low but stable levels. We attempted to sequence the genomes of the surviving passaged isolates but, given their low abundance, were unable to obtain sufficient depth of coverage for evolutionary analysis. In contrast, viral loads in Wolbachia-free control cells were consistently high during passaging. The surviving isolates passaged in the presence of Wolbachia exhibited a reduced ability to replicate even in Wolbachia-free cells. These experiments demonstrate the challenge for dengue in evolving resistance to Wolbachia-mediated blocking.}, } @article {pmid31186318, year = {2019}, author = {Chrostek, E and Gerth, M}, title = {Is Anopheles gambiae a Natural Host of Wolbachia?.}, journal = {mBio}, volume = {10}, number = {3}, pages = {}, pmid = {31186318}, issn = {2150-7511}, mesh = {Africa ; Animals ; Anopheles/*microbiology ; Female ; Genome, Bacterial ; Malaria/transmission ; Male ; Metagenomics ; Mosquito Vectors/*microbiology ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; *Symbiosis ; Wolbachia/*genetics/isolation & purification ; }, abstract = {Wolbachia (Alphaproteobacteria, Rickettsiales) is an intraovarially transmitted symbiont of insects able to exert striking phenotypes, including reproductive manipulations and pathogen blocking. These phenotypes make Wolbachia a promising tool to combat mosquito-borne diseases. Although Wolbachia is present in the majority of terrestrial arthropods, including many disease vectors, it was considered absent from Anopheles gambiae mosquitos, the main vectors of malaria in sub-Saharan Africa. In 2014, Wolbachia sequences were detected in A. gambiae samples collected in Burkina Faso. Subsequently, similar evidence came from collections all over Africa, revealing a high Wolbachia 16S rRNA sequence diversity, low abundance, and a lack of congruence between host and symbiont phylogenies. Here, we reanalyze and discuss recent evidence on the presence of Wolbachia sequences in A. gambiae. We find that although detected at increasing frequencies, the unusual properties of these Wolbachia sequences render them insufficient to diagnose natural infections in A. gambiae Future studies should focus on uncovering the origin of Wolbachia sequence variants in Anopheles and seeking sequence-independent evidence for this new symbiosis. Understanding the ecology of Anopheles mosquitos and their interactions with Wolbachia will be key in designing successful, integrative approaches to limit malaria spread. Although the prospect of using Wolbachia to fight malaria is intriguing, the newly discovered strains do not bring it closer to realization.IMPORTANCEAnopheles gambiae mosquitos are the main vectors of malaria, threatening around half of the world's population. The bacterial symbiont Wolbachia can interfere with disease transmission by other important insect vectors, but until recently, it was thought to be absent from natural A. gambiae populations. Here, we critically analyze the genomic, metagenomic, PCR, imaging, and phenotypic data presented in support of the presence of natural Wolbachia infections in A. gambiae We find that they are insufficient to diagnose Wolbachia infections and argue for the need of obtaining robust data confirming basic Wolbachia characteristics in this system. Determining the Wolbachia infection status of Anopheles is critical due to its potential to influence Anopheles population structure and Plasmodium transmission.}, } @article {pmid31176069, year = {2019}, author = {Caragata, EP and Tikhe, CV and Dimopoulos, G}, title = {Curious entanglements: interactions between mosquitoes, their microbiota, and arboviruses.}, journal = {Current opinion in virology}, volume = {37}, number = {}, pages = {26-36}, pmid = {31176069}, issn = {1879-6265}, support = {R01 AI078997/AI/NIAID NIH HHS/United States ; R21 AI136456/AI/NIAID NIH HHS/United States ; R01 AI141532/AI/NIAID NIH HHS/United States ; R01 AI081877/AI/NIAID NIH HHS/United States ; R01 AI121253/AI/NIAID NIH HHS/United States ; R21 AI131574/AI/NIAID NIH HHS/United States ; R01 AI061576/AI/NIAID NIH HHS/United States ; R01 AI122743/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/immunology/microbiology/virology ; Animals ; Anopheles/immunology/microbiology/virology ; Arbovirus Infections/immunology/microbiology/virology ; Arboviruses/pathogenicity ; Biological Control Agents/immunology ; Culex/immunology/microbiology/virology ; Dengue Virus/pathogenicity ; Host Microbial Interactions/*immunology ; Humans ; Immunity, Innate ; Microbial Interactions/*immunology ; Microbiota ; *Mosquito Vectors/immunology/microbiology/virology ; Virus Diseases/immunology/microbiology/virology ; Wolbachia/*immunology ; }, abstract = {Mosquitoes naturally harbor a diverse community of microorganisms that play a crucial role in their biology. Mosquito-microbiota interactions are abundant and complex. They can dramatically alter the mosquito immune response, and impede or enhance a mosquito's ability to transmit medically important arboviral pathogens. Yet critically, given the massive public health impact of arboviral disease, few such interactions have been well characterized. In this review, we describe the current state of knowledge of the role of microorganisms in mosquito biology, how microbial-induced changes to mosquito immunity moderate infection with arboviruses, cases of mosquito-microbial-virus interactions with a defined mechanism, and the molecular interactions that underlie the endosymbiotic bacterium Wolbachia's ability to block virus infection in mosquitoes.}, } @article {pmid31174466, year = {2019}, author = {Baião, GC and Schneider, DI and Miller, WJ and Klasson, L}, title = {The effect of Wolbachia on gene expression in Drosophila paulistorum and its implications for symbiont-induced host speciation.}, journal = {BMC genomics}, volume = {20}, number = {1}, pages = {465}, pmid = {31174466}, issn = {1471-2164}, support = {P 22634/FWF_/Austrian Science Fund FWF/Austria ; P 28255/FWF_/Austrian Science Fund FWF/Austria ; 2014-4353//Vetenskapsrådet/ ; P28255-B22//Austrian Science Fund/ ; }, mesh = {Amino Acids/metabolism ; Animals ; Carbohydrate Metabolism/genetics ; Drosophila/*genetics/metabolism/*microbiology ; Female ; *Genetic Speciation ; Immunity/genetics ; Lipid Metabolism/genetics ; Male ; Muscles/metabolism ; Pheromones/metabolism ; Proteolysis ; RNA-Seq ; Reproduction/genetics ; *Symbiosis ; Transcriptome ; Wolbachia/*physiology ; }, abstract = {BACKGROUND: The Neotropical fruit fly Drosophila paulistorum (Diptera: Drosophilidae) is a species complex in statu nascendi comprising six reproductively isolated semispecies, each harboring mutualistic Wolbachia strains. Although wild type flies of each semispecies are isolated from the others by both pre- and postmating incompatibilities, mating between semispecies and successful offspring development can be achieved once flies are treated with antibiotics to reduce Wolbachia titer. Here we use RNA-seq to study the impact of Wolbachia on D. paulistorum and investigate the hypothesis that the symbiont may play a role in host speciation. For that goal, we analyze samples of heads and abdomens of both sexes of the Amazonian, Centro American and Orinocan semispecies of D. paulistorum.

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

CONCLUSIONS: We report the first documentation of Ae. fluviatilis in the municipality of Saboeiro, northeastern Brazil. It is important to conduct additional entomological surveys to characterize the local Culicidae fauna. Ignoring the presence and dispersion of this vector could be a public health risk.}, } @article {pmid30807733, year = {2019}, author = {Bagheri, Z and Talebi, AA and Asgari, S and Mehrabadi, M}, title = {Wolbachia induce cytoplasmic incompatibility and affect mate preference in Habrobracon hebetor to increase the chance of its transmission to the next generation.}, journal = {Journal of invertebrate pathology}, volume = {163}, number = {}, pages = {1-7}, doi = {10.1016/j.jip.2019.02.005}, pmid = {30807733}, issn = {1096-0805}, mesh = {Animals ; Biological Control Agents ; Female ; Fertility ; *Host Microbial Interactions ; *Infectious Disease Transmission, Vertical ; Male ; Sex Ratio ; Sexual Behavior, Animal ; Symbiosis ; Wasps/*microbiology ; *Wolbachia/isolation & purification ; }, abstract = {Wolbachia are common intracellular bacteria that are generally found in arthropods, including a high proportion of insects and also some nematodes. This intracellular symbiont can affect sex ratio with a variety of reproductive anomalies in the host, including cytoplasmic incompatibility (CI) in haplodiploids. In this study, we questioned if the parasitoid wasp, Habrobracon hebetor (Hym.: Braconidae), an important biological control agent of many lepidopteran larvae, is infected with Wolbachia. To test this, DNA was extracted from adult insects and subjected to PCR using specific primers to Wolbachia target genes. The results showed a high rate of Wolbachia infection in this parasitoid wasp. To determine the biological function of Wolbachia in H. hebetor, we removed this bacterium from the wasps using antibiotic treatment (cured wasps). Results of crossing experiments revealed that Wolbachia induced CI in H. hebetor in which cured females crossed with infected males produced only males, while both male and female progeny were observed for other crosses. Also, we showed that the presence of Wolbachia in females increased fecundity and female offspring of this parasitoid wasp. The presence of Wolbachia in the males had no significant effect on fecundity and female production, but might have incurred costs. We also investigated the effect of Wolbachia on mate choice and found that Wolbachia affects mating behavior of H. hebetor. Together, we showed that Wolbachia induces CI in H. hebetor and affects host mating behavior in favor of its transmission. Wolbachia utilize these strategies to increase the frequency of infected females in the host population.}, } @article {pmid30798391, year = {2019}, author = {Singhal, K and Mohanty, S}, title = {Genome organisation and comparative genomics of four novel Wolbachia genome assemblies from Indian Drosophila host.}, journal = {Functional & integrative genomics}, volume = {19}, number = {4}, pages = {617-632}, pmid = {30798391}, issn = {1438-7948}, mesh = {Animals ; Bacterial Proteins/genetics/metabolism ; Drosophila/microbiology ; *Genome, Bacterial ; Host-Pathogen Interactions ; *Polymorphism, Genetic ; Symbiosis ; Wolbachia/*genetics/pathogenicity ; }, abstract = {Wolbachia has long been known to share an endosymbiotic relationship with its host as an obligate intracellular organism. Wolbachia diversity as different supergroups is found to be host-specific in most cases except a few, where the host species is seen to accommodate multiple strains. Besides, the Wolbachia genome must have undergone several changes in response to the evolving host genome in order to adapt and establish a strong association with its host, thus making a distinctive Wolbachia-host alliance. The present study focusses on four novel genome assembly and genome-wide sequence variations of Indian Wolbachia strains, i.e. wMel and wRi isolated from two different Drosophila hosts. The genome assembly has an average size of ~ 1.1 Mb and contains ~ 1100 genes, which is comparable with the previously sequenced Wolbachia genomes. The comparative genomics analysis of these genomes and sequence-wide comparison of some functionally significant genes, i.e. ankyrin repeats, Wsp and T4SS, highlight their sequence similarities and dissimilarities, further supporting the strain-specific association of Wolbachia to its host. Interestingly, some of the sequence variations are also found to be restricted to only Indian Wolbachia strains. Further analysis of prophage and their flanking regions in the Wolbachia genome reveals the presence of several functional genes which may assist the phage to reside inside the bacterial host, thus providing a trade-off for the endosymbiont-host association. Understanding this endosymbiont genome in different eco-geographical conditions has become imperative for the recent use of Wolbachia in medical entomology as a vector-control agent.}, } @article {pmid30796064, year = {2019}, author = {Newton, ILG and Slatko, BE}, title = {Symbiosis Comes of age at the 10[th] Biennial Meeting of Wolbachia Researchers.}, journal = {Applied and environmental microbiology}, volume = {85}, number = {8}, pages = {}, pmid = {30796064}, issn = {1098-5336}, support = {R21 AI137918/AI/NIAID NIH HHS/United States ; }, abstract = {Wolbachia pipientis is an alpha-proteobacterial, obligate intracellular microbe and arguably the most successful infection on our planet, colonizing 40-60% of insect species. Wolbachia are also present in most, but not all, filarial nematodes where they are obligate mutualists and are the targets for anti-filarial drug discovery. Although Wolbachia are related to important human pathogens they do not infect mammals, but instead are well known for their reproductive manipulations of insect populations, inducing the following phenotypes: male-killing, feminization, parthenogenesis induction, or cytoplasmic incompatibility (CI). The most common of these, CI, results in a sperm-egg incompatibility and increases the relative fecundity of infected females in a population. In the last decade, Wolbachia have also been shown to provide a benefit to insects, where the infection can inhibit RNA virus replication within the host. Wolbachia cannot be cultivated outside of host cells and no genetic tools are available in the symbiont, limiting approaches available to its study. This means that many questions fundamental to our understanding of Wolbachia basic biology remained unknown for decades. The tenth biennial international Wolbachia conference, "Wolbachia Evolution, Ecology, Genomics and Cell Biology: A Chronicle of the Most Ubiquitous Symbiont", was held on June 17-22, 2018, Salem, MA USA. In the review below we highlight the new science presented at the meeting, link it to prior efforts to answer these questions across the Wolbachia genus, and the importance to the field of symbiosis. The topics covered in this review are based on the presentations at the conference.}, } @article {pmid30762095, year = {2019}, author = {Martínez-Rodríguez, P and Rolán-Alvarez, E and Del Mar Pérez-Ruiz, M and Arroyo-Yebras, F and Carpena-Catoira, C and Carvajal-Rodríguez, A and Bella, JL}, title = {Geographic and Temporal Variation of Distinct Intracellular Endosymbiont Strains of Wolbachia sp. in the Grasshopper Chorthippus parallelus: a Frequency-Dependent Mechanism?.}, journal = {Microbial ecology}, volume = {77}, number = {4}, pages = {1036-1047}, pmid = {30762095}, issn = {1432-184X}, support = {CGL2016-75482-P//Ministerio de Ciencia y Tecnolog?a/ ; BFU2013-44635//Ministerio de Econom?a, Industria y Competitividad, Gobierno de Espa?a/ ; }, mesh = {Animals ; Biological Coevolution ; Computer Simulation ; Geography ; Grasshoppers/*microbiology ; Linear Models ; *Polymorphism, Genetic ; Seasons ; *Symbiosis ; Wolbachia/genetics/*physiology ; }, abstract = {Wolbachia is an intracellular endosymbiont that can produce a range of effects on host fitness, but the temporal dynamics of Wolbachia strains have rarely been experimentally evaluated. We compare interannual strain frequencies along a geographical region for understanding the forces that shape Wolbachia strain frequency in natural populations of its host, Chorthippus parallelus (Orthoptera, Acrididae). General linear models show that strain frequency changes significantly across geographical and temporal scales. Computer simulation allows to reject the compatibility of the observed patterns with either genetic drift or sampling errors. We use consecutive years to estimate total Wolbachia strain fitness. Our estimation of Wolbachia fitness is significant in most cases, within locality and between consecutive years, following a negatively frequency-dependent trend. Wolbachia spp. B and F strains show a temporal pattern of variation that is compatible with a negative frequency-dependent natural selection mechanism. Our results suggest that such a mechanism should be at least considered in future experimental and theoretical research strategies that attempt to understand Wolbachia biodiversity.}, } @article {pmid30761981, year = {2019}, author = {Showler, AJ and Kubofcik, J and Ricciardi, A and Nutman, TB}, title = {Differences in the Clinical and Laboratory Features of Imported Onchocerciasis in Endemic Individuals and Temporary Residents.}, journal = {The American journal of tropical medicine and hygiene}, volume = {100}, number = {5}, pages = {1216-1222}, pmid = {30761981}, issn = {1476-1645}, mesh = {Acute Disease ; Adult ; Animals ; Antibodies, Helminth/blood ; Asymptomatic Infections ; Chronic Disease ; Clinical Laboratory Techniques/*statistics & numerical data ; *Emigration and Immigration ; Endemic Diseases/*statistics & numerical data ; Eye Diseases/diagnosis/parasitology ; Female ; Humans ; Immunoglobulin G/blood ; Male ; Middle Aged ; Onchocerca volvulus ; Onchocerciasis/blood/*diagnosis ; }, abstract = {Many parasitic infections have different presenting features in endemic individuals (ENDs) and immunologically naive temporary residents (TRs). Temporary residents with loiasis often display acute symptoms and hypereosinophilia, in contrast to a parasite-induced subclinical state in chronically infected ENDs. Few studies have examined differences in ENDs and TRs infected with the related filarial parasite Onchocerca volvulus. We identified 40 TRs and 36 ENDs with imported onchocerciasis at the National Institutes of Health between 1976 and 2016. All study subjects received an extensive pretreatment medical history, physical examination, and laboratory investigations. We performed additional parasite-specific serologic testing on stored patient sera. Asymptomatic infection occurred in 12.5% of TRs and no ENDs (P = 0.06). Papular dermatitis was more common in TRs (47.5% versus 2.7%, P < 0.001), whereas more pigmentation changes occurred in ENDs (41.7% versus 15%, P = 0.01). Only endemic patients reported visual disturbance (13% versus 0%, P = 0.03). One TR (3.3%) had onchocercal eye disease, compared with 22.6% of ENDs (P = 0.053). Absolute eosinophil counts (AECs) were similar in ENDs and TRs (P = 0.5), and one-third of subjects had a normal AEC. Endemic individuals had higher filarial-specific IgG4 and were more likely to be positive for IgG4 antibodies to Ov-16. Temporary residents and ENDs with imported O. volvulus infection presented with different dermatologic manifestations; ocular involvement occurred almost exclusively in ENDs. Unlike Loa loa, clinical differences appear not to be eosinophil-mediated and may reflect chronicity, intensity of infection, or the presence of Wolbachia in O. volvulus.}, } @article {pmid30747079, year = {2019}, author = {Chegeni, TN and Fakhar, M}, title = {Promising Role of Wolbachia as Anti-parasitic Drug Target and Eco-Friendly Biocontrol Agent.}, journal = {Recent patents on anti-infective drug discovery}, volume = {14}, number = {1}, pages = {69-79}, doi = {10.2174/1574891X14666190211162403}, pmid = {30747079}, issn = {2212-4071}, mesh = {Animals ; Antiparasitic Agents/*pharmacology/*therapeutic use ; Humans ; Parasites/*drug effects ; Patents as Topic ; Wolbachia/*drug effects ; }, abstract = {BACKGROUND: Wolbachia is the most common endosymbiotic bacteria in insectborne parasites and it is the most common reproductive parasite in the world. Wolbachia has been found worldwide in numerous arthropod and parasite species, including insects, terrestrial isopods, spiders, mites and filarial nematodes. There is a complicated relationship between Wolbachia and its hosts and in some cases, they create a mutual relationship instead of a parasitic relationship. Some species are not able to reproduce in the absence of infection with Wolbachia. Thus, the use of existing strains of Wolbachia bacteria offers a potential strategy for the control of the population of mosquitoes and other pests and diseases.

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

CONCLUSIONS: This study revealed differences in the prevalence of Wolbachia and trypanosomes in G. p. palpalis according to HAT foci. The use of only one marker has underestimated the prevalence of Wolbachia, thus more markers in subsequent studies may improve its detection. The presence of Wolbachia seems to have no impact on the establishment of trypanosomes in G. p. palpalis. The tripartite association between tsetse, Wolbachia and trypanosomes varies according to studied areas. Studies aiming to evaluate the genetic polymorphism of Wolbachia and its density in tsetse flies could help to better understand this association.}, } @article {pmid30483601, year = {2018}, author = {Jeffries, CL and Lawrence, GG and Golovko, G and Kristan, M and Orsborne, J and Spence, K and Hurn, E and Bandibabone, J and Tantely, LM and Raharimalala, FN and Keita, K and Camara, D and Barry, Y and Wat'senga, F and Manzambi, EZ and Afrane, YA and Mohammed, AR and Abeku, TA and Hedge, S and Khanipov, K and Pimenova, M and Fofanov, Y and Boyer, S and Irish, SR and Hughes, GL and Walker, T}, title = {Novel Wolbachia strains in Anopheles malaria vectors from Sub-Saharan Africa.}, journal = {Wellcome open research}, volume = {3}, number = {}, pages = {113}, pmid = {30483601}, issn = {2398-502X}, support = {U01CK000512/ACL/ACL HHS/United States ; R21 AI124452/AI/NIAID NIH HHS/United States ; R21 AI138074/AI/NIAID NIH HHS/United States ; R21 AI129507/AI/NIAID NIH HHS/United States ; /WT_/Wellcome Trust/United Kingdom ; R01 AI123074/AI/NIAID NIH HHS/United States ; }, abstract = {Background: Wolbachia, a common insect endosymbiotic bacterium that can influence pathogen transmission and manipulate host reproduction, has historically been considered absent from the Anopheles (An.) genera, but has recently been found in An. gambiae s.l. populations in West Africa. As there are numerous Anopheles species that have the capacity to transmit malaria, we analysed a range of species across five malaria endemic countries to determine Wolbachia prevalence rates, characterise novel Wolbachia strains and determine any correlation between the presence of Plasmodium, Wolbachia and the competing bacterium Asaia. Methods: Anopheles adult mosquitoes were collected from five malaria-endemic countries: Guinea, Democratic Republic of the Congo (DRC), Ghana, Uganda and Madagascar, between 2013 and 2017. Molecular analysis was undertaken using quantitative PCR, Sanger sequencing, Wolbachia multilocus sequence typing (MLST) and high-throughput amplicon sequencing of the bacterial 16S rRNA gene. Results: Novel Wolbachia strains were discovered in five species: An. coluzzii, An. gambiae s.s., An. arabiensis, An. moucheti and An. species A, increasing the number of Anopheles species known to be naturally infected. Variable prevalence rates in different locations were observed and novel strains were phylogenetically diverse, clustering with Wolbachia supergroup B strains. We also provide evidence for resident strain variants within An. species A. Wolbachia is the dominant member of the microbiome in An. moucheti and An. species A but present at lower densities in An. coluzzii. Interestingly, no evidence of Wolbachia/Asaia co-infections was seen and Asaia infection densities were shown to be variable and location dependent. Conclusions: The important discovery of novel Wolbachia strains in Anopheles provides greater insight into the prevalence of resident Wolbachia strains in diverse malaria vectors. Novel Wolbachia strains (particularly high-density strains) are ideal candidate strains for transinfection to create stable infections in other Anopheles mosquito species, which could be used for population replacement or suppression control strategies.}, } @article {pmid30535314, year = {2019}, author = {Leimeister, CA and Schellhorn, J and Dörrer, S and Gerth, M and Bleidorn, C and Morgenstern, B}, title = {Prot-SpaM: fast alignment-free phylogeny reconstruction based on whole-proteome sequences.}, journal = {GigaScience}, volume = {8}, number = {3}, pages = {}, pmid = {30535314}, issn = {2047-217X}, mesh = {Amino Acid Sequence ; Animals ; Bacteria/classification ; Databases, Protein ; *Phylogeny ; Plants/classification ; Proteome/*chemistry ; Sequence Alignment/*methods ; *Software ; }, abstract = {Word-based or 'alignment-free' sequence comparison has become an active research area in bioinformatics. While previous word-frequency approaches calculated rough measures of sequence similarity or dissimilarity, some new alignment-free methods are able to accurately estimate phylogenetic distances between genomic sequences. One of these approaches is Filtered Spaced Word Matches. Here, we extend this approach to estimate evolutionary distances between complete or incomplete proteomes; our implementation of this approach is called Prot-SpaM. We compare the performance of Prot-SpaM to other alignment-free methods on simulated sequences and on various groups of eukaryotic and prokaryotic taxa. Prot-SpaM can be used to calculate high-quality phylogenetic trees for dozens of whole-proteome sequences in a matter of seconds or minutes and often outperforms other alignment-free approaches. The source code of our software is available through Github: https://github.com/jschellh/ProtSpaM.}, } @article {pmid30535231, year = {2019}, author = {Qi, LD and Sun, JT and Hong, XY and Li, YX}, title = {Diversity and Phylogenetic Analyses Reveal Horizontal Transmission of Endosymbionts Between Whiteflies and Their Parasitoids.}, journal = {Journal of economic entomology}, volume = {112}, number = {2}, pages = {894-905}, doi = {10.1093/jee/toy367}, pmid = {30535231}, issn = {1938-291X}, mesh = {Animals ; China ; *Hemiptera ; Phylogeny ; Symbiosis ; *Wolbachia ; }, abstract = {Endosymbionts are widely distributed among insects via intraspecific vertical transmission and interspecific horizontal transmission. Parasitoids have attracted considerable interest due to their possible role in the horizontal transmission of endosymbionts. Horizontal transmission of endosymbionts between whiteflies via parasitoids has been revealed in the laboratory. However, whether this occurs under field conditions remains unknown. Here, the diversity and phylogenetic relationships of endosymbionts in 1,350 whiteflies and 36 parasitoids that emerged from whitefly nymphs collected from three locations in Jiangsu Province of China were investigated. Only Rickettsia and Wolbachia were identified in both whiteflies and parasitoids, with an overall infection frequency of 22.67% in whiteflies and 16.67% in parasitoids for Wolbachia and of 12.15% in whiteflies and 25% in parasitoids for Rickettsia. Despite the distant relationship between whiteflies and their parasitoids, phylogenetic analyses revealed that the Rickettsia and Wolbachia individuals collected from the two types of organisms were grouped together. Furthermore, shared haplotypes were also identified, which was consistent with the horizontal transmission of endosymbionts between parasitoids and whiteflies. In addition, a parasitoid resistance-related symbiont, Hamiltonella, was detected in whiteflies at a 100% infection frequency, probably accounting for the relatively low parasitism of the whiteflies in the field. The factors affecting the infection frequency of the four secondary endosymbionts in whiteflies were also examined.}, } @article {pmid30534598, year = {2018}, author = {Chung, M and Munro, JB and Tettelin, H and Dunning Hotopp, JC}, title = {Using Core Genome Alignments To Assign Bacterial Species.}, journal = {mSystems}, volume = {3}, number = {6}, pages = {}, pmid = {30534598}, issn = {2379-5077}, support = {U19 AI110820/AI/NIAID NIH HHS/United States ; }, abstract = {With the exponential increase in the number of bacterial taxa with genome sequence data, a new standardized method to assign species designations is needed that is consistent with classically obtained taxonomic analyses. This is particularly acute for unculturable, obligate intracellular bacteria with which classically defined methods, like DNA-DNA hybridization, cannot be used, such as those in the Rickettsiales. In this study, we generated nucleotide-based core genome alignments for a wide range of genera with classically defined species, as well as those within the Rickettsiales. We created a workflow that uses the length, sequence identity, and phylogenetic relationships inferred from core genome alignments to assign genus and species designations that recapitulate classically obtained results. Using this method, most classically defined bacterial genera have a core genome alignment that is ≥10% of the average input genome length. Both Anaplasma and Neorickettsia fail to meet this criterion, indicating that the taxonomy of these genera should be reexamined. Consistently, genomes from organisms with the same species epithet have ≥96.8% identity of their core genome alignments. Additionally, these core genome alignments can be used to generate phylogenomic trees to identify monophyletic clades that define species and neighbor-network trees to assess recombination across different taxa. By these criteria, Wolbachia organisms are delineated into species different from the currently used supergroup designations, while Rickettsia organisms are delineated into 9 distinct species, compared to the current 27 species. By using core genome alignments to assign taxonomic designations, we aim to provide a high-resolution, robust method to guide bacterial nomenclature that is aligned with classically obtained results. IMPORTANCE With the increasing availability of genome sequences, we sought to develop and apply a robust, portable, and high-resolution method for the assignment of genera and species designations that can recapitulate classically defined taxonomic designations. Using cutoffs derived from the lengths and sequence identities of core genome alignments along with phylogenetic analyses, we sought to evaluate or reevaluate genus- and species-level designations for diverse taxa, with an emphasis on the order Rickettsiales, where species designations have been applied inconsistently. Our results indicate that the Rickettsia genus has an overabundance of species designations, that the current Anaplasma and Neorickettsia genus designations are both too broad and need to be divided, and that there are clear demarcations of Wolbachia species that do not align precisely with the existing supergroup designations.}, } @article {pmid30533772, year = {2018}, author = {Chung, M and Teigen, L and Libro, S and Bromley, RE and Kumar, N and Sadzewicz, L and Tallon, LJ and Foster, JM and Michalski, ML and Dunning Hotopp, JC}, title = {Multispecies Transcriptomics Data Set of Brugia malayi, Its Wolbachia Endosymbiont wBm, and Aedes aegypti across the B. malayi Life Cycle.}, journal = {Microbiology resource announcements}, volume = {7}, number = {18}, pages = {}, pmid = {30533772}, issn = {2576-098X}, support = {U19 AI110820/AI/NIAID NIH HHS/United States ; }, abstract = {Here, we present a comprehensive transcriptomics data set of Brugia malayi, its Wolbachia endosymbiont wBm, and its vector host. This study samples from 16 stages across the entire B. malayi life cycle, including stage 1 through 4 larvae, adult males and females, embryos, immature microfilariae, and mature microfilariae.}, } @article {pmid30533608, year = {2018}, author = {Newton, ILG and Sheehan, KB}, title = {Gateway Entry Vector Library of Wolbachia pipientis Candidate Effectors from Strain wMel.}, journal = {Microbiology resource announcements}, volume = {7}, number = {1}, pages = {}, pmid = {30533608}, issn = {2576-098X}, support = {R21 AI137918/AI/NIAID NIH HHS/United States ; }, abstract = {Wolbachia pipientis is an intracellular symbiont that modifies host biology using a type IV secretion system to inject bacterial effectors into the host cytoplasm. We utilized a bioinformatics approach to predict Wolbachia effectors and cloned the candidates into an entry vector, which can be utilized for subsequent analyses.}, } @article {pmid30503908, year = {2019}, author = {McLean, BJ and Dainty, KR and Flores, HA and O'Neill, SL}, title = {Differential suppression of persistent insect specific viruses in trans-infected wMel and wMelPop-CLA Aedes-derived mosquito lines.}, journal = {Virology}, volume = {527}, number = {}, pages = {141-145}, pmid = {30503908}, issn = {1096-0341}, support = {/WT_/Wellcome Trust/United Kingdom ; 102591/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Aedes/*microbiology/*virology ; Animals ; Cell Line ; Coinfection ; Flavivirus/*physiology ; Microbial Interactions ; Mosquito Vectors/microbiology/virology ; Orthobunyavirus/*physiology ; RNA, Viral/genetics/metabolism ; Species Specificity ; Virus Replication ; Wolbachia/*physiology ; }, abstract = {Wolbachia suppresses the replication of +ssRNA viruses such as dengue and Zika viruses in Aedes aegypti mosquitoes. However, the range of viruses affected by this endosymbiont is yet to be explored. Recently, novel insect-specific viruses (ISVs) have been described from numerous mosquito species and mosquito-derived cell lines. Cell-fusing agent virus (Flaviviridae) and Phasi Charoen-like virus (Bunyaviridae) persistently infect the Ae. aegypti cell line Aag2 which has been used for experimental studies with both the wMel and wMelPop-CLA strains. Wolbachia was found to restrict the replication of CFAV but not the PCLV infection in these lines. Furthermore, an additional Ae. albopictus cell line (RML-12) which contained either wMel or wMelPop-CLA was assessed. While no infectious +ssRNA or dsRNA viruses were detected, a PCLV infection was identified. These observations provide additional evidence to support that insect-specific, +ssRNA viruses can be suppressed in cell culture by Wolbachia but -ssRNA viruses may not.}, } @article {pmid30514847, year = {2018}, author = {Rahimi-Kaldeh, S and Ashouri, A and Bandani, A and Ris, N}, title = {Abiotic and biotic factors influence diapause induction in sexual and asexual strains of Trichogramma brassicae (Hym: Trichogrammatidae).}, journal = {Scientific reports}, volume = {8}, number = {1}, pages = {17600}, pmid = {30514847}, issn = {2045-2322}, mesh = {Animals ; Diapause, Insect/*physiology ; Photoperiod ; Reproduction, Asexual/*physiology ; Sexual Behavior, Animal/*physiology ; Temperature ; Wasps/*growth & development ; Wolbachia/pathogenicity ; }, abstract = {The effects of some abiotic (maternal photoperiod and offspring developmental temperature) and biotic (host quality during both maternal and offspring generations) factors on diapause induction were investigated for two sympatric strains of Trichogramma brassicae Bezdenko (Hym: Trichogrammatidae) differing by infection status with regard to Wolbachia. The mode of reproduction, developmental temperature, maternal photoperiod and the quality of the host significantly affected diapause induction. The highest percentage of diapausing individuals were observed with the sexual strain reared at 14 °C using a "high quality" host and after a long photophase during the maternal generation. Environment-by-Environment, as well as Genotype-by-Environment interactions, was observed. All these patterns were discussed with the goal of providing relevant protocols for the commercial mass-rearing of such biocontrol agents. A successful diapause in beneficial insects could affect the efficiency of mass rearing by increasing the duration of storage conditions based on a high percent emergence and providing a large number of individuals at the appropriate time in the field season.}, } @article {pmid30498246, year = {2018}, author = {Jeffries, CL and Tantely, LM and Raharimalala, FN and Hurn, E and Boyer, S and Walker, T}, title = {Diverse novel resident Wolbachia strains in Culicine mosquitoes from Madagascar.}, journal = {Scientific reports}, volume = {8}, number = {1}, pages = {17456}, pmid = {30498246}, issn = {2045-2322}, support = {//Wellcome Trust/United Kingdom ; }, mesh = {Alleles ; Animals ; *Biodiversity ; Culicidae/*microbiology ; DNA, Mitochondrial ; Female ; Geography ; Madagascar ; Mosquito Vectors/*microbiology ; Multilocus Sequence Typing ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Wolbachia/*classification/*genetics ; }, abstract = {Wolbachia endosymbiotic bacteria are widespread throughout insect species and Wolbachia transinfected in Aedes mosquito species has formed the basis for biocontrol programs as Wolbachia strains inhibit arboviral replication and can spread through populations. Resident strains in wild Culicine mosquito populations (the vectors of most arboviruses) requires further investigation given resident strains can also affect arboviral transmission. As Madagascar has a large diversity of both Culicine species and has had recent arboviral outbreaks, an entomology survey was undertaken, in five ecologically diverse sites, to determine the Wolbachia prevalence. We detected diverse novel resident Wolbachia strains within the Aedeomyia, Culex, Ficalbia, Mansonia and Uranotaenia genera. Wolbachia prevalence rates and strain characterisation through Sanger sequencing with multilocus sequence typing (MLST) and phylogenetic analysis revealed significant diversity and we detected co-infections with the environmentally acquired bacteria Asaia. Mosquitoes were screened for major arboviruses to investigate if any evidence could be provided for their potential role in transmission and we report the presence of Rift Valley fever virus in three Culex species: Culex tritaeniorhynchus, Culex antennatus and Culex decens. The implications of the presence of resident Wolbachia strains are discussed and how the discovery of novel strains can be utilized for applications in the development of biocontrol strategies.}, } @article {pmid30496431, year = {2019}, author = {Sakamoto, H and Suzuki, R and Nishizawa, N and Matsuda, T and Gotoh, T}, title = {Effects of Wolbachia/Cardinium Infection on the Mitochondrial Phylogeny of Oligonychus castaneae (Acari: Tetranychidae).}, journal = {Journal of economic entomology}, volume = {112}, number = {2}, pages = {883-893}, doi = {10.1093/jee/toy354}, pmid = {30496431}, issn = {1938-291X}, mesh = {Animals ; Bacteroidetes ; DNA, Mitochondrial ; Japan ; Male ; Phylogeny ; Symbiosis ; *Tetranychidae ; *Wolbachia ; }, abstract = {A wide range of invertebrates harbor intracellular endosymbiotic bacteria. Within these endosymbionts, Wolbachia and Cardinium, have been attracting particular attention because these bacteria frequently affect the genetic structure and genetic diversity of their hosts. They cause various reproductive alterations such as cytoplasmic incompatibility, parthenogenesis induction, male-killing, and feminization. Through these alterations, they also affect the maternally inherited organelles of their hosts. Mitochondrial DNA (mtDNA) can be used for molecular phylogenetic analysis of invertebrates. However, in Wolbachia- or Cardinium-infected invertebrates, phylogenetic trees based on mtDNA are often inconsistent with those based on nuclear DNA. In the present study, we determined the Wolbachia/Cardinium infection status of 45 populations of the mite, Oligonychus castaneae Ehara & Gotoh (Acari: Tetranychidae), collected throughout Japan. Then, we compared phylogenetic trees of O. castaneae based on both the cytochrome c oxidase subunit I (COI) gene of mtDNA and the 28S rRNA gene of nuclear DNA to clarify the effects of Wolbachia and/or Cardinium infection. We found 106 Wolbachia-infected individuals and 250 Cardinium-infected individuals in a total of 450 individuals, indicating an infection rate of 79%. No double-infected individuals were observed. In the 28S tree, almost all populations formed a single group. In the COI tree, O. castaneae formed four separate groups that more closely followed Wolbachia/Cardinium infection than geographic distribution. These results strongly suggest that the endosymbionts affected mitochondrial variation of O. castaneae.}, } @article {pmid30474731, year = {2019}, author = {Hosseinzadeh, S and Shams-Bakhsh, M and Mann, M and Fattah-Hosseini, S and Bagheri, A and Mehrabadi, M and Heck, M}, title = {Distribution and Variation of Bacterial Endosymbiont and "Candidatus Liberibacter asiaticus" Titer in the Huanglongbing Insect Vector, Diaphorina citri Kuwayama.}, journal = {Microbial ecology}, volume = {78}, number = {1}, pages = {206-222}, pmid = {30474731}, issn = {1432-184X}, support = {5300-163//California Citrus Research Board/ ; 2016-70016-24779//National Institute of Food and Agriculture/ ; 8062-22410-006-00-D//Agricultural Research Service/ ; }, mesh = {Animals ; Citrus/*microbiology/parasitology ; Endophytes/genetics/isolation & purification/*physiology ; Female ; Hemiptera/*microbiology/physiology ; Insect Vectors/microbiology/physiology ; Male ; Plant Diseases/*microbiology ; Rhizobiaceae/genetics/isolation & purification/*physiology ; *Symbiosis ; Wolbachia/genetics/isolation & purification/physiology ; }, abstract = {The Asian citrus psyllid (ACP), Diaphorina citri Kuwayama, is an economic insect pest in most citrus-growing regions and the vector of 'Candidatus Liberibacter asiaticus' (CLas), one of at least three known bacteria associated with Huanglongbing (HLB or citrus greening disease). D. citri harbors bacterial endosymbionts, including Wolbachia pipientis (strain Wolbachia wDi), 'Candidatus Carsonella ruddii,' and 'Candidatus Profftella armatura.' Many important functions of these bacteria can be inferred from their genome sequences, but their interactions with each other, CLas, and their D. citri host are poorly understood. In the present study, the titers of the endosymbionts in different tissues, in each sex, and in insects reared on healthy citrus (referred to as unexposed) and CLas-infected citrus (referred to as CLas-exposed) D. citri were investigated using real-time, quantitative PCR (qPCR) using two different quantitative approaches. Wolbachia and CLas were detected in all insect tissues. The titer of Wolbachia was higher in heads of CLas-exposed males as compared to unexposed males. In males and females, Wolbachia titer was highest in the Malpighian tubules. The highest titer of CLas was observed in the gut. Profftella and Carsonella titers were significantly reduced in the bacteriome of CLas-exposed males compared with that of unexposed males, but this effect was not observed in females. In ovaries of CLas-exposed females, the Profftella and Carsonella titers were increased as compared to non-exposed females. CLas appeared to influence the overall levels of the symbionts but did not drastically perturb the overall microbial community structure. In all the assessed tissues, CLas titer in males was significantly higher than that of females using absolute quantification. These data provide a better understanding of multi-trophic interactions regulating symbiont dynamics in the HLB pathosystem.}, } @article {pmid30470196, year = {2018}, author = {Doudoumis, V and Augustinos, A and Saridaki, A and Parker, A and Abd-Alla, AMM and Bourtzis, K and Tsiamis, G}, title = {Different laboratory populations similar bacterial profile? The case of Glossina palpalis gambiensis.}, journal = {BMC microbiology}, volume = {18}, number = {Suppl 1}, pages = {148}, pmid = {30470196}, issn = {1471-2180}, mesh = {Animals ; Bacteria/*classification/isolation & purification ; Enterobacteriaceae/genetics ; Female ; Gastrointestinal Tract/microbiology ; *Genetic Variation ; Male ; *Microbiota ; RNA, Ribosomal, 16S/genetics ; Symbiosis ; Tsetse Flies/*microbiology ; Wigglesworthia/genetics ; Wolbachia/genetics ; }, abstract = {BACKGROUND: Microbiota plays an important role in the biology, ecology and evolution of insects including tsetse flies. The bacterial profile of 3 Glossina palpalis gambiensis laboratory colonies was examined using 16S rRNA gene amplicon sequencing to evaluate the dynamics of the bacterial diversity within and between each G. p. gambiensis colony.

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

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

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

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

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

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

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

CONCLUSIONS: The ecologies and life-histories of the housefly and tsetse fly may significantly influence coevolution of MdSGHV and GpSGHV with their hosts. Although there are still many unanswered questions regarding the pathogenesis of SGHVs, and the extent to which microbiota influence expression of overt SGH symptoms, SGHVs are attractive 'explorers' to elucidate the immune responses of their hosts, and the transmission modes of other large DNA viruses.}, } @article {pmid30470185, year = {2018}, author = {Schneider, DI and Parker, AG and Abd-Alla, AM and Miller, WJ}, title = {High-sensitivity detection of cryptic Wolbachia in the African tsetse fly (Glossina spp.).}, journal = {BMC microbiology}, volume = {18}, number = {Suppl 1}, pages = {140}, pmid = {30470185}, issn = {1471-2180}, support = {P 22634/FWF_/Austrian Science Fund FWF/Austria ; P 28255/FWF_/Austrian Science Fund FWF/Austria ; }, mesh = {Animals ; Bacterial Outer Membrane Proteins/genetics ; Female ; In Situ Hybridization, Fluorescence/*methods ; Insect Vectors/microbiology ; Limit of Detection ; Male ; Polymerase Chain Reaction/*methods ; Sensitivity and Specificity ; Symbiosis ; Tsetse Flies/*microbiology ; Wolbachia/genetics/*isolation & purification ; }, abstract = {BACKGROUND: In African tsetse flies Glossina, spp. detection of bacterial symbionts such as Wolbachia is challenging since their prevalence and distribution are patchy, and natural symbiont titers can range at levels far below detection limit of standard molecular techniques. Reliable estimation of symbiont infection frequency, especially with regard to interrelations between symbionts and their potential impact on host biology, is of pivotal interest in the context of future applications for the control and eradication of Glossina-vectored African trypanosomosis. The presence or absence of symbionts is routinely screened with endpoint polymerase chain reaction (PCR), which has numerous advantages, but reaches its limits, when detecting infections at natural low titer. To not only determine presence of native tsetse symbionts but also to localize them to specific host tissues, fluorescence in situ hybridization (FISH) can be applied. However, classic FISH assays may not detect low-titer infections due to limitations in sensitivity.

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

CONCLUSION: All in all, in this study a radical computer-based pipeline is proposed that could be adopted under many other similar scenarios and pave the way for precision agriculture via optimized pest control.}, } @article {pmid30149795, year = {2018}, author = {Brown, AMV}, title = {Endosymbionts of Plant-Parasitic Nematodes.}, journal = {Annual review of phytopathology}, volume = {56}, number = {}, pages = {225-242}, doi = {10.1146/annurev-phyto-080417-045824}, pmid = {30149795}, issn = {1545-2107}, mesh = {Animals ; *Bacterial Physiological Phenomena ; Host-Parasite Interactions ; Nematoda/*microbiology ; Plant Diseases/parasitology/*prevention & control ; Plants/*parasitology ; *Symbiosis ; }, abstract = {Some of the most agriculturally important plant-parasitic nematodes (PPNs) harbor endosymbionts. Extensive work in other systems has shown that endosymbionts can have major effects on host virulence and biology. This review highlights the discovery, development, and diversity of PPN endosymbionts, incorporating inferences from genomic data. Cardinium, reported from five PPN hosts to date, is characterized by its presence in the esophageal glands and other tissues, with a discontinuous distribution across populations, and genomic data suggestive of horizontal gene exchange. Xiphinematobacter occurs in at least 27 species of dagger nematode in the ovaries and gut epithelial cells, where genomic data suggest it may serve in nutritional supplementation. Wolbachia, reported in just three PPNs, appears to have an ancient history in the Pratylenchidae and displays broad tissue distribution and genomic features intermediate between parasitic and reproductive groups. Finally, a model is described that integrates these insights to explain patterns of endosymbiont replacement.}, } @article {pmid30148833, year = {2018}, author = {Cerutti, F and Modesto, P and Rizzo, F and Cravero, A and Jurman, I and Costa, S and Giammarino, M and Mandola, ML and Goria, M and Radovic, S and Cattonaro, F and Acutis, PL and Peletto, S}, title = {The microbiota of hematophagous ectoparasites collected from migratory birds.}, journal = {PloS one}, volume = {13}, number = {8}, pages = {e0202270}, pmid = {30148833}, issn = {1932-6203}, mesh = {Animal Migration ; Animals ; Arthropods/*microbiology ; Bacteria/*isolation & purification ; Bird Diseases/*parasitology ; Birds/parasitology ; Computational Biology ; Ectoparasitic Infestations/parasitology/*veterinary ; Italy ; *Microbiota ; Molecular Typing ; Parasites/*microbiology ; RNA, Ribosomal, 16S ; Ticks/microbiology ; }, abstract = {Arthropod vectors are responsible for the transmission of human pathogens worldwide. Several arthropod species are bird ectoparasites, however, no study to date has characterized their microbiota as a whole. We sampled hematophagous ectoparasites that feed on migratory birds and performed 16S rRNA gene metabarcoding to characterize their microbial community. A total of 194 ectoparasites were collected from 115 avian hosts and classified into three groups: a) Hippoboscidae diptera; b) ticks; c) other arthropods. Metabarcoding showed that endosymbionts were the most abundant genera of the microbial community, including Wolbachia for Hippoboscidae diptera, Candidatus Midichloria for ticks, Wolbachia and Arsenophonus for the other arthropod group. Genera including pathogenic species were: Rickettsia, Borrelia, Coxiella, Francisella, Bartonella, Anaplasma. Co-infection with Borrelia-Rickettsia and Anaplasma-Rickettsia was also observed. A global overview of the microbiota of ectoparasites sampled from migratory birds was obtained with the use of 16S rRNA gene metabarcoding. A novel finding is the first identification of Rickettsia in the common swift louse fly, Crataerina pallida. Given their possible interaction with pathogenic viruses and bacteria, the presence of endosymbionts in arthropods merits attention. Finally, molecular characterization of genera, including both pathogenic and symbiont species, plays a pivotal role in the design of targeted molecular diagnostics.}, } @article {pmid30139962, year = {2019}, author = {Richardson, KM and Griffin, PC and Lee, SF and Ross, PA and Endersby-Harshman, NM and Schiffer, M and Hoffmann, AA}, title = {A Wolbachia infection from Drosophila that causes cytoplasmic incompatibility despite low prevalence and densities in males.}, journal = {Heredity}, volume = {122}, number = {4}, pages = {428-440}, pmid = {30139962}, issn = {1365-2540}, support = {R01 GM104325/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Biological Evolution ; Cytoplasm/*microbiology ; Drosophila/*genetics/*microbiology ; Female ; Fertility/genetics ; Male ; Phylogeny ; Reproduction ; Symbiosis/genetics ; Wolbachia/classification/genetics/*physiology ; }, abstract = {Wolbachia bacteria are common insect endosymbionts transmitted maternally and capable of spreading through insect populations by cytoplasmic incompatibility (CI) when infected males cause embryo death after mating with uninfected females. Selection in the Wolbachia endosymbiont occurs on female hosts and is expected to favour strong maternal transmission to female offspring, even at the cost of reduced CI. With maternal leakage, nuclear genes are expected to be selected to suppress cytoplasmic incompatibility caused by males while also reducing any deleterious effects associated with the infection. Here we describe a new type of Wolbachia strain from Drosophila pseudotakahashii likely to have arisen from evolutionary processes on host and/or Wolbachia genomes. This strain is often absent from adult male offspring, but always transmitted to females. It leads to males with low or non-detectable Wolbachia that nevertheless show CI. When detected in adult males, the infection has a low density relative to that in females, a phenomenon not previously seen in Wolbachia infections of Drosophila. This Wolbachia strain is common in natural populations, and shows reduced CI when older (infected) males are crossed. These patterns highlight that endosymbionts can have strong sex-specific effects and that high frequency Wolbachia strains persist through effects on female reproduction. Female-limited Wolbachia infections may be of applied interest if the low level of Wolbachia in males reduces deleterious fitness effects on the host.}, } @article {pmid30124918, year = {2018}, author = {Ali, H and Muhammad, A and Sanda Bala, N and Hou, Y}, title = {The Endosymbiotic Wolbachia and Host COI Gene Enables to Distinguish Between Two Invasive Palm Pests; Coconut Leaf Beetle, Brontispa longissima and Hispid Leaf Beetle, Octodonta nipae.}, journal = {Journal of economic entomology}, volume = {111}, number = {6}, pages = {2894-2902}, pmid = {30124918}, issn = {1938-291X}, mesh = {Animals ; Base Sequence ; Coleoptera/*classification/genetics/microbiology ; Electron Transport Complex IV/genetics ; Species Specificity ; Wolbachia/genetics ; }, abstract = {To elucidate taxonomic eminence of identical pest species is essential for many ecological and conservation studies. Without proficient skills, accurate molecular identification and characterization are laborious and time-consuming. The coconut leaf beetle, Brontispa longissima (Gestro) (Coleoptera: Chrysomelidae), is biologically and morphologically identical to hispid leaf beetle, Octodonta nipae (Maulik) (Coleoptera: Chrysomelidae), and is known as the most harming nuisances of palm cultivation worldwide. The present examination was to establish Wolbachia genotyping analysis along with host cytochrome oxidase subunit I (COI) gene for accurate identification between these individuals of the same family (Chrysomelidae). Here, we have cloned and sequenced a gene coding Wolbachia surface protein (wsp) and COI gene regions amplified from both species by polymerase chain reaction. The nucleotide sequences were directly determined (≈600 bp for wsp and ≈804 bp for COI) and aligned using the multiple alignment algorithms in the ESPript3 package and the MEGA5 program. Comparative sequence analysis indicated that the representative of wsp and COI sequences from these two beetles were highly variable. To ensure this bacterial variation, multilocus sequence typing (MLST) of bacterial genes was conducted, and the results vindicated the same trend of variations. Furthermore, the phylogenetic analysis also indicates that B. longissima and O. nipae being the two different species harbors two distinct Wolbachia Hertig and Burt (Rickettsiales: Anaplamataceae) supergroups B and A, respectively. The present outcomes quickly discriminate between these two species. Considering its simplicity and cost-effectiveness, it can be used as a diagnostic tool for discriminating such invasive species particularly B. longissima and O. nipae which has overlapping morphologic characters.}, } @article {pmid30113326, year = {2018}, author = {Showler, AJ and Nutman, TB}, title = {Imported onchocerciasis in migrants and travelers.}, journal = {Current opinion in infectious diseases}, volume = {31}, number = {5}, pages = {393-398}, pmid = {30113326}, issn = {1473-6527}, support = {Z01 AI000439/ImNIH/Intramural NIH HHS/United States ; }, mesh = {Animals ; Antibodies, Helminth/blood ; Communicable Diseases, Imported/*diagnosis/*drug therapy ; Diagnostic Tests, Routine/*methods ; *Disease Management ; Doxycycline/therapeutic use ; Filaricides/*therapeutic use ; Human Migration ; Humans ; Onchocerca volvulus/immunology ; Onchocerciasis/*diagnosis/*drug therapy ; Transients and Migrants ; Travel ; }, abstract = {PURPOSE OF REVIEW: With increasing international travel and mass global population migration, clinicians in nonendemic countries must be familiar with imported neglected tropical diseases including onchocerciasis, which is commonly known as 'river blindness'.

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

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

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

CONCLUSIONS: Because field females typically encompass a wide range of sizes while laboratory reared (and released) males typically fall into a narrow size range of large mosquitoes, these patterns can influence the success of release programmes which rely on cytoplasmic incompatibility to suppress populations and initiate replacement invasions. Releases could include some small males generated under low food or crowded conditions to counter this issue, although this would need to be weighed against issues associated with costs of producing males of various size classes.}, } @article {pmid30064911, year = {2018}, author = {Fukui, T and Kiuchi, T and Shoji, K and Kawamoto, M and Shimada, T and Katsuma, S}, title = {In vivo masculinizing function of the Ostrinia furnacalis Masculinizer gene.}, journal = {Biochemical and biophysical research communications}, volume = {503}, number = {3}, pages = {1768-1772}, doi = {10.1016/j.bbrc.2018.07.111}, pmid = {30064911}, issn = {1090-2104}, mesh = {Animals ; Female ; Insect Proteins/*genetics/metabolism ; Male ; Moths/embryology/*genetics ; Sex Chromosomes/*genetics/metabolism ; Sex Determination Processes/*genetics ; }, abstract = {The Masculinizer gene (Masc) encodes a CCCH tandem zinc finger protein essential for masculinization and dosage compensation in the silkworm Bombyx mori. Previously we identified a Masc orthologue from the crambid Ostrinia furnacalis (OfMasc) and observed its masculinizing activity in the B. mori cultured cell line BmN-4. However, the role of OfMasc in masculinization of O. furnacalis has not been assessed. In this study, we unexpectedly discovered that all of the male larvae that escaped from Wolbachia-induced embryonic male-killing by OfMasc cRNA injection expressed the female-type splicing variants of O. furnacalis doublesex (Ofdsx). To clarify the role of OfMasc in the masculinization process in vivo, we established a system to monitor both sex chromosome- and dsx splicing-based sexes from a single O. furnacalis embryo. Using this system, we investigated the effects of OfMasc knockdown in early embryos on Ofdsx splicing and found that depletion of OfMasc mRNA in male embryos induced the production of the female-type splicing variants of Ofdsx. This result indicates that OfMasc is required for masculinization in O. furnacalis, and that the Masc protein possesses masculinizing activity in an insect species that is phylogenetically distant from Bombycidae.}, } @article {pmid30061694, year = {2018}, author = {Fisher, ML and Watson, DW and Osborne, JA and Mochizuki, H and Breen, M and Schal, C}, title = {Growth kinetics of endosymbiont Wolbachia in the common bed bug, Cimex lectularius.}, journal = {Scientific reports}, volume = {8}, number = {1}, pages = {11444}, pmid = {30061694}, issn = {2045-2322}, support = {P30 ES025128/ES/NIEHS NIH HHS/United States ; NCHHU0017-13//U.S. Department of Housing and Urban Development (HUD)/International ; 2013-5-35 MBE//Alfred P. Sloan Foundation/International ; }, mesh = {Animals ; Bedbugs/*microbiology ; DNA/genetics ; Female ; Kinetics ; Larva/microbiology ; *Symbiosis ; Wolbachia/*growth & development ; }, abstract = {The common bed bug, Cimex lectularius harbors the endosymbiotic microorganism, Wolbachia (wCle), in a gonad-associated bacteriome as an obligate nutritional mutualist. The obligatory nature of this association suggests that all individuals in C. lectularius populations would be infected with wCle. However, studies spanning the past several decades have reported variation in both infection frequency and relative abundance of wCle in field-collected samples of bed bugs. Since the growth kinetics of wCle is poorly understood, the objective of this study was to quantify wCle over the life cycle of two strains of C. lectularius. Our results highlight that wCle is dynamic during bed bug development, changing relative to life stage, intermolt stage, and blood-fed status. These results suggest new hypotheses about the coordination of Wolbachia growth and regression with its host's physiology and endocrine events. The observed quantitative modulation of wCle during the bed bug life cycle and during periods of starvation may explain the disparities in wCle infections reported in field-collected C. lectularius.}, } @article {pmid30060072, year = {2018}, author = {Gillespie, JJ and Driscoll, TP and Verhoeve, VI and Rahman, MS and Macaluso, KR and Azad, AF}, title = {A Tangled Web: Origins of Reproductive Parasitism.}, journal = {Genome biology and evolution}, volume = {10}, number = {9}, pages = {2292-2309}, pmid = {30060072}, issn = {1759-6653}, support = {R01 AI017828/AI/NIAID NIH HHS/United States ; R01 AI122672/AI/NIAID NIH HHS/United States ; R01 AI126853/AI/NIAID NIH HHS/United States ; R21 AI126108/AI/NIAID NIH HHS/United States ; }, mesh = {Amino Acid Sequence ; Animals ; Arthropods/*microbiology/physiology ; Bacterial Proteins/chemistry/genetics ; Bacterial Toxins/chemistry/genetics ; Biological Evolution ; Female ; Male ; Models, Molecular ; Phylogeny ; Plasmids/genetics ; Reproduction ; Rickettsia felis/chemistry/*genetics/physiology ; Wolbachia/chemistry/*genetics/physiology ; }, abstract = {While typically a flea parasite and opportunistic human pathogen, the presence of Rickettsia felis (strain LSU-Lb) in the non-blood-feeding, parthenogenetically reproducing booklouse, Liposcelis bostrychophila, provides a system to ascertain factors governing not only host transitions but also obligate reproductive parasitism (RP). Analysis of plasmid pLbAR, unique to R. felis str. LSU-Lb, revealed a toxin-antitoxin module with similar features to prophage-encoded toxin-antitoxin modules utilized by parasitic Wolbachia strains to induce another form of RP, cytoplasmic incompatibility, in their arthropod hosts. Curiously, multiple deubiquitinase and nuclease domains of the large (3,841 aa) pLbAR toxin, as well the entire antitoxin, facilitated the detection of an assortment of related proteins from diverse intracellular bacteria, including other reproductive parasites. Our description of these remarkable components of the intracellular mobilome, including their presence in certain arthropod genomes, lends insight on the evolution of RP, while invigorating research on parasite-mediated biocontrol of arthropod-borne viral and bacterial pathogens.}, } @article {pmid30059498, year = {2018}, author = {Xue, L and Fang, X and Hyman, JM}, title = {Comparing the effectiveness of different strains of Wolbachia for controlling chikungunya, dengue fever, and zika.}, journal = {PLoS neglected tropical diseases}, volume = {12}, number = {7}, pages = {e0006666}, pmid = {30059498}, issn = {1935-2735}, support = {U01 GM097661/GM/NIGMS NIH HHS/United States ; }, mesh = {Aedes/*microbiology/physiology/virology ; Animals ; Chikungunya Fever/*transmission/virology ; Chikungunya virus/physiology ; Dengue/*transmission/virology ; Dengue Virus/physiology ; Female ; Humans ; Male ; Mosquito Control/*methods ; Mosquito Vectors/*microbiology/physiology/virology ; Pest Control, Biological/*methods ; Reproduction ; Wolbachia/classification/*physiology ; Zika Virus/physiology ; Zika Virus Infection/*transmission/virology ; }, abstract = {Once Aedes aegypti and Aedes albopictus mosquitoes that spread Chikungunya virus, dengue virus, and Zika virus are infected with Wolbachia, they have reduced egg laying rates, reduced transmission abilities, and shorter lifespans. Since most infected mosquitoes are only infectious in the last few days of their lives, shortening a mosquito's lifespan by a day or two can greatly reduce their abilities to spread mosquito-borne viral diseases, such as Chikungunya, dengue fever, and Zika. We developed a mathematical model to compare the effectiveness of the wMel and wAlbB strains of Wolbachia for controlling the spread of these viruses. The differences among the diseases, mosquitoes, and Wolbachia strains are captured by the model parameters for the mosquito-human transmission cycle. Moreover, the model accounts for the behavior changes of infectious population created by differences in the malaise caused by these viruses. We derived the effective and basic reproduction numbers for the model that are used to estimate the number of secondary infections from the infectious populations. In the same density of Wolbachia-free Aedes aegypti or Aedes albopictus mosquitoes, we observed that wMel and wAlbB strains of Wolbachia can reduce the transmission rates of these diseases effectively.}, } @article {pmid30056075, year = {2018}, author = {Ciuca, L and Simòn, F and Rinaldi, L and Kramer, L and Genchi, M and Cringoli, G and Acatrinei, D and Miron, L and Morchon, R}, title = {Seroepidemiological survey of human exposure to Dirofilaria spp. in Romania and Moldova.}, journal = {Acta tropica}, volume = {187}, number = {}, pages = {169-174}, doi = {10.1016/j.actatropica.2018.07.012}, pmid = {30056075}, issn = {1873-6254}, mesh = {Adolescent ; Adult ; Aged ; Aged, 80 and over ; Animals ; Antibodies, Helminth/*immunology ; Antigens, Helminth/*immunology ; Child ; Dirofilaria/*immunology ; Dirofilaria immitis/immunology ; Dirofilaria repens/immunology ; Dirofilariasis/*epidemiology/immunology ; Dog Diseases/epidemiology ; Dogs ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Male ; Middle Aged ; Moldova/epidemiology ; Prevalence ; Romania/epidemiology ; Seroepidemiologic Studies ; Surveys and Questionnaires ; Wolbachia/immunology ; Young Adult ; }, abstract = {The present study aimed to evaluate the extent of Dirofilaria immitis and D. repens exposure in humans from eastern and southern areas of Romania and central Moldova by serological methods. The serological screening was performed on a total of 450 serum samples (187 from Romania and 263 from Moldova). The sera were collected using a convenience sampling with the help of physicians from the hospitals of the study areas. All samples were analysed by a non-commercial ELISA test for the detection of IgG antibodies against adult somatic antigens of D. immitis and D. repens. The results showed a total of 49 (10.9%; 95% CI = 8.3-14.1%) individuals from Romania and Moldova with a positive response to IgG antibodies against both adult somatic antigens of D. immitis and D. repens. Specifically, 48 (10.7%; 95% CI = 8.0-14.0%) patients were positive for IgG-antibodies against adult somatic antigens of D. immitis, one (0.2%; 95% CI = 0.4-1.2%) against D. repens antigens, and four (0.9%; 95% CI = 0.4-3.3%). were positive for antigens of both parasites. At country level, out of 187 samples from Romania, 13 (6.9%; 95% CI = 4.1-11.5%) were positive for anti-D. immitis IgG with high exposure in the southern part of the country (Bucharest). Of the 263 people from Moldova, 36 (13.7%; 95% CI = 10.0-18.4%) were positive for D. immitis antigens from which three (1.1%, 95% CI = 0.4-3.3%) were positive for the antibodies against antigens of both parasites. Only one sample was found positive for anti-D. repens IgG. Positive IgG-ELISA results were confirmed by Western blot analysis. In addition, for further confirmation, a complementary ELISA was performed for anti-WSP IgG antibodies against Wolbachia endosymbionts. Our findings showed a noticeable exposure of humans from Romania and Moldova to Dirofilaria parasites. Serology can be useful for indicating exposure to Dirofilaria spp. in a healthy population in order to obtain useful data on the epidemiological scenario of human dirofilariosis in Eastern Europe.}, } @article {pmid30051873, year = {2018}, author = {Shi, M and White, VL and Schlub, T and Eden, JS and Hoffmann, AA and Holmes, EC}, title = {No detectable effect of Wolbachia wMel on the prevalence and abundance of the RNA virome of Drosophila melanogaster.}, journal = {Proceedings. Biological sciences}, volume = {285}, number = {1883}, pages = {}, pmid = {30051873}, issn = {1471-2954}, support = {R01 GM104325/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Australia ; Drosophila melanogaster/*microbiology/virology ; RNA Viruses/*physiology ; RNA, Viral/*analysis ; Transcriptome ; Wolbachia/*physiology ; }, abstract = {Wolbachia is an endosymbiotic bacterium that can block viral infections in arthropods, generating interest in its potential to control the spread of mosquito-borne disease. Drosophila melanogaster is model organism for Wolbachia infection, and the wMel strain of Wolbachia can improve host survival following viral infection. However, it is unclear whether wMel induces anti-viral blocking against the broader native virome of D. melanogaster, or whether the major effect of Wolbachia is a reduction in viral abundance rather than viral clearance. We examined the effect of Wolbachia on viral abundance by comparing the total transcriptome of wMel-positive and wMel-negative D. melanogaster populations sampled from six locations in Australia. In addition, we examined the impact of wMel on individual flies by obtaining transcriptome data from 20 wMel-positive and 20 wMel-negative D. melanogaster from the location (Melbourne) with highest density of wMel. These data revealed high viral abundance in both Wolbachia-positive and -negative populations and individuals. Notably, none of the viral species identified, representing RNA viruses from at least nine families/floating genera, showed evidence of protection by wMel. Although the viral loads of picorna-like viruses are reduced by wMel under experimental conditions, we observed no such effect here. These data show that D. melanogaster can harbour abundant RNA viruses regardless of its Wolbachia status and imply that the interaction between Wolbachia and viruses in nature is more complex than simple blocking.}, } @article {pmid30050351, year = {2018}, author = {Ta-Tang, TH and Crainey, JL and Post, RJ and Luz, SL and Rubio, JM}, title = {Mansonellosis: current perspectives.}, journal = {Research and reports in tropical medicine}, volume = {9}, number = {}, pages = {9-24}, pmid = {30050351}, issn = {1179-7282}, abstract = {Mansonellosis is a filarial disease caused by three species of filarial (nematode) parasites (Mansonella perstans, Mansonella streptocerca, and Mansonella ozzardi) that use humans as their main definitive hosts. These parasites are transmitted from person to person by bloodsucking females from two families of flies (Diptera). Biting midges (Ceratopogonidae) transmit all three species of Mansonella, but blackflies (Simuliidae) are also known to play a role in the transmission of M. ozzardi in parts of Latin America. M. perstans and M. streptocerca are endemic in western, eastern, and central Africa, and M. perstans is also present in the neotropical region from equatorial Brazil to the Caribbean coast. M. ozzardi has a patchy distribution in Latin America and the Caribbean. Mansonellosis infections are thought to have little pathogenicity and to be almost always asymptomatic, but occasionally causing itching, joint pains, enlarged lymph glands, and vague abdominal symptoms. In Brazil, M. ozzardi infections are also associated with corneal lesions. Diagnosis is usually performed by detecting microfilariae in peripheral blood or skin without any periodicity. There is no standard treatment at present for mansonellosis. The combination therapy of diethylcarbamazine plus mebendazole for M. perstans microfilaremia is presently one of the most widely used, but the use of ivermectin has also been proven to be very effective against microfilariae. Recently, doxycycline has shown excellent efficacy and safety when used as an antimicrobial against endosymbiotic Wolbachia bacteria harbored by some strains of M. perstans and M. ozzardi. Diethylcarbamazine and ivermectin have been used effectively to treat M. streptocerca infection. There are at present no estimates of the disease burden caused by mansonellosis, and thus its importance to many global health professionals and policy makers is presently limited to how it can interfere with diagnostic tools used in modern filarial disease control and elimination programs aimed at other species of filariae.}, } @article {pmid30046071, year = {2018}, author = {Ferguson, NM}, title = {Challenges and opportunities in controlling mosquito-borne infections.}, journal = {Nature}, volume = {559}, number = {7715}, pages = {490-497}, pmid = {30046071}, issn = {1476-4687}, support = {MR/R015600/1/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Animals ; Dengue/mortality/*prevention & control/transmission ; Gene Drive Technology ; Goals ; Humans ; Incidence ; Malaria/mortality/*prevention & control/transmission ; Mosquito Control/*methods ; Mosquito Vectors/genetics/microbiology ; Pest Control, Biological/methods ; Vaccines ; Wolbachia/pathogenicity ; }, abstract = {Mosquito-borne diseases remain a major cause of morbidity and mortality across the tropical regions. Despite much progress in the control of malaria, malaria-associated morbidity remains high, whereas arboviruses-most notably dengue-are responsible for a rising burden of disease, even in middle-income countries that have almost completely eliminated malaria. Here I discuss how new interventions offer the promise of considerable future reductions in disease burden. However, I emphasize that intervention programmes need to be underpinned by rigorous trials and quantitative epidemiological analyses. Such analyses suggest that the long-term goal of elimination is more feasible for dengue than for malaria, even if malaria elimination would offer greater overall health benefit to the public.}, } @article {pmid30032285, year = {2018}, author = {Ahmadi, S and Poorjavad, N}, title = {Behavioral and Biological Effects of Exposure to Tuta absoluta (Lepidoptera: Gelechiidae) Sex Pheromone on Several Trichogramma (Hymenoptera: Trichogrammatidae) Populations.}, journal = {Journal of economic entomology}, volume = {111}, number = {6}, pages = {2667-2675}, doi = {10.1093/jee/toy212}, pmid = {30032285}, issn = {1938-291X}, mesh = {Animals ; Female ; Male ; Moths/chemistry/*parasitology ; Sex Attractants/*pharmacology ; Sexual Behavior, Animal/*drug effects ; Wasps/microbiology/*physiology ; Wolbachia/physiology ; }, abstract = {Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) is one of the most destructive pests of tomato worldwide. Biological control of the pest using Trichogramma (Hymenoptera: Trichogrammatidae) wasps can be combined with other practices such as use of synthetic sex pheromones for mating disruption or mass trapping programs. In this study, effects of T. absoluta sex pheromone on behavioral responses and fertility life table parameters of Trichogramma evanescens Westwood (Hymenoptera: Trichogrammatidae), Wolbachia-uninfected (W-) Trichogramma brassicae Bezdenko (Hymenoptera: Trichogrammatidae), and Wolbachia-infected (W+) T. brassicae were investigated under laboratory conditions. Female wasps of T. evanescens and T. brassicae (W+) were attracted to the pest synthetic sex pheromone (around 62%) and calling virgin female moths (around 75%) and their responses were affected by the wasp age and temperature. Exposure to the host synthetic sex pheromone significantly reduced the percentage of adult emergence, longevity, and fecundity of female wasps, as well as the time spent to find a mate and duration of mating. However, tested populations were not affected similarly. Despite the laboratory tests, greenhouse experiments showed that the wasps were not caught in the traps baited with T. absoluta synthetic sex pheromone. Complementary studies are needed to precisely determine possible interference between pheromone application and releasing Trichogramma Westwood (Hymenoptera: Trichogrammatidae) wasps to achieve a successful integrated control of T. absoluta.}, } @article {pmid30025503, year = {2018}, author = {Zheng, B and Yu, J}, title = {Characterization of Wolbachia enhancing domain in mosquitoes with imperfect maternal transmission.}, journal = {Journal of biological dynamics}, volume = {12}, number = {1}, pages = {596-610}, doi = {10.1080/17513758.2018.1499969}, pmid = {30025503}, issn = {1751-3766}, mesh = {Animals ; Culicidae/*microbiology ; Dengue/*parasitology/*transmission ; Female ; *Infectious Disease Transmission, Vertical ; Models, Biological ; Pest Control, Biological ; Wolbachia/*physiology ; }, abstract = {A novel method to reduce the burden of dengue is to seed wild mosquitoes with Wolbachia-infected mosquitoes in dengue-endemic areas. Concerns in current mathematical models are to locate the Wolbachia introduction threshold. Our recent findings manifest that the threshold is highly dependent on the initial population size once Wolbachia infection alters the logistic control death rate of infected females. However, counting mosquitoes is beyond the realms of possibility. A plausible method is to monitor the infection frequency. We propose the concept of Wolbachia enhancing domain in which the infection frequency keeps increasing. A detailed description of the domain is presented. Our results suggest that both the initial population size and the infection frequency should be taken into account for optimal release strategies. Both Wolbachia fixation and extinction permit the oscillation of the infection frequency.}, } @article {pmid30021000, year = {2018}, author = {, }, title = {Correction: First detection of Wolbachia in the New Zealand biota.}, journal = {PloS one}, volume = {13}, number = {7}, pages = {e0201151}, pmid = {30021000}, issn = {1932-6203}, abstract = {[This corrects the article DOI: 10.1371/journal.pone.0195517.].}, } @article {pmid30020933, year = {2018}, author = {Moretti, R and Yen, PS and Houé, V and Lampazzi, E and Desiderio, A and Failloux, AB and Calvitti, M}, title = {Combining Wolbachia-induced sterility and virus protection to fight Aedes albopictus-borne viruses.}, journal = {PLoS neglected tropical diseases}, volume = {12}, number = {7}, pages = {e0006626}, pmid = {30020933}, issn = {1935-2735}, mesh = {Aedes/genetics/*microbiology/*physiology ; Animals ; Breeding ; Chikungunya Fever/transmission/virology ; Chikungunya virus/physiology ; Dengue/transmission/virology ; Dengue Virus/physiology ; Female ; Humans ; Infertility ; Male ; Mosquito Control/*methods ; Mosquito Vectors/genetics/*microbiology/*physiology ; Wolbachia/*physiology ; Zika Virus/physiology ; Zika Virus Infection/transmission/virology ; }, abstract = {Among the strategies targeting vector control, the exploitation of the endosymbiont Wolbachia to produce sterile males and/or invasive females with reduced vector competence seems to be promising. A new Aedes albopictus transinfection (ARwP-M) was generated by introducing wMel Wolbachia in the ARwP line which had been established previously by replacing wAlbA and wAlbB Wolbachia with the wPip strain. Various infection and fitness parameters were studied by comparing ARwP-M, ARwP and wild-type (SANG population) Ae. albopictus sharing the same genetic background. Moreover, the vector competence of ARwP-M related to chikungunya, dengue and zika viruses was evaluated in comparison with ARwP. ARwP-M showed a 100% rate of maternal inheritance of wMel and wPip Wolbachia. Survival, female fecundity and egg fertility did not show to differ between the three Ae. albopictus lines. Crosses between ARwP-M males and SANG females were fully unfertile regardless of male age while egg hatch in reverse crosses increased from 0 to about 17% with SANG males aging from 3 to 17 days. When competing with SANG males for SANG females, ARwP-M males induced a level of sterility significantly higher than that expected for an equal mating competitiveness (mean Fried index of 1.71 instead of 1). The overall Wolbachia density in ARwP-M females was about 15 fold higher than in ARwP, mostly due to the wMel infection. This feature corresponded to a strongly reduced vector competence for chikungunya and dengue viruses (in both cases, 5 and 0% rates of transmission at 14 and 21 days post infection) with respect to ARwP females. Results regarding Zika virus did not highlight significant differences between ARwP-M and ARwP. However, none of the tested ARwP-M females was capable at transmitting ZIKV. These findings are expected to promote the exploitation of Wolbachia to suppress the wild-type Ae. albopictus populations.}, } @article {pmid30006543, year = {2018}, author = {Karatepe, B and Aksoy, S and Karatepe, M}, title = {Investigation of Wolbachia spp. and Spiroplasma spp. in Phlebotomus species by molecular methods.}, journal = {Scientific reports}, volume = {8}, number = {1}, pages = {10616}, pmid = {30006543}, issn = {2045-2322}, support = {R01 AI051584/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; DNA, Bacterial/isolation & purification ; Phlebotomus/*microbiology ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S/genetics ; Spiroplasma/genetics/*isolation & purification ; Turkey ; Wolbachia/genetics/*isolation & purification ; }, abstract = {The aim of this study was to determine the presence of Wolbachia spp. and Spiroplasma spp. in natural populations of sand flies in Turkey by molecular methods. A total of 40 Phlebotomus specimens (19 female and 21 male) were used in this study. Genomic DNA from whole sand flies was isolated and Wolbachia spp. infection prevalence was investigated by using Wolbachia gene specific primer sets (wsp and GroEL). In addition, the DNA were analyzed for the presence of Spiroplasma infections utilizing bacterium specific 16 S rDNA PCR-amplification primers. Results of this analysis showed a Wolbachia infection prevalence of 70% (28/40). There was no sex-bias in infection prevalence, being 76% (16/21) and 63% (12/19) in males and females, respectively. Analysis of Spiroplasma infections indicated that 26% (5/19) of female sand flies were positive for infection, while none of the screened males (0/21) were positive. Of the 40 sand fly samples, only 2 were found to be positive for both Wolbachia spp. and Spiroplasma spp. The present study demonstrates the presence of Wolbachia and Spiroplasma infections in the natural sand fly populations in Turkey. This is the first report on Spiroplasma infection in the sand flies from Turkey.}, } @article {pmid29989657, year = {2018}, author = {Guo, Y and Hoffmann, AA and Xu, XQ and Zhang, X and Huang, HJ and Ju, JF and Gong, JT and Hong, XY}, title = {Wolbachia-induced apoptosis associated with increased fecundity in Laodelphax striatellus (Hemiptera: Delphacidae).}, journal = {Insect molecular biology}, volume = {27}, number = {6}, pages = {796-807}, doi = {10.1111/imb.12518}, pmid = {29989657}, issn = {1365-2583}, support = {//Medical Research Council/United Kingdom ; }, mesh = {Amino Acid Sequence ; Animals ; *Apoptosis ; Caspases/genetics/*metabolism ; Female ; Fertility ; Genetic Fitness ; Hemiptera/microbiology/*physiology ; Ovary/microbiology/physiology ; Wolbachia/*physiology ; }, abstract = {Wolbachia influence the fitness of their invertebrate hosts. They have effects on reproductive incompatibility and egg production. Although the former are well characterized, the mechanistic basis of the latter is unclear. Here, we investigate whether apoptosis, which has been implicated in fecundity in model insects, influences the interaction between fecundity and Wolbachia in the planthopper Laodelphax striatellus. Wolbachia-infected females produced about 30% more eggs than uninfected females. We used the terminal deoxyribonucleotidyl transferase (TDT)-mediated dUTP-digoxigenin nick end labeling staining to visualize apoptosis. Microscopic observations indicated that the Wolbachia strain wStri increased the number of ovarioles that contained apoptotic nurse cells in both young and aged adult females. The frequency of apoptosis was much higher in the infected females. The increased fecundity appeared to be a result of apoptosis of nurse cells, which provide nutrients to the growing oocytes. In addition, cell apoptosis inhibition by caspase messenger RNA interference in Wolbachia-infected L. striatellus markedly decreased egg numbers. Together, these data suggest that wStri might enhance fecundity by increasing the number of apoptotic cells in the ovaries in a caspase-dependent manner. Our findings establish a link between Wolbachia-induced apoptosis and egg production effects mediated by Wolbachia, although the way in which the endosymbiont influences caspase levels remains to be determined.}, } @article {pmid29981933, year = {2018}, author = {Ali, H and Muhammad, A and Bala, NS and Wang, G and Chen, Z and Peng, Z and Hou, Y}, title = {Genomic evaluations of Wolbachia and mtDNA in the population of coconut hispine beetle, Brontispa longissima (Coleoptera: Chrysomelidae).}, journal = {Molecular phylogenetics and evolution}, volume = {127}, number = {}, pages = {1000-1009}, doi = {10.1016/j.ympev.2018.07.003}, pmid = {29981933}, issn = {1095-9513}, mesh = {Animals ; Cocos/*parasitology ; Coleoptera/*genetics/*microbiology ; DNA, Mitochondrial/*genetics ; Asia, Eastern ; *Genomics ; Geography ; Haplotypes/genetics ; Likelihood Functions ; Multilocus Sequence Typing ; Phylogeny ; Polymorphism, Genetic ; Wolbachia/classification/*genetics ; }, abstract = {Wolbachia pipientis is a diverse, ubiquitous and most prevalent intracellular bacterial group of alpha-Proteobacteria that is concerned with many biological processes in arthropods. The coconut hispine beetle (CHB), Brontispa longissima (Gestro) is an economically important pest of palm cultivation worldwide. In the present study, we comprehensively surveyed the Wolbachia-infection prevalence and mitochondrial DNA (mtDNA) polymorphism in CHB from five different geographical locations, including China's Mainland and Taiwan, Vietnam, Thailand, Malaysia and Indonesia. A total of 540 sequences were screened in this study through three different genes, i.e., cytochrome oxidase subunit I (COI), Wolbachia outer surface protein (wsp) and multilocus sequencing type (MLST) genes. The COI genetic divergence ranges from 0.08% to 0.67%, and likewise, a significant genetic diversity (π = 0.00082; P = 0.049) was noted within and between all analyzed samples. In the meantime, ten different haplotypes (H) were characterized (haplotype diversity = 0.4379) from 21 different locations, and among them, H6 (46 individuals) have shown a maximum number of population clusters than others. Subsequently, Wolbachia-prevalence results indicated that all tested specimens of CHB were found positive (100%), which suggested that CHB was naturally infected with Wolbachia. Wolbachia sequence results (wsp gene) revealed a high level of nucleotide diversity (π = 0.00047) under Tajima's D test (P = 0.049). Meanwhile, the same trend of nucleotide diversity (π = 0.00041) was observed in Wolbachia concatenated MLST locus. Furthermore, phylogenetic analysis (wsp and concatenated MLST genes) revealed that all collected samples of CHB attributed to same Wolbachia B-supergroup. Our results strongly suggest that Wolbachia bacteria and mtDNA were highly concordant with each other and Wolbachia can affect the genetic structure and diversity within the CHB populations.}, } @article {pmid29971900, year = {2018}, author = {Truitt, AM and Kapun, M and Kaur, R and Miller, WJ}, title = {Wolbachia modifies thermal preference in Drosophila melanogaster.}, journal = {Environmental microbiology}, volume = {21}, number = {9}, pages = {3259-3268}, pmid = {29971900}, issn = {1462-2920}, support = {P 28255/FWF_/Austrian Science Fund FWF/Austria ; 0948041//National Science Foundation/ ; P28255-B22//Austrian Science Fund/ ; }, abstract = {Environmental variation can have profound and direct effects on fitness, fecundity, and host-symbiont interactions. Replication rates of microbes within arthropod hosts, for example, are correlated with incubation temperature but less is known about the influence of host-symbiont dynamics on environmental preference. Hence, we conducted thermal preference (Tp) assays and tested if infection status and genetic variation in endosymbiont bacterium Wolbachia affected temperature choice of Drosophila melanogaster. We demonstrate that isogenic flies infected with Wolbachia preferred lower temperatures compared with uninfected Drosophila. Moreover, Tp varied with respect to three investigated Wolbachia variants (wMel, wMelCS, and wMelPop). While uninfected individuals preferred 24.4°C, we found significant shifts of -1.2°C in wMel- and -4°C in flies infected either with wMelCS or wMelPop. We, therefore, postulate that Wolbachia-associated Tp variation within a host species might represent a behavioural accommodation to host-symbiont interactions and trigger behavioural self-medication and bacterial titre regulation by the host.}, } @article {pmid29968550, year = {2018}, author = {Yeap, HL and Endersby-Harshman, NM and Hoffmann, AA}, title = {The Effect of Nonrandom Mating on Wolbachia Dynamics: Implications for Population Replacement and Sterile Releases in Aedes Mosquitoes.}, journal = {The American journal of tropical medicine and hygiene}, volume = {99}, number = {3}, pages = {608-617}, pmid = {29968550}, issn = {1476-1645}, mesh = {Aedes/*microbiology ; Animals ; Dengue/*prevention & control ; Female ; Male ; Reproduction ; Wolbachia/*physiology ; }, abstract = {Wolbachia bacteria are known to cause deviations from random mating and affect sperm competition (SC) in some of their arthropod hosts. Because these effects could influence the effectiveness of Wolbachia in mosquito population replacement and suppression programs, we developed a theoretical framework to investigate them and we collected relevant data for the wMel infection in Aedes aegypti. Using incompatibility patterns as a measure of mating success of infected versus uninfected mosquitoes, we found some evidence that uninfected males sire more offspring than infected males. However, our theoretical framework suggests that this effect is unlikely to hamper Wolbachia invasion and has only minor effects on population suppression programs. Nevertheless, we suggest that mating effects and SC need to be monitored in an ongoing manner in release programs, given the possibility of ongoing selection for altered mating patterns.}, } @article {pmid29967922, year = {2019}, author = {Nesvorna, M and Bittner, V and Hubert, J}, title = {The Mite Tyrophagus putrescentiae Hosts Population-Specific Microbiomes That Respond Weakly to Starvation.}, journal = {Microbial ecology}, volume = {77}, number = {2}, pages = {488-501}, pmid = {29967922}, issn = {1432-184X}, support = {GA15-09038S//Grantová Agentura České Republiky/ ; }, mesh = {Acaridae/*microbiology/*physiology ; Animals ; Bacteria/classification/genetics/*isolation & purification ; Bacterial Physiological Phenomena ; Feeding Behavior ; Host Specificity ; *Microbiota ; }, abstract = {The effect of short-term nutrient deprivation was studied in five populations of the mite Tyrophagus putrescentiae with different microbiomes. The fresh weight, nutrient status, respiration, and population growth of the mites were observed for the five mite population-scale samples. The starvation caused the larvae and nymphs to be eliminated, resulting in a significant increase in the fresh weight of starved adult specimens. Three populations were negatively influenced by starvation, and the starved specimens were characterized by a decrease in nutrient status, respiration, and population growth. One population was not influenced or was slightly influenced by starvation, which had no effect on population growth or nutrient contents but caused a significant decrease in respiration. One population was positively influenced by starvation; the population growth increased in starved specimens, and starvation had no effect on respiration. Although starvation altered the bacterial profiles of the microbiomes, these differences were much smaller than those between the populations. The bacterial profiles of Staphylococcus, Bacillus, Kocuria, Brevibacterium, and unidentified Micrococcaceae and Enterobacteriaceae increased in starved specimens, whereas those of Bartonella and Solitalea-like genera were reduced in the starved mite populations. The profiles of the intracellular symbiont Cardinium decreased in the starved specimens, and the Wolbachia profile changes were dependent on the mite population. In mite populations, when the symbionts were rare, their profiles varied stochastically. Correlations between changes in the profiles of the bacterial taxa and mite fitness parameters, including nutrient status (lipids, proteins, saccharides, and glycogen contents), mite population growth, and respiration, were observed. Although the microbiomes were resistant to the perturbations caused by nutrition deficiency, the responses of the mites differed in terms of their population growth, respiration, and nutrient status.}, } @article {pmid29966368, year = {2018}, author = {Ross, PA and Hoffmann, AA}, title = {Continued Susceptibility of the wMel Wolbachia Infection in Aedes aegypti to Heat Stress Following Field Deployment and Selection.}, journal = {Insects}, volume = {9}, number = {3}, pages = {}, pmid = {29966368}, issn = {2075-4450}, abstract = {Aedes aegypti mosquitoes infected with the wMel strain of Wolbachia are being deployed to control the spread of arboviruses around the world through blockage of viral transmission. Blockage by Wolbachia in some scenarios may be affected by the susceptibility of wMel to cyclical heat stress during mosquito larval development. We therefore evaluated the potential to generate a heat-resistant strain of wMel in Ae. aegypti through artificial laboratory selection and through exposure to field temperatures across multiple generations. To generate an artificially selected strain, wMel-infected females reared under cyclical heat stress were crossed to wMel-infected males reared at 26 &deg;C. The low proportion of larvae that hatched founded the next generation, and this process was repeated for eight generations. The wMel heat-selected strain (wMel-HS) was similar to wMel (unselected) in its ability to induce cytoplasmic incompatibility and restore compatibility when larvae were reared under cyclical heat stress, but wMel-HS adults exhibited reduced Wolbachia densities at 26 &deg;C. To investigate the effects of field exposure, we compared the response of wMel-infected Ae. aegypti collected from Cairns, Australia where the infection has been established for seven years, to a wMel-infected population maintained in the laboratory for approximately 60 generations. Field and laboratory strains of wMel did not differ in their response to cyclical heat stress or in their phenotypic effects at 26 &deg;C. The capacity for the wMel infection in Ae. aegypti to adapt to high temperatures therefore appears limited, and alternative strains may need to be considered for deployment in environments where high temperatures are regularly experienced in mosquito breeding sites.}, } @article {pmid29957332, year = {2018}, author = {Ballesteros, C and Geary, JF and Mackenzie, CD and Geary, TG}, title = {Characterization of Divalent Metal Transporter 1 (DMT1) in Brugia malayi suggests an intestinal-associated pathway for iron absorption.}, journal = {International journal for parasitology. Drugs and drug resistance}, volume = {8}, number = {2}, pages = {341-349}, pmid = {29957332}, issn = {2211-3207}, mesh = {Animals ; Biological Transport ; Brugia malayi/genetics/growth & development/*metabolism ; Cation Transport Proteins/genetics/*metabolism ; *Host-Parasite Interactions ; Intestines/cytology/physiology ; Iron/*metabolism ; Iron Deficiencies ; Mice ; Microvilli/physiology ; Onchocerca volvulus/genetics/growth & development/metabolism ; Real-Time Polymerase Chain Reaction ; Saccharomyces cerevisiae/genetics/metabolism ; Sequence Alignment ; Wolbachia/metabolism ; }, abstract = {Lymphatic filariasis and onchocerciasis are neglected parasitic diseases which pose a threat to public health in tropical and sub-tropical regions. Strategies for control and elimination of these diseases by mass drug administration (MDA) campaigns are designed to reduce symptoms of onchocerciasis and transmission of both parasites to eventually eliminate the burden on public health. Drugs used for MDA are predominantly microfilaricidal, and prolonged rounds of treatment are required for eradication. Understanding parasite biology is crucial to unravelling the complex processes involved in host-parasite interactions, disease transmission, parasite immune evasion, and the emergence of drug resistance. In nematode biology, large gaps still exist in our understanding of iron metabolism, iron-dependent processes and their regulation. The acquisition of iron from the host is a crucial determinant of the success of a parasitic infection. Here we identify a filarial ortholog of Divalent Metal Transporter 1 (DMT1), a member of a highly conserved family of NRAMP proteins that play an essential role in the transport of ferrous iron in many species. We cloned and expressed the B. malayi NRAMP ortholog in the iron-deficient fet3fet4 strain of Saccharomyces cerevisiae, performed qPCR to estimate stage-specific expression, and localized expression of this gene by immunohistochemistry. Results from functional iron uptake assays showed that expression of this gene in the iron transport-deficient yeast strain significantly rescued growth in low-iron medium. DMT1 was highly expressed in adult female and male B. malayi and Onchocerca volvulus. Immunolocalization revealed that DMT1 is expressed in the intestinal brush border, lateral chords, and reproductive tissues of males and females, areas also inhabited by Wolbachia. We hypothesize based on our results that DMT1 in B. malayi functions as an iron transporter. The presence of this transporter in the intestine supports the hypothesis that iron acquisition by adult females requires oral ingestion and suggests that the intestine plays a functional role in at least some aspects of nutrient uptake.}, } @article {pmid29950416, year = {2018}, author = {Parry, R and Asgari, S}, title = {Aedes Anphevirus: an Insect-Specific Virus Distributed Worldwide in Aedes aegypti Mosquitoes That Has Complex Interplays with Wolbachia and Dengue Virus Infection in Cells.}, journal = {Journal of virology}, volume = {92}, number = {17}, pages = {}, pmid = {29950416}, issn = {1098-5514}, mesh = {Aedes/microbiology/*virology ; Animals ; Cell Line ; Chlorocebus aethiops ; Dengue Virus/physiology ; Evolution, Molecular ; Gene Expression Profiling/*methods ; Genome, Viral ; Host Specificity ; Humans ; Infectious Disease Transmission, Vertical/veterinary ; Insect Viruses/classification/physiology ; Mononegavirales/classification/*physiology ; Mosquito Vectors/microbiology/virology ; Phylogeny ; Sequence Analysis, RNA ; Vero Cells ; Virus Replication ; Wolbachia/*physiology ; }, abstract = {Insect-specific viruses (ISVs) of the yellow fever mosquito Aedes aegypti have been demonstrated to modulate transmission of arboviruses such as dengue virus (DENV) and West Nile virus by the mosquito. The diversity and composition of the virome of A. aegypti, however, remains poorly understood. In this study, we characterized Aedes anphevirus (AeAV), a negative-sense RNA virus from the order Mononegavirales AeAV identified from Aedes cell lines was infectious to both A. aegypti and Aedes albopictus cells but not to three mammalian cell lines. To understand the incidence and genetic diversity of AeAV, we assembled 17 coding-complete and two partial genomes of AeAV from available transcriptome sequencing (RNA-Seq) data. AeAV appears to transmit vertically and be present in laboratory colonies, wild-caught mosquitoes, and cell lines worldwide. Phylogenetic analysis of AeAV strains indicates that as the A. aegypti mosquito has expanded into the Americas and Asia-Pacific, AeAV has evolved into monophyletic African, American, and Asia-Pacific lineages. The endosymbiotic bacterium Wolbachia pipientis restricts positive-sense RNA viruses in A. aegypti Reanalysis of a small RNA library of A. aegypti cells coinfected with AeAV and Wolbachia produces an abundant RNA interference (RNAi) response consistent with persistent virus replication. We found Wolbachia enhances replication of AeAV compared to a tetracycline-cleared cell line, and AeAV modestly reduces DENV replication in vitro The results from our study improve understanding of the diversity and evolution of the virome of A. aegypti and adds to previous evidence that shows Wolbachia does not restrict a range of negative-strand RNA viruses.IMPORTANCE The mosquito Aedes aegypti transmits a number of arthropod-borne viruses (arboviruses), such as dengue virus and Zika virus. Mosquitoes also harbor insect-specific viruses that may affect replication of pathogenic arboviruses in their body. Currently, however, there are only a few insect-specific viruses described from A. aegypti in the literature. Here, we characterize a novel negative-strand virus, AeAV. Meta-analysis of A. aegypti samples showed that it is present in A. aegypti mosquitoes worldwide and is vertically transmitted. Wolbachia-transinfected mosquitoes are currently being used in biocontrol, as they effectively block transmission of several positive-sense RNA viruses in mosquitoes. Our results demonstrate that Wolbachia enhances the replication of AeAV and modestly reduces dengue virus replication in a cell line model. This study expands our understanding of the virome in A. aegypti as well as providing insight into the complexity of the Wolbachia virus restriction phenotype.}, } @article {pmid29947761, year = {2018}, author = {Faria, VG and Martins, NE and Schlötterer, C and Sucena, É}, title = {Readapting to DCV Infection without Wolbachia: Frequency Changes of Drosophila Antiviral Alleles Can Replace Endosymbiont Protection.}, journal = {Genome biology and evolution}, volume = {10}, number = {7}, pages = {1783-1791}, pmid = {29947761}, issn = {1759-6653}, support = {P 27630/FWF_/Austrian Science Fund FWF/Austria ; }, mesh = {Adaptation, Physiological ; Alleles ; Animals ; Drosophila melanogaster/genetics/*microbiology/physiology/*virology ; Genome, Insect ; Host-Pathogen Interactions ; Insect Viruses/*physiology ; Polymorphism, Genetic ; *Symbiosis ; Wolbachia/*physiology ; }, abstract = {There is now ample evidence that endosymbionts can contribute to host adaptation to environmental challenges. However, how endosymbiont presence affects the adaptive trajectory and outcome of the host is yet largely unexplored. In Drosophila, Wolbachia confers protection to RNA virus infection, an effect that differs between Wolbachia strains and can be targeted by selection. Adaptation to RNA virus infections is mediated by both Wolbachia and the host, raising the question of whether adaptive genetic changes in the host vary with the presence/absence of the endosymbiont. Here, we address this question using a polymorphic D. melanogaster population previously adapted to DCV infection for 35 generations in the presence of Wolbachia, from which we removed the endosymbiont and followed survival over the subsequent 20 generations of infection. After an initial severe drop, survival frequencies upon DCV selection increased significantly, as seen before in the presence of Wolbachia. Whole-genome sequencing, revealed that the major genes involved in the first selection experiment, pastrel and Ubc-E2H, continued to be selected in Wolbachia-free D. melanogaster, with the frequencies of protective alleles being closer to fixation in the absence of Wolbachia. Our results suggest that heterogeneity in Wolbachia infection status may be sufficient to maintain polymorphisms even in the absence of costs.}, } @article {pmid29938064, year = {2018}, author = {Amuzu, HE and Tsyganov, K and Koh, C and Herbert, RI and Powell, DR and McGraw, EA}, title = {Wolbachia enhances insect-specific flavivirus infection in Aedes aegypti mosquitoes.}, journal = {Ecology and evolution}, volume = {8}, number = {11}, pages = {5441-5454}, pmid = {29938064}, issn = {2045-7758}, abstract = {Mosquitoes transmit a diverse group of human flaviviruses including West Nile, dengue, yellow fever, and Zika viruses. Mosquitoes are also naturally infected with insect-specific flaviviruses (ISFs), a subgroup of the family not capable of infecting vertebrates. Although ISFs are not medically important, they are capable of altering the mosquito's susceptibility to flaviviruses and may alter host fitness. Wolbachia is an endosymbiotic bacterium of insects that when present in mosquitoes limits the replication of co-infecting pathogens, including flaviviruses. Artificially created Wolbachia-infected Aedes aegypti mosquitoes are being released into the wild in a series of trials around the globe with the hope of interrupting dengue and Zika virus transmission from mosquitoes to humans. Our work investigated the effect of Wolbachia on ISF infection in wild-caught Ae. aegypti mosquitoes from field release zones. All field mosquitoes were screened for the presence of ISFs using general degenerate flavivirus primers and their PCR amplicons sequenced. ISFs were found to be common and widely distributed in Ae. aegypti populations. Field mosquitoes consistently had higher ISF infection rates and viral loads compared to laboratory colony material indicating that environmental conditions may modulate ISF infection in Ae. aegypti. Surprisingly, higher ISF infection rates and loads were found in Wolbachia-infected mosquitoes compared to the Wolbachia-free mosquitoes. Our findings demonstrate that the symbiont is capable of manipulating the mosquito virome and that Wolbachia-mediated viral inhibition is not universal for flaviviruses. This may have implications for the Wolbachia-based DENV control strategy if ISFs confer fitness effects or alter mosquito susceptibility to other flaviviruses.}, } @article {pmid29931623, year = {2019}, author = {Arai, H and Hirano, T and Akizuki, N and Abe, A and Nakai, M and Kunimi, Y and Inoue, MN}, title = {Multiple Infection and Reproductive Manipulations of Wolbachia in Homona magnanima (Lepidoptera: Tortricidae).}, journal = {Microbial ecology}, volume = {77}, number = {1}, pages = {257-266}, pmid = {29931623}, issn = {1432-184X}, mesh = {Animals ; Anti-Bacterial Agents/pharmacology ; *Bacterial Physiological Phenomena ; Cytoplasm ; DNA, Bacterial/analysis ; Female ; Larva/microbiology ; Male ; Moths/drug effects/*genetics/*microbiology ; Phenotype ; Pupa/microbiology ; Reproduction/*physiology ; Sex Ratio ; Symbiosis ; *Wolbachia/classification/genetics/physiology ; }, abstract = {Endosymbiotic bacterium Wolbachia interacts with host in either a mutualistic or parasitic manner. Wolbachia is frequently identified in various arthropod species, and to date, Wolbachia infections have been detected in different insects. Here, we found a triple Wolbachia infection in Homona magnanima, a serious tea pest, and investigated the effects of three infecting Wolbachia strains (wHm-a, -b, and -c) on the host. Starting with the triple-infected host line (W[abc]), which was collected in western Tokyo in 1999 and maintained in laboratory, we established an uninfected line (W[-]) and three singly infected lines (W[a], W[b], and W[c]) using antibiotics. Mating experiments with the host lines revealed that only wHm-b induced cytoplasmic incompatibility (CI) in H. magnanima, with the intensities of CI different between the W[b] and W[abc] lines. Regarding mutualistic effects, wHm-c shortened larval development time and increased pupal weight in both the W[c] and W[abc] lines to the same extent, whereas no distinct phenotype was observed in lines singly infected with wHm-a. Based on quantitative PCR analysis, Wolbachia density in the W[a] line was higher than in the other host lines (p < 0.01, n = 10). Wolbachia density in the W[b] line was also higher than in the W[c] and W[abc] lines, while no difference was observed between the W[c] and W[abc] lines. These results indicate that the difference in the CI intensity between a single or multiple infection may be attributed to the difference in wHm-b density. However, no correlation was observed between mutualistic effects and Wolbachia density.}, } @article {pmid29908171, year = {2018}, author = {Goindin, D and Cannet, A and Delannay, C and Ramdini, C and Gustave, J and Atyame, C and Vega-Rúa, A}, title = {Screening of natural Wolbachia infection in Aedes aegypti, Aedes taeniorhynchus and Culex quinquefasciatus from Guadeloupe (French West Indies).}, journal = {Acta tropica}, volume = {185}, number = {}, pages = {314-317}, doi = {10.1016/j.actatropica.2018.06.011}, pmid = {29908171}, issn = {1873-6254}, mesh = {Aedes/*microbiology ; Animals ; Culex/*microbiology ; Guadeloupe ; Mosquito Control/methods ; Wolbachia/genetics/*isolation & purification ; }, abstract = {Guadeloupe islands are threatened by several mosquito-borne viruses such as Dengue, Chikungunya, Zika and West Nile virus. It appears essential to look for alternative mosquito control methods such as the incompatible insect technique (ITT) aiming at sterilizing wild females by inundative releases of incompatible males. Before considering the implementation of such a strategy, the characterization of genetic diversity of the endocellular bacterium Wolbachia regarding the local mosquito populations is a critical issue. Here, for the first time, we describe the prevalence and diversity of Wolbachia in natural populations of three mosquito species from Guadeloupe: Aedes aegypti, Aedes taeniorhynchus and Culex quinquefasciatus. The detection of Wolbachia in natural Ae. aegypti, Ae. taeniorhynchus and Cx. quinquefasciatus populations was conducted by studying Wolbachia 16S ribosomal RNA gene using a TaqMan quantitative real-time PCR and results were confirmed by conventional PCR and sequencing. In addition, molecular typing of wPip strains in Cx. quinquefasciatus was done by PCR-RFLP. We did not find Wolbachia infection in any of Ae. aegypti and Ae. taeniorhynchus studied populations. Natural Wolbachia infection was detected in Cx. quinquefasciatus with prevalence varying from 79.2% to 95.8%. In addition, no polymorphism was found between the Wolbachia strains infecting Cx. quinquefasciatus specimens, all carrying an infection from the same Wolbachia genetic wPip-I group. These results pave the way for the evaluation of the feasibility of IIT programs to fight against these medically-important mosquito species in Guadeloupe.}, } @article {pmid29901734, year = {2018}, author = {Gloria-Soria, A and Chiodo, TG and Powell, JR}, title = {Lack of Evidence for Natural Wolbachia Infections in Aedes aegypti (Diptera: Culicidae).}, journal = {Journal of medical entomology}, volume = {55}, number = {5}, pages = {1354-1356}, pmid = {29901734}, issn = {1938-2928}, support = {R01 AI101112/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/*microbiology ; Animals ; *Wolbachia ; }, abstract = {Wolbachia is a genus of endosymbiotic bacteria that infects 66% of all insect species. Its major impact on insects is in reproduction: sterility, production of one sex, and/or parthenogenesis. Another effect was discovered when the disease-transmitting mosquito, Aedes aegypti Linnaeus (Diptera: Culicidae), was infected with Wolbachia isolated from Drosophila: infected female mosquitoes became less capable of transmitting diseases such as dengue fever and chikungunya. This has led to releases of Ae. aegypti carrying Wolbachia in an attempt to control disease. An open question is whether there are natural Wolbachia infections of this mosquito. We assayed DNA from 2,663 Ae. aegypti from 27 countries on six continents, 230 from laboratory strains, and 72 Aedes mascarensis MacGregor (Diptera: Culicidae) for presence of Wolbachia DNA. Within the limits of our polymerase chain reaction-based assay, we found no evidence of Wolbachia, suggesting that natural infections of this endosymbiont are unlikely to occur throughout the worldwide distribution of Ae. aegypti.}, } @article {pmid29897678, year = {2019}, author = {Uribe-Alvarez, C and Chiquete-Félix, N and Morales-García, L and Bohórquez-Hernández, A and Delgado-Buenrostro, NL and Vaca, L and Peña, A and Uribe-Carvajal, S}, title = {Wolbachia pipientis grows in Saccharomyces cerevisiae evoking early death of the host and deregulation of mitochondrial metabolism.}, journal = {MicrobiologyOpen}, volume = {8}, number = {4}, pages = {e00675}, pmid = {29897678}, issn = {2045-8827}, mesh = {Animals ; Bacterial Proteins/genetics/metabolism ; Host-Pathogen Interactions ; Insecta/*microbiology/physiology ; Mitochondria/*metabolism ; Oxidative Phosphorylation ; Saccharomyces cerevisiae/chemistry/*metabolism ; Wolbachia/*growth & development ; }, abstract = {Wolbachia sp. has colonized over 70% of insect species, successfully manipulating host fertility, protein expression, lifespan, and metabolism. Understanding and engineering the biochemistry and physiology of Wolbachia holds great promise for insect vector-borne disease eradication. Wolbachia is cultured in cell lines, which have long duplication times and are difficult to manipulate and study. The yeast strain Saccharomyces cerevisiae W303 was used successfully as an artificial host for Wolbachia wAlbB. As compared to controls, infected yeast lost viability early, probably as a result of an abnormally high mitochondrial oxidative phosphorylation activity observed at late stages of growth. No respiratory chain proteins from Wolbachia were detected, while several Wolbachia F1 F0 -ATPase subunits were revealed. After 5 days outside the cell, Wolbachia remained fully infective against insect cells.}, } @article {pmid29893631, year = {2018}, author = {Vasconcelos, EJR and Billeter, SA and Jett, LA and Meinersmann, RJ and Barr, MC and Diniz, PPVP and Oakley, BB}, title = {Assessing Cat Flea Microbiomes in Northern and Southern California by 16S rRNA Next-Generation Sequencing.}, journal = {Vector borne and zoonotic diseases (Larchmont, N.Y.)}, volume = {18}, number = {9}, pages = {491-499}, doi = {10.1089/vbz.2018.2282}, pmid = {29893631}, issn = {1557-7759}, mesh = {Animals ; Bacteria/classification/genetics/*isolation & purification ; California/epidemiology ; Cat Diseases/epidemiology/parasitology ; Cats ; Ctenocephalides/*microbiology ; DNA, Bacterial/genetics ; RNA, Bacterial/*genetics ; RNA, Ribosomal, 16S/*genetics ; }, abstract = {Flea-borne diseases (FBDs) impact both human and animal health worldwide. Because adult fleas are obligately hematophagous and can harbor potential pathogens, fleas act as ectoparasites of vertebrates, as well as zoonotic disease vectors. Cat fleas (Ctenocephalides felis) are important vectors of two zoonotic bacterial genera listed as priority pathogens by the National Institute of Allergy and Infectious Diseases (NIAID-USA): Bartonella spp. and Rickettsia spp., causative agents of bartonelloses and rickettsioses, respectively. In this study, we introduce the first microbiome analysis of C. felis samples from California, determining the presence and abundance of relevant pathogenic genera by characterizing the cat flea microbiome through 16S rRNA next-generation sequencing (16S-NGS). Samples from both northern (NoCal) and southern (SoCal) California were assessed to expand current knowledge regarding FBDs in the state. We identified Rickettsia and Bartonella, as well as the endosymbiont Wolbachia, as the most abundant genera, followed by less abundant taxa. In comparison to our previous study screening Californian cat fleas for rickettsiae using PCR/digestion/sequencing of the ompB gene, the 16S-NGS approach applied herein showed a 95% level of agreement in detecting Rickettsia spp. There was no overall difference in microbiome diversity between NoCal and SoCal samples. Bacterial taxa identified by 16S-NGS in this study may help to improve epidemiological investigations, pathogen surveillance efforts, and clinical diagnostics of FBDs in California and elsewhere.}, } @article {pmid29891919, year = {2018}, author = {Balvín, O and Roth, S and Talbot, B and Reinhardt, K}, title = {Co-speciation in bedbug Wolbachia parallel the pattern in nematode hosts.}, journal = {Scientific reports}, volume = {8}, number = {1}, pages = {8797}, pmid = {29891919}, issn = {2045-2322}, mesh = {Animals ; Bedbugs/*microbiology ; Biosynthetic Pathways/genetics ; Biotin/biosynthesis ; *Genetic Speciation ; *Symbiosis ; Wolbachia/*classification/genetics/*isolation & purification/metabolism ; }, abstract = {Wolbachia bacteria, vertically transmitted intracellular endosymbionts, are associated with two major host taxa in which they show strikingly different symbiotic modes. In some taxa of filarial nematodes, where Wolbachia are strictly obligately beneficial to the host, they show complete within- and among-species prevalence as well as co-phylogeny with their hosts. In arthropods, Wolbachia usually are parasitic; if beneficial effects occurs, they can be facultative or obligate, related to host reproduction. In arthropods, the prevalence of Wolbachia varies within and among taxa, and no co-speciation events are known. However, one arthropod species, the common bedbug Cimex lectularius was recently found to be dependent on the provision of biotin and riboflavin by Wolbachia, representing a unique case of Wolbachia providing nutritional and obligate benefits to an arthropod host, perhaps even in a mutualistic manner. Using the presence of presumably functional biotin gene copies, our study demonstrates that the obligate relationship is maintained at least in 10 out of 15 species of the genera Cimex and Paracimex. The remaining five species harboured Wolbachia as well, demonstrating the first known case of 100% prevalence of Wolbachia among higher arthropod taxa. Moreover, we show the predicted co-cladogenesis between Wolbachia and their bedbug hosts, also as the first described case of Wolbachia co-speciation in arthropods.}, } @article {pmid29890648, year = {2018}, author = {Badawi, M and Moumen, B and Giraud, I and Grève, P and Cordaux, R}, title = {Investigating the Molecular Genetic Basis of Cytoplasmic Sex Determination Caused by Wolbachia Endosymbionts in Terrestrial Isopods.}, journal = {Genes}, volume = {9}, number = {6}, pages = {}, pmid = {29890648}, issn = {2073-4425}, abstract = {In animals, sexual differences between males and females are usually determined by sex chromosomes. Alternatively, sex may also be determined by vertically transmitted intracellular microbial endosymbionts. The best known cytoplasmic sex manipulative endosymbiont is Wolbachia which can, for instance, feminize genetic males into phenotypic females in the terrestrial isopod Armadillidium vulgare. However, the molecular genetic basis of cytoplasmic sex determination is unknown. To identify candidate genes of feminization induced by Wolbachia strain wVulC from A. vulgare, we sequenced the genome of Wolbachia strain wCon from Cylisticus convexus, the most closely related known Wolbachia strain to wVulC that does not induce feminization, and compared it to the wVulC genome. Then, we performed gene expression profiling of the 216 resulting wVulC candidate genes throughout host developmental stages in A. vulgare and the heterologous host C. convexus. We identified a set of 35 feminization candidate genes showing differential expression during host sexual development. Interestingly, 27 of the 35 genes are present in the f element, which is a piece of a feminizing Wolbachia genome horizontally transferred into the nuclear genome of A. vulgare and involved in female sex determination. Assuming that the molecular genetic basis of feminization by Wolbachia and the f element is the same, the 27 genes are candidates for acting as master sex determination genes in A. vulgare females carrying the f element.}, } @article {pmid29876068, year = {2018}, author = {Zhang, L and Yun, Y and Hu, G and Peng, Y}, title = {Insights into the bacterial symbiont diversity in spiders.}, journal = {Ecology and evolution}, volume = {8}, number = {10}, pages = {4899-4906}, pmid = {29876068}, issn = {2045-7758}, abstract = {Most spiders are natural enemies of pests, and it is beneficial for the biological control of pests to learn the relationships between symbionts and their spider hosts. Research on the bacterial communities of insects has been conducted recently, but only a few studies have addressed the bacterial communities of spiders. To obtain a complete overview of the microbial communities of spiders, we examined eight species of spider (Pirata subpiraticus, Agelena difficilis, Artema atlanta, Nurscia albofasciata, Agelena labyrinthica, Ummeliata insecticeps, Dictis striatipes, and Hylyphantes graminicola) with high-throughput sequencing based on the V3 and V4 regions of the 16S rRNA gene. The bacterial communities of the spider samples were dominated by five types of endosymbionts, Wolbachia, Cardinium, Rickettsia, Spiroplasma, and Rickettsiella. The dominant OTUs (operational taxonomic units) from each of the five endosymbionts were analyzed, and the results showed that different spider species were usually dominated by special OTUs. In addition to endosymbionts, Pseudomonas, Sphingomonas, Acinetobacter, Novosphingobium, Aquabacterium, Methylobacterium, Brevundimonas, Rhizobium, Bradyrhizobium, Citrobacter, Arthrobacter, Pseudonocardia, Microbacterium, Lactobacillus, and Lactococcus were detected in spider samples in our study. Moreover, the abundance of Sphingomonas, Methylobacterium, Brevundimonas, and Rhizobium in the spider D. striatipes was significantly higher (p < .05) than the bacterial abundance of these species in seven other spider species. These findings suggest that same as in insects, co-infection of multiple types of endosymbionts is common in the hosts of the Araneae order, and other bacterial taxa also exist in spiders besides the endosymbionts.}, } @article {pmid29868291, year = {2018}, author = {Li, J and Wang, N and Liu, Y and Qiu, S}, title = {Proteomics of Nasonia vitripennis and the effects of native Wolbachia infection on N. vitripennis.}, journal = {PeerJ}, volume = {6}, number = {}, pages = {e4905}, pmid = {29868291}, issn = {2167-8359}, abstract = {BACKGROUND: Nasonia vitripennis, a parasitic wasp, is a good model organism to study developmental and evolutionary genetics and to evaluate the interactions between insect hosts and their symbionts. Wolbachia may be the most prevalent endosymbiont among insect species due to their special ability to improve the fitness of the infected hosts. Transinfection of bacteria or fungi could substantially alter the expression of host immune system components. However, few studies have focused on the effects of native Wolbachia infection. Accordingly, in this study, we evaluated the proteomics of N. vitripennis following Wolbachia infection.

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

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

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

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

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

TRIAL REGISTRATION: ClinicalTrials.gov, identifier: NCT03055585 . Registered on 14 February 2017.}, } @article {pmid33601807, year = {2018}, author = {Noor-Shazleen-Husnie, MM and Emelia, O and Ahmad-Firdaus, MS and Zainol-Ariffin, P and Aishah-Hani, A}, title = {Detection of Wolbachia in wild mosquito populations from selected areas in Peninsular Malaysia by loop-mediated isothermal amplification (LAMP) technique.}, journal = {Tropical biomedicine}, volume = {35}, number = {2}, pages = {330-346}, pmid = {33601807}, issn = {2521-9855}, abstract = {Wolbachia, a naturally endosymbiotic bacteria, has shown its potential as one of biological control agents for vector borne diseases. Due to large number of mosquitoes involved in Wolbachia screening, Loop-mediated isothermal amplification (LAMP) is recommended as a convenient and time-saving technique. This study aimed to evaluate a newly developed LAMP assay for detection of Wolbachia by targeting 16S rDNA gene in samples of wild mosquito populations. The LAMP products were confirmed by colorimetric detection using hydroxy naphthol blue (HNB), digestion with RsaI restriction enzyme and gel electrophoresis. The restriction enzyme digestion of PCR products was performed to differentiate between Wolbachia supergroups A and B. Out of 765 mosquito samples tested, 349 (45.6%) and 237 (31%) of the samples were positive for LAMP and PCR techniques respectively. The prevalence of Wolbachia detected in mosquitoes was significantly higher using LAMP as compared to PCR. There is significant association between numbers of mosquitoes positive with Wolbachia detected using LAMP and PCR (χ[2]=61.31; df=1; p < 0.05) with a kappa (κ) value of 0.27 and Phi value, 0.283. This study highlighted the potential of LAMP as a sensitive, specific and rapid tool for screening of Wolbachia in mosquitoes, thus it presents as an alternative to PCR-based assays.}, } @article {pmid29851312, year = {2018}, author = {Dahmani, M and Tahir, D and Cabre, O and Raoult, D and Fenollar, F and Davoust, B and Mediannikov, O}, title = {Prevalence of Anaplasmataceae and Filariidae species in unowned and military dogs in New Caledonia.}, journal = {Veterinary medicine and science}, volume = {4}, number = {2}, pages = {140-149}, pmid = {29851312}, issn = {2053-1095}, mesh = {Anaplasmataceae/physiology ; Anaplasmataceae Infections/epidemiology/parasitology/*veterinary ; Animals ; Dog Diseases/*epidemiology/parasitology ; Dogs ; Female ; Filariasis/epidemiology/parasitology/*veterinary ; Filarioidea/physiology ; Male ; Military Personnel ; New Caledonia/epidemiology ; Ownership ; Prevalence ; }, abstract = {Dogs are competent reservoir hosts of several zoonotic agents, including Filariidae nematodes and Anaplasmataceae family bacteria. The latter family unites human and veterinary pathogens (Anaplasma, Ehrlichia and Neorickettsia bacteria) with Wolbachia, some of which are obligatory endosymbionts of pathogenic filarial nematodes. The epidemiology of Anaplasmataceae and Filariidae species infecting dogs living in kennels in New Caledonia was studied. 64 EDTA blood samples were screened for the presence of Anaplasmataceae and filarial nematodes. Molecular study was conducted using primers and probe targeting the of 23S rRNA long fragment of Anaplasmataceae species. Next, all blood sample was screened for the presence of Filariidae species targeting the primers and probe targeting the COI gene, as well as primers targeting the COI and 5S rRNA genes of all filarial worms. Anaplasma platys was identified in 8/64 (12.5, 95% confidence interval [CI]: 4.4-20.6%) and Wolbachia endosymbiont of Dirofilaria immitis in 8/64 (12.5%, CI: 4.4-20.6%). Filariidae species investigation was performed and showed that 11/64 (17.2%, CI: 7.9-26.4%) dogs were infected with D. immitis, whereas, 2/64 (3.1%, CI: 0.0-7.3%) were infected with Acanthocheilonema reconditum. Finally, we checked the occurrence of co-infection between Anaplasmataceae and Filariidae species. Co-occurrence with Wolbachia endosymbiont of D. immitis was observed in seven dogs, one dog was co-infected with A. platys and A. reconditum and another was co-infected with Wolbachia endosymbiont of D. immitis and A. reconditum. These results are the first report of Anaplasmataceae and Filariidae occurring in dogs in New Caledonia.}, } @article {pmid29851149, year = {2018}, author = {Ote, M and Yamamoto, D}, title = {The Wolbachia protein TomO interacts with a host RNA to induce polarization defects in Drosophila oocytes.}, journal = {Archives of insect biochemistry and physiology}, volume = {99}, number = {1}, pages = {e21475}, doi = {10.1002/arch.21475}, pmid = {29851149}, issn = {1520-6327}, mesh = {Animals ; Bacterial Proteins/*genetics/metabolism ; Body Patterning ; Drosophila simulans/*embryology/*microbiology ; Embryo, Nonmammalian/microbiology ; Embryonic Development ; Oocytes/growth & development ; RNA/metabolism ; Wolbachia/genetics/*physiology ; }, abstract = {Wolbachia is an endosymbiont prevalent in arthropods. To maximize its transmission thorough the female germline, Wolbachia induces in infected hosts male-to-female transformation, male killing, parthenogenesis, and cytoplasmic incompatibility, depending on the host species and Wolbachia strain involved. However, the molecular mechanisms underlying these host manipulations by Wolbachia remain largely unknown. The Wolbachia strain wMel, an inhabitant of Drosophila melanogaster, impairs host oogenesis only when transplanted into a heterologous host, for example, Drosophila simulans. We found that egg polarity defects induced by wMel infection in D. simulans can be recapitulated in the natural host D. melanogaster by transgenic overexpression of a variant of the Wolbachia protein Toxic manipulator of oogenesis (TomO), TomOwMel[∆HS] , in the female germline. RNA immunoprecipitation assays demonstrated that TomO physically associates with orb mRNA, which, as a result, fails to interact with the translation repressor Cup. This leads to precocious translation of Orb, a posterior determinant, and thereby to the misspecification of oocytes and accompanying polarity defects. We propose that the ability of TomO to bind to orb mRNA might provide a means for Wolbachia to enter the oocyte located at the posterior end of the egg chamber, thereby accomplishing secure maternal transmission thorough the female germline.}, } @article {pmid29845544, year = {2018}, author = {O'Neill, SL}, title = {The Use of Wolbachia by the World Mosquito Program to Interrupt Transmission of Aedes aegypti Transmitted Viruses.}, journal = {Advances in experimental medicine and biology}, volume = {1062}, number = {}, pages = {355-360}, doi = {10.1007/978-981-10-8727-1_24}, pmid = {29845544}, issn = {0065-2598}, mesh = {Aedes/*microbiology/virology ; Animals ; Australia ; Humans ; Mosquito Control ; Mosquito Vectors/*microbiology/virology ; Virus Diseases/transmission/virology ; *Virus Physiological Phenomena ; Viruses/genetics ; Wolbachia/*physiology ; }, abstract = {The biological control of mosquito transmission by the use of the naturally occurring insect-specific bacterial endosymbiont Wolbachia has been successfully tested in small field trials. The approach has been translated successfully to larger field sites in Townsville, Australia and expanded to more than 10 countries through the Eliminate Dengue Program. The broader application of the program beyond limiting the transmission of dengue and including other Aedes aegypti borne mosquitoes has seen the program growing into a global not-for-profit initiative to be known as the World Mosquito Program.}, } @article {pmid29843598, year = {2018}, author = {Perry, KD and Baker, GJ and Powis, KJ and Kent, JK and Ward, CM and Baxter, SW}, title = {Cryptic Plutella species show deep divergence despite the capacity to hybridize.}, journal = {BMC evolutionary biology}, volume = {18}, number = {1}, pages = {77}, pmid = {29843598}, issn = {1471-2148}, support = {UA00146//University of Adelaide/International ; DAS00094//Grains Research and Development Corporation/International ; DAS00155//Grains Research and Development Corporation/International ; DAS00155//Grains Research and Development Corporation/International ; DAS00155//Grains Research and Development Corporation/International ; DP120100047//Australian Research Council/International ; FT140101303//Australian Research Council/International ; }, mesh = {Animals ; Australia ; Biological Assay ; Crosses, Genetic ; DNA, Mitochondrial/genetics ; Female ; Fertility ; *Genetic Variation ; Genetics, Population ; Geography ; Haplotypes/genetics ; Heterozygote ; *Hybridization, Genetic/drug effects ; Insecticide Resistance/drug effects/genetics ; Insecticides/toxicity ; Likelihood Functions ; Male ; Mitochondria/genetics ; Moths/*genetics/microbiology ; Phylogeny ; Species Specificity ; Sympatry ; Wolbachia/drug effects/physiology ; }, abstract = {BACKGROUND: Understanding genomic and phenotypic diversity among cryptic pest taxa has important implications for the management of pests and diseases. The diamondback moth, Plutella xylostella L., has been intensively studied due to its ability to evolve insecticide resistance and status as the world's most destructive pest of brassicaceous crops. The surprise discovery of a cryptic species endemic to Australia, Plutella australiana Landry & Hebert, raised questions regarding the distribution, ecological traits and pest status of the two species, the capacity for gene flow and whether specific management was required. Here, we collected Plutella from wild and cultivated brassicaceous plants from 75 locations throughout Australia and screened 1447 individuals to identify mtDNA lineages and Wolbachia infections. We genotyped genome-wide SNP markers using RADseq in coexisting populations of each species. In addition, we assessed reproductive compatibility in crossing experiments and insecticide susceptibility phenotypes using bioassays.

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

CONCLUSIONS: Wolbachia infection leads to small sex-dependent changes on the flight tones of Ae. aegypti, but it does not seem to prevent the stereotyped harmonic interaction between males and females. Therefore, when released in the natural habitat to breed with native individuals, Wolbachia-harboring individuals shall be fit enough to meet the criteria of acoustically-related mating behavior and promote bacteria dispersion effectively.}, } @article {pmid29468305, year = {2018}, author = {Ramalho, MO and Vieira, AS and Pereira, MC and Moreau, CS and Bueno, OC}, title = {Transovarian Transmission of Blochmannia and Wolbachia Endosymbionts in the Neotropical Weaver Ant Camponotus textor (Hymenoptera, Formicidae).}, journal = {Current microbiology}, volume = {75}, number = {7}, pages = {866-873}, pmid = {29468305}, issn = {1432-0991}, support = {007343/2014-00//CAPES Foundation/ ; 157837/2015-7//Conselho Nacional de Desenvolvimento Científico e Tecnológico/ ; }, mesh = {Animals ; Ants/growth & development/*microbiology ; Enterobacteriaceae/classification/genetics/*isolation & purification/physiology ; Female ; In Situ Hybridization, Fluorescence ; Ovary/growth & development/microbiology ; Ovum/growth & development/microbiology ; *Symbiosis ; Wolbachia/classification/genetics/*isolation & purification/physiology ; }, abstract = {Camponotus is a hyper-diverse ant genus that is associated with the obligate endosymbiont Blochmannia, and often also with Wolbachia, but morphological studies on the location of these bacteria in the queen's ovaries during oogenesis remain limited. In the present study, we used the Neotropical weaver ant Camponotus textor to characterize the ovary using histology (HE) techniques, and to document the location of Blochmannia and Wolbachia during oogenesis through fluorescence in situ hybridization (FISH). This is the first morphological report of these two bacteria in the same host with polytrophic meroistic ovaries and reveals that Blochmannia is found inside late-stage oocytes and Wolbachia is associated with the nuclei of the nurse cells. Our results provide insights into the developmental sequence of when these bacteria reach the egg, with Blochmannia establishing itself in the egg first, and Wolbachia only reaching the egg shortly before completing egg development. Studies such as this provide understanding about the mechanisms and timing of the establishment of these endosymbionts in the host.}, } @article {pmid29468014, year = {2018}, author = {Zu Dohna, H and Houry, C and Kambris, Z}, title = {A comparative analysis of Wolbachia-induced host reproductive phenotypes reveals transition rate heterogeneity.}, journal = {Ecology and evolution}, volume = {8}, number = {4}, pages = {1945-1953}, pmid = {29468014}, issn = {2045-7758}, abstract = {The endosymbiotic bacterium Wolbachia infects a wide range of arthropods and their relatives. It is an intracellular parasite transmitted through the egg from mother to offspring. Wolbachia can spread and persist through various means of host reproductive manipulation. How these different mechanisms of host manipulation evolved in Wolbachia is unclear. Which host reproductive phenotype is most likely to be ancestral and whether evolutionary transitions between some host phenotypes are more common than others remain unanswered questions. Recent studies have revealed multiple cases where the same Wolbachia strain can induce different reproductive phenotypes in different hosts, raising the question to what degree the induced host phenotype should be regarded as a trait of Wolbachia. In this study, we constructed a phylogenetic tree of Wolbachia and analyzed the patterns of host phenotypes along that tree. We were able to detect a phylogenetic signal of host phenotypes on the Wolbachia tree, indicating that the induced host phenotype can be regarded as a Wolbachia trait. However, we found no clear support for the previously stated hypothesis that cytoplasmic incompatibility is ancestral to Wolbachia in arthropods. Our analysis provides evidence for heterogeneous transition rates between host phenotypes.}, } @article {pmid29467241, year = {2018}, author = {Kamath, AD and Deehan, MA and Frydman, HM}, title = {Polar cell fate stimulates Wolbachia intracellular growth.}, journal = {Development (Cambridge, England)}, volume = {145}, number = {6}, pages = {}, pmid = {29467241}, issn = {1477-9129}, support = {R56 AI097589/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Drosophila/*microbiology ; Female ; Fluorescent Antibody Technique ; In Situ Hybridization, Fluorescence ; Oogenesis/physiology ; Ovary/cytology/microbiology ; Polar Bodies/*microbiology ; Symbiosis/*physiology ; Viral Tropism/*physiology ; Wolbachia/*pathogenicity ; }, abstract = {Bacteria are crucial partners in the development and evolution of vertebrates and invertebrates. A large fraction of insects harbor Wolbachia, bacterial endosymbionts that manipulate host reproduction to favor their spreading. Because they are maternally inherited, Wolbachia are under selective pressure to reach the female germline and infect the offspring. However, Wolbachia infection is not limited to the germline. Somatic cell types, including stem cell niches, have higher Wolbachia loads compared with the surrounding tissue. Here, we show a novel Wolbachia tropism to polar cells (PCs), specialized somatic cells in the Drosophila ovary. During oogenesis, all stages of PC development are easily visualized, facilitating the investigation of the kinetics of Wolbachia intracellular growth. Wolbachia accumulation is triggered by particular events of PC morphogenesis, including differentiation from progenitors and between stages 8 and 9 of oogenesis. Moreover, induction of ectopic PC fate is sufficient to promote Wolbachia accumulation. We found that Wolbachia PC tropism is evolutionarily conserved across most Drosophila species, but not in Culex mosquitos. These findings highlight the coordination of endosymbiont tropism with host development and cell differentiation.}, } @article {pmid29466376, year = {2018}, author = {Yadav, S and Frazer, J and Banga, A and Pruitt, K and Harsh, S and Jaenike, J and Eleftherianos, I}, title = {Endosymbiont-based immunity in Drosophila melanogaster against parasitic nematode infection.}, journal = {PloS one}, volume = {13}, number = {2}, pages = {e0192183}, pmid = {29466376}, issn = {1932-6203}, support = {R01 AI110675/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Drosophila melanogaster/*immunology/microbiology/parasitology ; Host-Parasite Interactions ; Nematoda/*physiology ; *Symbiosis ; Wolbachia/isolation & purification ; }, abstract = {Associations between endosymbiotic bacteria and their hosts represent a complex ecosystem within organisms ranging from humans to protozoa. Drosophila species are known to naturally harbor Wolbachia and Spiroplasma endosymbionts, which play a protective role against certain microbial infections. Here, we investigated whether the presence or absence of endosymbionts affects the immune response of Drosophila melanogaster larvae to infection by Steinernema carpocapsae nematodes carrying or lacking their mutualistic Gram-negative bacteria Xenorhabdus nematophila (symbiotic or axenic nematodes, respectively). We find that the presence of Wolbachia alone or together with Spiroplasma promotes the survival of larvae in response to infection with S. carpocapsae symbiotic nematodes, but not against axenic nematodes. We also find that Wolbachia numbers are reduced in Spiroplasma-free larvae infected with axenic compared to symbiotic nematodes, and they are also reduced in Spiroplasma-containing compared to Spiroplasma-free larvae infected with axenic nematodes. We further show that S. carpocapsae axenic nematode infection induces the Toll pathway in the absence of Wolbachia, and that symbiotic nematode infection leads to increased phenoloxidase activity in D. melanogaster larvae devoid of endosymbionts. Finally, infection with either type of nematode alters the metabolic status and the fat body lipid droplet size in D. melanogaster larvae containing only Wolbachia or both endosymbionts. Our results suggest an interaction between Wolbachia endosymbionts with the immune response of D. melanogaster against infection with the entomopathogenic nematodes S. carpocapsae. Results from this study indicate a complex interplay between insect hosts, endosymbiotic microbes and pathogenic organisms.}, } @article {pmid29447357, year = {2019}, author = {Jewell, NP and Dufault, S and Cutcher, Z and Simmons, CP and Anders, KL}, title = {Analysis of cluster-randomized test-negative designs: cluster-level methods.}, journal = {Biostatistics (Oxford, England)}, volume = {20}, number = {2}, pages = {332-346}, pmid = {29447357}, issn = {1468-4357}, support = {R56 AI134724/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/microbiology ; Animals ; Biostatistics/*methods ; Dengue/prevention & control ; Humans ; *Models, Statistical ; Mosquito Vectors ; *Random Allocation ; Randomized Controlled Trials as Topic/*methods ; Wolbachia/pathogenicity ; }, abstract = {Intervention trials of vector control methods often require community level randomization with appropriate inferential methods. For many interventions, the possibility of confounding due to the effects of health-care seeking behavior on disease ascertainment remains a concern. The test-negative design, a variant of the case-control method, was introduced to mitigate this issue in the assessment of the efficacy of influenza vaccination (measured at an individual level) on influenza infection. Here, we introduce a cluster-randomized test-negative design that includes randomization of the intervention at a group level. We propose several methods for estimation and inference regarding the relative risk (RR). The inferential methods considered are based on the randomization distribution induced by permuting intervention assignment across two sets of randomly selected clusters. The motivating example is a current study of the efficacy of randomized releases of Wolbachia-infected Aedes aegypti mosquitoes to reduce the incidence of dengue in Yogyakarta City, Indonesia. Estimation and inference techniques are assessed through a simulation study.}, } @article {pmid29443908, year = {2018}, author = {Lim, EXY and Lee, WS and Madzokere, ET and Herrero, LJ}, title = {Mosquitoes as Suitable Vectors for Alphaviruses.}, journal = {Viruses}, volume = {10}, number = {2}, pages = {}, pmid = {29443908}, issn = {1999-4915}, mesh = {Alphavirus/*physiology ; Alphavirus Infections/*transmission/*virology ; Animals ; Coinfection ; Culicidae/*virology ; Humans ; Mosquito Vectors/*virology ; Mutation ; RNA, Viral ; }, abstract = {Alphaviruses are arthropod-borne viruses and are predominantly transmitted via mosquito vectors. This vector preference by alphaviruses raises the important question of the determinants that contribute to vector competence. There are several tissue barriers of the mosquito that the virus must overcome in order to establish a productive infection. Of importance are the midgut, basal lamina and the salivary glands. Infection of the salivary glands is crucial for virus transmission during the mosquito's subsequent bloodfeed. Other factors that may contribute to vector competence include the microflora and parasites present in the mosquito, environmental conditions, the molecular determinants of the virus to adapt to the vector, as well as the effect of co-infection with other viruses. Though mosquito innate immunity is a contributing factor to vector competence, it will not be discussed in this review. Detailed understanding of these factors will be instrumental in minimising transmission of alphaviral diseases.}, } @article {pmid29435771, year = {2018}, author = {Zélé, F and Weill, M and Magalhães, S}, title = {Identification of spider-mite species and their endosymbionts using multiplex PCR.}, journal = {Experimental & applied acarology}, volume = {74}, number = {2}, pages = {123-138}, pmid = {29435771}, issn = {1572-9702}, support = {FCT-ANR//BIA-EVF/0013/2012//Faculdade de Ciências e Tecnologia (FCT), Universidade de Lisboa (PT)/ ; FCT-TUBITAK/0001/2014//Faculdade de Ciências e Tecnologia (FCT), Universidade de Lisboa (PT)/ ; SFRH/BPD/125020/2016//Faculdade de Ciências e Tecnologia (FCT), Universidade de Lisboa (PT)/ ; }, mesh = {Animals ; Bacteria/*isolation & purification ; Bacterial Physiological Phenomena ; Female ; Male ; *Microbiota ; Multiplex Polymerase Chain Reaction/economics/*methods ; Portugal ; Spain ; *Symbiosis ; Tetranychidae/*classification/genetics/microbiology ; }, abstract = {Spider mites of the genus Tetranychidae are severe crop pests. In the Mediterranean a few species coexist, but they are difficult to identify based on morphological characters. Additionally, spider mites often harbour several species of endosymbiotic bacteria, which may affect the biology of their hosts. Here, we propose novel, cost-effective, multiplex diagnostic methods allowing a quick identification of spider-mite species as well as of the endosymbionts they carry. First, we developed, and successfully multiplexed in a single PCR, primers to identify Tetranychus urticae, T. evansi and T. ludeni, some of the most common tetranychids found in southwest Europe. Moreover, we demonstrated that this method allows detecting multiple species in a single pool, even at low frequencies (up to 1/100), and can be used on entire mites without DNA extraction. Second, we developed another set of primers to detect spider-mite endosymbionts, namely Wolbachia, Cardinium and Rickettsia in a multiplex PCR, along with a generalist spider-mite primer to control for potential failure of DNA amplification in each PCR. Overall, our method represents a simple, cost-effective and reliable method to identify spider-mite species and their symbionts in natural field populations, as well as to detect contaminations in laboratory rearings. This method may easily be extended to other species.}, } @article {pmid29435238, year = {2018}, author = {Zhang, YK and Yang, K and Zhu, YX and Hong, XY}, title = {Symbiont-conferred reproduction and fitness benefits can favour their host occurrence.}, journal = {Ecology and evolution}, volume = {8}, number = {3}, pages = {1626-1633}, pmid = {29435238}, issn = {2045-7758}, abstract = {Double infections of Wolbachia and Spiroplasma are frequent in natural populations of Tetranychus truncatus, a polyphagous mite species that has been a dominant species in China since 2009. However, little is known about the causes and ecological importance of such coexistences. In this study, we established T. truncatus strains with different infection types and then inferred the impact of the two endosymbionts on host reproduction and fitness. Double infection induced cytoplasmic incompatibility, which was demonstrated by reduction in egg hatchability of incompatible crosses. However, doubly infected females produced more eggs relative to other strains. Wolbachia and Spiroplasma did not affect host survival, whereas doubly infected females and males developed faster than other strains. Such reproduction and fitness benefits provided by double infections may be associated with the lower densities of each symbiont, and the quantitative results also confirmed competition between Wolbachia and Spiroplasma in doubly infected females. These symbiont-conferred beneficial effects maintain stable prevalence of the symbionts and also help drive T. truncatus outbreaks in combination with other environmental factors.}, } @article {pmid29429122, year = {2018}, author = {Campo-Duarte, DE and Vasilieva, O and Cardona-Salgado, D and Svinin, M}, title = {Optimal control approach for establishing wMelPop Wolbachia infection among wild Aedes aegypti populations.}, journal = {Journal of mathematical biology}, volume = {76}, number = {7}, pages = {1907-1950}, pmid = {29429122}, issn = {1432-1416}, mesh = {Aedes/*microbiology/pathogenicity ; Animals ; Computational Biology ; Computer Simulation ; Dengue/prevention & control/transmission ; Female ; Host Microbial Interactions/physiology ; Humans ; Male ; Mathematical Concepts ; *Models, Biological ; Mosquito Vectors/*microbiology/pathogenicity ; Pest Control, Biological/*methods/statistics & numerical data ; Population Dynamics/statistics & numerical data ; Wolbachia/pathogenicity/*physiology ; }, abstract = {Wolbachia-based biocontrol has recently emerged as a potential method for prevention and control of dengue and other vector-borne diseases. Major vector species, such as Aedes aegypti females, when deliberately infected with Wolbachia become less capable of getting viral infections and transmitting the virus to human hosts. In this paper, we propose an explicit sex-structured population model that describes an interaction of uninfected (wild) male and female mosquitoes and those deliberately infected with wMelPop strain of Wolbachia in the same locality. This particular strain of Wolbachia is regarded as the best blocker of dengue and other arboviral infections. However, wMelPop strain of Wolbachia also causes the loss of individual fitness in Aedes aegypti mosquitoes. Our model allows for natural introduction of the decision (or control) variable, and we apply the optimal control approach to simulate wMelPop Wolbachia infestation of wild Aedes aegypti populations. The control action consists in continuous periodic releases of mosquitoes previously infected with wMelPop strain of Wolbachia in laboratory conditions. The ultimate purpose of control is to find a tradeoff between reaching the population replacement in minimum time and with minimum cost of the control effort. This approach also allows us to estimate the number of Wolbachia-carrying mosquitoes to be released in day-by-day control action. The proposed method of biological control is safe to human health, does not contaminate the environment, does not make harm to non-target species, and preserves their interaction with mosquitoes in the ecosystem.}, } @article {pmid29428509, year = {2018}, author = {Sahoo, RK and Lohman, DJ and Wahlberg, N and Müller, CJ and Brattström, O and Collins, SC and Peggie, D and Aduse-Poku, K and Kodandaramaiah, U}, title = {Evolution of Hypolimnas butterflies (Nymphalidae): Out-of-Africa origin and Wolbachia-mediated introgression.}, journal = {Molecular phylogenetics and evolution}, volume = {123}, number = {}, pages = {50-58}, doi = {10.1016/j.ympev.2018.02.001}, pmid = {29428509}, issn = {1095-9513}, mesh = {Africa ; Animals ; Base Sequence ; Bayes Theorem ; Biodiversity ; *Biological Evolution ; Butterflies/*genetics/*microbiology ; DNA, Mitochondrial/genetics ; Haplotypes/genetics ; Larva/physiology ; Likelihood Functions ; Mitochondria/genetics ; Phylogeny ; Phylogeography ; Sequence Analysis, DNA ; Wolbachia/*physiology ; }, abstract = {Hypolimnas butterflies (Nymphalidae), commonly known as eggflies, are a popular model system for studying a wide range of ecological questions including mimicry, polymorphism, wing pattern evolution, and Wolbachia-host interactions. The lack of a time-calibrated phylogeny for this group has precluded understanding its evolutionary history. We reconstruct a species-level phylogeny using a nine gene dataset and estimate species divergence times. Based on the resulting tree, we investigate the taxon's historical biogeography, examine the evolution of host plant preferences, and test the hypothesis that the endosymbiotic bacterium Wolbachia mediates gene transfer between species. Our analyses indicate that the species are grouped within three strongly supported, deeply divergent clades. However, relationships among these three clades are uncertain. In addition, many Hypolimnas species are not monophyletic or monophyletic with weak support, suggesting widespread incomplete lineage sorting and/or introgression. Biogeographic analysis strongly indicates that the genus diverged from its ancestor in Africa and subsequently dispersed to Asia; the strength of this result is not affected by topological uncertainties. While the larvae of African species feed almost exclusively on Urticaceae, larvae of species found further east often feed on several additional families. Interestingly, we found an identical mitochondrial haplotype in two Hypolimnas species, H. bolina and H. alimena, and a strong association between this mitotype and the Wolbachia strain wBol1a. Future investigations should explore the plausibility of Wolbachia-mediated introgression between species.}, } @article {pmid29396201, year = {2018}, author = {Ritchie, SA and van den Hurk, AF and Smout, MJ and Staunton, KM and Hoffmann, AA}, title = {Mission Accomplished? We Need a Guide to the 'Post Release' World of Wolbachia for Aedes-borne Disease Control.}, journal = {Trends in parasitology}, volume = {34}, number = {3}, pages = {217-226}, doi = {10.1016/j.pt.2017.11.011}, pmid = {29396201}, issn = {1471-5007}, mesh = {Aedes/*microbiology/*virology ; Animals ; Mosquito Vectors/*microbiology/*virology ; Pest Control, Biological/standards ; RNA Viruses/*physiology ; Wolbachia/*physiology ; }, abstract = {Historically, sustained control of Aedes aegypti, the vector of dengue, chikungunya, yellow fever, and Zika viruses, has been largely ineffective. Subsequently, two novel 'rear and release' control strategies utilizing mosquitoes infected with Wolbachia are currently being developed and deployed widely. In the incompatible insect technique, male Aedes mosquitoes, infected with Wolbachia, suppress populations through unproductive mating. In the transinfection strategy, both male and female Wolbachia-infected Ae. aegypti mosquitoes rapidly infect the wild population with Wolbachia, blocking virus transmission. It is critical to monitor the long-term stability of Wolbachia in host populations, and also the ability of this bacterium to continually inhibit virus transmission. Ongoing release and monitoring programs must be future-proofed should political support weaken when these vectors are successfully controlled.}, } @article {pmid29390142, year = {2018}, author = {Zélé, F and Santos, I and Olivieri, I and Weill, M and Duron, O and Magalhães, S}, title = {Endosymbiont diversity and prevalence in herbivorous spider mite populations in South-Western Europe.}, journal = {FEMS microbiology ecology}, volume = {94}, number = {4}, pages = {}, doi = {10.1093/femsec/fiy015}, pmid = {29390142}, issn = {1574-6941}, mesh = {Animals ; Bacteroidetes/classification/genetics/*isolation & purification ; Biological Evolution ; Europe ; Herbivory ; Microbiota/genetics ; Phylogeny ; Prevalence ; RNA, Ribosomal, 16S/genetics ; Rickettsia/classification/genetics/*isolation & purification ; Symbiosis ; Tetranychidae/*microbiology ; Wolbachia/classification/genetics/*isolation & purification ; }, abstract = {Bacterial endosymbionts are known as important players of the evolutionary ecology of their hosts. However, their distribution, prevalence and diversity are still largely unexplored. To this aim, we investigated infections by the most common bacterial reproductive manipulators in herbivorous spider mites of South-Western Europe. Across 16 populations belonging to three Tetranychus species, Wolbachia was the most prevalent (ca. 61%), followed by Cardinium (12%-15%), while only few individuals were infected by Rickettsia (0.9%-3%), and none carried Arsenophonus or Spiroplasma. These endosymbionts are here reported for the first time in Tetranychus evansi and Tetranychus ludeni, and showed variable infection frequencies between and within species, with several cases of coinfections. Moreover, Cardinium was more prevalent in Wolbachia-infected individuals, which suggests facilitation between these symbionts. Finally, sequence comparisons revealed no variation of the Wolbachia wsp and Rickettsia gtlA genes, but some diversity of the Cardinium 16S rRNA, both between and within populations of the three mite species. Some of the Cardinium sequences identified belonged to distantly-related clades, and the lack of association between these sequences and spider mite mitotypes suggests repeated host switching of Cardinium. Overall, our results reveal a complex community of symbionts in this system, opening the path for future studies.}, } @article {pmid29375803, year = {2018}, author = {Thongsripong, P and Chandler, JA and Green, AB and Kittayapong, P and Wilcox, BA and Kapan, DD and Bennett, SN}, title = {Mosquito vector-associated microbiota: Metabarcoding bacteria and eukaryotic symbionts across habitat types in Thailand endemic for dengue and other arthropod-borne diseases.}, journal = {Ecology and evolution}, volume = {8}, number = {2}, pages = {1352-1368}, pmid = {29375803}, issn = {2045-7758}, support = {P20 RR018727/RR/NCRR NIH HHS/United States ; U54 AI065359/AI/NIAID NIH HHS/United States ; }, abstract = {Vector-borne diseases are a major health burden, yet factors affecting their spread are only partially understood. For example, microbial symbionts can impact mosquito reproduction, survival, and vectorial capacity, and hence affect disease transmission. Nonetheless, current knowledge of mosquito-associated microbial communities is limited. To characterize the bacterial and eukaryotic microbial communities of multiple vector species collected from different habitat types in disease endemic areas, we employed next-generation 454 pyrosequencing of 16S and 18S rRNA amplicon libraries, also known as metabarcoding. We investigated pooled whole adult mosquitoes of three medically important vectors, Aedes aegypti, Ae. albopictus, and Culex quinquefasciatus, collected from different habitats across central Thailand where we previously characterized mosquito diversity. Our results indicate that diversity within the mosquito microbiota is low, with the majority of microbes assigned to one or a few taxa. Two of the most common eukaryotic and bacterial genera recovered (Ascogregarina and Wolbachia, respectively) are known mosquito endosymbionts with potentially parasitic and long evolutionary relationships with their hosts. Patterns of microbial composition and diversity appeared to differ by both vector species and habitat for a given species, although high variability between samples suggests a strong stochastic element to microbiota assembly. In general, our findings suggest that multiple factors, such as habitat condition and mosquito species identity, may influence overall microbial community composition, and thus provide a basis for further investigations into the interactions between vectors, their microbial communities, and human-impacted landscapes that may ultimately affect vector-borne disease risk.}, } @article {pmid29370307, year = {2018}, author = {Ant, TH and Herd, CS and Geoghegan, V and Hoffmann, AA and Sinkins, SP}, title = {The Wolbachia strain wAu provides highly efficient virus transmission blocking in Aedes aegypti.}, journal = {PLoS pathogens}, volume = {14}, number = {1}, pages = {e1006815}, pmid = {29370307}, issn = {1553-7374}, support = {MC_PC_15087/MRC_/Medical Research Council/United Kingdom ; 202888/Z/16/Z/WT_/Wellcome Trust/United Kingdom ; MC_UU_12014/8/MRC_/Medical Research Council/United Kingdom ; /WT_/Wellcome Trust/United Kingdom ; EP-D-16-021/EPA/EPA/United States ; }, mesh = {Aedes/*microbiology/*virology ; Alphavirus Infections/microbiology/transmission/virology ; Animals ; Antibiosis/*physiology ; *Dengue Virus/pathogenicity ; Female ; Infectious Disease Transmission, Vertical/*prevention & control/veterinary ; Inheritance Patterns ; Male ; Pest Control, Biological ; Semliki forest virus/pathogenicity/physiology ; Viral Load ; Wolbachia/*physiology ; }, abstract = {Introduced transinfections of the inherited bacteria Wolbachia can inhibit transmission of viruses by Aedes mosquitoes, and in Ae. aegypti are now being deployed for dengue control in a number of countries. Only three Wolbachia strains from the large number that exist in nature have to date been introduced and characterized in this species. Here novel Ae. aegypti transinfections were generated using the wAlbA and wAu strains. In its native Ae. albopictus, wAlbA is maintained at lower density than the co-infecting wAlbB, but following transfer to Ae. aegypti the relative strain density was reversed, illustrating the strain-specific nature of Wolbachia-host co-adaptation in determining density. The wAu strain also reached high densities in Ae. aegypti, and provided highly efficient transmission blocking of dengue and Zika viruses. Both wAu and wAlbA were less susceptible than wMel to density reduction/incomplete maternal transmission resulting from elevated larval rearing temperatures. Although wAu does not induce cytoplasmic incompatibility (CI), it was stably combined with a CI-inducing strain as a superinfection, and this would facilitate its spread into wild populations. Wolbachia wAu provides a very promising new option for arbovirus control, particularly for deployment in hot tropical climates.}, } @article {pmid29363888, year = {2018}, author = {Liu, QQ and Zhang, TS and Li, CX and Gu, JW and Hou, JB and Dong, H}, title = {Decision-making in a bisexual line and a thelytokous Wolbachia-infected line of Trichogramma dendrolimi Matsumura (Hymenoptera: Trichogrammatidae) regarding behavior toward their hosts.}, journal = {Pest management science}, volume = {74}, number = {7}, pages = {1720-1727}, doi = {10.1002/ps.4867}, pmid = {29363888}, issn = {1526-4998}, mesh = {Animals ; Decision Making ; Female ; Insect Control ; Moths/*parasitology ; *Pest Control, Biological ; Wasps/*microbiology/*physiology ; Wolbachia/*physiology ; }, abstract = {BACKGROUND: The use of thelytokous Wolbachia-infected Trichogramma (parasitic wasps) has long been considered as a way to enhance the efficacy of biocontrol. However, Wolbachia can affect the host physiology. We compared decision-making between bisexual and thelytokous Wolbachia-infected lines of Trichogramma dendrolimi Matsumura regarding behavior toward fresh and old eggs of Corcyra cephalonica at 25 ± 1 °C and 70 ± 5% relative humidity.

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

CONCLUSION: Wolbachia affected the fitness of T. dendrolimi negatively. The potential of the thelytokous line as a biocontrol agent would not be as good as that of the bisexual line when decision-making only is considered. Therefore, further evaluations need to be carried out before the thelytokous line can be used in practical biocontrol. © 2018 Society of Chemical Industry.}, } @article {pmid29358725, year = {2018}, author = {Schmidt, TL and Filipović, I and Hoffmann, AA and Rašić, G}, title = {Fine-scale landscape genomics helps explain the slow spatial spread of Wolbachia through the Aedes aegypti population in Cairns, Australia.}, journal = {Heredity}, volume = {120}, number = {5}, pages = {386-395}, pmid = {29358725}, issn = {1365-2540}, mesh = {Aedes/*microbiology ; Animals ; Australia ; Female ; Genetics, Population ; *Genomics ; Humans ; Larva ; Male ; Polymorphism, Single Nucleotide/*genetics ; Wolbachia/*genetics/physiology ; }, abstract = {The endosymbiotic bacterium Wolbachia suppresses the capacity for arbovirus transmission in the mosquito Aedes aegypti, and can spread spatially through wild mosquito populations following local introductions. Recent introductions in Cairns, Australia have demonstrated slower than expected spatial spread. Potential reasons for this include: (i) barriers to Ae. aegypti dispersal; (ii) higher incidence of long-range dispersal; and (iii) intergenerational loss of Wolbachia. We investigated these three potential factors using genome-wide single-nucleotide polymorphisms (SNPs) and an assay for the Wolbachia infection wMel in 161 Ae. aegypti collected from Cairns in 2015. We detected a small but significant barrier effect of Cairns highways on Ae. aegypti dispersal using distance-based redundancy analysis and patch-based simulation analysis. We detected a pair of putative full-siblings in ovitraps 1312 m apart, indicating long-distance female movement likely mediated by human transport. We also found a pair of full-siblings of different infection status, indicating intergenerational loss of Wolbachia in the field. These three factors are all expected to contribute to the slow spread of Wolbachia through Ae. aegypti populations, though from our results it is unclear whether Wolbachia loss and long-distance movement are sufficiently common to reduce the speed of spatial spread appreciably. Our findings inform the strategic deployment of Wolbachia-infected mosquitoes during releases, and show how parameter estimates from laboratory studies may differ from those estimated using field data. Our landscape genomics approach can be extended to other host/symbiont systems that are being considered for biocontrol.}, } @article {pmid29358578, year = {2018}, author = {Bonneau, M and Atyame, C and Beji, M and Justy, F and Cohen-Gonsaud, M and Sicard, M and Weill, M}, title = {Culex pipiens crossing type diversity is governed by an amplified and polymorphic operon of Wolbachia.}, journal = {Nature communications}, volume = {9}, number = {1}, pages = {319}, pmid = {29358578}, issn = {2041-1723}, mesh = {Amino Acid Sequence ; Animals ; Bacterial Proteins/*genetics/metabolism ; Base Sequence ; Crosses, Genetic ; Culex/genetics/*microbiology ; Drosophila melanogaster/genetics/*microbiology ; Female ; *Genome, Bacterial ; Host Specificity ; Male ; *Operon ; Polymorphism, Genetic ; Sequence Alignment ; Sequence Homology, Amino Acid ; Symbiosis/genetics ; Toxin-Antitoxin Systems/genetics ; Wolbachia/*genetics/metabolism ; }, abstract = {Culex pipiens mosquitoes are infected with Wolbachia (wPip) that cause an important diversity of cytoplasmic incompatibilities (CIs). Functional transgenic studies have implicated the cidA-cidB operon from wPip and its homolog in wMel in CI between infected Drosophila males and uninfected females. However, the genetic basis of the CI diversity induced by different Wolbachia strains was unknown. We show here that the remarkable diversity of CI in the C. pipiens complex is due to the presence, in all tested wPip genomes, of several copies of the cidA-cidB operon, which undergoes diversification through recombination events. In 183 isofemale lines of C. pipiens collected worldwide, specific variations of the cidA-cidB gene repertoires are found to match crossing types. The diversification of cidA-cidB is consistent with the hypothesis of a toxin-antitoxin system in which the gene cidB co-diversifies with the gene cidA, particularly in putative domains of reciprocal interactions.}, } @article {pmid29351633, year = {2018}, author = {Lindsey, ARI and Rice, DW and Bordenstein, SR and Brooks, AW and Bordenstein, SR and Newton, ILG}, title = {Evolutionary Genetics of Cytoplasmic Incompatibility Genes cifA and cifB in Prophage WO of Wolbachia.}, journal = {Genome biology and evolution}, volume = {10}, number = {2}, pages = {434-451}, pmid = {29351633}, issn = {1759-6653}, support = {P30 DK058404/DK/NIDDK NIH HHS/United States ; R01 AI132581/AI/NIAID NIH HHS/United States ; R21 HD086833/HD/NICHD NIH HHS/United States ; T32 GM080178/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Arthropods/*microbiology/physiology ; Evolution, Molecular ; *Genes, Viral ; Male ; Prophages/*genetics ; Reproduction ; Symbiosis ; Transcriptome ; Wolbachia/genetics/physiology/*virology ; }, abstract = {The bacterial endosymbiont Wolbachia manipulates arthropod reproduction to facilitate its maternal spread through host populations. The most common manipulation is cytoplasmic incompatibility (CI): Wolbachia-infected males produce modified sperm that cause embryonic mortality, unless rescued by embryos harboring the same Wolbachia. The genes underlying CI, cifA and cifB, were recently identified in the eukaryotic association module of Wolbachia's prophage WO. Here, we use transcriptomic and genomic approaches to address three important evolutionary facets of the cif genes. First, we assess whether or not cifA and cifB comprise a classic toxin-antitoxin operon in wMel and show that the two genes exhibit striking, transcriptional differences across host development. They can produce a bicistronic message despite a predicted hairpin termination element in their intergenic region. Second, cifA and cifB strongly coevolve across the diversity of phage WO. Third, we provide new domain and functional predictions across homologs within Wolbachia, and show that amino acid sequences vary substantially across the genus. Finally, we investigate conservation of cifA and cifB and find frequent degradation and loss of the genes in strains that no longer induce CI. Taken together, we demonstrate that cifA and cifB exhibit complex transcriptional regulation in wMel, provide functional annotations that broaden the potential mechanisms of CI induction, and report recurrent erosion of cifA and cifB in non-CI strains, thus expanding our understanding of the most widespread form of reproductive parasitism.}, } @article {pmid29348597, year = {2018}, author = {Aljayyoussi, G and Tyrer, HE and Ford, L and Sjoberg, H and Pionnier, N and Waterhouse, D and Davies, J and Gamble, J and Metuge, H and Cook, DAN and Steven, A and Sharma, R and Guimaraes, AF and Clare, RH and Cassidy, A and Johnston, KL and Myhill, L and Hayward, L and Wanji, S and Turner, JD and Taylor, MJ and Ward, SA}, title = {Author Correction: Short-Course, High-Dose Rifampicin Achieves Wolbachia Depletion Predictive of Curative Outcomes in Preclinical Models of Lymphatic Filariasis and Onchocerciasis.}, journal = {Scientific reports}, volume = {8}, number = {1}, pages = {1384}, doi = {10.1038/s41598-018-19723-1}, pmid = {29348597}, issn = {2045-2322}, support = {MR/L018756/1/MRC_/Medical Research Council/United Kingdom ; }, abstract = {A correction to this article has been published and is linked from the HTML version of this paper. The error has been fixed in the paper.}, } @article {pmid29346449, year = {2018}, author = {Chrostek, E and Teixeira, L}, title = {Within host selection for faster replicating bacterial symbionts.}, journal = {PloS one}, volume = {13}, number = {1}, pages = {e0191530}, pmid = {29346449}, issn = {1932-6203}, mesh = {Animals ; Drosophila melanogaster/*microbiology ; Genes, Bacterial ; *Host-Pathogen Interactions ; *Symbiosis ; Virulence/genetics ; Wolbachia/genetics/*growth & development/pathogenicity ; }, abstract = {Wolbachia is a widespread, intracellular symbiont of arthropods, able to induce reproductive distortions and antiviral protection in insects. Wolbachia can also be pathogenic, as is the case with wMelPop, a virulent variant of the endosymbiont of Drosophila melanogaster. An extensive genomic amplification of the 20kb region encompassing eight Wolbachia genes, called Octomom, is responsible for wMelPop virulence. The Octomom copy number in wMelPop can be highly variable between individual D. melanogaster flies, even when comparing siblings arising from a single female. Moreover, Octomom copy number can change rapidly between generations. These data suggest an intra-host variability in Octomom copy number between Wolbachia cells. Since wMelPop Wolbachia with different Octomom copy numbers grow at different rates, we hypothesized that selection could act on this intra-host variability. Here we tested if total Octomom copy number changes during the lifespan of individual Drosophila hosts, revealing selection for different Wolbachia populations. We performed a time course analysis of Octomom amplification in flies whose mothers were controlled for Octomom copy number. We show that despite the Octomom copy number being relatively stable it increases slightly throughout D. melanogaster adult life. This indicates that there is selection acting on the intra-host variation in the Octomom copy number over the lifespan of individual hosts. This within host selection for faster replicating bacterial symbionts may be in conflict with between host selection against highly pathogenic Wolbachia.}, } @article {pmid29336504, year = {2018}, author = {Osei-Amo, S and Hussein, M and Asad, S and Hugo, L and Asgari, S}, title = {Wolbachia-induced transcription factor GATA4 suppresses ovary-specific genes blastoderm-specific protein 25D and imaginal disc growth factor.}, journal = {Insect molecular biology}, volume = {27}, number = {3}, pages = {295-304}, doi = {10.1111/imb.12371}, pmid = {29336504}, issn = {1365-2583}, mesh = {Aedes/*genetics/*microbiology ; Animals ; Blastoderm/growth & development ; Female ; GATA4 Transcription Factor/*genetics/metabolism ; Imaginal Discs/growth & development ; Insect Proteins/*genetics/metabolism ; Ovary/metabolism ; Up-Regulation ; Wolbachia/*physiology ; }, abstract = {The endosymbiotic bacterium Wolbachia infects a wide array of insect hosts and has been implicated in a range of biological modifications as a consequence of its infection. Previously, it was shown that the transcription factor GATA4 was significantly induced in Wolbachia wMelPop-CLA strain infected Aedes aegypti whole mosquitoes and cells. Here, we provide evidence that this induction also occurs in mosquito ovaries where the ovary-specific genes blastoderm-specific protein 25D (Bsg25D) and imaginal disc growth factor (Disc) are suppressed by Wolbachia. We further demonstrate that transcriptional depletion of GATA4 results in upregulation of both genes and conversely its overexpression leads to downregulation of the genes, suggesting that Wolbachia-induced GATA4 plays a suppressive regulatory role with regards to Bsg25D and Disc expression in mosquito ovaries. When the Disc gene was silenced in mosquitoes, we did not observe any difference in the number of mature ovarian follicles developed between treatment groups. However, we did find a significant delay in the hatching of eggs that had been laid by Disc knockdown mosquitoes.}, } @article {pmid29334340, year = {2019}, author = {Amala, M and Rajamanikandan, S and Prabhu, D and Surekha, K and Jeyakanthan, J}, title = {Identification of anti-filarial leads against aspartate semialdehyde dehydrogenase of Wolbachia endosymbiont of Brugia malayi: combined molecular docking and molecular dynamics approaches.}, journal = {Journal of biomolecular structure & dynamics}, volume = {37}, number = {2}, pages = {394-410}, doi = {10.1080/07391102.2018.1427633}, pmid = {29334340}, issn = {1538-0254}, mesh = {Amino Acid Sequence ; Animals ; Anthelmintics/*chemistry/pharmacology ; Aspartate-Semialdehyde Dehydrogenase/antagonists & inhibitors/*chemistry ; Binding Sites ; Brugia malayi/*enzymology ; Catalytic Domain ; Chemical Phenomena ; Drug Discovery/methods ; Enzyme Inhibitors/*chemistry/pharmacology ; Humans ; Hydrogen Bonding ; Hydrophobic and Hydrophilic Interactions ; *Molecular Docking Simulation ; *Molecular Dynamics Simulation ; Protein Binding ; }, abstract = {Lymphatic filariasis is a debilitating vector borne parasitic disease that infects human lymphatic system by nematode Brugia malayi. Currently available anti-filarial drugs are effective only on the larval stages of parasite. So far, no effective drugs are available for humans to treat filarial infections. In this regard, aspartate semialdehyde dehydrogenase (ASDase) in lysine biosynthetic pathway from Wolbachia endosymbiont Brugia malayi represents an attractive therapeutic target for the development of novel anti-filarial agents. In this present study, molecular modeling combined with molecular dynamics simulations and structure-based virtual screening were performed to identify potent lead molecules against ASDase. Based on Glide score, toxicity profile, binding affinity and mode of interactions with the ASDase, five potent lead molecules were selected. The molecular docking and dynamics results revealed that the amino acid residues Arg103, Asn133, Cys134, Gln161, Ser164, Lys218, Arg239, His246, and Asn321 plays a crucial role in effective binding of Top leads into the active site of ASDase. The stability of the ASDase-lead complexes was confirmed by running the 30 ns molecular dynamics simulations. The pharmacokinetic properties of the identified lead molecules are in the acceptable range. Furthermore, density functional theory and binding free energy calculations were performed to rank the lead molecules. Thus, the identified lead molecules can be used for the development of anti-filarial agents to combat the pathogenecity of Brugia malayi.}, } @article {pmid29330177, year = {2018}, author = {Zhu, YX and Song, YL and Zhang, YK and Hoffmann, AA and Zhou, JC and Sun, JT and Hong, XY}, title = {Incidence of Facultative Bacterial Endosymbionts in Spider Mites Associated with Local Environments and Host Plants.}, journal = {Applied and environmental microbiology}, volume = {84}, number = {6}, pages = {}, pmid = {29330177}, issn = {1098-5336}, mesh = {Animals ; Bacteroidetes/*physiology ; China ; Environment ; Solanum lycopersicum/growth & development ; Glycine max/growth & development ; Spiroplasma/*physiology ; *Symbiosis ; Tetranychidae/*microbiology/physiology ; Wolbachia/*physiology ; Zea mays/growth & development ; }, abstract = {Spider mites are frequently associated with multiple endosymbionts whose infection patterns often exhibit spatial and temporal variation. However, the association between endosymbiont prevalence and environmental factors remains unclear. Here, we surveyed endosymbionts in natural populations of the spider mite, Tetranychus truncatus, in China, screening 935 spider mites from 21 localities and 12 host plant species. Three facultative endosymbiont lineages, Wolbachia, Cardinium, and Spiroplasma, were detected at different infection frequencies (52.5%, 26.3%, and 8.6%, respectively). Multiple endosymbiont infections were observed in most local populations, and the incidence of individuals with the Wolbachia-Spiroplasma coinfection was higher than expected from the frequency of each infection within a population. Endosymbiont infection frequencies exhibited associations with environmental factors: Wolbachia infection rates increased at localities with higher annual mean temperatures, while Cardinium and Spiroplasma infection rates increased at localities from higher altitudes. Wolbachia was more common in mites from Lycopersicon esculentum and Glycine max compared to those from Zea mays This study highlights that host-endosymbiont interactions may be associated with environmental factors, including climate and other geographically linked factors, as well as the host's food plant.IMPORTANCE The aim of this study was to examine the incidence of endosymbiont distribution and the infection patterns in spider mites. The main findings are that multiple endosymbiont infections were more common than expected and that endosymbiont infection frequencies were associated with environmental factors. This work highlights that host-endosymbiont interactions need to be studied within an environmental and geographic context.}, } @article {pmid29311247, year = {2018}, author = {Mann, M and Fattah-Hosseini, S and Ammar, ED and Stange, R and Warrick, E and Sturgeon, K and Shatters, R and Heck, M}, title = {Diaphorina citri Nymphs Are Resistant to Morphological Changes Induced by "Candidatus Liberibacter asiaticus" in Midgut Epithelial Cells.}, journal = {Infection and immunity}, volume = {86}, number = {4}, pages = {}, pmid = {29311247}, issn = {1098-5522}, mesh = {Animals ; Epithelial Cells/*microbiology ; Female ; Gastrointestinal Microbiome ; Hemiptera/*microbiology ; In Situ Hybridization, Fluorescence ; Male ; Microscopy, Confocal ; Nymph/*microbiology ; Oxidative Stress ; Phenotype ; Rhizobiaceae/*physiology ; }, abstract = {"Candidatus Liberibacter asiaticus" is the causative bacterium associated with citrus greening disease. "Ca Liberibacter asiaticus" is transmitted by Diaphorina citri more efficiently when it is acquired by nymphs rather than adults. Why this occurs is not known. We compared midguts of D. citri insects reared on healthy or "Ca Liberibacter asiaticus"-infected citrus trees using quantitative PCR, confocal microscopy, and mitochondrial superoxide staining for evidence of oxidative stress. Consistent with its classification as propagative, "Ca Liberibacter asiaticus" titers were higher in adults than in nymphs. Our previous work showed that adult D. citri insects have basal levels of karyorrhexis (fragmentation of the nucleus) in midgut epithelial cells, which is increased in severity and frequency in response to "Ca Liberibacter asiaticus." Here, we show that nymphs exhibit lower levels of early-stage karyorrhexis than adults and are refractory to the induction of advanced karyorrhexis by "Ca Liberibacter asiaticus" in the midgut epithelium. MitoSox Red staining showed that guts of infected adults, particularly males, experienced oxidative stress in response to "Ca Liberibacter asiaticus." A positive correlation between the titers of "Ca Liberibacter asiaticus" and the Wolbachia endosymbiont was observed in adult and nymph midguts, suggesting an interplay between these bacteria during development. We hypothesize that the resistance of the nymph midgut to late-stage karyorrhexis through as yet unknown molecular mechanisms benefits "Ca Liberibacter asiaticus" for efficient invasion of midgut epithelial cells, which may be a factor explaining the developmental dependency of "Ca Liberibacter asiaticus" acquisition by the vector.}, } @article {pmid29308790, year = {2018}, author = {Sharma, R and Aljayyoussi, G and Tyrer, HE and Gamble, J and Hayward, L and Guimaraes, AF and Davies, J and Waterhouse, D and Cook, DAN and Myhill, LJ and Clare, RH and Cassidy, A and Steven, A and Johnston, KL and Ford, L and Turner, JD and Ward, SA and Taylor, MJ}, title = {Corrigendum: Minocycline as a re-purposed anti-Wolbachia macrofilaricide: superiority compared with doxycycline regimens in a murine infection model of human lymphatic filariasis.}, journal = {Scientific reports}, volume = {8}, number = {}, pages = {46934}, doi = {10.1038/srep46934}, pmid = {29308790}, issn = {2045-2322}, abstract = {This corrects the article DOI: 10.1038/srep23458.}, } @article {pmid29303740, year = {2018}, author = {De Ambrogi, M}, title = {Highlights from the 66th annual ASTMH meeting.}, journal = {The Lancet. Infectious diseases}, volume = {18}, number = {1}, pages = {32}, doi = {10.1016/S1473-3099(17)30717-X}, pmid = {29303740}, issn = {1474-4457}, mesh = {Antimalarials/*therapeutic use ; Arbovirus Infections/*prevention & control ; Disease Eradication ; *Disease Management ; Drug Therapy, Combination/methods ; Global Health ; HIV Infections/*drug therapy/prevention & control ; Humans ; Lassa Fever/diagnosis/*pathology/therapy ; Malaria/*drug therapy ; Mosquito Control/*methods ; Randomized Controlled Trials as Topic ; Treatment Outcome ; Wolbachia/growth & development ; }, } @article {pmid29301977, year = {2018}, author = {Dunning Hotopp, JC and Klasson, L}, title = {The Complexities and Nuances of Analyzing the Genome of Drosophila ananassae and Its Wolbachia Endosymbiont.}, journal = {G3 (Bethesda, Md.)}, volume = {8}, number = {1}, pages = {373-374}, pmid = {29301977}, issn = {2160-1836}, support = {R01 CA206188/CA/NCI NIH HHS/United States ; }, mesh = {Animals ; Drosophila/genetics ; Drosophila melanogaster ; Genome ; Retroelements ; Wolbachia/*genetics ; }, abstract = {In "Retrotransposons Are the Major Contributors to the Expansion of the Drosophila ananassae Muller F Element," Leung et al. (2017) improved contigs attributed to the Muller F element from the original CAF1 assembly, and used them to conclude that most of the sequence expansion of the fourth chromosome of D. ananassae is due to a higher transposon load than previously thought, but is not due to Wolbachia DNA integrations. While we do not disagree with the first conclusion, the authors base their second conclusion on the lack of homology detected between their improved CAF1 genome assembly attributed to D. ananassae and reference Wolbachia genomes. While the consensus CAF1 genome assembly lacks any sequence similarity to the reference genome of the Wolbachia endosymbiont of Drosophila melanogaster (wMel), numerous studies from multiple laboratories provide experimental support for a large lateral/horizontal gene transfer (LGT) of a Wolbachia genome into this D. ananassae line. As such, we strongly suspect that the original whole genome assembly was either constructed after the removal of all Wolbachia reads, or that Wolbachia sequences were directly removed from the contigs in the CAF1 assembly. Hence, Leung et al. (2017) could not have identified the Wolbachia LGT using the CAF1 assembly. This manuscript by Leung et al. (2017) highlights that an assembly of the Wolbachia sequence reads and their mate pairs was erroneously attributed solely to the Wolbachia endosymbiont, albeit before we understood the extent of LGT in D. ananassae As such, we recommend that the sequences deposited at the National Center for Biotechnology Information (NCBI) under PRJNA13365 should not be attributed to Wolbachia endosymbiont of D. ananassae, but should have their taxonomy reclassified by NCBI as "Unclassified sequences." As our knowledge about genome biology improves, we need to reconsider and reanalyze earlier genomes removing the prejudice introduced from now defunct paradigms.}, } @article {pmid29300732, year = {2018}, author = {Specht, S and Pfarr, KM and Arriens, S and Hübner, MP and Klarmann-Schulz, U and Koschel, M and Sternberg, S and Martin, C and Ford, L and Taylor, MJ and Hoerauf, A}, title = {Combinations of registered drugs reduce treatment times required to deplete Wolbachia in the Litomosoides sigmodontis mouse model.}, journal = {PLoS neglected tropical diseases}, volume = {12}, number = {1}, pages = {e0006116}, pmid = {29300732}, issn = {1935-2735}, mesh = {Animals ; Anti-Bacterial Agents/*administration & dosage ; Disease Models, Animal ; Drug Therapy, Combination/methods ; Filariasis/*drug therapy ; Filarioidea/*microbiology ; Fluoroquinolones/administration & dosage ; Mice ; Moxifloxacin ; Rifampin/administration & dosage/analogs & derivatives ; Tetracyclines/administration & dosage ; Time Factors ; Treatment Outcome ; Wolbachia/*drug effects ; }, abstract = {Filarial parasites can be targeted by antibiotic treatment due to their unique endosymbiotic relationship with Wolbachia bacteria. This finding has led to successful treatment strategies in both, human onchocerciasis and lymphatic filariasis. A 4-6 week treatment course using doxycycline results in long-term sterility and safe macrofilaricidal activity in humans. However, current treatment times and doxycycline contraindications in children and pregnant women preclude widespread administration of doxycycline in public health control programs; therefore, the search for shorter anti-wolbachial regimens is a focus of ongoing research. We have established an in vivo model for compound screening, using mice infected with Litomosoides sigmodontis. We could show that gold standard doxycycline treatment did not only deplete Wolbachia, it also resulted in a larval arrest. In this model, combinations of registered antibiotics were tested for their anti-wolbachial activity. Administration of rifamycins in combination with doxycycline for 7 days successfully depleted Wolbachia by > 2 log (>99% reduction) and thus resulted in a significant reduction of the treatment duration. Using a triple combination of a tetracycline (doxycycline or minocycline), a rifamycin and a fluoroquinolone (moxifloxacin) led to an even greater shortening of the treatment time. Testing all double combinations that could be derived from the triple combinations revealed that the combination of rifapentine (15mg/kg) and moxifloxacin (2 x 200mg/kg) showed the strongest reduction of treatment time in intraperitoneal and also oral administration routes. The rifapentine plus moxifloxacin combination was equivalent to the triple combination with additional doxycycline (>99% Wolbachia reduction). These investigations suggest that it is possible to shorten anti-wolbachial treatment times with combination treatments in order to achieve the target product profile (TPP) requirements for macrofilaricidal drugs of no more than 7-10 days of treatment.}, } @article {pmid29297702, year = {2018}, author = {Schultz, MJ and Connor, JH and Frydman, HM}, title = {Group B Wolbachia Strain-Dependent Inhibition of Arboviruses.}, journal = {DNA and cell biology}, volume = {37}, number = {1}, pages = {2-6}, pmid = {29297702}, issn = {1557-7430}, mesh = {Aedes/virology ; Animals ; Arboviruses/*physiology ; Culicidae/virology ; Dengue Virus/physiology ; Phylogeny ; Virus Replication/physiology ; Wolbachia/*virology ; Zika Virus/physiology ; }, abstract = {Mosquito-borne viruses, including Zika virus (ZIKV) and dengue virus (DENV), are global threats that continue to infect millions annually. Historically, efforts to combat the spread of these diseases have sought to eradicate the mosquito population. This has had limited success. Recent efforts to combat the spread of these diseases have targeted the mosquito population and the mosquito's ability to transmit viruses by altering the mosquito's microbiome. The introduction of particular strains of Wolbachia bacteria into mosquitos suppresses viral growth and blocks disease transmission. This novel strategy is being tested worldwide to reduce DENV and has early indications of success. The Wolbachia genus comprised divergent strains that are divided in major phylogenetic clades termed supergroups. All Wolbachia field trials currently utilize supergroup A Wolbachia in Aedes aegypti mosquitos to limit virus transmission. Here we discuss our studies of Wolbachia strains not yet used in virus control strategies but that show strong potential to reduce ZIKV replication. These strains are important opportunities in the search for novel tools to reduce the levels of mosquito-borne viruses and provide additional models for mechanistic studies.}, } @article {pmid29297293, year = {2017}, author = {Gruntenko, NЕ and Ilinsky, YY and Adonyeva, NV and Burdina, EV and Bykov, RA and Menshanov, PN and Rauschenbach, IY}, title = {Various Wolbachia genotypes differently influence host Drosophila dopamine metabolism and survival under heat stress conditions.}, journal = {BMC evolutionary biology}, volume = {17}, number = {Suppl 2}, pages = {252}, pmid = {29297293}, issn = {1471-2148}, mesh = {Alkaline Phosphatase/metabolism ; Animals ; Arylalkylamine N-Acetyltransferase/metabolism ; Dihydroxyphenylalanine/pharmacology ; Dopamine/*metabolism ; Drosophila Proteins/metabolism ; Drosophila melanogaster/enzymology/genetics/*microbiology ; Female ; Genotype ; *Hot Temperature ; Male ; *Stress, Physiological/drug effects ; Survival Analysis ; Wolbachia/drug effects/*genetics ; }, abstract = {BACKGROUND: One of the most widespread prokaryotic symbionts of invertebrates is the intracellular bacteria of Wolbachia genus which can be found in about 50% of insect species. Wolbachia causes both parasitic and mutualistic effects on its host that include manipulating the host reproductive systems in order to increase their transmission through the female germline, and increasing the host fitness. One of the mechanisms, promoting adaptation in biological organisms, is a non-specific neuroendocrine stress reaction. In insects, this reaction includes catecholamines, dopamine, serotonin and octopamine, which act as neurotransmitters, neuromodulators and neurohormones. The level of dopamine metabolism correlates with heat stress resistance in Drosophila adults.

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

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

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

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

CONCLUSIONS: In this study we show variation in Wolbachia-mediated DENV blocking in Aedes aegypti that may arise from genetic contributions and environmental influences on the mosquito-Wolbachia association. This suggests that Wolbachia-mediated blocking may have the ability to evolve through time or be expressed differentially across environments. The long-term efficacy of Wolbachia in the field will be dependent on the stability of blocking. Understanding the mechanism of blocking will be necessary for successful development of strategies that counter the emergence of evolved resistance or variation in its expression under diverse field conditions.}, } @article {pmid29279375, year = {2018}, author = {Carrington, LB and Tran, BCN and Le, NTH and Luong, TTH and Nguyen, TT and Nguyen, PT and Nguyen, CVV and Nguyen, HTC and Vu, TT and Vo, LT and Le, DT and Vu, NT and Nguyen, GT and Luu, HQ and Dang, AD and Hurst, TP and O'Neill, SL and Tran, VT and Kien, DTH and Nguyen, NM and Wolbers, M and Wills, B and Simmons, CP}, title = {Field- and clinically derived estimates of Wolbachia-mediated blocking of dengue virus transmission potential in Aedes aegypti mosquitoes.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {115}, number = {2}, pages = {361-366}, pmid = {29279375}, issn = {1091-6490}, support = {//Wellcome Trust/United Kingdom ; }, mesh = {Aedes/microbiology/*virology ; Animals ; Dengue/blood/transmission/*virology ; Dengue Virus/*physiology ; Humans ; Logistic Models ; Mosquito Vectors/microbiology/*virology ; Pest Control, Biological/methods ; Time Factors ; Viremia/blood/virology ; Wolbachia/*physiology ; }, abstract = {The wMel strain of Wolbachia can reduce the permissiveness of Aedes aegypti mosquitoes to disseminated arboviral infections. Here, we report that wMel-infected Ae. aegypti (Ho Chi Minh City background), when directly blood-fed on 141 viremic dengue patients, have lower dengue virus (DENV) transmission potential and have a longer extrinsic incubation period than their wild-type counterparts. The wMel-infected mosquitoes that are field-reared have even greater relative resistance to DENV infection when fed on patient-derived viremic blood meals. This is explained by an increased susceptibility of field-reared wild-type mosquitoes to infection than laboratory-reared counterparts. Collectively, these field- and clinically relevant findings support the continued careful field-testing of wMel introgression for the biocontrol of Ae. aegypti-born arboviruses.}, } @article {pmid29274498, year = {2018}, author = {Kajtoch, Ł and Montagna, M and Wanat, M}, title = {Species delimitation within the Bothryorrhynchapion weevils: Multiple evidence from genetics, morphology and ecological associations.}, journal = {Molecular phylogenetics and evolution}, volume = {120}, number = {}, pages = {354-363}, doi = {10.1016/j.ympev.2017.12.022}, pmid = {29274498}, issn = {1095-9513}, mesh = {Animals ; DNA/chemistry/isolation & purification/metabolism ; Electron Transport Complex IV/classification/genetics ; Female ; Genetic Variation ; Male ; Peptide Elongation Factor 1/classification/genetics ; Phylogeny ; Sequence Analysis, DNA ; Weevils/*classification/genetics/parasitology ; Wolbachia/genetics/physiology ; }, abstract = {Curculionidae is a hyperdiverse group of beetles, whose taxonomy and phylogenetics are still poorly understood, especially at the genus level. The latest work on the evolution of Apionini showed a noticeable "mess" in the subtribe Oxystomatina, where most of the morphology-based genera were found to be polyphyletic or paraphyletic. These discrepancies between classical taxonomy and molecular phylogenetics implied the need for further taxonomic revision of these groups. Here, we used sets of morphological, molecular and ecological characters to verify the taxonomic statuses and disentangle the phylogenetic relations among the Bothryorrhynchapion apionids, which are classified as a subgenus of Cyanapion. Morphological data including morphometrics, and multilocus molecular analyses confirmed the monophyly of the Bothryorrhynchapion and species statuses of five species. The morphological analyses showed that Cyanapion (Bothryorrhynchapion) protractum (Sharp, 1891) from the southeast Palaearctic is a synonym of C. (B.) gyllenhalii (Kirby). Moreover, ecological features (host plant use and presence/absence of the endosymbiotic bacteria Wolbachia) helped to unravel the relations among the examined weevils. The speciation of Bothryorrhynchapion apionids was probably affected by allopatric distribution, shifts in the preferred host plants (Vicia sp. or Lathyrus sp.) of sympatric taxa, and infection by different strains of Wolbachia. The paper presents the first comprehensive description of the species' morphology, biology and ecology, and includes a key to the species.}, } @article {pmid29257089, year = {2017}, author = {Poff, KE and Stever, H and Reil, JB and Seabourn, P and Ching, AJ and Aoki, S and Logan, M and Michalski, JR and Santamaria, J and Adams, JW and Eiben, JA and Yew, JY and Ewing, CP and Magnacca, KN and Bennett, GM}, title = {The Native Hawaiian Insect Microbiome Initiative: A Critical Perspective for Hawaiian Insect Evolution.}, journal = {Insects}, volume = {8}, number = {4}, pages = {}, pmid = {29257089}, issn = {2075-4450}, abstract = {Insects associate with a diversity of microbes that can shape host ecology and diversity by providing essential biological and adaptive services. For most insect groups, the evolutionary implications of host-microbe interactions remain poorly understood. Geographically discrete areas with high biodiversity offer powerful, simplified model systems to better understand insect-microbe interactions. Hawaii boasts a diverse endemic insect fauna (~6000 species) characterized by spectacular adaptive radiations. Despite this, little is known about the role of bacteria in shaping this diversity. To address this knowledge gap, we inaugurate the Native Hawaiian Insect Microbiome Initiative (NHIMI). The NHIMI is an effort intended to develop a framework for informing evolutionary and biological studies in Hawaii. To initiate this effort, we have sequenced the bacterial microbiomes of thirteen species representing iconic, endemic Hawaiian insect groups. Our results show that native Hawaiian insects associate with a diversity of bacteria that exhibit a wide phylogenetic breadth. Several groups show predictable associations with obligate microbes that permit diet specialization. Others exhibit unique ecological transitions that are correlated with shifts in their microbiomes (e.g., transition to carrion feeding from plant-feeding in Nysius wekiuicola). Finally, some groups, such as the Hawaiian Drosophila, have relatively diverse microbiomes with a conserved core of bacterial taxa across multiple species and islands.}, } @article {pmid29250466, year = {2017}, author = {Šochová, E and Husník, F and Nováková, E and Halajian, A and Hypša, V}, title = {Arsenophonus and Sodalis replacements shape evolution of symbiosis in louse flies.}, journal = {PeerJ}, volume = {5}, number = {}, pages = {e4099}, pmid = {29250466}, issn = {2167-8359}, abstract = {Symbiotic interactions between insects and bacteria are ubiquitous and form a continuum from loose facultative symbiosis to greatly intimate and stable obligate symbiosis. In blood-sucking insects living exclusively on vertebrate blood, obligate endosymbionts are essential for hosts and hypothesized to supplement B-vitamins and cofactors missing from their blood diet. The role and distribution of facultative endosymbionts and their evolutionary significance as seeds of obligate symbioses are much less understood. Here, using phylogenetic approaches, we focus on the Hippoboscidae phylogeny as well as the stability and dynamics of obligate symbioses within this bloodsucking group. In particular, we demonstrate a new potentially obligate lineage of Sodalis co-evolving with the Olfersini subclade of Hippoboscidae. We also show several likely facultative Sodalis lineages closely related to Sodalis praecaptivus (HS strain) and suggest repeated acquisition of novel symbionts from the environment. Similar to Sodalis, Arsenophonus endosymbionts also form both obligate endosymbiotic lineages co-evolving with their hosts (Ornithomyini and Ornithoica groups) as well as possibly facultative infections incongruent with the Hippoboscidae phylogeny. Finally, we reveal substantial diversity of Wolbachia strains detected in Hippoboscidae samples falling into three supergroups: A, B, and the most common F. Altogether, our results prove the associations between Hippoboscoidea and their symbiotic bacteria to undergo surprisingly dynamic, yet selective, evolutionary processes strongly shaped by repeated endosymbiont replacements. Interestingly, obligate symbionts only originate from two endosymbiont genera, Arsenophonus and Sodalis, suggesting that the host is either highly selective about its future obligate symbionts or that these two lineages are the most competitive when establishing symbioses in louse flies.}, } @article {pmid29248525, year = {2018}, author = {Huang, M and Luo, J and Hu, L and Zheng, B and Yu, J}, title = {Assessing the efficiency of Wolbachia driven Aedes mosquito suppression by delay differential equations.}, journal = {Journal of theoretical biology}, volume = {440}, number = {}, pages = {1-11}, doi = {10.1016/j.jtbi.2017.12.012}, pmid = {29248525}, issn = {1095-8541}, mesh = {Aedes/*microbiology/parasitology ; Animals ; Female ; Infertility/*microbiology ; Male ; Models, Biological ; *Models, Theoretical ; Mosquito Vectors ; Pest Control, Biological/*methods ; Wolbachia/*physiology ; }, abstract = {To suppress wild population of Aedes mosquitoes, the primary transmission vector of life-threatening diseases such as dengue, malaria, and Zika, an innovative strategy is to release male mosquitoes carrying the bacterium Wolbachia into natural areas to drive female sterility by cytoplasmic incompatibility. We develop a model of delay differential equations, incorporating the strong density restriction in the larval stage, to assess the delicate impact of life table parameters on suppression efficiency. Through mathematical analysis, we find the sufficient and necessary condition for global stability of the complete suppression state. This condition, combined with the experimental data for Aedes albopictus population in Guangzhou, helps us predict a large range of releasing intensities for suppression success. In particular, we find that if the number of released infected males is no less than four times the number of mosquitoes in wild areas, then the mosquito density in the peak season can be reduced by 95%. We introduce an index to quantify the dependence of suppression efficiency on parameters. The invariance of some quantitative properties of the index values under various perturbations of the same parameter justifies the applicability of this index, and the robustness of our modeling approach. The index yields a ranking of the sensitivity of all parameters, among which the adult mortality has the highest sensitivity and is considerably more sensitive than the natural larvae mortality.}, } @article {pmid29234308, year = {2017}, author = {Chrostek, E and Pelz-Stelinski, K and Hurst, GDD and Hughes, GL}, title = {Horizontal Transmission of Intracellular Insect Symbionts via Plants.}, journal = {Frontiers in microbiology}, volume = {8}, number = {}, pages = {2237}, pmid = {29234308}, issn = {1664-302X}, support = {R21 AI124452/AI/NIAID NIH HHS/United States ; R21 AI129507/AI/NIAID NIH HHS/United States ; U01CK000512/ACL/ACL HHS/United States ; }, abstract = {Experimental evidence is accumulating that endosymbionts of phytophagous insects may transmit horizontally via plants. Intracellular symbionts known for manipulating insect reproduction and altering fitness (Rickettsia, Cardinium, Wolbachia, and bacterial parasite of the leafhopper Euscelidius variegatus) have been found to travel from infected insects into plants. Other insects, either of the same or different species can acquire the symbiont from the plant through feeding, and in some cases transfer it to their progeny. These reports prompt many questions regarding how intracellular insect symbionts are delivered to plants and how they affect them. Are symbionts passively transported along the insect-plant-insect path, or do they actively participate in the process? How widespread are these interactions? How does symbiont presence influence the plant? And what conditions are required for the new infection to establish in an insect? From an ecological, evolutionary, and applied perspective, this mode of horizontal transmission could have profound implications if occurring frequently enough or if new stable symbiont infections are established. Transmission of symbionts through plants likely represents an underappreciated means of infection, both in terms of symbiont epidemiology and the movement of symbionts to new host species.}, } @article {pmid29234159, year = {2018}, author = {Zug, R and Hammerstein, P}, title = {Evolution of reproductive parasites with direct fitness benefits.}, journal = {Heredity}, volume = {120}, number = {3}, pages = {266-281}, pmid = {29234159}, issn = {1365-2540}, mesh = {Animals ; Biological Evolution ; Female ; Fertility ; *Genetic Fitness ; Male ; *Models, Genetic ; Parasites/*physiology ; *Reproduction ; *Symbiosis ; }, abstract = {Maternally inherited symbionts such as Wolbachia have long been seen mainly as reproductive parasites, with deleterious effects on host fitness. It is becoming clear, however, that, frequently, these symbionts also have beneficial effects on host fitness, either along with reproductive parasitism or not. Using the examples of cytoplasmic incompatibility (CI) and male-killing (MK), we here analyze the effect of direct fitness benefits on the evolution of reproductive parasites. By means of a simple theoretical framework, we synthesize and extend earlier modeling approaches for CI and MK, which usually ignore fitness benefits. Moreover, our framework is not restricted to a particular mechanism underlying the fitness benefit (e.g., protection against pathogens). We derive invasion conditions and equilibrium frequencies for the different infection scenarios. Our results demonstrate the importance of a symbiont's "effective fecundity" (i.e., the product of the relative fecundity of an infected female and her transmission efficiency) for a symbiont's invasion success. In particular, we adopt the concept of effective fecundity to scenarios where CI and MK co-occur in one host population. We confirm that direct fitness benefits substantially facilitate the invasion and spread of infections (for example, by lowering or removing the invasion threshold) or even make invasion possible in the first place (for example, if reproductive parasitism is weak or absent). Finally, we discuss the role of direct fitness benefits in long-term evolutionary dynamics of reproductive phenotypes and highlight their potential to resolve genetic conflicts between maternally inherited symbionts and their hosts.}, } @article {pmid29216317, year = {2017}, author = {Fraser, JE and De Bruyne, JT and Iturbe-Ormaetxe, I and Stepnell, J and Burns, RL and Flores, HA and O'Neill, SL}, title = {Novel Wolbachia-transinfected Aedes aegypti mosquitoes possess diverse fitness and vector competence phenotypes.}, journal = {PLoS pathogens}, volume = {13}, number = {12}, pages = {e1006751}, pmid = {29216317}, issn = {1553-7374}, support = {/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Aedes/growth & development/*microbiology/physiology/virology ; Animals ; Communicable Disease Control/methods ; Culex/microbiology ; Dengue Virus/isolation & purification/physiology ; Drosophila melanogaster/microbiology ; Drosophila simulans/microbiology ; Female ; Fertility ; Infectious Disease Transmission, Vertical/*veterinary ; Male ; Mosquito Control/methods ; Mosquito Vectors/*microbiology/physiology/virology ; Organ Specificity ; Ovary/microbiology/physiology ; RNA, Viral/isolation & purification ; Salivary Glands/microbiology/physiology ; Sex Characteristics ; Species Specificity ; Survival Analysis ; Viral Tropism ; Wolbachia/*growth & development/isolation & purification ; }, abstract = {Wolbachia pipientis from Drosophila melanogaster (wMel) is an endosymbiotic bacterium that restricts transmission of human pathogenic flaviviruses and alphaviruses, including dengue, Zika, and chikungunya viruses, when introduced into the mosquito vector Aedes aegypti. To date, wMel-infected Ae. aegypti have been released in field trials in 5 countries to evaluate the effectiveness of this strategy for disease control. Despite the success in establishing wMel-infected mosquitoes in wild populations, and the well-characterized antiviral capabilities of wMel, transinfecting different or additional Wolbachia strains into Ae. aegypti may improve disease impact, and perhaps more importantly, could provide a strategy to account for the possible evolution of resistant arboviruses. Here, we report the successful transinfection of Ae. aegypti with the Wolbachia strains wMelCS (D. melanogaster), wRi (D. simulans) and wPip (Culex quinquefasciatus) and assess the effects on Ae. aegypti fitness, cytoplasmic incompatibility, tissue tropism and pathogen blocking in a laboratory setting. The results demonstrate that wMelCS provides a similar degree of protection against dengue virus as wMel following an infectious blood meal, and significantly reduces viral RNA levels beyond that of wMel following a direct challenge with infectious virus in mosquitoes, with no additional fitness cost to the host. The protection provided by wRi is markedly weaker than that of wMelCS, consistent with previous characterisations of these lines in Drosophila, while wPip was found to substantially reduce the fitness of Ae. aegypti. Thus, we determine wMelCS as a key candidate for further testing in field-relevant fitness tests and viremic blood feeding challenges in a clinical setting to determine if it may represent an alternative Wolbachia strain with more desirable attributes than wMel for future field testing.}, } @article {pmid29215202, year = {2018}, author = {Rock, DI and Smith, AH and Joffe, J and Albertus, A and Wong, N and O'Connor, M and Oliver, KM and Russell, JA}, title = {Context-dependent vertical transmission shapes strong endosymbiont community structure in the pea aphid, Acyrthosiphon pisum.}, journal = {Molecular ecology}, volume = {27}, number = {8}, pages = {2039-2056}, doi = {10.1111/mec.14449}, pmid = {29215202}, issn = {1365-294X}, mesh = {Animals ; Aphids/genetics/*microbiology ; Coinfection/genetics/*microbiology ; Coxiellaceae/genetics/pathogenicity ; *Ecology ; Host Specificity/genetics ; Microbiota/genetics ; Pisum sativum/parasitology ; Serratia/genetics/pathogenicity ; Symbiosis/*genetics ; }, abstract = {Animal-associated microbiomes are often comprised of structured, multispecies communities, with particular microbes showing trends of co-occurrence or exclusion. Such structure suggests variable community stability, or variable costs and benefits-possibilities with implications for symbiont-driven host adaptation. In this study, we performed systematic screening for maternally transmitted, facultative endosymbionts of the pea aphid, Acyrthosiphon pisum. Sampling across six locales, with up to 5 years of collection in each, netted significant and consistent trends of community structure. Co-infections between Serratia symbiotica and Rickettsiella viridis were more common than expected, while Rickettsia and X-type symbionts colonized aphids with Hamiltonella defensa more often than expected. Spiroplasma co-infected with other endosymbionts quite rarely, showing tendencies to colonize as a single species monoculture. Field estimates of maternal transmission rates help to explain our findings: while Serratia and Rickettsiella improved each other's transmission, Spiroplasma reduced transmission rates of co-infecting endosymbionts. In summary, our findings show that North American pea aphids harbour recurring combinations of facultative endosymbionts. Common symbiont partners play distinct roles in pea aphid biology, suggesting the creation of "generalist" aphids receiving symbiont-based defence against multiple ecological stressors. Multimodal selection, at the host level, may thus partially explain our results. But more conclusively, our findings show that within-host microbe interactions, and their resulting impacts on transmission rates, are an important determinant of community structure. Widespread distributions of heritable symbionts across plants and invertebrates hint at the far-reaching implications for these findings, and our work further shows the benefits of symbiosis research within a natural context.}, } @article {pmid29214346, year = {2018}, author = {Singhal, K and Mohanty, S}, title = {Comparative genomics reveals the presence of putative toxin-antitoxin system in Wolbachia genomes.}, journal = {Molecular genetics and genomics : MGG}, volume = {293}, number = {2}, pages = {525-540}, pmid = {29214346}, issn = {1617-4623}, mesh = {Amino Acid Sequence ; Animals ; Bacterial Proteins/classification/genetics ; Drosophila/microbiology ; Drosophila melanogaster/microbiology ; Genes, Bacterial/genetics ; Genetic Variation ; Genome, Bacterial/*genetics ; Genomics/*methods ; Host-Pathogen Interactions ; Phylogeny ; Sequence Homology, Amino Acid ; Toxin-Antitoxin Systems/*genetics ; Wolbachia/*genetics/isolation & purification/physiology ; }, abstract = {Multiple toxin-antitoxin (TA) systems are housed in different locations within the bacterial genome and are known to be associated with various cellular processes and stress-related adaptation. In endosymbionts, although, the TA system has scarce occurrence but studies have highlighted its presence in enhancing host-symbiont interactions. Wolbachia, an obligate endosymbiont, has recently been proposed as a biocontrol agent which may be helpful in controlling vector-borne diseases. There are reports suggesting the role of TA system in inducing cytoplasmic incompatibility in case of Wolbachia, however, the underlying mechanism is still not known. The present study, therefore, aims at exploring the diversity of TA system in four novel (sourced from India) and three reference genomes (NCBI) of Wolbachia strains. Interestingly, we found several putative toxins and antitoxins of RelEB family of Type II TA system in these Wolbachia genomes. The results show wMel genome possessed more number of putative TA loci than wRi genome. In addition, searching through the other sequenced Wolbachia genomes in NCBI, a complete absence of TA system was observed in Wolbachia-infected nematodes. The sequence-wide analysis of all the putative RelEB proteins present amongst the Wolbachia endosymbiont and within the free-living bacterial genomes reveal strain-specific similarities and conserved sequences. However, large amount of sequence diversity was observed between Wolbachia and free-living bacteria. Understanding this sequence variation may help shed light on the differences between these two forms of bacteria and could also explain their niche preferences.}, } @article {pmid29194776, year = {2018}, author = {Kubiak, K and Sielawa, H and Chen, W and Dzika, E}, title = {Endosymbiosis and its significance in dermatology.}, journal = {Journal of the European Academy of Dermatology and Venereology : JEADV}, volume = {32}, number = {3}, pages = {347-354}, doi = {10.1111/jdv.14721}, pmid = {29194776}, issn = {1468-3083}, mesh = {Animals ; *Host-Pathogen Interactions ; Humans ; Insecta/microbiology ; Skin/*microbiology/*parasitology/virology ; Skin Diseases/*microbiology/*parasitology/virology ; *Symbiosis ; Wolbachia/physiology ; }, abstract = {Proposed at the beginning of the twentieth century to explain the origin of eukaryotic organelles from prokaryotes, endosymbiosis is now medically defined by various interaction patterns between microorganisms and their residing hosts, best exemplified by the bacterial endosymbiont Wolbachia identified in arthropods and filarial nematodes, which can influence normal development, reproduction, survival and transmission of the hosts. Based on the transmission modes, vertical or horizontal, and the function of the endosymbionts, the host-symbiont dependence can be divided into primary or secondary. In dermatology, the role of endosymbionts in skin ectoparasitosis has aroused great interests in the past years. Riesia pediculicola is a primary bacterial endosymbiont in body lice Pediculus humanus, and supplement their hosts with vitamin B, especially pantothenic acid. In cimicosis, the Gram-negative Wolbachia can synthesize biotin and riboflavin, which are crucial for the growth and reproduction of the bedbug Cimex lectularius. In human demodicosis and rosacea, further study is required to prove the pathogenic role of the Gram-negative bacteria Bacillus oleronius or the Gram-positive bacteria Bacillus cereus demonstrated in the Demodex mites. The high infection rate of adult female ticks Ixodes ricinus with the Gram-negative bacteria Midichloria mitochondrii present in the mitochondria in diverse ovarian cells, with the high seroprevalence rate in tick-exposed subjects, raises the possibility that this non-pathogenic endosymbiont may play a role in immune response and successful transmission of the tick-borne pathogen. The anaerobic protozoan Trichomonas vaginalis and bacteria Mycoplasma hominis are two obligate parasites in the urogenital epithelium, with partially overlapping symptoms. Intracellular localization of Mycoplasma hominis can avoid host immune response and penetration of antibiotics, while Trichomonas vaginalis infected with Mycoplasma hominis seems to have a higher cytopathic activity and amoeboid transformation rate. Further study on the biology and pathogenesis of different endosymbionts in dermatological parasitosis will help for the development of new treatment modalities.}, } @article {pmid33592944, year = {2017}, author = {Vythilingam, I and Wan-Yusoff, WS}, title = {Dengue vector control in Malaysia: Are we moving in the right direction?.}, journal = {Tropical biomedicine}, volume = {34}, number = {4}, pages = {746-758}, pmid = {33592944}, issn = {2521-9855}, abstract = {Dengue is a major public health problem across more than 123 countries. Vector control has been the hallmark of the dengue control programme in many countries in Southeast Asia since there are no anti-dengue drugs available, and the most recent dengue vaccine is partly efficacious. House-to-house larval surveys, source reduction, larviciding, fogging, ULV which have been carried out since the inception of the dengue control programme in the 1970s are no longer practicable and need to be augmented by more targeted but less ambitious outbreak responses that focus on a few tools that might justify expense of deployment. However, according to recent reports these tools have not really been evaluated for their effectiveness in dengue control. Novel techniques such the release of genetically modified mosquitoes (RIDL) and the use of the bacterium Wolbachia to control the populations of the Ae. aegypti are still under trial. In this review proactive methods to detect epidemics have been suggested. Tools based on adult mosquitoes is an important strategy for dengue vector surveillance and control. The outbreak response may be more efficient when timely vector control measures are implemented after the immediate detection of an infected mosquito.}, } @article {pmid31014588, year = {2017}, author = {González-Álvarez, VH and Fernández de Mera, IG and Cabezas-Cruz, A and de la Fuente, J and Ortega-Morales, AI and Almazán, C}, title = {Molecular survey of Rickettsial organisms in ectoparasites from a dog shelter in Northern Mexico.}, journal = {Veterinary parasitology, regional studies and reports}, volume = {10}, number = {}, pages = {143-148}, doi = {10.1016/j.vprsr.2017.10.005}, pmid = {31014588}, issn = {2405-9390}, mesh = {Animals ; Base Sequence ; DNA, Bacterial/*genetics ; Dog Diseases/epidemiology/*parasitology ; Dogs ; Ectoparasitic Infestations/epidemiology/parasitology/*veterinary ; Housing, Animal ; Phthiraptera/microbiology ; Phylogeny ; Polymerase Chain Reaction ; RNA, Bacterial/genetics ; RNA, Ribosomal, 16S/genetics ; Rickettsia/*genetics/isolation & purification ; Siphonaptera/microbiology ; Species Specificity ; Ticks/microbiology ; }, abstract = {The objective of this study was to screen and identify rickettsial organisms in ectoparasites collected from dogs in a shelter in Gomez Palacio, Durango, Mexico. One hundred dogs were inspected for ectoparasites. All the dogs were parasitized with Rhipicephalus sanguineus ticks, three with Heterodoxus spiniger lice and one with Ctenocephalides felis fleas. DNA was extracted from the ectoparasites found on each dog, and PCR with the primers for the Anaplasmataceae 16S rRNA and citrate synthase gltA genes were performed. Eight DNA samples obtained from ticks, three from lice and one from fleas were positive to 16S rRNA. Only one sample from C. felis and one from H. spiniger were positive to gltA. Sequence analysis of amplified products from C. felis showed identity to Rickettsia felis, Wolbachia pipientis, and Wolbachia spp., while a sequence from H. spiniger showed identity to Wolbachia spp. Herein we report the molecular detection of R. felis, W. pipientis, and Wolbachia spp. in C. felis and H. spiniger in northern Mexico. These results contribute to the knowledge of the microorganisms present in ectoparasites from dogs in Mexico.}, } @article {pmid29188381, year = {2017}, author = {Rahimi-Kaldeh, S and Ashouri, A and Bandani, A and Tomioka, K}, title = {The effect of Wolbachia on diapause, fecundity, and clock gene expression in Trichogramma brassicae (Hymenoptera: Trichogrammatidae).}, journal = {Development genes and evolution}, volume = {227}, number = {6}, pages = {401-410}, pmid = {29188381}, issn = {1432-041X}, mesh = {Animals ; CLOCK Proteins/*genetics ; *Diapause ; Female ; Gene Expression ; Insect Proteins/*genetics ; Photoperiod ; RNA, Messenger ; Wasps/*genetics/*physiology ; Wolbachia/*physiology ; }, abstract = {The short day lengths of late summer in moderate regions are used to induce diapause in various insects. Many studies have shown the maternal effect of photoperiod on diapause induction of Trichogramma wasps, but there is no study to show the relationship between photoperiodic regimes and clock genes in these useful biological control agents. Here, we investigated the role of photoperiods on diapause, fecundity, and clock gene expression (clk, cyc, cry2, per, and timeout) in asexual and sexual Trichogramma brassicae as a model insect to find any differences between two strains. Asexual strain was infected by Wolbachia, an endosymbiont bacterium. The diapause percentage was significantly higher under short days (8 h in sexual and 12 h in the asexual T. brassicae), although the diapause percentage of the sexual strain was significantly higher than the asexual one in all the photoperiods. The ANOVA revealed no significant changes between different photoperiods in the clock gene expression in the sexual strain but significant photoperiodic changes in clk, cyc, and timeout in the asexual strain. Our results showed that the mRNA levels of clock genes of asexual T. brassicae were significantly lower than those of sexual strain. The fecundity was significantly higher in the asexual strain. These results suggest that Wolbachia infection makes disturbance on the clock gene expression which consequently reduces the percentage of diapause but increases the fecundity in asexual T. brassicae.}, } @article {pmid29187976, year = {2017}, author = {Conner, WR and Blaxter, ML and Anfora, G and Ometto, L and Rota-Stabelli, O and Turelli, M}, title = {Genome comparisons indicate recent transfer of wRi-like Wolbachia between sister species Drosophila suzukii and D. subpulchrella.}, journal = {Ecology and evolution}, volume = {7}, number = {22}, pages = {9391-9404}, pmid = {29187976}, issn = {2045-7758}, support = {R01 GM104325/GM/NIGMS NIH HHS/United States ; }, abstract = {Wolbachia endosymbionts may be acquired by horizontal transfer, by introgression through hybridization between closely related species, or by cladogenic retention during speciation. All three modes of acquisition have been demonstrated, but their relative frequency is largely unknown. Drosophila suzukii and its sister species D. subpulchrella harbor Wolbachia, denoted wSuz and wSpc, very closely related to wRi, identified in California populations of D. simulans. However, these variants differ in their induced phenotypes: wRi causes significant cytoplasmic incompatibility (CI) in D. simulans, but CI has not been detected in D. suzukii or D. subpulchrella. Our draft genomes of wSuz and wSpc contain full-length copies of 703 of the 734 single-copy genes found in wRi. Over these coding sequences, wSuz and wSpc differ by only 0.004% (i.e., 28 of 704,883 bp); they are sisters relative to wRi, from which each differs by 0.014%-0.015%. Using published data from D. melanogaster, Nasonia wasps and Nomada bees to calibrate relative rates of Wolbachia versus host nuclear divergence, we conclude that wSuz and wSpc are too similar-by at least a factor of 100-to be plausible candidates for cladogenic transmission. These three wRi-like Wolbachia, which differ in CI phenotype in their native hosts, have different numbers of orthologs of genes postulated to contribute to CI; and the CI loci differ at several nucleotides that may account for the CI difference. We discuss the general problem of distinguishing alternative modes of Wolbachia acquisition, focusing on the difficulties posed by limited knowledge of variation in absolute and relative rates of molecular evolution for host nuclear genomes, mitochondria, and Wolbachia.}, } @article {pmid29186405, year = {2018}, author = {Bleidorn, C and Gerth, M}, title = {A critical re-evaluation of multilocus sequence typing (MLST) efforts in Wolbachia.}, journal = {FEMS microbiology ecology}, volume = {94}, number = {1}, pages = {}, doi = {10.1093/femsec/fix163}, pmid = {29186405}, issn = {1574-6941}, mesh = {Genetic Markers/genetics ; Genetic Variation/genetics ; Genome, Bacterial/*genetics ; Multilocus Sequence Typing/*methods ; Phylogeny ; Wolbachia/*genetics ; }, abstract = {Wolbachia (Alphaproteobacteria, Rickettsiales) is the most common, and arguably one of the most important inherited symbionts. Molecular differentiation of Wolbachia strains is routinely performed with a set of five multilocus sequence typing (MLST) markers. However, since its inception in 2006, the performance of MLST in Wolbachia strain typing has not been assessed objectively. Here, we evaluate the properties of Wolbachia MLST markers and compare it to 252 other single copy loci present in the genome of most Wolbachia strains. Specifically, we investigated how well MLST performs at strain differentiation, at reflecting genetic diversity of strains, and as phylogenetic marker. We find that MLST loci are outperformed by other loci at all tasks they are currently employed for, and thus that they do not reflect the properties of a Wolbachia strain very well. We argue that whole genome typing approaches should be used for Wolbachia typing in the future. Alternatively, if few loci approaches are necessary, we provide a characterisation of 252 single copy loci for a number a criteria, which may assist in designing specific typing systems or phylogenetic studies.}, } @article {pmid29183717, year = {2018}, author = {Dorigatti, I and McCormack, C and Nedjati-Gilani, G and Ferguson, NM}, title = {Using Wolbachia for Dengue Control: Insights from Modelling.}, journal = {Trends in parasitology}, volume = {34}, number = {2}, pages = {102-113}, pmid = {29183717}, issn = {1471-5007}, support = {MR/R015600/1/MRC_/Medical Research Council/United Kingdom ; U01 GM110721/GM/NIGMS NIH HHS/United States ; }, mesh = {Aedes/*microbiology ; Animals ; *Biological Control Agents ; Dengue/*prevention & control/transmission ; Dengue Virus/*physiology ; Insect Vectors/microbiology ; *Models, Theoretical ; Wolbachia/*physiology ; }, abstract = {Dengue is the most common arboviral infection of humans, responsible for a substantial disease burden across the tropics. Traditional insecticide-based vector-control programmes have limited effectiveness, and the one licensed vaccine has a complex and imperfect efficacy profile. Strains of the bacterium Wolbachia, deliberately introduced into Aedes aegyptimosquitoes, have been shown to be able to spread to high frequencies in mosquito populations in release trials, and mosquitoes infected with these strains show markedly reduced vector competence. Thus, Wolbachia represents an exciting potential new form of biocontrol for arboviral diseases, including dengue. Here, we review how mathematical models give insight into the dynamics of the spread of Wolbachia, the potential impact of Wolbachia on dengue transmission, and we discuss the remaining challenges in evaluation and development.}, } @article {pmid29181449, year = {2017}, author = {Mann, E and Stouthamer, CM and Kelly, SE and Dzieciol, M and Hunter, MS and Schmitz-Esser, S}, title = {Transcriptome Sequencing Reveals Novel Candidate Genes for Cardinium hertigii-Caused Cytoplasmic Incompatibility and Host-Cell Interaction.}, journal = {mSystems}, volume = {2}, number = {6}, pages = {}, pmid = {29181449}, issn = {2379-5077}, abstract = {Cytoplasmic incompatibility (CI) is an intriguing, widespread, symbiont-induced reproductive failure that decreases offspring production of arthropods through crossing incompatibility of infected males with uninfected females or with females infected with a distinct symbiont genotype. For years, the molecular mechanism of CI remained unknown. Recent genomic, proteomic, biochemical, and cell biological studies have contributed to understanding of CI in the alphaproteobacterium Wolbachia and implicate genes associated with the WO prophage. Besides a recently discovered additional lineage of alphaproteobacterial symbionts only moderately related to Wolbachia, Cardinium (Bacteroidetes) is the only other symbiont known to cause CI, and genomic evidence suggests that it has very little homology with Wolbachia and evolved this phenotype independently. Here, we present the first transcriptomic study of the CI Cardinium strain cEper1, in its natural host, Encarsia suzannae, to detect important CI candidates and genes involved in the insect-Cardinium symbiosis. Highly expressed transcripts included genes involved in manipulating ubiquitination, apoptosis, and host DNA. Female-biased genes encoding ribosomal proteins suggest an increase in general translational activity of Cardinium in female wasps. The results confirm previous genomic analyses that indicated that Wolbachia and Cardinium utilize different genes to induce CI, and transcriptome patterns further highlight expression of some common pathways that these bacteria use to interact with the host and potentially cause this enigmatic and fundamental manipulation of host reproduction. IMPORTANCE The majority of insects carry maternally inherited intracellular bacteria that are important in their hosts' biology, ecology, and evolution. Some of these bacterial symbionts cause a reproductive failure known as cytoplasmic incompatibility (CI). In CI, the mating of symbiont-infected males and uninfected females produces few or no daughters. The CI symbiont then spreads and can have a significant impact on the insect host population. Cardinium, a bacterial endosymbiont of the parasitoid wasp Encarsia in the Bacteroidetes, is the only bacterial lineage known to cause CI outside the Alphaproteobacteria, where Wolbachia and another recently discovered CI symbiont reside. Here, we sought insight into the gene expression of a CI-inducing Cardinium strain in its natural host, Encarsia suzannae. Our study provides the first insights into the Cardinium transcriptome and provides support for the hypothesis that Wolbachia and Cardinium target similar host pathways with distinct and largely unrelated sets of genes.}, } @article {pmid29178391, year = {2018}, author = {Otten, C and Brilli, M and Vollmer, W and Viollier, PH and Salje, J}, title = {Peptidoglycan in obligate intracellular bacteria.}, journal = {Molecular microbiology}, volume = {107}, number = {2}, pages = {142-163}, pmid = {29178391}, issn = {1365-2958}, support = {/WT_/Wellcome Trust/United Kingdom ; MR/N501840/1/MRC_/Medical Research Council/United Kingdom ; 101824/Z/13/Z/WT_/Wellcome Trust/United Kingdom ; }, mesh = {Anaplasma marginale/classification/genetics/immunology/metabolism ; Animals ; *Bacteria/classification/genetics/immunology/metabolism ; Cell Wall/metabolism ; Chlamydia/classification/genetics/immunology/metabolism ; Cytoplasm/metabolism ; Genome, Bacterial/genetics ; *Host Microbial Interactions ; Humans ; Immunity, Innate/immunology ; Intracellular Space/*microbiology ; Orientia tsutsugamushi/classification/genetics/immunology/metabolism ; Peptidoglycan/chemistry/*genetics/*metabolism ; Phylogeny ; Wolbachia/classification/genetics/immunology/metabolism ; }, abstract = {Peptidoglycan is the predominant stress-bearing structure in the cell envelope of most bacteria, and also a potent stimulator of the eukaryotic immune system. Obligate intracellular bacteria replicate exclusively within the interior of living cells, an osmotically protected niche. Under these conditions peptidoglycan is not necessarily needed to maintain the integrity of the bacterial cell. Moreover, the presence of peptidoglycan puts bacteria at risk of detection and destruction by host peptidoglycan recognition factors and downstream effectors. This has resulted in a selective pressure and opportunity to reduce the levels of peptidoglycan. In this review we have analysed the occurrence of genes involved in peptidoglycan metabolism across the major obligate intracellular bacterial species. From this comparative analysis, we have identified a group of predicted 'peptidoglycan-intermediate' organisms that includes the Chlamydiae, Orientia tsutsugamushi, Wolbachia and Anaplasma marginale. This grouping is likely to reflect biological differences in their infection cycle compared with peptidoglycan-negative obligate intracellular bacteria such as Ehrlichia and Anaplasma phagocytophilum, as well as obligate intracellular bacteria with classical peptidoglycan such as Coxiella, Buchnera and members of the Rickettsia genus. The signature gene set of the peptidoglycan-intermediate group reveals insights into minimal enzymatic requirements for building a peptidoglycan-like sacculus and/or division septum.}, } @article {pmid29169377, year = {2017}, author = {Pagès, N and Muñoz-Muñoz, F and Verdún, M and Pujol, N and Talavera, S}, title = {First detection of Wolbachia-infected Culicoides (Diptera: Ceratopogonidae) in Europe: Wolbachia and Cardinium infection across Culicoides communities revealed in Spain.}, journal = {Parasites & vectors}, volume = {10}, number = {1}, pages = {582}, pmid = {29169377}, issn = {1756-3305}, support = {FAU2008-0019//INIA/ ; }, mesh = {Animals ; Bacterial Infections/*epidemiology/microbiology ; Bacteroidetes/genetics/*isolation & purification ; Bluetongue/transmission ; Ceratopogonidae/classification/*microbiology ; Disease Vectors ; Europe/epidemiology ; Gram-Negative Bacterial Infections/epidemiology/microbiology ; Insect Control ; Insect Vectors/*microbiology ; Prevalence ; Spain/epidemiology ; Symbiosis ; Wolbachia/genetics/*isolation & purification ; }, abstract = {BACKGROUND: Biting midges of the genus Culicoides (Diptera: Ceratopogonidae) transmit pathogens that cause important diseases. No effective technique has been found to properly control either Culicoides spp. abundance or their likelihood to transmit pathogens. Endosymbionts, particularly Wolbachia, represent powerful alternatives to control arthropods of health interest. In arthropods, Wolbachia can reduce vector fitness and vector's pathogen transmission capacity, thus being a potential target for population reduction and replacement strategies.

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

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

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

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

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

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

CONCLUSIONS: Although there are not enough cases to indicate statistical significance, the results strongly suggest that including doxycycline into canine heartworm treatment protocols decreases post-treatment complications and mortality in naturally infected clinical cases.}, } @article {pmid29141766, year = {2018}, author = {Sarwar, MS and Jahan, N and Shahbaz, F}, title = {Molecular Detection and Characterization of Wolbachia pipientis from Culex quinquefasciatus Collected from Lahore, Pakistan.}, journal = {The American journal of tropical medicine and hygiene}, volume = {98}, number = {1}, pages = {154-161}, pmid = {29141766}, issn = {1476-1645}, mesh = {Animals ; Culex/*microbiology ; DNA, Bacterial/genetics ; Electrophoresis, Agar Gel ; Female ; Male ; Ovary/microbiology ; Pakistan ; Phylogeny ; Polymerase Chain Reaction ; Sequence Analysis, DNA ; Wolbachia/*genetics ; }, abstract = {The gram-negative, pleomorphic endosymbiont Wolbachia is known to infect a large number of insects and other arthropods naturally. This bacterium modifies the host biology, mainly causing reproductive alterations including feminization, death of male, parthenogenesis, and importantly cytoplasmic incompatibility. Wolbachia-induced cytoplasmic incompatibility results in nonviable offspring and vector population suppression. In addition, this bacterium rapidly spreads and propagates within the host population. This study is the first report on Wolbachia detection and characterization from Culex quinquefasciatus collected from Lahore, Pakistan. For this purpose, mosquito adults were collected from different localities of Lahore and identified at the species level. A total of 145 pairs of ovaries were individually subjected to DNA isolation, and polymerase chain reaction amplification of three (wsp, 16S rRNA, and ftsZ) genes were investigated. In all, 128 females were found positive, representing 82.3% infection rate. The phylogenetic analysis indicated that the detected endosymbiont had 100% homology with Wolbachia pipientis wPip strain and supergroup B. The detection of the local strain of Wolbachia (wPip) will be useful in investigating its potential for the control of dengue vector (Aedes aegypti) and reducing dengue transmission in Pakistan.}, } @article {pmid29132425, year = {2017}, author = {Zhang, D and Zhang, M and Wu, Y and Gilles, JRL and Yamada, H and Wu, Z and Xi, Z and Zheng, X}, title = {Establishment of a medium-scale mosquito facility: optimization of the larval mass-rearing unit for Aedes albopictus (Diptera: Culicidae).}, journal = {Parasites & vectors}, volume = {10}, number = {1}, pages = {569}, pmid = {29132425}, issn = {1756-3305}, support = {R01 AI080597/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/genetics/growth & development/microbiology/*physiology ; Animals ; Female ; Larva/growth & development/physiology ; Life Cycle Stages ; Male ; Pupa/growth & development/physiology ; Reproduction ; Wolbachia/physiology ; }, abstract = {BACKGROUND: Standardized larval rearing units for mosquito production are essential for the establishment of a mass-rearing facility. Two larval rearing units, developed respectively by the Guangzhou Wolbaki Biotech Co. Ltd. (Wolbaki) and Insect Pest Control Laboratory, Joint FAO/IAEA Division of Nuclear Techniques in Food and Agriculture (FAO/IAEA-IPCL), are tested to assess their potential uses to mass-rear the larval stages of Aedes albopictus in support of the establishment of a medium-scale mosquito facility for the application of mosquito genetic control strategies.

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

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

CONCLUSIONS: The current study has indicated that both the Wol-unit and IAEA-unit are suitable for larvae mass-rearing for Ae. albopictus. However, the IAEA-unit, with higher male production and less space required compared to the Wol-unit, is recommended to be used in support of the establishment of a medium-sized mosquito facility.}, } @article {pmid29125253, year = {2017}, author = {Tmimi, FZ and Bkhache, M and Mounaji, K and Failloux, AB and Sarih, M}, title = {First report of the endobacteria Wolbachia in natural populations of Culex pipiens in Morocco.}, journal = {Journal of vector ecology : journal of the Society for Vector Ecology}, volume = {42}, number = {2}, pages = {349-351}, doi = {10.1111/jvec.12275}, pmid = {29125253}, issn = {1948-7134}, mesh = {Animals ; Culex/*microbiology ; Morocco ; Wolbachia/*physiology ; }, } @article {pmid29122518, year = {2017}, author = {Frentiu, FD}, title = {Lipids and Pathogen Blocking by Wolbachia.}, journal = {Trends in parasitology}, volume = {33}, number = {12}, pages = {916-917}, doi = {10.1016/j.pt.2017.10.007}, pmid = {29122518}, issn = {1471-5007}, support = {MC_PC_15087/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Aedes/*microbiology/*virology ; Animals ; Dengue/microbiology/prevention & control/transmission/virology ; Dengue Virus/physiology ; Disease Transmission, Infectious/*prevention & control ; Lipids/*physiology ; Microbial Interactions/*physiology ; Mosquito Vectors/microbiology/virology ; Wolbachia/*metabolism/*virology ; Zika Virus/physiology ; Zika Virus Infection/microbiology/prevention & control/transmission/virology ; }, abstract = {Mosquito-borne viruses are major human pathogens. Introducing Wolbachia into mosquitoes could reduce disease burdens because these bacteria block virus transmission. How Wolbachia does this is unclear, but new data show that modulation of host-cell lipids is critical.}, } @article {pmid29116011, year = {2017}, author = {Tsai, CH and Chen, TH and Lin, C and Shu, PY and Su, CL and Teng, HJ}, title = {The impact of temperature and Wolbachia infection on vector competence of potential dengue vectors Aedes aegypti and Aedes albopictus in the transmission of dengue virus serotype 1 in southern Taiwan.}, journal = {Parasites & vectors}, volume = {10}, number = {1}, pages = {551}, pmid = {29116011}, issn = {1756-3305}, mesh = {Aedes/anatomy & histology/*microbiology/physiology/*virology ; Animals ; Dengue/epidemiology/transmission/virology ; Dengue Virus/*physiology ; Female ; Mosquito Vectors/*microbiology/physiology/*virology ; Salivary Glands/virology ; Taiwan/epidemiology ; Temperature ; Thorax ; Viral Load ; Virus Replication ; Wolbachia/isolation & purification/*physiology ; }, abstract = {BACKGROUND: We evaluated the impact of temperature and Wolbachia infection on vector competence of the local Aedes aegypti and Ae. albopictus populations of southern Taiwan in the laboratory.

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

CONCLUSIONS: In southern Taiwan, Ae. aegypti is the main vector of dengue and Ae. albopictus has a non-significant role in the transmission of dengue virus due to the high prevalence of Wolbachia infection in the local mosquito population of southern Taiwan.}, } @article {pmid29114401, year = {2017}, author = {Robinson, KM and Hawkins, AS and Santana-Cruz, I and Adkins, RS and Shetty, AC and Nagaraj, S and Sadzewicz, L and Tallon, LJ and Rasko, DA and Fraser, CM and Mahurkar, A and Silva, JC and Dunning Hotopp, JC}, title = {Aligner optimization increases accuracy and decreases compute times in multi-species sequence data.}, journal = {Microbial genomics}, volume = {3}, number = {9}, pages = {e000122}, pmid = {29114401}, issn = {2057-5858}, support = {R01 CA206188/CA/NCI NIH HHS/United States ; U19 AI110820/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Brugia malayi/genetics ; Chromosome Mapping ; Data Accuracy ; Databases, Genetic ; Datasets as Topic ; Genome, Human ; High-Throughput Nucleotide Sequencing ; Humans ; Metagenomics ; Plasmodium falciparum/genetics ; Sequence Alignment/*methods ; *Sequence Analysis, DNA ; *Software ; Time Factors ; Wolbachia/genetics ; }, abstract = {As sequencing technologies have evolved, the tools to analyze these sequences have made similar advances. However, for multi-species samples, we observed important and adverse differences in alignment specificity and computation time for bwa- mem (Burrows-Wheeler aligner-maximum exact matches) relative to bwa-aln. Therefore, we sought to optimize bwa-mem for alignment of data from multi-species samples in order to reduce alignment time and increase the specificity of alignments. In the multi-species cases examined, there was one majority member (i.e. Plasmodium falciparum or Brugia malayi) and one minority member (i.e. human or the Wolbachia endosymbiont wBm) of the sequence data. Increasing bwa-mem seed length from the default value reduced the number of read pairs from the majority sequence member that incorrectly aligned to the reference genome of the minority sequence member. Combining both source genomes into a single reference genome increased the specificity of mapping, while also reducing the central processing unit (CPU) time. In Plasmodium, at a seed length of 18 nt, 24.1 % of reads mapped to the human genome using 1.7±0.1 CPU hours, while 83.6 % of reads mapped to the Plasmodium genome using 0.2±0.0 CPU hours (total: 107.7 % reads mapping; in 1.9±0.1 CPU hours). In contrast, 97.1 % of the reads mapped to a combined Plasmodium-human reference in only 0.7±0.0 CPU hours. Overall, the results suggest that combining all references into a single reference database and using a 23 nt seed length reduces the computational time, while maximizing specificity. Similar results were found for simulated sequence reads from a mock metagenomic data set. We found similar improvements to computation time in a publicly available human-only data set.}, } @article {pmid29114059, year = {2017}, author = {Gomes, FM and Hixson, BL and Tyner, MDW and Ramirez, JL and Canepa, GE and Alves E Silva, TL and Molina-Cruz, A and Keita, M and Kane, F and Traoré, B and Sogoba, N and Barillas-Mury, C}, title = {Effect of naturally occurring Wolbachia in Anopheles gambiae s.l. mosquitoes from Mali on Plasmodium falciparum malaria transmission.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {114}, number = {47}, pages = {12566-12571}, pmid = {29114059}, issn = {1091-6490}, support = {ZIA AI000947/ImNIH/Intramural NIH HHS/United States ; ZIA AI001062/ImNIH/Intramural NIH HHS/United States ; }, mesh = {Animals ; Anopheles/*microbiology/parasitology ; Female ; *Host-Parasite Interactions ; Host-Pathogen Interactions ; Insect Vectors/*microbiology/parasitology ; Malaria, Falciparum/epidemiology/parasitology/pathology/*transmission ; Mali/epidemiology ; Oocysts/pathogenicity/physiology ; Phylogeny ; Plasmodium falciparum/*microbiology ; RNA, Ribosomal, 16S/genetics ; Severity of Illness Index ; Sporozoites/pathogenicity/physiology ; Wolbachia/classification/*genetics/isolation & purification ; }, abstract = {A naturally occurring Wolbachia strain (wAnga-Mali) was identified in mosquitoes of the Anopheles gambiae complex collected in the Malian villages of Dangassa and Kenieroba. Phylogenetic analysis of the nucleotide sequence of two 16S rRNA regions showed that wAnga-Mali clusters with Wolbachia strains from supergroup A and has the highest homology to a Wolbachia strain isolated from cat fleas (Ctenocephalides). wAnga-Mali is different from two Wolbachia strains previously reported in A. gambiae from Burkina Faso (wAnga_VK5_STP and wAnga_VK5_3.1a). Quantitative analysis of Wolbachia and Plasmodium sporozoite infection in field-collected mosquitoes indicates that the prevalence and intensity of Plasmodium falciparum sporozoite infection is significantly lower in Wolbachia-infected females. The presence of Wolbachia in females from a laboratory Anopheles coluzzii (A. gambiae, M form) colony experimentally infected with P. falciparum (NF54 strain) gametocyte cultures slightly enhanced oocyst infection. However, Wolbachia infection significantly reduced the prevalence and intensity of sporozoite infection, as observed in the field. This indicates that wAnga-Mali infection does not limit early stages of Plasmodium infection in the mosquito, but it has a strong deleterious effect on sporozoites and reduces malaria transmission.}, } @article {pmid29106651, year = {2018}, author = {Sosa, EJ and Burguener, G and Lanzarotti, E and Defelipe, L and Radusky, L and Pardo, AM and Marti, M and Turjanski, AG and Fernández Do Porto, D}, title = {Target-Pathogen: a structural bioinformatic approach to prioritize drug targets in pathogens.}, journal = {Nucleic acids research}, volume = {46}, number = {D1}, pages = {D413-D418}, pmid = {29106651}, issn = {1362-4962}, mesh = {Amino Acid Sequence ; Anti-Infective Agents/*chemistry/pharmacology ; Binding Sites ; Communicable Diseases/drug therapy ; Computational Biology/*methods ; *Databases, Factual ; Drug Discovery ; *Genome, Bacterial ; *Genome, Fungal ; *Genome, Helminth ; *Genome, Protozoan ; Humans ; Internet ; Metabolic Networks and Pathways/drug effects/genetics ; Models, Molecular ; Molecular Targeted Therapy ; Protein Binding ; Protein Conformation, alpha-Helical ; Protein Conformation, beta-Strand ; Protein Interaction Domains and Motifs ; Sequence Alignment ; Sequence Homology, Amino Acid ; Software ; }, abstract = {Available genomic data for pathogens has created new opportunities for drug discovery and development to fight them, including new resistant and multiresistant strains. In particular structural data must be integrated with both, gene information and experimental results. In this sense, there is a lack of an online resource that allows genome wide-based data consolidation from diverse sources together with thorough bioinformatic analysis that allows easy filtering and scoring for fast target selection for drug discovery. Here, we present Target-Pathogen database (http://target.sbg.qb.fcen.uba.ar/patho), designed and developed as an online resource that allows the integration and weighting of protein information such as: function, metabolic role, off-targeting, structural properties including druggability, essentiality and omic experiments, to facilitate the identification and prioritization of candidate drug targets in pathogens. We include in the database 10 genomes of some of the most relevant microorganisms for human health (Mycobacterium tuberculosis, Mycobacterium leprae, Klebsiella pneumoniae, Plasmodium vivax, Toxoplasma gondii, Leishmania major, Wolbachia bancrofti, Trypanosoma brucei, Shigella dysenteriae and Schistosoma Smanosoni) and show its applicability. New genomes can be uploaded upon request.}, } @article {pmid29099918, year = {2017}, author = {Chung, M and Small, ST and Serre, D and Zimmerman, PA and Dunning Hotopp, JC}, title = {Draft genome sequence of the Wolbachia endosymbiont of Wuchereria bancrofti wWb.}, journal = {Pathogens and disease}, volume = {75}, number = {9}, pages = {}, pmid = {29099918}, issn = {2049-632X}, support = {U19 AI110820/AI/NIAID NIH HHS/United States ; T32 AI007024/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Cluster Analysis ; Genes, rRNA ; *Genome, Bacterial ; Open Reading Frames ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; RNA, Ribosomal, 23S/genetics ; RNA, Ribosomal, 5S/genetics ; *Sequence Analysis, DNA ; Wolbachia/*genetics/isolation & purification ; Wuchereria bancrofti/*microbiology ; }, abstract = {The draft genome assembly of the Wolbachia endosymbiont of Wuchereria bancrofti (wWb) consists of 1060 850 bp in 100 contigs and contains 961 ORFs, with a single copy of the 5S rRNA, 16S rRNA and 23S rRNA and each of the 34 tRNA genes. Phylogenetic core genome analyses show wWb to cluster with other strains in supergroup D of the Wolbachia phylogeny, while being most closely related to the Wolbachia endosymbiont of Brugia malayi strain TRS (wBm). The wWb and wBm genomes share 779 orthologous clusters with wWb having 101 unclustered genes and wBm having 23 unclustered genes. The higher number of unclustered genes in the wWb genome likely reflects the fragmentation of the draft genome.}, } @article {pmid29099491, year = {2018}, author = {Pan, X and Pike, A and Joshi, D and Bian, G and McFadden, MJ and Lu, P and Liang, X and Zhang, F and Raikhel, AS and Xi, Z}, title = {The bacterium Wolbachia exploits host innate immunity to establish a symbiotic relationship with the dengue vector mosquito Aedes aegypti.}, journal = {The ISME journal}, volume = {12}, number = {1}, pages = {277-288}, pmid = {29099491}, issn = {1751-7370}, support = {R01 AI080597/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/immunology/*microbiology ; Animals ; *Immunity, Innate ; Mosquito Vectors/immunology/*microbiology ; Symbiosis/*physiology ; Toll-Like Receptors/metabolism ; Wolbachia/*physiology ; }, abstract = {A host's immune system plays a central role in shaping the composition of the microbiota and, in return, resident microbes influence immune responses. Symbiotic associations of the maternally transmitted bacterium Wolbachia occur with a wide range of arthropods. It is, however, absent from the dengue and Zika vector mosquito Aedes aegypti in nature. When Wolbachia is artificially forced to form symbiosis with this new mosquito host, it boosts the basal immune response and enhances the mosquito's resistance to pathogens, including dengue, Zika virus and malaria parasites. The mechanisms involved in establishing a symbiotic relationship between Wolbachia and A. aegypti, and the long-term outcomes of this interaction, are not well understood. Here, we have demonstrated that both the immune deficiency (IMD) and Toll pathways are activated by the Wolbachia strain wAlbB upon its introduction into A. aegypti. Silencing the Toll and IMD pathways via RNA interference reduces the wAlbB load. Notably, wAlbB induces peptidoglycan recognition protein (PGRP)-LE expression in the carcass of A. aegypti, and its silencing results in a reduction of symbiont load. Using transgenic mosquitoes with stage-specific induction of the IMD and Toll pathways, we have shown that elevated wAlbB infection in these mosquitoes is maintained via maternal transmission. These results indicate that host innate immunity is utilized to establish and promote host-microbial symbiosis. Our results will facilitate a long-term projection of the stability of the Wolbachia-A. aegypti mosquito system that is being developed to control dengue and Zika virus transmission to humans.}, } @article {pmid29095113, year = {2018}, author = {Moghadam, NN and Thorshauge, PM and Kristensen, TN and de Jonge, N and Bahrndorff, S and Kjeldal, H and Nielsen, JL}, title = {Strong responses of Drosophila melanogaster microbiota to developmental temperature.}, journal = {Fly}, volume = {12}, number = {1}, pages = {1-12}, pmid = {29095113}, issn = {1933-6942}, mesh = {Acetobacter/growth & development/isolation & purification ; Animals ; Drosophila melanogaster/*growth & development/*microbiology ; Female ; Gastrointestinal Microbiome ; Leuconostoc/growth & development/isolation & purification ; Male ; Temperature ; Wolbachia/growth & development/isolation & purification ; }, abstract = {Physiological responses to changes in environmental conditions such as temperature may partly arise from the resident microbial community that integrates a wide range of bio-physiological aspects of the host. In the present study, we assessed the effect of developmental temperature on the thermal tolerance and microbial community of Drosophila melanogaster. We also developed a bacterial transplantation protocol in order to examine the possibility of reshaping the host bacterial composition and assessed its influence on the thermotolerance phenotype. We found that the temperature during development affected thermal tolerance and the microbial composition of male D. melanogaster. Flies that developed at low temperature (13°C) were the most cold resistant and showed the highest abundance of Wolbachia, while flies that developed at high temperature (31°C) were the most heat tolerant and had the highest abundance of Acetobacter. In addition, feeding newly eclosed flies with bacterial suspensions from intestines of flies developed at low temperatures changed the heat tolerance of recipient flies. However, we were not able to link this directly to a change in the host bacterial composition.}, } @article {pmid29094498, year = {2019}, author = {Pierzynowska, K and Skowron Volponi, M and Węgrzyn, G}, title = {Multiple factors correlating with wing malformations in the population of Parnassius apollo (Lepidoptera: Papilionidae) restituted from a low number of individuals: A mini review.}, journal = {Insect science}, volume = {26}, number = {3}, pages = {380-387}, doi = {10.1111/1744-7917.12554}, pmid = {29094498}, issn = {1744-7917}, support = {530-L140-D242-17-1A//University of Gdansk/ ; }, mesh = {Animals ; *Butterflies ; Conservation of Natural Resources ; *Endangered Species ; Insect Proteins/genetics ; Laccase/metabolism ; Wings, Animal/*abnormalities ; Wolbachia ; Yersinia ; }, abstract = {The Apollo butterfly, Parnassius apollo (Linnaeus), was common in Europe over 100 years ago, but currently it is considered as near threatened. Different conservation programs have promoted the persistence of this species; however, it is still endangered. An example of such programs was the action devoted to reestablish the Apollo butterfly population in Pieniny National Park (Poland) from only 20-30 individuals which had survived till the last decade of the 20th century. This reintroduction has been successful; however, unexpected developmental problems appeared. Butterflies with deformed or reduced wings became frequent in the population living in the natural habitat, and particularly among those reared under seminatural conditions (in the same environment, but fenced by a net). Until recently, reasons for these malformations remained unknown. However, reports published during last months indicated that there are genetic, biochemical, and microbiological factors contributing to this phenomenon. In the malformed individuals, lesions in the wingless gene and dysfunctions of laccase 1 and 2 were found to be significantly more frequent than in normal insects. A large fraction of butterflies with deformed or reduced wings was devoid of the prokaryotic symbiont Wolbachia, which was present in most normal individuals. Moreover, Yersinia pseudotuberculosis (Pfeiffer) Smith and Thal, and Serratia sp., bacteria pathogenic to insects, were detected in the biological material from both normal and malformed butterflies from this population. These findings are summarized and discussed in this review, in the light of conservation of insects and restitution of their populations from a low number of individuals.}, } @article {pmid29093481, year = {2017}, author = {Karakuş, M and Karabey, B and Orçun Kalkan, Ş and Özdemir, G and Oğuz, G and Erişöz Kasap, Ö and Alten, B and Töz, S and Özbel, Y}, title = {Midgut Bacterial Diversity of Wild Populations of Phlebotomus (P.) papatasi, the Vector of Zoonotic Cutaneous Leishmaniasis (ZCL) in Turkey.}, journal = {Scientific reports}, volume = {7}, number = {1}, pages = {14812}, pmid = {29093481}, issn = {2045-2322}, mesh = {Animals ; Bacteria/genetics/*isolation & purification ; Female ; Gastrointestinal Microbiome ; Humans ; Insect Vectors/*microbiology ; Leishmaniasis, Cutaneous/*epidemiology ; Male ; Phlebotomus/*microbiology ; Proteobacteria/genetics/isolation & purification ; Turkey/epidemiology ; Wolbachia/genetics/isolation & purification ; }, abstract = {Phlebotomine sand flies are hematophagous insects that harbor bacterial, viral and parasitic agents like Bartonella sp., Phleboviruses and Leishmania spp., respectively. There are few reports on bacterial microbiota of Phlebotomus (P.) papatasi but no data available for natural populations of Turkey, where leishmaniasis is endemic. Therefore, we aimed to investigate the midgut bacterial flora of different populations of P. papatasi. Sand flies were collected from different towns (Karaburun, Urla, Ayvacik and Başçayır) located in the western part of Turkey. Laboratory reared P. papatasi were included in the study as an insectarium population. After sterile washing steps, sand flies were dissected and guts were separated. Three pools, (males, unfed females and blood-fed females) were generated for each population. Prokaryotic 16 S rRNA gene was amplified and DGGE was performed. Fourteen different organisms belonging to two Phylum (Proteobactericea and Furmicutes) were identified according to sequence results in the studied pools. The presence of Wolbachia sp. was shown for the first time in the wild-caught sand fly populations of Turkey. This is the first report of gut bacterial flora of wild-caught P. papatasi collected in an endemic area for leishmaniasis in Turkey. Microbiome profiling of wild-caught sand flies will be of great help in the investigating of possible vector control candidates for paratransgenic control approach.}, } @article {pmid29093474, year = {2017}, author = {Kaur, R and Siozios, S and Miller, WJ and Rota-Stabelli, O}, title = {Insertion sequence polymorphism and genomic rearrangements uncover hidden Wolbachia diversity in Drosophila suzukii and D. subpulchrella.}, journal = {Scientific reports}, volume = {7}, number = {1}, pages = {14815}, pmid = {29093474}, issn = {2045-2322}, support = {P 22634/FWF_/Austrian Science Fund FWF/Austria ; P 28255/FWF_/Austrian Science Fund FWF/Austria ; }, mesh = {Animals ; DNA Transposable Elements ; Drosophila/*microbiology/physiology ; Fertility ; Multilocus Sequence Typing ; Phylogeny ; *Polymorphism, Genetic ; Wolbachia/classification/*genetics ; }, abstract = {Ability to distinguish between closely related Wolbachia strains is crucial for understanding the evolution of Wolbachia-host interactions and the diversity of Wolbachia-induced phenotypes. A useful model to tackle these issues is the Drosophila suzukii - Wolbachia association. D. suzukii, a destructive insect pest, harbor a non-CI inducing Wolbachia 'wSuz' closely related to the strong CI-inducing wRi strain. Multi locus sequence typing (MLST) suggests presence of genetic homogeneity across wSuz strains infecting European and American D. suzukii populations, although different Wolbachia infection frequencies and host fecundity levels have been observed in both populations. Currently, it is not clear if these differences are due to cryptic wSuz polymorphism, host background, geographical factors or a combination of all of them. Here, we have identified geographical diversity in wSuz in D. suzukii populations from different continents using a highly diagnostic set of markers based on insertion sequence (IS) site polymorphism and genomic rearrangements (GR). We further identified inter-strain diversity between Wolbachia infecting D. suzukii and its sister species D. subpulchrella (wSpc). Based on our results, we speculate that discernible wSuz variants may associate with different observed host phenotypes, a hypothesis that demands future investigation. More generally, our results demonstrate the utility of IS and GRs in discriminating closely related Wolbachia strains.}, } @article {pmid29084217, year = {2017}, author = {Dodson, BL and Andrews, ES and Turell, MJ and Rasgon, JL}, title = {Wolbachia effects on Rift Valley fever virus infection in Culex tarsalis mosquitoes.}, journal = {PLoS neglected tropical diseases}, volume = {11}, number = {10}, pages = {e0006050}, pmid = {29084217}, issn = {1935-2735}, support = {R01 AI116636/AI/NIAID NIH HHS/United States ; R21 AI128918/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Culex/*microbiology/*virology ; Host-Pathogen Interactions ; Insect Vectors/microbiology/virology ; Mosquito Control ; Pest Control, Biological/*methods ; Rift Valley fever virus/*physiology ; Wolbachia/*physiology ; }, abstract = {Innovative tools are needed to alleviate the burden of mosquito-borne diseases, and strategies that target the pathogen are being considered. A possible tactic is the use of Wolbachia, a maternally inherited, endosymbiotic bacterium that can (but does not always) suppress diverse pathogens when introduced to naive mosquito species. We investigated effects of somatic Wolbachia (strain wAlbB) infection on Rift Valley fever virus (RVFV) in Culex tarsalis mosquitoes. When compared to Wolbachia-uninfected mosquitoes, there was no significant effect of Wolbachia infection on RVFV infection, dissemination, or transmission frequencies, nor on viral body or saliva titers. Within Wolbachia-infected mosquitoes, there was a modest negative correlation between RVFV body titers and Wolbachia density, suggesting that Wolbachia may slightly suppress RVFV in a density-dependent manner in this mosquito species. These results are contrary to previous work in the same mosquito species, showing Wolbachia-induced enhancement of West Nile virus infection rates. Taken together, these results highlight the importance of exploring the breadth of pathogen modulations induced by Wolbachia.}, } @article {pmid29080766, year = {2017}, author = {Diosdado, A and Gómez, PJ and Morchón, R and Simón, F and González-Miguel, J}, title = {Interaction between Wolbachia and the fibrinolytic system as a possible pathological mechanism in cardiopulmonary dirofilariosis.}, journal = {Veterinary parasitology}, volume = {247}, number = {}, pages = {64-69}, doi = {10.1016/j.vetpar.2017.10.001}, pmid = {29080766}, issn = {1873-2550}, mesh = {Amino Acid Sequence ; Animals ; Dirofilaria immitis/*microbiology ; Dirofilariasis/*parasitology ; Dog Diseases/*parasitology ; Dogs ; Fibrinolysin/metabolism ; Fibrinolysis ; Host-Parasite Interactions ; Humans ; Membrane Proteins/genetics/*metabolism ; Models, Molecular ; Recombinant Proteins ; Sequence Alignment/veterinary ; Symbiosis ; Tissue Plasminogen Activator/*metabolism ; Wolbachia/*physiology ; }, abstract = {Dirofilaria immitis is a mosquito-borne parasite that produces an inflammatory process in the wall of the blood vessels of its definitive host during cardiopulmonary dirofilariosis, known as proliferative endarteritis. Parasite antigens participate in the appearance of this inflammatory event, among other mechanisms through the over-activation of the host fibrinolytic system. Since Wolbachia, endosymbiont bacteria of filarial nematodes, is released into the vertebrate host when worms die, the aim of this work was to analyse the interaction between this bacteria and the host fibrinolytic system to complete the study of this part of the host-parasite relationships. For that purpose, the recombinant form of the major Wolbachia surface protein (rWSP) was cloned, sequenced and expressed and then, its ability to bind plasminogen and enhance the generation of plasmin was assessed. We demonstrated that rWSP is a conserved antigen within the family Onchocercidae with ability to bind plasminogen and stimulate plasmin generation in a tissue-plasminogen activator (t-PA) and lysine residues of the rWSP-dependent manner. These results indicate that the recruitment of plasminogen by Wolbachia and the possible excess of plasmin generated could contribute to exacerbate the pathological events occurred at the vascular level during cardiopulmonary dirofilariosis, as well as in other diseases caused by filarial nematodes that harbour Wolbachia, when the bacteria is released after the death of the worms.}, } @article {pmid29078351, year = {2017}, author = {Turner, JD and Sharma, R and Al Jayoussi, G and Tyrer, HE and Gamble, J and Hayward, L and Priestley, RS and Murphy, EA and Davies, J and Waterhouse, D and Cook, DAN and Clare, RH and Cassidy, A and Steven, A and Johnston, KL and McCall, J and Ford, L and Hemingway, J and Ward, SA and Taylor, MJ}, title = {Albendazole and antibiotics synergize to deliver short-course anti-Wolbachia curative treatments in preclinical models of filariasis.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, volume = {114}, number = {45}, pages = {E9712-E9721}, pmid = {29078351}, issn = {1091-6490}, support = {MR/L018756/1/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Albendazole/*pharmacology ; Animals ; Anti-Bacterial Agents/*pharmacology ; Benzimidazoles/pharmacology ; Brugia malayi/microbiology ; Drug Synergism ; Female ; Filariasis/*drug therapy ; Male ; Mice ; Mice, Inbred BALB C ; Minocycline/pharmacology ; Rifampin/pharmacology ; Wolbachia/*drug effects ; }, abstract = {Elimination of filariasis requires a macrofilaricide treatment that can be delivered within a 7-day period. Here we have identified a synergy between the anthelmintic albendazole (ABZ) and drugs depleting the filarial endosymbiont Wolbachia, a proven macrofilaricide target, which reduces treatment from several weeks to 7 days in preclinical models. ABZ had negligible effects on Wolbachia but synergized with minocycline or rifampicin (RIF) to deplete symbionts, block embryogenesis, and stop microfilariae production. Greater than 99% Wolbachia depletion following 7-day combination of RIF+ABZ also led to accelerated macrofilaricidal activity. Thus, we provide preclinical proof-of-concept of treatment shortening using antibiotic+ABZ combinations to deliver anti-Wolbachia sterilizing and macrofilaricidal effects. Our data are of immediate public health importance as RIF+ABZ are registered drugs and thus immediately implementable to deliver a 1-wk macrofilaricide. They also suggest that novel, more potent anti-Wolbachia drugs under development may be capable of delivering further treatment shortening, to days rather than weeks, if combined with benzimidazoles.}, } @article {pmid29076805, year = {2018}, author = {Lourenço-de-Oliveira, R and Marques, JT and Sreenu, VB and Atyame Nten, C and Aguiar, ERGR and Varjak, M and Kohl, A and Failloux, AB}, title = {Culex quinquefasciatus mosquitoes do not support replication of Zika virus.}, journal = {The Journal of general virology}, volume = {99}, number = {2}, pages = {258-264}, pmid = {29076805}, issn = {1465-2099}, support = {G0801822/MRC_/Medical Research Council/United Kingdom ; MC_UU_12014/12/MRC_/Medical Research Council/United Kingdom ; MC_UU_12014/8/MRC_/Medical Research Council/United Kingdom ; U01 AI115595/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Culex/*virology ; Female ; Mosquito Vectors/*virology ; Phenotype ; *Virus Replication ; Wolbachia/*physiology ; Zika Virus/*physiology ; Zika Virus Infection/*transmission/virology ; }, abstract = {The rapid spread of Zika virus (ZIKV) in the Americas raised many questions about the role of Culex quinquefasciatus mosquitoes in transmission, in addition to the key role played by the vector Aedes aegypti. Here we analysed the competence of Cx. quinquefasciatus (with or without Wolbachia endosymbionts) for a ZIKV isolate. We also examined the induction of RNA interference pathways after viral challenge and the production of small virus-derived RNAs. We did not observe any infection nor such small virus-derived RNAs, regardless of the presence or absence of Wolbachia. Thus, Cx. quinquefasciatus does not support ZIKV replication and Wolbachia is not involved in producing this phenotype. In short, these mosquitoes are very unlikely to play a role in transmission of ZIKV.}, } @article {pmid29076641, year = {2017}, author = {Bhattacharya, T and Newton, ILG}, title = {Mi Casa es Su Casa: how an intracellular symbiont manipulates host biology.}, journal = {Environmental microbiology}, volume = {}, number = {}, pages = {}, pmid = {29076641}, issn = {1462-2920}, support = {R21 AI121849/AI/NIAID NIH HHS/United States ; }, abstract = {Wolbachia pipientis, the most common intracellular infection on the planet, infects 40% of insects as well as nematodes, isopods and arachnids. Wolbachia are obligately intracellular and challenging to study; there are no genetic tools for manipulating Wolbachia nor can they be cultured outside of host cells. Despite these roadblocks, the research community has defined a set of Wolbachia loci involved in host interaction: Wolbachia effectors. Through the use of Drosophila genetics, surrogate systems and biochemistry, the field has begun to define the toolkit Wolbachia use for host manipulation. Below we review recent findings identifying these Wolbachia effectors and point to potential, as yet uncharacterized, links between known phenotypes induced by Wolbachia infection and predicted effectors.}, } @article {pmid29074653, year = {2017}, author = {Ward, CM and Baxter, SW}, title = {Draft Genome Assembly of a Wolbachia Endosymbiont of Plutella australiana.}, journal = {Genome announcements}, volume = {5}, number = {43}, pages = {}, pmid = {29074653}, issn = {2169-8287}, abstract = {Wolbachia spp. are endosymbiotic bacteria that infect around 50% of arthropods and cause a broad range of effects, including manipulating host reproduction. Here, we present the annotated draft genome assembly of Wolbachia strain wAus, which infects Plutella australiana, a cryptic ally of the major Brassica pest Plutella xylostella (diamondback moth).}, } @article {pmid29071168, year = {2017}, author = {Zhang, L and Zhang, G and Yun, Y and Peng, Y}, title = {Bacterial community of a spider, Marpiss magister (Salticidae).}, journal = {3 Biotech}, volume = {7}, number = {6}, pages = {371}, pmid = {29071168}, issn = {2190-572X}, abstract = {Arthropods are associated with various microorganisms which confer benefits to their hosts. Recently, research has been conducted on bacterial communities of insects to provide an insight into the potential interactions of the symbiotic bacteria and their hosts. Spiders are interesting to study as they are perceived to be natural enemies of pests. The effect of endosymbionts on spiders has been reported, but little is known about the overall bacterial communities present in spiders. Here, we report on the characterization of bacterial communities present in the whole body of the spider Marpiss magister using Illumina sequencing of 16S rRNA amplicons. Our study shows that the most abundant phyla of bacteria included Proteobacteria, Tenericutes, Bacteroidetes and Actinobacteria. At the genus level, the most abundant genera included Rickettsia, Wolbachia, Spiroplasma, and Cardinium. Besides these dominant endosymbionts, our study also showed the existence of bacteria in the genera Arthrobacter, Novosphingobium, Acinetobacter, Pseudomonas, Aquabacterium and Sphingomonas at an abundance ranging from 0.65 to 0.84%, and the existence of bacterial in genera Lactobacillus, Sphingobium, Methylobacterium, Bradyrhizobium, Propionibacterium, Brevundimonas, Achromobacter, Microbacterium, Corynebacterium, and Flavobacterium at a slightly lower abundance ranging from 0.1 to 0.5%. Therefore, our finding indicates that endosymbionts are not the only microbiota present in the spider M. magister, and other bacterial taxa also exist in its bacterial community.}, } @article {pmid29067008, year = {2017}, author = {Zhukova, M and Sapountzis, P and Schiøtt, M and Boomsma, JJ}, title = {Diversity and Transmission of Gut Bacteria in Atta and Acromyrmex Leaf-Cutting Ants during Development.}, journal = {Frontiers in microbiology}, volume = {8}, number = {}, pages = {1942}, pmid = {29067008}, issn = {1664-302X}, support = {323085/ERC_/European Research Council/International ; }, abstract = {The social Hymenoptera have distinct larval and adult stages separated by metamorphosis, which implies striking remodeling of external and internal body structures during the pupal stage. This imposes challenges to gut symbionts as existing cultures are lost and may or may not need to be replaced. To elucidate the extent to which metamorphosis interrupts associations between bacteria and hosts, we analyzed changes in gut microbiota during development and traced the transmission routes of dominant symbionts from the egg to adult stage in the leaf-cutting ants Acromyrmex echinatior and Atta cephalotes, which are both important functional herbivores in the New World tropics. Bacterial density remained similar across the developmental stages of Acromyrmex, but Atta brood had very low bacterial prevalences suggesting that bacterial gut symbionts are not actively maintained. We found that Wolbachia was the absolute dominant bacterial species across developmental stages in Acromyrmex and we confirmed that Atta lacks Wolbachia also in the immature stages, and had mostly Mollicutes bacteria in the adult worker guts. Wolbachia in Acromyrmex appeared to be transovarially transmitted similar to transmission in solitary insects. In contrast, Mollicutes were socially transmitted from old workers to newly emerged callows. We found that larval and pupal guts of both ant species contained Pseudomonas and Enterobacter bacteria that are also found in fungus gardens, but hardly or not in adult workers, suggesting they are beneficial only for larval growth and development. Our results reveal that transmission pathways for bacterial symbionts may be very different both between developmental stages and between sister genera and that identifying the mechanisms of bacterial acquisition and loss will be important to clarify their putative mutualistic functions.}, } @article {pmid29066721, year = {2017}, author = {He, S and Zhang, X and Liang, J and Tang, S}, title = {Multiscale modelling the effects of CI genetic evolution in mosquito population on the control of dengue fever.}, journal = {Scientific reports}, volume = {7}, number = {1}, pages = {13895}, pmid = {29066721}, issn = {2045-2322}, mesh = {Animals ; Culicidae/*genetics/microbiology/*physiology ; Cytoplasm/*genetics ; Dengue/*prevention & control/transmission ; Evolution, Molecular ; Female ; Gene Frequency ; Genotype ; Male ; *Models, Genetic ; Mosquito Control/*methods ; Reproduction/*genetics ; Wolbachia/genetics ; }, abstract = {Endosymbiotic Wolbachia bacteria are widely applied for the control of dengue fever by manipulating the reproductive mechanism of mosquitoes, including maternal inheritance and cytoplasmic incompatibility (CI). CI means that the offsprings from the matings between Wolbachia infected males and uninfected females can not be hatched. At present, CI effect is assumed as a constant in most of dynamic systems for the spread of Wolbachia. However, their spread may arouse the evolution of mosquitoes to resist CI. Thus, a multiscale model combining a birth-pulse model with a gene-induced discrete model for the frequencies of alleles is proposed to describe the spread of Wolbachia in mosquito population with resistance allele of CI. The main results indicate that the strategy of population eradication can not be realized, while the strategy of population replacement may be realized with the success of sensitive or resistance allele. If appropriate Wolbachia strains can not be selected, then there is a high probability of the failure of population replacement. Moreover, Wolbachia-induced parameters may arouse the catastrophic shifts among stable states of the model. In addition, the demographic parameters and Wolbachia-induced parameters may affect the level and the speed of population replacement and the density of uninfected mosquitoes.}, } @article {pmid29065795, year = {2017}, author = {Al-Abd, NM and Nor, ZM and Junaid, QO and Mansor, M and Hasan, MS and Kassim, M}, title = {Antifilarial activity of caffeic acid phenethyl ester on Brugia pahangi in vitro and in vivo.}, journal = {Pathogens and global health}, volume = {111}, number = {7}, pages = {388-394}, pmid = {29065795}, issn = {2047-7732}, mesh = {Animals ; Brugia pahangi/*drug effects ; Caffeic Acids/administration & dosage/*therapeutic use ; Dose-Response Relationship, Drug ; Filariasis/*drug therapy/parasitology ; Gerbillinae ; Humans ; Male ; Phenylethyl Alcohol/administration & dosage/*analogs & derivatives/therapeutic use ; Wolbachia/drug effects ; }, abstract = {Lymphatic filariasis (LF) is a vector borne disease caused by parasitic worms such as Wuchereria bancrofti, Brugia malayi and B. timori, which are transmitted by mosquitoes. Current therapeutics to treat LF are mainly microfilarcidal, and lack activity against adult worms. This set back, poses a challenge for the control and elimination of filariasis. Thus, in this study the activities of caffeic acid phenethyl ester (CAPE) against the filarial worm B. pahangi and its bacterial endosymbiont, Wolbachia were evaluated. Different concentrations (2, 5, 10, 15, 20 μg/ml) of CAPE were used to assess its effects on motility, viability and microfilarial (mf) production of B. pahangi in vitro. Anti-Wolbachial activity of CAPE was measured in worms by quantification of Wolbachial wsp gene copy number using real-time polymerase chain reaction. Our findings show that CAPE was found to significantly reduce adult worm motility, viability, and mf release both in vitro and in vivo. 20 μg/ml of CAPE halts the release of mf in vitro by day 6 of post treatment. Also, the number of adult worms recovered in vivo were reduced significantly during and after treatment with 50 mg/kg of CAPE relative to control drugs, diethylcarbamazine and doxycycline. Real time PCR based on the Wolbachia ftsZ gene revealed a significant reduction in Wolbachia copy number upon treatment. Anti-Wolbachia and antifilarial properties of CAPE require further investigation as an alternative strategy to treat LF.}, } @article {pmid29063344, year = {2018}, author = {Hornok, S and Ágh, N and Takács, N and Kontschán, J and Hofmann-Lehmann, R}, title = {Haematospirillum and insect Wolbachia DNA in avian blood.}, journal = {Antonie van Leeuwenhoek}, volume = {111}, number = {3}, pages = {479-483}, doi = {10.1007/s10482-017-0961-0}, pmid = {29063344}, issn = {1572-9699}, support = {115854//OTKA/ ; 108571//OTKA/ ; }, mesh = {Alphaproteobacteria/*genetics ; Animals ; Birds/*blood ; *DNA, Bacterial ; Female ; *Genes, Insect ; Male ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA ; Wolbachia/*genetics ; }, abstract = {In this study, blood samples of 259 Acrocephalus sp. warblers were molecularly analysed for Anaplasmataceae and Rhodospirillaceae based on PCR amplification of 16S rRNA gene fragments. One bird blood sample (from Reed Warbler, Acrocephalus scirpaceus) yielded a sequence with 99.8% identity to Haematospirillum jordaniae. This is the first molecular evidence for the occurrence of this species in the blood of any vertebrate other than human. Another bird blood sample (from Marsh Warbler: Acrocephalus palustris) yielded a Wolbachia sequence, closely related to a moth endosymbiont with 99.8% identity. A nematode origin of Wolbachia DNA detected here in avian blood can be excluded, because results of phylogenetic analysis showed its closest alignment with insect wolbachiae. This is the first finding of insect Wolbachia DNA in the circulatory system of birds, which can be explained either by the inoculation of wolbachiae by blood-sucking vectors, or passing of Wolbachia DNA from the gut into the blood of this insectivorous bird species.}, } @article {pmid29053633, year = {2017}, author = {Skaljac, M and Kanakala, S and Zanic, K and Puizina, J and Pleic, IL and Ghanim, M}, title = {Diversity and Phylogenetic Analyses of Bacterial Symbionts in Three Whitefly Species from Southeast Europe.}, journal = {Insects}, volume = {8}, number = {4}, pages = {}, pmid = {29053633}, issn = {2075-4450}, abstract = {Bemisia tabaci (Gennadius), Trialeurodes vaporariorum (Westwood), and Siphoninus phillyreae (Haliday) are whitefly species that harm agricultural crops in many regions of the world. These insects live in close association with bacterial symbionts that affect host fitness and adaptation to the environment. In the current study, we surveyed the infection of whitefly populations in Southeast Europe by various bacterial symbionts and performed phylogenetic analyses on the different symbionts detected. Arsenophonus and Hamiltonella were the most prevalent symbionts in all three whitefly species. Rickettsia was found to infect mainly B. tabaci, while Wolbachia mainly infected both B. tabaci and S. phillyreae. Furthermore, Cardinium was rarely found in the investigated whitefly populations, while Fritschea was never found in any of the whitefly species tested. Phylogenetic analyses revealed a diversity of several symbionts (e.g., Hamiltonella, Arsenophonus, Rickettsia), which appeared in several clades. Reproductively isolated B. tabaci and T. vaporariorum shared the same (or highly similar) Hamiltonella and Arsenophonus, while these symbionts were distinctive in S. phillyreae. Interestingly, Arsenophonus from S. phillyreae did not cluster with any of the reported sequences, which could indicate the presence of Arsenophonus, not previously associated with whiteflies. In this study, symbionts (Wolbachia, Rickettsia, and Cardinium) known to infect a wide range of insects each clustered in the same clades independently of the whitefly species. These results indicate horizontal transmission of bacterial symbionts between reproductively isolated whitefly species, a mechanism that can establish new infections that did not previously exist in whiteflies.}, } @article {pmid29042606, year = {2017}, author = {Signor, S}, title = {Population genomics of Wolbachia and mtDNA in Drosophila simulans from California.}, journal = {Scientific reports}, volume = {7}, number = {1}, pages = {13369}, pmid = {29042606}, issn = {2045-2322}, mesh = {California ; DNA, Mitochondrial/*genetics ; Drosophila simulans/*microbiology ; Evolution, Molecular ; Gene Dosage ; *Genetics, Population ; *Genomics/methods ; Linkage Disequilibrium ; Phylogeny ; Polymorphism, Single Nucleotide ; Wolbachia/classification/*genetics ; }, abstract = {Wolbachia pipientis is an intracellular endosymbiont infecting many arthropods and filarial nematodes. Little is known about the short-term evolution of Wolbachia or its interaction with its host. Wolbachia is maternally inherited, resulting in co-inheritance of mitochondrial organelles such as mtDNA. Here I explore the evolution of Wolbachia, and the relationship between Wolbachia and mtDNA, using a large inbred panel of Drosophila simulans. I compare this to the only other large population genomic Wolbachia dataset from D. melanogaster. I find reduced diversity relative to expectation in both Wolbachia and mtDNA, but only mtDNA shows evidence of a recent selective sweep or population bottleneck. I estimate Wolbachia and mtDNA titre in each genotype, and I find considerable variation in both phenotypes, despite low genetic diversity in Wolbachia and mtDNA. A phylogeny of Wolbachia and of mtDNA suggest a recent origin of the infection derived from a single origin. Using Wolbachia and mtDNA titre as a phenotype, I perform the first association analysis using this phenotype with the nuclear genome and find several implicated regions, including one which contains four CAAX-box protein processing genes. CAAX-box protein processing can be an important part of host-pathogen interactions in other systems, suggesting interesting directions for future research.}, } @article {pmid29036401, year = {2017}, author = {Santos-Garcia, D and Silva, FJ and Morin, S and Dettner, K and Kuechler, SM}, title = {The All-Rounder Sodalis: A New Bacteriome-Associated Endosymbiont of the Lygaeoid Bug Henestaris halophilus (Heteroptera: Henestarinae) and a Critical Examination of Its Evolution.}, journal = {Genome biology and evolution}, volume = {9}, number = {10}, pages = {2893-2910}, pmid = {29036401}, issn = {1759-6653}, mesh = {Animals ; DNA, Bacterial ; Databases, Factual ; Enterobacteriaceae/*classification/genetics/*physiology/ultrastructure ; *Evolution, Molecular ; Genome Size ; Genome, Bacterial ; Heteroptera/*microbiology ; Metabolic Networks and Pathways ; *Phylogeny ; Pseudogenes ; Sequence Analysis, DNA ; *Symbiosis ; }, abstract = {Hemipteran insects are well-known in their ability to establish symbiotic relationships with bacteria. Among them, heteropteran insects present an array of symbiotic systems, ranging from the most common gut crypt symbiosis to the more restricted bacteriome-associated endosymbiosis, which have only been detected in members of the superfamily Lygaeoidea and the family Cimicidae so far. Genomic data of heteropteran endosymbionts are scarce and have merely been analyzed from the Wolbachia endosymbiont in bed bug and a few gut crypt-associated symbionts in pentatomoid bugs. In this study, we present the first detailed genomic analysis of a bacteriome-associated endosymbiont of a phytophagous heteropteran, present in the seed bug Henestaris halophilus (Hemiptera: Heteroptera: Lygaeoidea). Using phylogenomics and genomics approaches, we have assigned the newly characterized endosymbiont to the Sodalis genus, named as Candidatus Sodalis baculum sp. nov. strain kilmister. In addition, our findings support the reunification of the Sodalis genus, currently divided into six different genera. We have also conducted comparative analyses between 15 Sodalis species that present different genome sizes and symbiotic relationships. These analyses suggest that Ca. Sodalis baculum is a mutualistic endosymbiont capable of supplying the amino acids tyrosine, lysine, and some cofactors to its host. It has a small genome with pseudogenes but no mobile elements, which indicates middle-stage reductive evolution. Most of the genes in Ca. Sodalis baculum are likely to be evolving under purifying selection with several signals pointing to the retention of the lysine/tyrosine biosynthetic pathways compared with other Sodalis.}, } @article {pmid29021316, year = {2017}, author = {Kageyama, D and Yoshimura, K and Sugimoto, TN and Katoh, TK and Watada, M}, title = {Maternally transmitted non-bacterial male killer in Drosophila biauraria.}, journal = {Biology letters}, volume = {13}, number = {10}, pages = {}, pmid = {29021316}, issn = {1744-957X}, mesh = {Animals ; Anti-Bacterial Agents/pharmacology ; Drosophila/*physiology ; Female ; Japan ; Male ; Maternal Inheritance ; Phenotype ; Polymerase Chain Reaction ; Sex Ratio ; Symbiosis ; Tetracycline/pharmacology ; }, abstract = {A maternally inherited, all-female trait is widely found among arthropods, which is caused by bacterial endosymbionts such as Wolbachia, Rickettsia, Spiroplasma and Cardinium We discovered a single female of Drosophila biauraria, collected from Tomakomai, Hokkaido, Japan, that produced all-female offspring. This all-female trait was maternally inherited in the iso-female line (SP12F) by backcrossing with males of a normal line (SP11-20) with a 1 : 1 sex ratio derived from the same population. The all-female trait was not affected by tetracycline treatment performed for two consecutive generations. However, the microinjection of filter-sterilized homogenate of SP12F females into SP11-20 females established all-female matrilines. Our data suggest the role of transmissible agents, most likely viruses, but not bacteria or protists, as the possible cause of the all-female phenotype, which is likely to be achieved by killing of male embryos because egg hatch rates of SP12F were nearly half those of SP11-20. This is the first report in Diptera to demonstrate a maternally inherited virus-like element as the cause of the male-killing phenotype in D. biauraria.}, } @article {pmid28991902, year = {2017}, author = {de Oliveira, S and Villela, DAM and Dias, FBS and Moreira, LA and Maciel de Freitas, R}, title = {How does competition among wild type mosquitoes influence the performance of Aedes aegypti and dissemination of Wolbachia pipientis?.}, journal = {PLoS neglected tropical diseases}, volume = {11}, number = {10}, pages = {e0005947}, pmid = {28991902}, issn = {1935-2735}, mesh = {Aedes/*microbiology ; Animals ; Host-Pathogen Interactions ; Larva ; Pest Control, Biological/methods ; Population Density ; Species Specificity ; Survival Analysis ; Wolbachia/*physiology ; }, abstract = {BACKGROUND: Wolbachia has been deployed in several countries to reduce transmission of dengue, Zika and chikungunya viruses. During releases, Wolbachia-infected females are likely to lay their eggs in local available breeding sites, which might already be colonized by local Aedes sp. mosquitoes. Therefore, there is an urgent need to estimate the deleterious effects of intra and interspecific larval competition on mosquito life history traits, especially on the duration of larval development time, larval mortality and adult size.

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

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

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

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

CONCLUSION: To the best of our knowledge this is the first study on Wolbachia screening from Northeast part of India and also first report of natural Wolbachia infection from colonised Anopheles stephensi species. The current understanding on distribution of Wolbachia strains naturally present within insect species from this geographical region should aid future Wolbachia mediated vector control strategies.}, } @article {pmid28932814, year = {2017}, author = {Simhadri, RK and Fast, EM and Guo, R and Schultz, MJ and Vaisman, N and Ortiz, L and Bybee, J and Slatko, BE and Frydman, HM}, title = {The Gut Commensal Microbiome of Drosophila melanogaster Is Modified by the Endosymbiont Wolbachia.}, journal = {mSphere}, volume = {2}, number = {5}, pages = {}, pmid = {28932814}, issn = {2379-5042}, abstract = {Endosymbiotic Wolbachia bacteria and the gut microbiome have independently been shown to affect several aspects of insect biology, including reproduction, development, life span, stem cell activity, and resistance to human pathogens, in insect vectors. This work shows that Wolbachia bacteria, which reside mainly in the fly germline, affect the microbial species present in the fly gut in a lab-reared strain. Drosophila melanogaster hosts two main genera of commensal bacteria-Acetobacter and Lactobacillus. Wolbachia-infected flies have significantly reduced titers of Acetobacter. Sampling of the microbiome of axenic flies fed with equal proportions of both bacteria shows that the presence of Wolbachia bacteria is a significant determinant of the composition of the microbiome throughout fly development. However, this effect is host genotype dependent. To investigate the mechanism of microbiome modulation, the effect of Wolbachia bacteria on Imd and reactive oxygen species pathways, the main regulators of immune response in the fly gut, was measured. The presence of Wolbachia bacteria does not induce significant changes in the expression of the genes for the effector molecules in either pathway. Furthermore, microbiome modulation is not due to direct interaction between Wolbachia bacteria and gut microbes. Confocal analysis shows that Wolbachia bacteria are absent from the gut lumen. These results indicate that the mechanistic basis of the modulation of composition of the microbiome by Wolbachia bacteria is more complex than a direct bacterial interaction or the effect of Wolbachia bacteria on fly immunity. The findings reported here highlight the importance of considering the composition of the gut microbiome and host genetic background during Wolbachia-induced phenotypic studies and when formulating microbe-based disease vector control strategies. IMPORTANCEWolbachia bacteria are intracellular bacteria present in the microbiome of a large fraction of insects and parasitic nematodes. They can block mosquitos' ability to transmit several infectious disease-causing pathogens, including Zika, dengue, chikungunya, and West Nile viruses and malaria parasites. Certain extracellular bacteria present in the gut lumen of these insects can also block pathogen transmission. However, our understanding of interactions between Wolbachia and gut bacteria and how they influence each other is limited. Here we show that the presence of Wolbachia strain wMel changes the composition of gut commensal bacteria in the fruit fly. Our findings implicate interactions between bacterial species as a key factor in determining the overall composition of the microbiome and thus reveal new paradigms to consider in the development of disease control strategies.}, } @article {pmid28913925, year = {2018}, author = {Blackwood, JC and Vargas, R and Fauvergue, X}, title = {A cascade of destabilizations: Combining Wolbachia and Allee effects to eradicate insect pests.}, journal = {The Journal of animal ecology}, volume = {87}, number = {1}, pages = {59-72}, doi = {10.1111/1365-2656.12756}, pmid = {28913925}, issn = {1365-2656}, mesh = {Animals ; *Genetic Fitness ; Insecta/microbiology/*physiology ; Models, Biological ; *Pest Control, Biological ; Population Density ; Wolbachia/*physiology ; }, abstract = {The management of insect pests has long been dominated by the use of chemical insecticides, with the aim of instantaneously killing enough individuals to limit their damage. To minimize unwanted consequences, environmentally friendly approaches have been proposed that utilize biological control and take advantage of intrinsic demographic processes to reduce pest populations. We address the feasibility of a novel pest management strategy based on the release of insects infected with Wolbachia, which causes cytoplasmic incompatibilities in its host population, into a population with a pre-existing Allee effect. We hypothesize that the transient decline in population size caused by a successful invasion of Wolbachia can bring the population below its Allee threshold and, consequently, trigger extinction. We develop a stochastic population model that accounts for Wolbachia-induced cytoplasmic incompatibilities in addition to an Allee effect arising from mating failures at low population densities. Using our model, we identify conditions under which cytoplasmic incompatibilities and Allee effects successfully interact to drive insect pest populations towards extinction. Based on our results, we delineate control strategies based on introductions of Wolbachia-infected insects. We extend this analysis to evaluate control strategies that implement successive introductions of two incompatible Wolbachia strains. Additionally, we consider methods that combine Wolbachia invasion with mating disruption tactics to enhance the pre-existing Allee effect. We demonstrate that Wolbachia-induced cytoplasmic incompatibility and the Allee effect act independently from one another: the Allee effect does not modify the Wolbachia invasion threshold, and cytoplasmic incompatibilities only have a marginal effect on the Allee threshold. However, the interaction of these two processes can drive even large populations to extinction. The success of this method can be amplified by the introduction of multiple Wolbachia cytotypes as well as the addition of mating disruption. Our study extends the existing literature by proposing the use of Wolbachia introductions to capitalize on pre-existing Allee effects and consequently eradicate insect pests. More generally, it highlights the importance of transient dynamics, and the relevance of manipulating a cascade of destabilizatons for pest management.}, } @article {pmid28906572, year = {2018}, author = {Leggewie, M and Krumkamp, R and Badusche, M and Heitmann, A and Jansen, S and Schmidt-Chanasit, J and Tannich, E and Becker, SC}, title = {Culex torrentium mosquitoes from Germany are negative for Wolbachia.}, journal = {Medical and veterinary entomology}, volume = {32}, number = {1}, pages = {115-120}, doi = {10.1111/mve.12270}, pmid = {28906572}, issn = {1365-2915}, mesh = {Animals ; Culex/*microbiology ; Female ; Germany ; Species Specificity ; West Nile virus/*isolation & purification ; Wolbachia/*isolation & purification ; }, abstract = {Wolbachia (Rickettsiales: Anaplasmataceae) infects a wide range of arthropods, including several mosquito species. The bacterium is known to induce a plethora of phenotypes in its host, examples being the reproductive phenotype cytoplasmic incompatibility or resistance against infection with arboviruses. The latter is especially relevant when assessing the vector competence of mosquito species for emerging arboviruses. Thus, knowledge of Wolbachia infection status is important for the assessment of vector competence. To facilitate Wolbachia screening in mosquito populations, a quantitative polymerase chain reaction (qPCR) assay was developed to enable high-throughput analysis of mosquito samples. Using this assay, the Wolbachia infection status of the two most common Culex mosquito species in Germany, Culex pipiens biotype pipiens Linnaeus (Diptera: Culicidae) and Culex torrentium Martini (Diptera: Culicidae), was assessed. About 93% of all tested C. pipiens biotype pipiens individuals were positive for Wolbachia, whereas none of the C. torrentium samples was found to be infected. Furthermore, other applications of the qPCR assay were explored by assessing a potential link between the levels of Wolbachia and West Nile virus (WNV) infections in German C. pipiens biotype pipiens mosquitoes. No relationship was found between the two variables, indicating that a Wolbachia-induced antiviral phenotype in this mosquito population is not exclusively attributable to the general level of bacterial infection.}, } @article {pmid28905230, year = {2017}, author = {Huber, D and Reil, I and Duvnjak, S and Jurković, D and Lukačević, D and Pilat, M and Beck, A and Mihaljević, Ž and Vojta, L and Polkinghorne, A and Beck, R}, title = {Molecular detection of Anaplasma platys, Anaplasma phagocytophilum and Wolbachia sp. but not Ehrlichia canis in Croatian dogs.}, journal = {Parasitology research}, volume = {116}, number = {11}, pages = {3019-3026}, pmid = {28905230}, issn = {1432-1955}, mesh = {Anaplasma/classification/*isolation & purification ; Anaplasma phagocytophilum/genetics/*isolation & purification ; Anaplasmosis/epidemiology/*microbiology ; Animals ; Base Sequence ; Croatia/epidemiology ; Dog Diseases/epidemiology/*microbiology ; Dogs ; Ehrlichia canis/*isolation & purification ; Ehrlichiosis/veterinary ; Phylogeny ; Polymerase Chain Reaction/veterinary ; RNA, Bacterial ; RNA, Ribosomal, 16S/genetics ; Rhipicephalus sanguineus/microbiology ; Rickettsiaceae Infections/epidemiology/microbiology/*veterinary ; Wolbachia/genetics/*isolation & purification ; }, abstract = {The bacteria Anaplasma platys, Anaplasma phagocytophilum and Ehrlichia canis are tick-borne agents that cause canine vector-borne disease. The prevalence of these pathogens in South Eastern Europe is unknown with the exception of an isolated case of A. platys detected in a dog imported into Germany from Croatia. To gain a better insight into their presence and prevalence, PCR-based screening for these bacterial pathogens was performed on domesticated dogs from different regions of Croatia. Blood samples from 1080 apparently healthy dogs from coastal and continental parts of Croatia as well as tissue samples collected from 63 deceased dogs with a history of anaemia and thrombocytopenia were collected for molecular screening by an Anaplasmataceae-specific 16S rRNA conventional PCR. Positive samples were confirmed using a second Anaplasmataceae-specific PCR assay with the PCR product sequenced for the purpose of bacterial species identification. All sequenced isolates were georeferenced and a kernel intensity estimator was used to identify clusters of greater case intensity. 42/1080 (3.8%; CI 2.7-5.0) of the healthy dogs were PCR positive for bacteria in the Anaplasmataceae. Sequencing of the 16S rRNA gene amplified from these positive samples revealed the presence of A. platys in 2.5% (CI 1.6-3.4%, 27 dogs), A. phagocytophilum in 0.3% (CI 0-0.6%, 3 dogs) and a Wolbachia endosymbiont in 1.1% (CI 0.4-1.6%, 12 dogs) of dogs screened in this study. Necropsied dogs were free from infection. Notably, no evidence of E. canis infection was found in any animal. This survey represents a rare molecular study of Anaplasmataceae in dogs in South Eastern Europe, confirming the presence of A. platys and A. phagocytophilum but not E. canis. The absence of E. canis was surprising given it has been described in all other Mediterranean countries surveyed and raises questions over the regional vector capacity of the Rhipicephalus sanguineus tick.}, } @article {pmid28904344, year = {2017}, author = {Geoghegan, V and Stainton, K and Rainey, SM and Ant, TH and Dowle, AA and Larson, T and Hester, S and Charles, PD and Thomas, B and Sinkins, SP}, title = {Perturbed cholesterol and vesicular trafficking associated with dengue blocking in Wolbachia-infected Aedes aegypti cells.}, journal = {Nature communications}, volume = {8}, number = {1}, pages = {526}, pmid = {28904344}, issn = {2041-1723}, support = {/WT_/Wellcome Trust/United Kingdom ; 202888/Z/16/Z/WT_/Wellcome Trust/United Kingdom ; MC_PC_15087/MRC_/Medical Research Council/United Kingdom ; MC_UU_12014/8/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Aedes/*metabolism/*microbiology/virology ; Animals ; Cholesterol/*metabolism ; Cytoplasmic Vesicles/*metabolism ; Dengue/*transmission/virology ; Dengue Virus/*physiology ; Humans ; Wolbachia/*physiology ; }, abstract = {Wolbachia are intracellular maternally inherited bacteria that can spread through insect populations and block virus transmission by mosquitoes, providing an important approach to dengue control. To better understand the mechanisms of virus inhibition, we here perform proteomic quantification of the effects of Wolbachia in Aedes aegypti mosquito cells and midgut. Perturbations are observed in vesicular trafficking, lipid metabolism and in the endoplasmic reticulum that could impact viral entry and replication. Wolbachia-infected cells display a differential cholesterol profile, including elevated levels of esterified cholesterol, that is consistent with perturbed intracellular cholesterol trafficking. Cyclodextrins have been shown to reverse lipid accumulation defects in cells with disrupted cholesterol homeostasis. Treatment of Wolbachia-infected Ae. aegypti cells with 2-hydroxypropyl-β-cyclodextrin restores dengue replication in Wolbachia-carrying cells, suggesting dengue is inhibited in Wolbachia-infected cells by localised cholesterol accumulation. These results demonstrate parallels between the cellular Wolbachia viral inhibition phenotype and lipid storage genetic disorders. Wolbachia infection of mosquitoes can block dengue virus infection and is tested in field trials, but the mechanism of action is unclear. Using proteomics, Geoghegan et al. here identify effects of Wolbachia on cholesterol homeostasis and dengue virus replication in Aedes aegypti.}, } @article {pmid28902190, year = {2017}, author = {Lindsey, ARI and Stouthamer, R}, title = {The effects of outbreeding on a parasitoid wasp fixed for infection with a parthenogenesis-inducing Wolbachia symbiont.}, journal = {Heredity}, volume = {119}, number = {6}, pages = {411-417}, pmid = {28902190}, issn = {1365-2540}, mesh = {Animals ; Biological Coevolution ; Crosses, Genetic ; Female ; Fertility ; Heterozygote ; Homozygote ; Male ; Models, Genetic ; *Parthenogenesis ; *Sex Ratio ; Symbiosis ; Wasps/*genetics/microbiology ; *Wolbachia ; }, abstract = {Trichogramma wasps can be rendered asexual by infection with the maternally inherited symbiont Wolbachia. Previous studies indicate the Wolbachia strains infecting Trichogramma wasps are host-specific, inferred by failed horizontal transfer of Wolbachia to novel Trichogramma hosts. Additionally, Trichogramma can become dependent upon their Wolbachia infection for the production of female offspring, leaving them irreversibly asexual, further linking host and symbiont. We hypothesized Wolbachia strains infecting irreversibly asexual, resistant to horizontal transfer Trichogramma would show adaptation to a particular host genetic background. To test this, we mated Wolbachia-dependent females with males from a Wolbachia-naïve population to create heterozygous wasps. We measured sex ratios and fecundity, a proxy for Wolbachia fitness, produced by heterozygous wasps, and by their recombinant offspring. We find a heterozygote advantage, resulting in higher fitness for Wolbachia, as wasps will produce more offspring without any reduction in the proportion of females. While recombinant wasps did not differ in total fecundity after 10 days, recombinants produced fewer offspring early on, leading to an increased female-biased sex ratio for the whole brood. Despite the previously identified barriers to horizontal transfer of Wolbachia to and from Trichogramma pretiosum, there were no apparent barriers for Wolbachia to induce parthenogenesis in these non-native backgrounds. This is likely due to the route of infection being introgression rather than horizontal transfer, and possibly the co-evolution of Wolbachia with the mitochondria rather than the nuclear genome. These results help to elucidate the mechanisms by which Wolbachia adapt to hosts and the evolution of host-symbiont phenotypes.}, } @article {pmid28892518, year = {2017}, author = {Rohrscheib, CE and Frentiu, FD and Horn, E and Ritchie, FK and van Swinderen, B and Weible, MW and O'Neill, SL and Brownlie, JC}, title = {Response to: Comment on Rohrscheib et al. 2016 "Intensity of mutualism breakdown is determined by temperature not amplification of Wolbachia genes".}, journal = {PLoS pathogens}, volume = {13}, number = {9}, pages = {e1006521}, pmid = {28892518}, issn = {1553-7374}, mesh = {Genome, Bacterial ; Phylogeny ; *Symbiosis ; Temperature ; Wolbachia/*genetics ; }, } @article {pmid28892498, year = {2017}, author = {Chrostek, E and Teixeira, L}, title = {Comment on Rohrscheib et al. 2016 "Intensity of mutualism breakdown is determined by temperature not amplification of Wolbachia genes".}, journal = {PLoS pathogens}, volume = {13}, number = {9}, pages = {e1006540}, pmid = {28892498}, issn = {1553-7374}, } @article {pmid28878066, year = {2017}, author = {Gebiola, M and Giorgini, M and Kelly, SE and Doremus, MR and Ferree, PM and Hunter, MS}, title = {Cytological analysis of cytoplasmic incompatibility induced by Cardinium suggests convergent evolution with its distant cousin Wolbachia.}, journal = {Proceedings. Biological sciences}, volume = {284}, number = {1862}, pages = {}, pmid = {28878066}, issn = {1471-2954}, mesh = {Aneuploidy ; Animals ; *Cytophagaceae ; Cytoplasm/*microbiology ; Female ; Male ; Mitosis ; Reproduction ; Symbiosis ; Wasps/*microbiology ; Wolbachia ; }, abstract = {Cytoplasmic incompatibility (CI) is a conditional sterility in numerous arthropods that is caused by inherited, intracellular bacteria such as Wolbachia Matings between males carrying CI-inducing Wolbachia and uninfected females, or between males and females infected with different Wolbachia strains, result in progeny that die during very early embryogenesis. Multiple studies in diploid (Drosophila) and haplodiploid (Nasonia) insects have shown that CI-Wolbachia cause a failure of the paternally derived chromatin from resolving into distinct chromosomes. This leads to the formation of chromatin bridges and other mitotic defects as early as the first mitotic division, and to early mitotic arrest. It is currently unknown if CI-inducing symbionts other than Wolbachia affect similar cellular processes. Here, we investigated CI caused by an unrelated bacterium, Cardinium, which naturally infects a parasitic wasp, Encarsia suzannae CI crosses in this host-symbiont system resulted in early mitotic defects including asynchrony of paternal and maternal chromosome sets as they enter mitosis, chromatin bridges and improper chromosome segregation that spanned across multiple mitotic divisions, triggering embryonic death through accumulated aneuploidy. We highlight small differences with CI-Wolbachia, which could be due to the underlying CI mechanism or host-specific effects. Our results suggest a convergence of CI-related cellular phenotypes between these two unrelated symbionts.}, } @article {pmid33592927, year = {2017}, author = {Teo, CHJ and Lim, PKC and Voon, K and Mak, JW}, title = {Detection of dengue viruses and Wolbachia in Aedes aegypti and Aedes albopictus larvae from four urban localities in Kuala Lumpur, Malaysia.}, journal = {Tropical biomedicine}, volume = {34}, number = {3}, pages = {583-597}, pmid = {33592927}, issn = {2521-9855}, abstract = {Dengue fever (DF) is currently one of the most important mosquito-borne diseases that affects humans. Dengue fever (DF) and dengue hemorrhagic fever (DHF) are caused by four serotypes of dengue viruses (DENV-1 to DENV-4). The main vector transmitting dengue is Aedes aegypti while Aedes albopictus acts as a secondary vector. As treatment is unavailable and the first dengue vaccine approved in Mexico, Dengvaxia® has yet to be accepted worldwide, prevention of the disease relies heavily on surveillance and control of mosquito vectors. A transgene driver, Wolbachia was found to limit the transmission of dengue virus in Aedes mosquitoes. Wolbachia alone was able to inhibit viral replication, dissemination and transmission in A. aeygpti mosquitoes in experimental studies. In A. albopictus, Wolbachia did not affect the replication of dengue virus but was able to reduce the viral infection of mosquito salivary glands and limit transmission. Studies on Wolbachia have all been carried out in adult Aedes mosquitoes, hence this study was conducted to determine the presence of dengue virus serotypes and Wolbachia in A. aegypti and A. albopictus larvae collected from ovitraps in four localities in Kuala Lumpur viz. Happy Gardens, IMU Bukit Jalil, Ampang and Taman Yarl. Another objective of this study was to determine the association between dengue virus serotypes and the presence of Wolbachia in A. aegypti and A. albopictus larvae. A total of 300 mosquito larvae was collected; 99 (Happy Gardens), 85 (Bukit Jalil), 73 (Ampang) and 43 (Taman Yarl). Out of 300 larvae collected, 284 were identified as A. albopictus and 16 others were identified as A. aegypti. Of the 284 A. albopictus larvae collected, 211 (74.3%) and 73 (25.7%) were found to be negative and positive for dengue virus respectively. The dengue serotypes detected were 2 DENV-2 (2.7%), 58 DENV-3 (79.5%) and 13 DENV-4 (17.8%). DENV-1 was not detected in any of the A. albopictus larvae. For A. aegypti, out of 16 A. aegypti larvae collected, 12 (75%) were found to be negative and 4 (25%) were positive for DENV-2. For the detection of Wolbachia in A. albopictus, 71 out of 284 (25%) and 213 (75%) larvae were found to be positive and negative for Wolbachia respectively. For A. aegypti, 4 (25%) and 12 (75%) out of 16 larvae were positive and negative for Wolbachia respectively. This is the first report of Wolbachia in A. albopictus and A. aegypti larvae in Malaysia. A chisquare test analysis to determine the association between dengue virus and Wolbachia in A. albopictus and A. aegypti larvae collected from the four localities in Kuala Lumpur showed that there was no association (χ[2] = 3.080; df = 1; P > 0.05).}, } @article {pmid28856446, year = {2018}, author = {Bliman, PA and Aronna, MS and Coelho, FC and da Silva, MAHB}, title = {Ensuring successful introduction of Wolbachia in natural populations of Aedes aegypti by means of feedback control.}, journal = {Journal of mathematical biology}, volume = {76}, number = {5}, pages = {1269-1300}, pmid = {28856446}, issn = {1432-1416}, mesh = {Aedes/*microbiology ; Animals ; Arbovirus Infections/prevention & control/transmission ; Computer Simulation ; Dengue/*prevention & control/transmission ; Female ; Humans ; Male ; Mathematical Concepts ; Models, Biological ; Mosquito Vectors/*microbiology ; Pest Control, Biological/*methods/statistics & numerical data ; Wolbachia/pathogenicity/*physiology ; }, abstract = {The control of the spread of dengue fever by introduction of the intracellular parasitic bacterium Wolbachia in populations of the vector Aedes aegypti, is presently one of the most promising tools for eliminating dengue, in the absence of an efficient vaccine. The success of this operation requires locally careful planning to determine the adequate number of individuals carrying the Wolbachia parasite that need to be introduced into the natural population. The introduced mosquitoes are expected to eventually replace the Wolbachia-free population and guarantee permanent protection against the transmission of dengue to human. In this study, we propose and analyze a model describing the fundamental aspects of the competition between mosquitoes carrying Wolbachia and mosquitoes free of the parasite. We then use feedback control techniques to devise an introduction protocol that is proved to guarantee that the population converges to a stable equilibrium where the totality of mosquitoes carry Wolbachia.}, } @article {pmid28855327, year = {2017}, author = {Sullivan, W}, title = {Wolbachia, bottled water, and the dark side of symbiosis.}, journal = {Molecular biology of the cell}, volume = {28}, number = {18}, pages = {2343-2346}, pmid = {28855327}, issn = {1939-4586}, mesh = {Animals ; Humans ; Symbiosis/*physiology ; }, abstract = {Obligate endosymbiosis is operationally defined when loss or removal of the endosymbiont from the host results in the death of both. Whereas these relationships are typically viewed as mutualistic, molecular and cellular analysis reveals numerous instances in which these symbiotic relationships are established by alternative, nonmutualistic strategies. The endosymbiont usurps or integrates into core host processes, creating a need where none previously existed. Here I discuss examples of these addictive symbiotic relationships and how they are a likely outcome of all complex evolving systems.}, } @article {pmid28854601, year = {2017}, author = {Rice, DW and Sheehan, KB and Newton, ILG}, title = {Large-Scale Identification of Wolbachia pipientis Effectors.}, journal = {Genome biology and evolution}, volume = {9}, number = {7}, pages = {1925-1937}, pmid = {28854601}, issn = {1759-6653}, mesh = {Animals ; Bacterial Proteins/genetics/*metabolism ; Computational Biology/*methods ; Drosophila melanogaster/genetics/metabolism/*microbiology ; Gene Expression Regulation, Bacterial ; Genetic Association Studies ; Host-Pathogen Interactions ; Phylogeny ; Saccharomyces cerevisiae/genetics/growth & development ; Symbiosis ; Wolbachia/genetics/*metabolism ; }, abstract = {Wolbachia pipientis is an intracellular symbiont of arthropods well known for the reproductive manipulations induced in the host and, more recently, for the ability of Wolbachia to block virus replication in insect vectors. Since Wolbachia cannot yet be genetically manipulated, and due to the constraints imposed when working with an intracellular symbiont, little is known about mechanisms used by Wolbachia for host interaction. Here we employed a bioinformatics pipeline and identified 163 candidate effectors, potentially secreted by Wolbachia into the host cell. A total of 84 of these candidates were then subjected to a screen of growth defects induced in yeast upon heterologous expression which identified 14 top candidates likely secreted by Wolbachia. These predicted secreted effectors may function in concert as we find that their native expression is correlated and is highly upregulated at specific time points during Drosophila development. In addition, the evolutionary histories of some of these predicted effectors are also correlated, suggesting they may function together, or in the same pathway, during host infection. Similarly, most of these predicted effectors are limited to one or two Wolbachia strains-perhaps reflecting shared evolutionary history and strain specific functions in host manipulation. Identification of these Wolbachia candidate effectors is the first step in dissecting the mechanisms of symbiont-host interaction in this important system.}, } @article {pmid28845255, year = {2017}, author = {Vi, TT and Kien, DT and Wills, B and Simmons, CP}, title = {A29 Wolbachia for dengue control; will dengue viruses evolve resistance?.}, journal = {Virus evolution}, volume = {3}, number = {Suppl 1}, pages = {}, doi = {10.1093/ve/vew036.028}, pmid = {28845255}, issn = {2057-1577}, } @article {pmid28842593, year = {2017}, author = {Ojha, A and Sinha, DK and Padmakumari, AP and Bentur, JS and Nair, S}, title = {Bacterial Community Structure in the Asian Rice Gall Midge Reveals a Varied Microbiome Rich in Proteobacteria.}, journal = {Scientific reports}, volume = {7}, number = {1}, pages = {9424}, pmid = {28842593}, issn = {2045-2322}, mesh = {Animals ; *Biodiversity ; Diptera/*microbiology ; Metagenome ; Metagenomics/methods ; *Microbiota ; Proteobacteria/*classification/*genetics ; RNA, Ribosomal, 16S/genetics ; }, abstract = {The Asian rice gall midge (ARGM) has emerged as a model gall forming pest of rice. The ARGM infestation of rice results in failure of panicle formation and economic loss. Understanding the molecular basis of ARGM-rice interactions is very crucial in order to control this devastating pest of rice. The current investigation was devised to identify bacterial communities present in the ARGM and in addition the bacterial diversity in the maggots during their interaction with susceptible or resistant rice varieties. Sequencing of 16S rRNA bacterial gene (V3-V4 region) revealed differences in the microflora of the ARGM maggots feeding on susceptible or resistant rice hosts. Results revealed that Wolbachia was the predominant bacterium in pupae and adults while Pseudomonas was predominant in maggots. Further, we observed that members of proteobacteria were predominant across all the samples. There was high species diversity in maggots isolated from susceptible rice and a high representation of unclassified bacteria in maggots isolated from resistant rice. This is the first study that reports variation of microbiome of the ARGM, based on host phenotype from which it was isolated, and results suggest that these variations could have an important role in host's susceptibility.}, } @article {pmid28841917, year = {2017}, author = {Almeida, F and Suesdek, L}, title = {Effects of Wolbachia on ovarian apoptosis in Culex quinquefasciatus (Say, 1823) during the previtellogenic and vitellogenic periods.}, journal = {Parasites & vectors}, volume = {10}, number = {1}, pages = {398}, pmid = {28841917}, issn = {1756-3305}, mesh = {Animals ; *Apoptosis ; Culex/*microbiology/*physiology ; Female ; Microscopy, Confocal ; Oocytes/microbiology/pathology ; Oogenesis ; Ovary/cytology/microbiology/ultrastructure ; Vitellogenesis ; Wolbachia/*physiology ; }, abstract = {BACKGROUND: Apoptosis is programmed cell death that ordinarily occurs in ovarian follicular cells in various organisms. In the best-studied holometabolous insect, Drosophila, this kind of cell death occurs in all three cell types found in the follicles, sometimes leading to follicular atresia and egg degeneration. On the other hand, egg development, quantity and viability in the mosquito Culex quinquefasciatus are disturbed by the infection with the endosymbiont Wolbachia. Considering that Wolbachia alters reproductive traits, we hypothesised that such infection would also alter the apoptosis in the ovarian cells of this mosquito. The goal of this study was to comparatively describe the occurrence of apoptosis in Wolbachia-infected and uninfected ovaries of Cx. quinquefasciatus during oogenesis and vitellogenesis. For this, we recorded under confocal microscopy the occurrence of apoptosis in all three cell types of the ovarian follicle. In the first five days of adult life we observed oogenesis and, after a blood meal, the initiation step of vitellogenesis.

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

CONCLUSIONS: Our data support the idea that vertical transmission of DENV is rare in nature, even in the absence of Wolbachia. Indeed, we observed that vertical transmission rates were low even when the midgut barrier was bypassed, which might help to explain why we only observed a trend towards lower vertical transmission rates in the presence of Wolbachia. Nevertheless, the low prevalence of disseminated DENV infection and lower DENV load in the ovaries supports the hypothesis that the presence of Wolbachia in Ae. aegypti would have an effect on the vertical transmission of DENV in the field.}, } @article {pmid28618183, year = {2017}, author = {Hussain, M and Akutse, KS and Ravindran, K and Lin, Y and Bamisile, BS and Qasim, M and Dash, CK and Wang, L}, title = {Effects of different temperature regimes on survival of Diaphorina citri and its endosymbiotic bacterial communities.}, journal = {Environmental microbiology}, volume = {19}, number = {9}, pages = {3439-3449}, doi = {10.1111/1462-2920.13821}, pmid = {28618183}, issn = {1462-2920}, mesh = {Animals ; China ; Citrus/*parasitology ; Halomonadaceae/genetics/*growth & development ; Hemiptera/*microbiology ; Hot Temperature ; Real-Time Polymerase Chain Reaction ; Symbiosis/*physiology ; Wolbachia/genetics/*growth & development ; }, abstract = {The Asian citrus psyllid, Diaphorina citri, is a major pest of citrus and vector of citrus greening (huanglongbing) in Asian. In our field-collected psyllid samples, we discovered that Fuzhou (China) and Faisalabad (Pakistan), populations harbored an obligate primary endosymbiont Candidatus Carsonella (gen. nov.) with a single species, Candidatus Carsonella ruddii (sp. nov.) and a secondary endosymbiont, Wolbachia surface proteins (WSP) which are intracellular endosymbionts residing in the bacteriomes. Responses of these symbionts to different temperatures were examined and their host survival assessed. Diagnostic PCR assays showed that the endosymbionts infection rates were not significantly reduced in both D. citri populations after 24 h exposure to cold or heat treatments. Although quantitative PCR assays showed significant reduction of WSP relative densities at 40°C for 24 h, a substantial decrease occurred as the exposure duration increased beyond 3 days. Under the same temperature regimes, Ca. C. ruddii density was initially less affected during the first exposure day, but rapidly reduced at 3-5 days compared to WSP. However, the mortality of the psyllids increased rapidly as exposure time to heat treatment increased. The responses of the two symbionts to unfavorable temperature regimes highlight the complex host-symbionts interactions between D. citri and its associated endosymbionts.}, } @article {pmid28617844, year = {2017}, author = {Bhattacharya, T and Newton, ILG and Hardy, RW}, title = {Wolbachia elevates host methyltransferase expression to block an RNA virus early during infection.}, journal = {PLoS pathogens}, volume = {13}, number = {6}, pages = {e1006427}, pmid = {28617844}, issn = {1553-7374}, support = {P40 OD018537/OD/NIH HHS/United States ; R01 AI090077/AI/NIAID NIH HHS/United States ; R01 GM084947/GM/NIGMS NIH HHS/United States ; R21 AI121849/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; DNA (Cytosine-5-)-Methyltransferases/genetics/*metabolism ; Drosophila Proteins/genetics/*metabolism ; Drosophila melanogaster/*enzymology/microbiology/physiology/*virology ; Host-Pathogen Interactions ; Sindbis Virus/*physiology ; Symbiosis ; Virus Replication ; Wolbachia/*physiology ; }, abstract = {Wolbachia pipientis is an intracellular endosymbiont known to confer host resistance against RNA viruses in insects. However, the causal mechanism underlying this antiviral defense remains poorly understood. To this end, we have established a robust arthropod model system to study the tripartite interaction involving Sindbis virus and Wolbachia strain wMel within its native host, Drosophila melanogaster. By leveraging the power of Drosophila genetics and a parallel, highly tractable D. melanogaster derived JW18 cell culture system, we determined that in addition to reducing infectious virus production, Wolbachia negatively influences Sindbis virus particle infectivity. This is further accompanied by reductions in viral transcript and protein levels. Interestingly, unchanged ratio of proteins to viral RNA copies suggest that Wolbachia likely does not influence the translational efficiency of viral transcripts. Additionally, expression analyses of candidate host genes revealed D. melanogaster methyltransferase gene Mt2 as an induced host factor in the presence of Wolbachia. Further characterization of viral resistance in Wolbachia-infected flies lacking functional Mt2 revealed partial recovery of virus titer relative to wild-type, accompanied by complete restoration of viral RNA and protein levels, suggesting that Mt2 acts at the stage of viral genome replication. Finally, knockdown of Mt2 in Wolbachia uninfected JW18 cells resulted in increased virus infectivity, thus demonstrating its previously unknown role as an antiviral factor against Sindbis virus. In conclusion, our findings provide evidence supporting the role of Wolbachia-modulated host factors towards RNA virus resistance in arthropods, alongside establishing Mt2's novel antiviral function against Sindbis virus in D. melanogaster.}, } @article {pmid28612849, year = {2017}, author = {Hansen, AK and Skidmore, IH}, title = {Psyllids, It's What's on the Inside That Counts: Community Cross Talk Facilitates Prophage Interactions.}, journal = {mSphere}, volume = {2}, number = {3}, pages = {}, pmid = {28612849}, issn = {2379-5042}, abstract = {Despite the availability of massive microbial community data sets (e.g., metagenomes), there is still a lack of knowledge on what molecular mechanisms facilitate cross talk between microbes and prophage within a community context. A study published in mSphere by Jain and colleagues (M. Jain, L. A. Fleites, and D. W. Gabriel, mSphere 2:e00171-17, 2017, https://doi.org/10.1128/mSphereDirect.00171-17) reports on an intriguing new twist of how a prophage of the bacterium "Candidatus Liberibacter asiaticus" may have its lytic cycle suppressed partly because of a protein that is expressed by a cooccurring bacterium, Wolbachia. Both of these microbes coexist along with other microbial tenants inside their sap-feeding insect host, a psyllid. Although these results are still preliminary and alternative hypotheses need to be tested, these results suggest an interesting new dimension on how regulation of microbial genomes occurs in a community context.}, } @article {pmid28611731, year = {2017}, author = {Crainey, JL and Hurst, J and Lamberton, PHL and Cheke, RA and Griffin, CE and Wilson, MD and de Araújo, CPM and Basáñez, MG and Post, RJ}, title = {The Genomic Architecture of Novel Simulium damnosum Wolbachia Prophage Sequence Elements and Implications for Onchocerciasis Epidemiology.}, journal = {Frontiers in microbiology}, volume = {8}, number = {}, pages = {852}, pmid = {28611731}, issn = {1664-302X}, support = {/WT_/Wellcome Trust/United Kingdom ; 77615/MRC_/Medical Research Council/United Kingdom ; }, abstract = {Research interest in Wolbachia is growing as new discoveries and technical advancements reveal the public health importance of both naturally occurring and artificial infections. Improved understanding of the Wolbachia bacteriophages (WOs) WOcauB2 and WOcauB3 [belonging to a sub-group of four WOs encoding serine recombinases group 1 (sr1WOs)], has enhanced the prospect of novel tools for the genetic manipulation of Wolbachia. The basic biology of sr1WOs, including host range and mode of genomic integration is, however, still poorly understood. Very few sr1WOs have been described, with two such elements putatively resulting from integrations at the same Wolbachia genome loci, about 2 kb downstream from the FtsZ cell-division gene. Here, we characterize the DNA sequence flanking the FtsZ gene of wDam, a genetically distinct line of Wolbachia isolated from the West African onchocerciasis vector Simulium squamosum E. Using Roche 454 shot-gun and Sanger sequencing, we have resolved >32 kb of WO prophage sequence into three contigs representing three distinct prophage elements. Spanning ≥36 distinct WO open reading frame gene sequences, these prophage elements correspond roughly to three different WO modules: a serine recombinase and replication module (sr1RRM), a head and base-plate module and a tail module. The sr1RRM module contains replication genes and a Holliday junction recombinase and is unique to the sr1 group WOs. In the extreme terminal of the tail module there is a SpvB protein homolog-believed to have insecticidal properties and proposed to have a role in how Wolbachia parasitize their insect hosts. We propose that these wDam prophage modules all derive from a single WO genome, which we have named here sr1WOdamA1. The best-match database sequence for all of our sr1WOdamA1-predicted gene sequences was annotated as of Wolbachia or Wolbachia phage sourced from an arthropod. Clear evidence of exchange between sr1WOdamA1 and other Wolbachia WO phage sequences was also detected. These findings provide insights into how Wolbachia could affect a medically important vector of onchocerciasis, with potential implications for future control methods, as well as supporting the hypothesis that Wolbachia phages do not follow the standard model of phage evolution.}, } @article {pmid28609446, year = {2017}, author = {Murray, RL and Herridge, EJ and Ness, RW and Bussière, LF}, title = {Are sex ratio distorting endosymbionts responsible for mating system variation among dance flies (Diptera: Empidinae)?.}, journal = {PloS one}, volume = {12}, number = {6}, pages = {e0178364}, pmid = {28609446}, issn = {1932-6203}, mesh = {Animals ; Biological Evolution ; Diptera/classification/*microbiology ; Female ; Host-Pathogen Interactions ; Linear Models ; Male ; Reproduction ; Rickettsia/*physiology ; Sex Ratio ; Species Specificity ; Spiroplasma/*physiology ; *Symbiosis ; Wolbachia/*physiology ; }, abstract = {Maternally inherited bacterial endosymbionts are common in many arthropod species. Some endosymbionts cause female-biased sex ratio distortion in their hosts that can result in profound changes to a host's mating behaviour and reproductive biology. Dance flies (Diptera: Empidinae) are well known for their unusual reproductive biology, including species with female-specific ornamentation and female-biased lek-like swarming behaviour. The cause of the repeated evolution of female ornaments in these flies remains unknown, but is probably associated with female-biased sex ratios in individual species. In this study we assessed whether dance flies harbour sex ratio distorting endosymbionts that might have driven these mating system evolutionary changes. We measured the incidence and prevalence of infection by three endosymbionts that are known to cause female-biased sex ratios in other insect hosts (Wolbachia, Rickettsia and Spiroplasma) across 20 species of dance flies. We found evidence of widespread infection by all three symbionts and variation in sex-specific prevalence across the taxa sampled. However, there was no relationship between infection prevalence and adult sex ratio measures and no evidence that female ornaments are associated with high prevalences of sex-biased symbiont infections. We conclude that the current distribution of endosymbiont infections is unlikely to explain the diversity in mating systems among dance fly species.}, } @article {pmid28608866, year = {2017}, author = {Jain, M and Fleites, LA and Gabriel, DW}, title = {A Small Wolbachia Protein Directly Represses Phage Lytic Cycle Genes in "Candidatus Liberibacter asiaticus" within Psyllids.}, journal = {mSphere}, volume = {2}, number = {3}, pages = {}, pmid = {28608866}, issn = {2379-5042}, abstract = {Huanglongbing (HLB) is a severe disease of citrus caused by an uncultured alphaproteobacterium "Candidatus Liberibacter asiaticus" and transmitted by Asian citrus psyllids (Diaphorina citri). Two prophage genomes, SC1 and SC2, integrated in "Ca. Liberibacter asiaticus" strain UF506 were described previously, and very similar prophages are found resident in the majority of "Ca. Liberibacter asiaticus" strains described worldwide. The SC1 lytic cycle is marked by upregulation of prophage late genes, including a functional holin (SC1_gp110); these late genes are activated when "Ca. Liberibacter asiaticus" is in planta, but not when infecting the psyllid host. We previously reported that the holin promoter is strongly and constitutively active in Liberibacter crescens (a cultured proxy for uncultured "Ca. Liberibacter asiaticus") but is suppressed in a dose-dependent manner by crude aqueous extracts from D. citri applied exogenously. Here we report that the suppressor activity of the crude psyllid extract was heat labile and abolished by proteinase K treatment, indicating a proteinaceous repressor and of a size smaller than 30 kDa. The repressor was affinity captured from D. citri aqueous extracts using biotinylated holin promoter DNA immobilized on magnetic beads and subjected to liquid chromatography-tandem mass spectrometry (LC-MS/MS). Protein database interrogation was used to identify a small DNA-binding protein encoded by a gene carried by Wolbachia strain wDi, a resident endosymbiont of D. citri as the repressor. The in vitro-translated Wolbachia repressor protein was able to penetrate L. crescens cells, bind to "Ca. Liberibacter asiaticus" promoter DNA, and partially suppress holin promoter-driven β-glucuronidase (GUS) activity, indicating potential involvement of an additional interacting partner(s) or posttranslational modification(s) for complete suppression. Expression of the Wolbachia repressor protein appeared to be constitutive irrespective of "Ca. Liberibacter asiaticus" infection status of the insect host. IMPORTANCE Host acquisition of a new microbial species can readily perturb the dynamics of preexisting microbial associations. Molecular cross talk between microbial associates may be necessary for efficient resource allocation and enhanced survival. Classic examples involve quorum sensing (QS), which detects population densities and is both used and coopted to control expression of bacterial genes, including host adaptation factors. We report that a 56-amino-acid repressor protein made by the resident psyllid endosymbiont Wolbachia can enter cells of Liberibacter crescens, a cultured proxy for the uncultured psyllid endosymbiont "Ca. Liberibacter asiaticus" and repress "Ca. Liberibacter asiaticus" phage lytic cycle genes. Such repression in "Ca. Liberibacter asiaticus" may be critical to survival of both endosymbionts, since phage-mediated lysis would likely breach the immunogenic threshold of the psyllid, invoking a systemic and nonspecific innate immune reaction.}, } @article {pmid28606944, year = {2017}, author = {Emerson, KJ and Glaser, RL}, title = {Cytonuclear Epistasis Controls the Density of Symbiont Wolbachia pipientis in Nongonadal Tissues of Mosquito Culex quinquefasciatus.}, journal = {G3 (Bethesda, Md.)}, volume = {7}, number = {8}, pages = {2627-2635}, pmid = {28606944}, issn = {2160-1836}, mesh = {Animals ; Chromosome Mapping ; Culex/*microbiology ; *Epistasis, Genetic ; Female ; Gonads/*microbiology ; Inheritance Patterns/genetics ; Male ; Quantitative Trait Loci/genetics ; Symbiosis/*genetics ; Wolbachia/*genetics/*growth & development ; }, abstract = {Wolbachia pipientis, a bacterial symbiont infecting arthropods and nematodes, is vertically transmitted through the female germline and manipulates its host's reproduction to favor infected females. Wolbachia also infects somatic tissues where it can cause nonreproductive phenotypes in its host, including resistance to viral pathogens. Wolbachia-mediated phenotypes are strongly associated with the density of Wolbachia in host tissues. Little is known, however, about how Wolbachia density is regulated in native or heterologous hosts. Here, we measure the broad-sense heritability of Wolbachia density among families in field populations of the mosquito Culex pipiens, and show that densities in ovary and nongonadal tissues of females in the same family are not correlated, suggesting that Wolbachia density is determined by distinct mechanisms in the two tissues. Using introgression analysis between two different strains of the closely related species C. quinquefasciatus, we show that Wolbachia densities in ovary tissues are determined primarily by cytoplasmic genotype, while densities in nongonadal tissues are determined by both cytoplasmic and nuclear genotypes and their epistatic interactions. Quantitative-trait-locus mapping identified two major-effect quantitative-trait loci in the C. quinquefasciatus genome explaining a combined 23% of variance in Wolbachia density, specifically in nongonadal tissues. A better understanding of how Wolbachia density is regulated will provide insights into how Wolbachia density can vary spatiotemporally in insect populations, leading to changes in Wolbachia-mediated phenotypes such as viral pathogen resistance.}, } @article {pmid28603242, year = {2017}, author = {Torres-Román, JS and Bazalar-Palacios, J and Ruiz, EF and Avilez, JL and Rodríguez-Morales, AJ}, title = {New strategies for the eradication of Aedes aegypti: what challenges do we face in Latin America?.}, journal = {Le infezioni in medicina}, volume = {25}, number = {2}, pages = {193-194}, pmid = {28603242}, issn = {2532-8689}, mesh = {*Aedes/genetics/microbiology ; Animals ; Animals, Genetically Modified ; Arbovirus Infections/prevention & control/transmission ; Endemic Diseases ; Humans ; *Insect Vectors ; Latin America ; Mosquito Control/economics/*methods ; Pest Control, Biological/economics/*methods ; Wolbachia/physiology ; }, abstract = {Not available.}, } @article {pmid28600214, year = {2017}, author = {Globisch, D and Eubanks, LM and Shirey, RJ and Pfarr, KM and Wanji, S and Debrah, AY and Hoerauf, A and Janda, KD}, title = {Validation of onchocerciasis biomarker N-acetyltyramine-O-glucuronide (NATOG).}, journal = {Bioorganic & medicinal chemistry letters}, volume = {27}, number = {15}, pages = {3436-3440}, pmid = {28600214}, issn = {1464-3405}, mesh = {Animals ; Biomarkers/urine ; Chromatography, Liquid/methods ; Glucuronides/metabolism/*urine ; Humans ; Limit of Detection ; Mass Spectrometry/*methods ; Metabolomics/methods ; Neglected Diseases/metabolism/*urine ; Onchocerca volvulus/*isolation & purification/metabolism ; Onchocerciasis/metabolism/*urine ; Tyramine/*analogs & derivatives/metabolism/urine ; }, abstract = {The Neglected Tropical Disease onchocerciasis is a parasitic disease. Despite many control programmes by the World Health Organization (WHO), large communities in West and Central Africa are still affected. Besides logistic challenges during biannual mass drug administration, the lack of a robust, point-of-care diagnostic is limiting successful eradication of onchocerciasis. Towards the implementation of a non-invasive and point-of-care diagnostic, we have recently reported the discovery of the biomarker N-acetyltyramine-O-glucuronide (NATOG) in human urine samples using a metabolomics-mining approach. NATOG's biomarker value was enhanced during an investigation in a rodent model. Herein, we further detail the specificity of NATOG in active onchocerciasis infections as well as the co-infecting parasites Loa loa and Mansonella perstans. Our results measured by liquid chromatography coupled with mass spectrometry (LC-MS) reveal elevated NATOG values in mono- and co-infection samples only in the presence of the nematode Onchocerca volvulus. Metabolic pathway investigation of l-tyrosine/tyramine in all investigated nematodes uncovered an important link between the endosymbiotic bacterium Wolbachia and O. volvulus for the biosynthesis of NATOG. Based on these extended studies, we suggest NATOG as a biomarker for tracking active onchocerciasis infections and provide a threshold concentration value of NATOG for future diagnostic tool development.}, } @article {pmid28599659, year = {2017}, author = {Duchemin, JB and Mee, PT and Lynch, SE and Vedururu, R and Trinidad, L and Paradkar, P}, title = {Zika vector transmission risk in temperate Australia: a vector competence study.}, journal = {Virology journal}, volume = {14}, number = {1}, pages = {108}, pmid = {28599659}, issn = {1743-422X}, mesh = {Animals ; Australia ; Climate ; Disease Transmission, Infectious ; Gastrointestinal Tract/*virology ; Humans ; Mosquito Vectors/*virology ; RNA, Viral/*analysis/genetics ; Real-Time Polymerase Chain Reaction ; Risk Assessment ; Saliva/*virology ; Zika Virus/genetics/*isolation & purification ; Zika Virus Infection/transmission ; }, abstract = {BACKGROUND: Zika virus is an emerging pathogen of global importance. It has been responsible for recent outbreaks in the Americas and in the Pacific region. This study assessed five different mosquito species from the temperate climatic zone in Australia and included Aedes albopictus as a potentially invasive species.

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

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

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

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

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

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

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

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

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

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

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

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

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

CONCLUSION: Wolbachia infections were found only among four mosquito species in Sri Lanka: Aedes albopictus, Culex quinquefasciatus, Armigeres subalbatus and Mansonia uniformis. Sequence data showed high haplotype diversity among the Wolbachia strains.}, } @article {pmid28486424, year = {2017}, author = {Fenner, J and Seltzer, J and Peyton, S and Sullivan, H and Tolson, P and Walsh, RP and Hill, J and Counterman, BA}, title = {Demographic Variation of Wolbachia Infection in the Endangered Mitchell's Satyr Butterfly.}, journal = {Insects}, volume = {8}, number = {2}, pages = {}, pmid = {28486424}, issn = {2075-4450}, abstract = {The Mitchell's satyr, Neonympha mitchellii, is an endangered species that is limited to highly isolated habitats in the northern and southern United States. Conservation strategies for isolated endangered species often implement captive breeding and translocation programs for repopulation. However, these programs risk increasing the spread of harmful pathogens, such as the bacterial endosymbiont Wolbachia. Wolbachia can manipulate the host's reproduction leading to incompatibilities between infected and uninfected hosts. This study uses molecular methods to screen for Wolbachia presence across the distribution of the Mitchell's satyr and its subspecies, St. Francis satyr, which are both federally listed as endangered and are considered two of the rarest butterflies in North America. The screens confirmed the presence of Wolbachia in the northern and newly discovered southern populations of the Mitchell's satyr, but not in the St. Francis satyr population. These results combined with previous reports of Wolbachia in N. mitchellii, highlight that Wolbachia infection varies both geographically and temporally in satyr populations. The temporal variance shows the importance of continued monitoring of Wolbachia infection during conservation programs. To reduce the risk of reproductive incompatibilities, it is advised that all individuals collected for conservation purposes be screened for Wolbachia and recommended to avoid the use of infected individuals for captive breeding and translocation programs.}, } @article {pmid28477267, year = {2017}, author = {Toribio-Fernández, R and Bella, JL and Martínez-Rodríguez, P and Funkhouser-Jones, LJ and Bordenstein, SR and Pita, M}, title = {Chromosomal localization of Wolbachia inserts in the genomes of two subspecies of Chorthippus parallelus forming a Pyrenean hybrid zone.}, journal = {Chromosome research : an international journal on the molecular, supramolecular and evolutionary aspects of chromosome biology}, volume = {25}, number = {3-4}, pages = {215-225}, pmid = {28477267}, issn = {1573-6849}, support = {P30 DK058404/DK/NIDDK NIH HHS/United States ; R21 HD086833/HD/NICHD NIH HHS/United States ; }, mesh = {Animals ; *Chromosomes, Bacterial ; *Genome, Insect ; Grasshoppers/*genetics ; Heterochromatin ; *Hybridization, Genetic ; In Situ Hybridization, Fluorescence ; Male ; *Mutagenesis, Insertional ; *Polytene Chromosomes ; Sequence Analysis, DNA ; Wolbachia/*genetics ; }, abstract = {Wolbachia are endosymbiotic bacteria of arthropods and nematodes that can manipulate the reproduction of various host organisms to facilitate their own maternal transmission. Moreover, Wolbachia's presence in host germ cells may contribute to the many cases of lateral gene transfer from Wolbachia to host genomes that have been described. A previous study in Chorthippus parallelus, a well-known orthopteroid forming a hybrid zone in the Pyrenees, identified Wolbachia sequences from two major supergroups in the genomes of infected and uninfected Chorthippus parallelus parallelus (Cpp) and Chorthippus parallelus erythropus (Cpe) subspecies. In this study, we map the Wolbachia genomic inserts to specific regions on the chromosomes of Cpp and Cpe by fluorescent in situ hybridization (FISH) using tyramides to increase the accuracy and detection of these insertions. Additionally, we consider some of the possible roles that these bacterial inserts play in the organization and function of the grasshopper genome, as well as how they can serve as markers for phylogenetic relationships of these organisms.}, } @article {pmid28471518, year = {2017}, author = {Li, S and Zhou, C and Chen, G and Zhou, Y}, title = {Bacterial microbiota in small brown planthopper populations with different rice viruses.}, journal = {Journal of basic microbiology}, volume = {57}, number = {7}, pages = {590-596}, doi = {10.1002/jobm.201700004}, pmid = {28471518}, issn = {1521-4028}, mesh = {Animals ; Bacteria/classification/genetics/*isolation & purification ; Hemiptera/*microbiology/virology ; High-Throughput Nucleotide Sequencing ; Insect Vectors/*microbiology/virology ; *Microbiota/genetics ; Oryza ; Plant Viruses/isolation & purification/physiology ; RNA, Ribosomal, 16S ; *Symbiosis ; Tenuivirus/isolation & purification/physiology ; Wolbachia/genetics/isolation & purification ; }, abstract = {The small brown planthopper (SBPH) is an important virus vector, transmitting Rice stripe virus (RSV), and Rice black-streaked dwarf virus (RBSDV). Insect symbionts play an essential role in the insect fitness, however, it is still unclear about their contributions to viral transmission by SBPH. Here, we investigated endosymbiont communities in non-viruliferous, RSV-infected, and RBSDV-infected SBPH populations using Illumina 16S rRNA gene MiSeq sequencing. In total, 281,803 effective sequences of the 16S rRNA gene were generated from different samples. Sequence analysis revealed the percentages of these bacterial groups in different SBPH populations on several taxonomic levels ranging from phyla to genera. The extremely consistent bacterial diversity and abundance indicated that RSV or RBSDV infection did not affect the composition and abundance of symbionts in SBPH. It was notable that Wolbachia was dominant in all populations. The symbiosis between Wolbachia and SBPH might be potentially studied and utilized to control pest SBPH in the future.}, } @article {pmid28464440, year = {2017}, author = {Martinez, J and Tolosana, I and Ok, S and Smith, S and Snoeck, K and Day, JP and Jiggins, FM}, title = {Symbiont strain is the main determinant of variation in Wolbachia-mediated protection against viruses across Drosophila species.}, journal = {Molecular ecology}, volume = {26}, number = {15}, pages = {4072-4084}, pmid = {28464440}, issn = {1365-294X}, support = {//Wellcome Trust/United Kingdom ; 281668/ERC_/European Research Council/International ; }, mesh = {Animals ; *Disease Resistance ; Drosophila/genetics/*microbiology/virology ; Genome, Bacterial ; Genome, Insect ; Phenotype ; *Symbiosis ; Viruses/pathogenicity ; Wolbachia/*genetics ; }, abstract = {Wolbachia is a common heritable bacterial symbiont in insects. Its evolutionary success lies in the diverse phenotypic effects it has on its hosts coupled to its propensity to move between host species over evolutionary timescales. In a survey of natural host-symbiont associations in a range of Drosophila species, we found that 10 of 16 Wolbachia strains protected their hosts against viral infection. By moving Wolbachia strains between host species, we found that the symbiont genome had a much greater influence on the level of antiviral protection than the host genome. The reason for this was that the level of protection depended on the density of the symbiont in host tissues, and Wolbachia rather than the host-controlled density. The finding that virus resistance and symbiont density are largely under the control of symbiont genes in this system has important implications both for the evolution of these traits and for public health programmes using Wolbachia to prevent mosquitoes from transmitting disease.}, } @article {pmid28446677, year = {2017}, author = {Schultz, MJ and Isern, S and Michael, SF and Corley, RB and Connor, JH and Frydman, HM}, title = {Variable Inhibition of Zika Virus Replication by Different Wolbachia Strains in Mosquito Cell Cultures.}, journal = {Journal of virology}, volume = {91}, number = {14}, pages = {}, pmid = {28446677}, issn = {1098-5514}, support = {R01 AI099210/AI/NIAID NIH HHS/United States ; R21 NS101151/NS/NINDS NIH HHS/United States ; R56 AI097589/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes ; Animals ; *Antibiosis ; Cell Line ; Protein Biosynthesis ; RNA, Viral/biosynthesis ; Transcription, Genetic ; Viral Nonstructural Proteins/biosynthesis ; *Virus Replication ; Wolbachia/*physiology ; Zika Virus/*physiology ; }, abstract = {Mosquito-borne arboviruses are a major source of human disease. One strategy to reduce arbovirus disease is to reduce the mosquito's ability to transmit virus. Mosquito infection with the bacterial endosymbiont Wolbachia pipientis wMel is a novel strategy to reduce Aedes mosquito competency for flavivirus infection. However, experiments investigating cyclic environmental temperatures have shown a reduction in maternal transmission of wMel, potentially weakening the integration of this strain into a mosquito population relative to that of other Wolbachia strains. Consequently, it is important to investigate additional Wolbachia strains. All Zika virus (ZIKV) suppression studies are limited to the wMel Wolbachia strain. Here we show ZIKV inhibition by two different Wolbachia strains: wAlbB (isolated from Aedes albopictus mosquitoes) and wStri (isolated from the planthopper Laodelphax striatellus) in mosquito cells. Wolbachia strain wStri inhibited ZIKV most effectively. Single-cycle infection experiments showed that ZIKV RNA replication and nonstructural protein 5 translation were reduced below the limits of detection in wStri-containing cells, demonstrating early inhibition of virus replication. ZIKV replication was rescued when Wolbachia was inhibited with a bacteriostatic antibiotic. We observed a partial rescue of ZIKV growth when Wolbachia-infected cells were supplemented with cholesterol-lipid concentrate, suggesting competition for nutrients as one of the possible mechanisms of Wolbachia inhibition of ZIKV. Our data show that wAlbB and wStri infection causes inhibition of ZIKV, making them attractive candidates for further in vitro mechanistic and in vivo studies and future vector-centered approaches to limit ZIKV infection and spread.IMPORTANCE Zika virus (ZIKV) has swiftly spread throughout most of the Western Hemisphere. This is due in large part to its replication in and spread by a mosquito vector host. There is an urgent need for approaches that limit ZIKV replication in mosquitoes. One exciting approach for this is to use a bacterial endosymbiont called Wolbachia that can populate mosquito cells and inhibit ZIKV replication. Here we show that two different strains of Wolbachia, wAlbB and wStri, are effective at repressing ZIKV in mosquito cell lines. Repression of virus growth is through the inhibition of an early stage of infection and requires actively replicating Wolbachia Our findings further the understanding of Wolbachia viral inhibition and provide novel tools that can be used in an effort to limit ZIKV replication in the mosquito vector, thereby interrupting the transmission and spread of the virus.}, } @article {pmid28439127, year = {2017}, author = {Becking, T and Giraud, I and Raimond, M and Moumen, B and Chandler, C and Cordaux, R and Gilbert, C}, title = {Diversity and evolution of sex determination systems in terrestrial isopods.}, journal = {Scientific reports}, volume = {7}, number = {1}, pages = {1084}, pmid = {28439127}, issn = {2045-2322}, mesh = {Animals ; *Evolution, Molecular ; Gene Expression Profiling ; *Genetic Variation ; Isopoda/classification/*genetics ; Phylogeny ; *Sex Characteristics ; }, abstract = {Sex determination systems are highly variable in many taxa, sometimes even between closely related species. Yet the number and direction of transitions between these systems have seldom been characterized, and the underlying mechanisms are still poorly understood. Here we generated transcriptomes for 19 species of terrestrial isopod crustaceans, many of which are infected by Wolbachia bacterial endosymbionts. Using 88 single-copy orthologous genes, we reconstructed a fully resolved and dated phylogeny of terrestrial isopods. An original approach involving crossings of sex-reversed individuals allowed us to characterize the heterogametic systems of five species (one XY/XX and four ZW/ZZ). Mapping of these and previously known heterogametic systems onto the terrestrial isopod phylogeny revealed between 3 and 13 transitions of sex determination systems during the evolution of these taxa, most frequently from female to male heterogamety. Our results support that WW individuals are viable in many species, suggesting sex chromosomes are at an incipient stage of their evolution. Together, these data are consistent with the hypothesis that nucleo-cytoplasmic conflicts generated by Wolbachia endosymbionts triggered recurrent turnovers of sex determination systems in terrestrial isopods. They further establish terrestrial isopods as a model to study evolutionary transitions in sex determination systems and pave the way to molecularly characterize these systems.}, } @article {pmid28435138, year = {2017}, author = {Baldridge, G and Higgins, L and Witthuhn, B and Markowski, T and Baldridge, A and Armien, A and Fallon, A}, title = {Proteomic analysis of a mosquito host cell response to persistent Wolbachia infection.}, journal = {Research in microbiology}, volume = {168}, number = {7}, pages = {609-625}, pmid = {28435138}, issn = {1769-7123}, support = {R01 AI081322/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/genetics/metabolism/*microbiology/virology ; Amino Acids/metabolism ; Animals ; Carbohydrate Metabolism ; Cell Line ; Cytoskeletal Proteins ; Energy Metabolism ; Flavivirus/genetics/physiology ; Insect Proteins/genetics/*metabolism ; Lipid Metabolism ; Membrane Proteins/metabolism ; Nucleotides/metabolism ; Proteome ; Proteomics ; Secondary Metabolism ; Signal Transduction ; Wolbachia/isolation & purification/*metabolism/virology ; }, abstract = {Wolbachia pipientis, an obligate intracellular bacterium associated with arthropods and filarial worms, is a target for filarial disease treatment and provides a gene drive agent for insect vector population suppression/replacement. We compared proteomes of Aedes albopictus mosquito C/wStr1 cells persistently infected with Wolbachia strain wStr, relative to uninfected C7-10 control cells. Among approximately 2500 proteins, iTRAQ data identified 815 differentially abundant proteins. As functional classes, energy and central intermediary metabolism proteins were elevated in infected cells, while suppressed proteins with roles in host DNA replication, transcription and translation suggested that Wolbachia suppresses pathways that support host cell growth and proliferation. Vacuolar ATPase subunits were strongly elevated, consistent with high densities of Wolbachia contained individually within vacuoles. Other differential level proteins had roles in ROS neutralization, protein modification/degradation and signaling, including hypothetical proteins whose functions in Wolbachia infection can potentially be manipulated by RNAi interference or transfection. Detection of flavivirus proteins supports further analysis of poorly understood, insect-specific flaviviruses and their potential interactions with Wolbachia, particularly in mosquitoes transinfected with Wolbachia. This study provides a framework for future attempts to manipulate pathways in insect cell lines that favor production of Wolbachia for eventual genetic manipulation, transformation and transinfection of vector species.}, } @article {pmid28426794, year = {2017}, author = {Monnin, D and Kremer, N and Desouhant, E and Vavre, F}, title = {Impact of Wolbachia on oxidative stress sensitivity in the parasitic wasp Asobara japonica.}, journal = {PloS one}, volume = {12}, number = {4}, pages = {e0175974}, pmid = {28426794}, issn = {1932-6203}, mesh = {Animals ; Host-Pathogen Interactions ; *Oxidative Stress ; Symbiosis ; Wasps/*microbiology ; Wolbachia/*physiology ; }, abstract = {The oxidative homeostasis is the balance between reactive oxygen species and antioxidant molecules. In addition to be considered as a key factor underlying life-history traits evolution, the oxidative homeostasis has been shown to be involved in many host-symbiont associations. Previous studies suggest an interaction between the bacterial endosymbiont Wolbachia and the oxidative homeostasis of some insect hosts. This interaction is likely to exert a strong influence on the host evolution, as it has been proposed in the wasp Asobara tabida, whose dependence upon Wolbachia is due to the evolutionary loss of its ability to regulate the oxidative homeostasis in the absence of the symbiont. Although such cases of complete dependence are rare, cases of insects having lost only a part of their autonomy over the control of the oxidative homeostasis might be more common. If so, one can expect that insects having coevolved with Wolbachia will be more sensitive to oxidative stress when cured of their symbionts. We tested this hypothesis by studying the effects of an experimentally-induced oxidative stress on various life-history traits of Asobara japonica, a species closely related to A. tabida. For most of the life-history traits studied, the sensitivity of the wasps to oxidative stress did not correlate with their infection status. The only exception was the parasitic success. However, contrarily to our expectation, the sensitivity to oxidative stress was increased, rather than decreased, when Wolbachia was present. This result suggests that Wolbachia does not participate to mitigate oxidative stress in A. japonica, and that on the contrary its presence might still be costly in stressful environments.}, } @article {pmid30721615, year = {2017}, author = {Bogacheva, AS and Shaikevich, EV and Rakova, VM and Ganushkina, LA}, title = {[(THE FAUNA OF BLOODSUCKING MOSQUITOES IN THE NIZHNY NOVGOROD REGION, THEIR INFECTION WITH DIROFILARIA AND ENDOSYMBIOTIC BACTERIA)].}, journal = {Meditsinskaia parazitologiia i parazitarnye bolezni}, volume = {1}, number = {1}, pages = {43-47}, pmid = {30721615}, issn = {0025-8326}, mesh = {Aedes/microbiology/parasitology ; Animals ; DNA, Helminth/genetics ; Dirofilaria immitis/microbiology/pathogenicity ; Dirofilaria repens/microbiology/pathogenicity ; Dirofilariasis/*microbiology/*parasitology ; Dogs ; Humans ; Mosquito Vectors/genetics/*microbiology/*parasitology ; *Symbiosis ; Wolbachia/isolation & purification/pathogenicity ; }, abstract = {The fauna of bloodsucking mosquitoes in the Nizhny Novgorod Region is represented by 11 species from 5 genera of the family Culicidae. During 2014-2015, the predominant species were Ochlerotatus cantans and Aedes cinereus mosqui- toes in both a population aggregate and woodland. The infected mosquitoes accounted for 1.3% of their total number and were registered only in the village of Fokino. The investigators identified two human pathogenic nematode species: Diro- filaria immits and Dirofilaria repens (0.9% and 0.4% respectively). The effective carriers of Dirofilaria in the examined area can be Ae.cinereus and Och.cantans as only these species were found to have an invasive stage of the parasite. The symbiotic bacterium Wolbachia was detected in the mosquitoes that were not infected with dirofilariasis. This is the first study in Russia to investigate the effects of Wolbachia on the susceptibility of dirofilariasis vectors to infection.}, } @article {pmid30387572, year = {2016}, author = {Rakova, VM}, title = {DIROFILARIASIS: CURRENT ASPECTS OF STUDIES.}, journal = {Meditsinskaia parazitologiia i parazitarnye bolezni}, volume = {4}, number = {4}, pages = {48-52}, pmid = {30387572}, issn = {0025-8326}, mesh = {Animals ; Dirofilaria/classification/*physiology ; Dirofilariasis/*epidemiology/*parasitology/therapy ; Europe/epidemiology ; *Geographic Information Systems ; Humans ; }, abstract = {The review touches upon the main directions and some of the most important results of an investigation of dirofilariasis the species of the genus Dirofilaria, their spread, and hosts. It describes cases of human infection with Candidatus Dirofilaria hongkongensis, a study of the prevalence of filariasis using a geographic information system, data on the contamination of Dirofilaria in Europe, and current views on the endosymbiotic relations of Dirofilaria with the Wolbachia bacterium.}, } @article {pmid32260037, year = {2016}, author = {}, title = {Short news.}, journal = {Virologie (Montrouge, France)}, volume = {20}, number = {4}, pages = {248-252}, doi = {10.1684/vir.2016.0662}, pmid = {32260037}, issn = {1267-8694}, } @article {pmid29491905, year = {2016}, author = {Humphreys, DP and Rundle, HD and Dyer, KA}, title = {Patterns of reproductive isolation in the Drosophila subquinaria complex: can reinforced premating isolation cascade to other species?.}, journal = {Current zoology}, volume = {62}, number = {2}, pages = {183-191}, pmid = {29491905}, issn = {1674-5507}, abstract = {The reinforcement of premating barriers due to reduced hybrid fitness in sympatry may cause secondary sexual isolation within a species as a by-product. Consistent with this, in the fly Drosophila subquinaria, females that are sympatric with D. recens mate at very low rates not only with D. recens, but also with conspecific D. subquinaria males from allopatry. Here, we ask if these effects of reinforcement cascade more broadly to affect sexual isolation with other closely related species. We assay reproductive isolation of these species with D. transversa and find that choosy D. subquinaria females from the region sympatric with D. recens discriminate strongly against male D. transversa, whereas D. subquinaria from the allopatric region do not. This increased sexual isolation cannot be explained by natural selection to avoid mating with this species, as they are allopatric in geographic range and we do not identify any intrinsic postzygotic isolation between D. subquinaria and D. transversa. Variation in epicuticular hydrocarbons, which are used as mating signals in D. subquinaria, follow patterns of premating isolation: D. transversa and allopatric D. subquinaria are most similar to each other and differ from sympatric D. subquinaria, and those of D. recens are distinct from the other two species. We suggest that the secondary effects of reinforcement may cascade to strengthen reproductive isolation with other species that were not a target of selection. These effects may enhance the divergence that occurs in allopatry to help explain why some species are already sexually isolated upon secondary contact.}, } @article {pmid29588003, year = {2015}, author = {Lees, RS and Gilles, JR and Hendrichs, J and Vreysen, MJ and Bourtzis, K}, title = {Back to the future: the sterile insect technique against mosquito disease vectors.}, journal = {Current opinion in insect science}, volume = {10}, number = {}, pages = {156-162}, doi = {10.1016/j.cois.2015.05.011}, pmid = {29588003}, issn = {2214-5753}, abstract = {With the global burden of mosquito-borne diseases increasing, and some conventional vector control tools losing effectiveness, the sterile insect technique (SIT) is a potential new tool in the arsenal. Equipment and protocols have been developed and validated for efficient mass-rearing, irradiation and release of Aedines and Anophelines that could be useful for several control approaches. Assessment of male quality is becoming more sophisticated, and several groups are well advanced in pilot site selection and population surveillance. It will not be long before SIT feasibility has been evaluated in various settings. Until perfect sexing mechanisms exist, combination of Wolbachia-induced phenotypes, such as cytoplasmic incompatibility and pathogen interference, and irradiation may prove to be the safest solution for population suppression.}, } @article {pmid32846693, year = {2015}, author = {Johnson, KN}, title = {Bacteria and antiviral immunity in insects.}, journal = {Current opinion in insect science}, volume = {8}, number = {}, pages = {97-103}, doi = {10.1016/j.cois.2015.01.008}, pmid = {32846693}, issn = {2214-5753}, abstract = {Tripartite interactions between viruses, bacteria and hosts can have significant implications for the outcome of infections. In insects recent studies have shown that these interactions can greatly influence the transmission of vector borne disease, including transmission of dengue virus. Mosquito gut bacteria modulate arbovirus infection of the mosquito host and specific bacterial isolates have been identified that negatively impact the outcome of infection. In addition, an endosymbiotic bacterium Wolbachia, which is very common in insects, protects flies and mosquitoes from virus infection. This antiviral protection has implications for both naturally infected insects and for use in disease control. This review summarises the current state of knowledge for each of these interactions with an emphasis on natural interactions.}, } @article {pmid32669893, year = {2014}, author = {Higazi, TB and Geary, TG and Mackenzie, CD}, title = {Chemotherapy in the treatment, control, and elimination of human onchocerciasis.}, journal = {Research and reports in tropical medicine}, volume = {5}, number = {}, pages = {77-93}, pmid = {32669893}, issn = {1179-7282}, abstract = {Onchocerciasis treatment is one of the most positive stories in tropical medicine although major challenges remain to reaching the ultimate goal of disease elimination. Such challenges are to be expected when the therapeutic goal is to kill and safely remove a large multistage, efficient, metazoan infectious agent such as Onchocerca volvulus that has an exceptionally complicated relationship with its host. Successful control of onchocerciasis has often been hampered by host reactions following chemotherapy, that can sometimes cause significant tissue pathology. Presence of other filariae, particularly Loa loa, in endemic onchocerciasis-treatment areas also poses severe problems due to adverse reactions caused by drug-induced death of the coincident microfilariae of this usually clinically benign species. Although ivermectin has been very successful, there is a need to enhance the progress toward elimination of onchocerciasis; new drugs and their efficient use are keys to this. The permanent absence of Onchocerca microfilaridermia, defined as the lack of resurgence of skin microfilarial loads after treatment, is the ultimate characteristic of a useful new chemotherapeutic agent. Several drugs are under investigation to achieve this, including the reassessment of currently available and previously tested agents, such as the antibiotic, doxycycline, which targets the adult parasites through its anti-Wolbachia endosymbiont activity. Flubendazole, a benzimidazole derivative approved for treatment of human gastrointestinal nematodes, is also being considered for repurposing as a macrofilaricide to aid in the achievement of eradication. The managerial challenges existing at the population level also need to be addressed; these include drug-distribution fatigue, the need to include noncompliant people, civil unrest in endemic areas, political cross-border issues, restrictions of age and pregnancy, and complications due to integration with other treatment programs. It is likely that a panel of chemotherapeutic options, new and old, supported by strong and effective distribution systems will be the best way to address challenges of treatment and elimination of this infection. Future research should also address management of treatment and control, and consider how new treatment paradigms can be incorporated to meet time lines set for global elimination by 2025.}, } @article {pmid28565520, year = {1999}, author = {Shoemaker, DD and Katju, V and Jaenike, J}, title = {WOLBACHIA AND THE EVOLUTION OF REPRODUCTIVE ISOLATION BETWEEN DROSOPHILA RECENS AND DROSOPHILA SUBQUINARIA.}, journal = {Evolution; international journal of organic evolution}, volume = {53}, number = {4}, pages = {1157-1164}, doi = {10.1111/j.1558-5646.1999.tb04529.x}, pmid = {28565520}, issn = {1558-5646}, abstract = {Endosymbiotic bacteria of the genus Wolbachia are widespread among insects and in many cases cause cytoplasmic incompatibility in crosses between infected males and uninfected females. Such findings have been used to argue that Wolbachia have played an important role in insect speciation. Theoretical models, however, indicate that Wolbachia alone are unlikely to lead to stable reproductive isolation between two formerly conspecific populations. Here we analyze the components of reproductive isolation between Drosophila recens, which is infected with Wolbachia, and its uninfected sister species Drosophila subquinaria. Laboratory pairings demonstrated that gene flow via matings between D. recens females and D. subquinaria males is hindered by behavioral isolation. Matings readily occurred in the reciprocal cross (D. quinaria females × D. recens males), but very few viable progeny were produced. The production of viable hybrids via this route was restored by antibiotic curing of D. recens of their Wolbachia symbionts, indicating that hybrid offspring production is greatly reduced by cytoplasmic incompatibility in the crosses involving infected D. recens males. Thus, behavioral isolation and Wolbachia-induced cytoplasmic incompatibility act as complementary asymmetrical isolating mechanisms between these two species. In accordance with Haldane's rule, hybrid females were fertile, whereas hybrid males invariably were sterile. Levels of mtDNA variation in D. recens are much lower than in either D. subquinaria or D. falleni, neither of which is infected with Wolbachia. The low haplotype diversity in D. recens is likely due to an mtDNA sweep associated with the spread of Wolbachia. Nevertheless, the existence of several mtDNA haplotypes in this species indicates that Wolbachia have been present as a potential isolating mechanism for a substantial period of evolutionary time. Finally, we argue that although Wolbachia by themselves are unlikely to bring about speciation, they can increase the rate of speciation in insects.}, } @article {pmid28565514, year = {1999}, author = {Belshaw, R and Quicke, DLJ and Völkl, W and Godfray, HCJ}, title = {MOLECULAR MARKERS INDICATE RARE SEX IN A PREDOMINANTLY ASEXUAL PARASITOID WASP.}, journal = {Evolution; international journal of organic evolution}, volume = {53}, number = {4}, pages = {1189-1199}, doi = {10.1111/j.1558-5646.1999.tb04532.x}, pmid = {28565514}, issn = {1558-5646}, abstract = {The parasitoid wasp genus Lysiphlebus (Hymenoptera: Braconidae: Aphidiinae) contains a taxonomically poorly resolved group of both sexual (arrhenotokous) species and asexual (thelytokous) clones. Maximum-parsimony and maximum-likelihood analyses of mitochondrial DNA sequence data from specimens collected across Western Europe showed that asexuality, which does not appear to be caused by the bacterium Wolbachia, is concentrated in two geographically widespread lineages, the older of which diverged from the closest extant sexual taxa approximately 0.5 million years ago. However, the DNA sequences of a nuclear intron (elongation factor-1α) showed no congruence with this pattern, and a much higher frequency of heterozygotes with very high allelic diversity was observed among the asexual females compared to that among females from the sexual species. This pattern is consistent with maternally inherited asexuality coupled with a history of rare sex with members of several closely related sexual populations or species. Our observations reinforce recent arguments that rare sex may be more important for the persistence of otherwise asexual lineages than hitherto appreciated.}, } @article {pmid28568785, year = {1997}, author = {Poinsot, D and Merçot, H}, title = {WOLBACHIA INFECTION IN DROSOPHILA SIMULANS: DOES THE FEMALE HOST BEAR A PHYSIOLOGICAL COST?.}, journal = {Evolution; international journal of organic evolution}, volume = {51}, number = {1}, pages = {180-186}, doi = {10.1111/j.1558-5646.1997.tb02399.x}, pmid = {28568785}, issn = {1558-5646}, abstract = {Fitness traits of three Drosophila simulans strains infected by endocellular bacteria belonging to the genus Wolbachia have been compared with those of replicate stocks previously cured from the infection by an antibiotic treatment. The traits measured were development time, egg-to-adult viability, egg hatch, productivity, fecundity, and the number of functional ovarioles. Individuals of the first strain were bi-infected by two Wolbachia variants, wHa and wNo. The second strain was infected by wHa, the third one by wNo. The Wolbachia studied here cause cytoplasmic incompatibility (CI), a high embryonic mortality (70% to > 90%) when an infected male is crossed with an uninfected female. Three generations after antibiotic treatment, we observed in all strains a significant drop in productivity in the cured stocks. This drop was not due to antibiotic toxicity and was associated with the loss of the Wolbachia. However the effect had disappeared in two of the three strains five generations after treatment, and could not be found in the third strain in a third measurement carried out 14 generations after treatment. The temporary nature of the productivity difference indicates that Wolbachia do not enhance productivity in infected strains. On the other hand, in all traits measured, our results show the absence of any negative effects of the Wolbachia on their host. This could be explained when considering Wolbachia evolution, as maternally transmitted parasites bear a strong selective pressure not to harm their female host. However, CI would allow the bacteria to be maintained even when harming the female. The apparent absence of deleterious effects caused by these Wolbachia might result from a trade-off, where a relatively low bacteria density would advantage the Wolbachia by suppressing any deleterious effects on the female host, at the cost of a weaker maternal transmission rate of the infection.}, } @article {pmid28568927, year = {1996}, author = {Orr, MR}, title = {LIFE-HISTORY ADAPTATION AND REPRODUCTIVE ISOLATION IN A GRASSHOPPER HYBRID ZONE.}, journal = {Evolution; international journal of organic evolution}, volume = {50}, number = {2}, pages = {704-716}, doi = {10.1111/j.1558-5646.1996.tb03880.x}, pmid = {28568927}, issn = {1558-5646}, abstract = {Patterns of life-history adaptation and reproductive isolation were investigated in the acridid grasshoppers Melanoplus sanguinipes and M. devastator, which hybridize along an altitudinal gradient in the Sierra Nevada of California. Melanoplus sanguinipes females crossed with M. devastator males produced eggs that were approximately half as viable as eggs from other crosses. Diminished viability was not attributable either to infection by Wolbachia pipientis or to failure of sperm transfer. When offered an opportunity to choose a mate, females from all populations discriminated against males of the other species, whereas in no-choice tests measuring copulation duration only females from the tails of the clines showed preferences. Melanoplus sanguinipes, found at high elevations where the growing season is short, exhibited faster egg hatch, faster larval development, smaller adult body sizes, and smaller clutch sizes than M. devastator. Melanoplus devastator, from California's Central Valley, endured a hot and dry summer in a reproductive diapause that was absent in M. sanguinipes. Clines in reproductive diapause and clutch size coincided with the region of reproductive incompatibility. Development time, body size, and hatch time also changed across the hybrid zone, but the regions of largest transitions in these traits were either difficult to locate using the limited populations studied here or were not coincident with the zone's center. A method is described for combining ecological and phylogenetic analyses to address the unknown issue of whether life-history divergence has conributed to reproductive isolation in this system.}, } @article {pmid28568404, year = {1994}, author = {Turelli, M}, title = {EVOLUTION OF INCOMPATIBILITY-INDUCING MICROBES AND THEIR HOSTS.}, journal = {Evolution; international journal of organic evolution}, volume = {48}, number = {5}, pages = {1500-1513}, doi = {10.1111/j.1558-5646.1994.tb02192.x}, pmid = {28568404}, issn = {1558-5646}, abstract = {In many insect species, males infected with microbes related to Wolbachia pipientis are "incompatible" with uninfected females. Crosses between infected males and uninfected females produce significantly fewer adult progeny than the other three possible crosses. The incompatibility-inducing microbes are usually maternally transmitted. Thus, incompatibility tends to confer a reproductive advantage on infected females in polymorphic populations, allowing these infections to spread. This paper analyzes selection on parasite and host genes that affect such incompatibility systems. Selection among parasite variants does not act directly on the level of incompatibility with uninfected females. In fact, selection favors rare parasite variants that increase the production of infected progeny by infected mothers, even if these variants reduce incompatibility with uninfected females. However, productivity-reducing parasites that cause partial incompatibility with hosts harboring alternative variants can be favored once they become sufficiently abundant locally. Thus, they may spread spatially by a process analogous to the spread of underdominant chromosome rearrangements. The dynamics of modifier alleles in the host are more difficult to predict, because such alleles will occur in both infected and uninfected individuals. Nevertheless, the relative fecundity of infected females compared to uninfected females, the efficiency of maternal transmission and the mutual compatibility of infected individuals all tend to increase under within-population selection on both host and parasite genes. In addition, selection on host genes favors increased compatibility between infected males and uninfected females. Although vertical transmission tends to harmonize host and parasite evolution, competition among parasite variants will tend to maintain incompatibility.}, } @article {pmid28568258, year = {1994}, author = {Prout, T}, title = {SOME EVOLUTIONARY POSSIBILITIES FOR A MICROBE THAT CAUSES INCOMPATIBILITY IN ITS HOST.}, journal = {Evolution; international journal of organic evolution}, volume = {48}, number = {3}, pages = {909-911}, doi = {10.1111/j.1558-5646.1994.tb01371.x}, pmid = {28568258}, issn = {1558-5646}, } @article {pmid28568710, year = {1993}, author = {Kambhampati, S and Rai, KS and Burgun, SJ}, title = {UNIDIRECTIONAL CYTOPLASMIC INCOMPATIBILITY IN THE MOSQUITO, AEDES ALBOPICTUS.}, journal = {Evolution; international journal of organic evolution}, volume = {47}, number = {2}, pages = {673-677}, doi = {10.1111/j.1558-5646.1993.tb02121.x}, pmid = {28568710}, issn = {1558-5646}, } @article {pmid28421042, year = {2017}, author = {Novakova, E and Woodhams, DC and Rodríguez-Ruano, SM and Brucker, RM and Leff, JW and Maharaj, A and Amir, A and Knight, R and Scott, J}, title = {Mosquito Microbiome Dynamics, a Background for Prevalence and Seasonality of West Nile Virus.}, journal = {Frontiers in microbiology}, volume = {8}, number = {}, pages = {526}, pmid = {28421042}, issn = {1664-302X}, abstract = {Symbiotic microbial communities augment host phenotype, including defense against pathogen carriage and infection. We sampled the microbial communities in 11 adult mosquito host species from six regions in southern Ontario, Canada over 3 years. Of the factors examined, we found that mosquito species was the largest driver of the microbiota, with remarkable phylosymbiosis between host and microbiota. Seasonal shifts of the microbiome were consistently repeated over the 3-year period, while region had little impact. Both host species and seasonal shifts in microbiota were associated with patterns of West Nile virus (WNV) in these mosquitoes. The highest prevalence of WNV, with a seasonal spike each year in August, was in the Culex pipiens/restuans complex, and high WNV prevalence followed a decrease in relative abundance of Wolbachia in this species. Indeed, mean temperature, but not precipitation, was significantly correlated with Wolbachia abundance. This suggests that at higher temperatures Wolbachia abundance is reduced leading to greater susceptibility to WNV in the subsequent generation of C. pipiens/restuans hosts. Different mosquito genera harbored significantly different bacterial communities, and presence or abundance of Wolbachia was primarily associated with these differences. We identified several operational taxonomic units (OTUs) of Wolbachia that drive overall microbial community differentiation among mosquito taxa, locations and timepoints. Distinct Wolbachia OTUs were consistently found to dominate microbiomes of Cx. pipiens/restuans, and of Coquilletidia perturbans. Seasonal fluctuations of several other microbial taxa included Bacillus cereus, Enterococcus, Methylobacterium, Asaia, Pantoea, Acinetobacter johnsonii, Pseudomonas, and Mycoplasma. This suggests that microbiota may explain some of the variation in vector competence previously attributed to local environmental processes, especially because Wolbachia is known to affect carriage of viral pathogens.}, } @article {pmid28420428, year = {2017}, author = {Armoo, S and Doyle, SR and Osei-Atweneboana, MY and Grant, WN}, title = {Significant heterogeneity in Wolbachia copy number within and between populations of Onchocerca volvulus.}, journal = {Parasites & vectors}, volume = {10}, number = {1}, pages = {188}, pmid = {28420428}, issn = {1756-3305}, mesh = {Animals ; *DNA Copy Number Variations ; Ecotype ; Forests ; *Genetic Heterogeneity ; *Genome, Bacterial ; Genome, Helminth ; Grassland ; High-Throughput Nucleotide Sequencing ; Ivermectin/pharmacology ; Onchocerca volvulus/genetics/*microbiology ; Real-Time Polymerase Chain Reaction ; Wolbachia/*genetics ; }, abstract = {BACKGROUND: Wolbachia are intracellular bacteria found in arthropods and several filarial nematode species. The filarial Wolbachia have been proposed to be involved in the immunopathology associated with onchocerciasis. Higher Wolbachia-to-nematode ratios have been reported in the savannah-ecotype compared to the forest-ecotype, and have been interpreted as consistent with a correlation between Wolbachia density and disease severity. However, factors such as geographic stratification and ivermectin drug exposure can lead to significant genetic heterogeneity in the nematode host populations, so we investigated whether Wolbachia copy number variation is also associated with these underlying factors.

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

CONCLUSIONS: While the scale of transcriptional changes associated with wFlu infection might be small, the scope is rather large, which confirms that native Wolbachia infections maintain intricate molecular relationships with their mosquito hosts even after lengthy periods of co-evolution. We have also identified several potential means through which wFlu infection might influence Plasmodium infection in Ae. fluviatilis, and these genes should form the basis of future investigation into the enhancement of Plasmodium by Wolbachia.}, } @article {pmid28032096, year = {2016}, author = {Simsek, S and Ciftci, AT}, title = {Serological and Molecular Detection of Dirofilaria Species in Stray Dogs and Investigation of Wolbachia DNA by PCR in Turkey.}, journal = {Journal of arthropod-borne diseases}, volume = {10}, number = {4}, pages = {445-453}, pmid = {28032096}, issn = {2322-1984}, abstract = {BACKGROUND: Dirofilaria immitis and Dirofilaria repens are the most common species of filarial nematodes described in the dogs. A single-step multiplex PCR was applied to detect and differentiate simultaneously and unequivocally D. immitis and D. repens on DNA extracted from canine peripheral blood and besides to detect the seroprevalance of D. immitis by ELISA in Elazig Province, Turkey. A PCR detection of the Wolbachia, which plays an important role in D. immitis biology and contributes to the inflammatory pathology of the heartworm, was also applied for the first time in Turkey.

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

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

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

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

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

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

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

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

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

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

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

CONCLUSION: Aedes aegypti queenslandensis are genomically indistinguishable from the type form, which points to these forms freely interbreeding at least in Australia and Singapore. Based on our findings, it is unlikely that the presence of very pale Ae. aegypti will affect the success of Aedes control programs based on Wolbachia-infected, sterile or RIDL mosquitoes.}, } @article {pmid27802941, year = {2016}, author = {Mayor, S}, title = {Sixty seconds on . . . Wolbachia.}, journal = {BMJ (Clinical research ed.)}, volume = {355}, number = {}, pages = {i5847}, doi = {10.1136/bmj.i5847}, pmid = {27802941}, issn = {1756-1833}, mesh = {Aedes/*microbiology/virology ; Animals ; Chikungunya Fever/transmission ; Dengue/transmission ; Humans ; Insect Vectors/*microbiology/virology ; Virus Diseases/*transmission ; Virus Replication/physiology ; Wolbachia/*physiology ; Zika Virus Infection/transmission ; }, } @article {pmid27802919, year = {2017}, author = {Budachetri, K and Williams, J and Mukherjee, N and Sellers, M and Moore, F and Karim, S}, title = {The microbiome of neotropical ticks parasitizing on passerine migratory birds.}, journal = {Ticks and tick-borne diseases}, volume = {8}, number = {1}, pages = {170-173}, pmid = {27802919}, issn = {1877-9603}, support = {P20 GM103476/GM/NIGMS NIH HHS/United States ; P20 RR016476/RR/NCRR NIH HHS/United States ; }, mesh = {*Animal Migration ; Animals ; Bacteria/genetics/*isolation & purification ; Bird Diseases/*parasitology ; DNA, Bacterial/genetics ; *Microbiota ; Passeriformes/*parasitology ; Tick Infestations/parasitology/*veterinary ; }, abstract = {Seasonal migration of passerine birds between temperate North America and tropical Central and South America is an ecological phenomenon. Migration of birds has been associated with the introduction of ectoparasites like ticks or tick-borne pathogens across the avian migration routes. In this study, the microbial diversity was determined in the ticks and bird DNA samples using 454 pyrosequencing of bacterial 16S rRNA gene. Tick DNA samples showed the dominance of genera Lactococcus, Francisella, Raoultella, Wolbachia and Rickettsia across all the ticks, but birds DNA did not share common microbial diversity with ticks. Furthermore, "Candidatus Rickettsia amblyommii" infection in the 91 ticks collected off the songbirds was also quantified by qPCR assay. Interestingly, "Candidatus R. amblyommii" was tested positive in 24 ticks (26% infection), and infection varied from as low as three copies to thousands of copies, but bird blood samples showed no amplification. Our results provide evidence that songbirds serve as transport carrier for immature ticks, and less likely to be a reservoir for "Candidatus R. amblyommii".}, } @article {pmid27718295, year = {2016}, author = {Coon, KL and Brown, MR and Strand, MR}, title = {Mosquitoes host communities of bacteria that are essential for development but vary greatly between local habitats.}, journal = {Molecular ecology}, volume = {25}, number = {22}, pages = {5806-5826}, pmid = {27718295}, issn = {1365-294X}, support = {R01 AI106892/AI/NIAID NIH HHS/United States ; T32 GM007103/GM/NIGMS NIH HHS/United States ; }, mesh = {Aedes/*microbiology ; Animals ; Bacteria/*classification ; Ecosystem ; *Microbiota ; RNA, Ribosomal, 16S/genetics ; Southeastern United States ; }, abstract = {Mosquitoes are insects of interest because several species vector disease-causing pathogens to humans and other vertebrates. We previously reported that mosquitoes from long-term laboratory cultures require living bacteria in their gut to develop, but development does not depend on particular species of bacteria. Here, we focused on three distinct but interrelated areas of study to better understand the role of bacteria in mosquito development by studying field and laboratory populations of Aedes aegypti, Aedes albopictus and Culex quinquefasciatus from the southeastern United States. Sequence analysis of bacterial 16S rRNA gene amplicons showed that bacterial community composition differed substantially in larvae from different collection sites, whereas larvae from the same site shared similarities. Although previously unknown to be infected by Wolbachia, results also indicated that Ae. aegypti from one field site hosted a dual infection. Regardless of collection site or factors like Wolbachia infection, however, each mosquito species required living bacteria in their digestive tract to develop. Results also identified several concerns in using antibiotics to eliminate the bacterial community in larvae in order to study its developmental consequences. Altogether, our results indicate that several mosquito species require living bacteria for development. We also hypothesize these species do not rely on particular bacteria because larvae do not reliably encounter the same bacteria in the aquatic habitats they develop in.}, } @article {pmid27784636, year = {2016}, author = {Puggioli, A and Calvitti, M and Moretti, R and Bellini, R}, title = {wPip Wolbachia contribution to Aedes albopictus SIT performance: Advantages under intensive rearing.}, journal = {Acta tropica}, volume = {164}, number = {}, pages = {473-481}, doi = {10.1016/j.actatropica.2016.10.014}, pmid = {27784636}, issn = {1873-6254}, mesh = {Aedes/*physiology ; Animals ; Female ; Infertility/physiopathology ; Male ; Mosquito Vectors/*physiology ; Pupa ; Reproduction/*drug effects ; Sexual Behavior, Animal/*drug effects ; Wolbachia/*physiology ; }, abstract = {As a part of a project aiming at the suppression of the mosquito vector Aedes albopictus, a specific Ae. albopictus line producing sterile males, ARwP, was tested for its suitability to intense rearing conditions compatible with mass production and field release. This line was developed by the Italian National Agency for New Technologies, Energy and Sustainable Economic Development thanks to the artificial infection with a heterologous Wolbachia strain, resulting in a bidirectional incompatibility pattern with wild-type Ae. albopictus. ARwP was reared under Standard Operating Procedures at the Centro Agricoltura Ambiente and compared with a wild-type strain in terms of time of pupation onset, production of male pupae in the following 24h and mechanical sexing efficacy. Mating competitiveness of ARwP males was also evaluated in comparison with irradiated wild-type males in large field enclosures. ARwP males demonstrated a significantly shorter time of pupation onset, a higher rate of production of male pupae in the following 24h and a lower percentage of residual contaminant females when applying mechanical sexing procedures. In addition, ARwP males were more efficient than wild-types in competing for wild-type females in large enclosures, thus inducing a level of sterility significantly higher than that expected for an equal mating competitiveness. These results encourage the use of this Ae. albopictus strain as suppression tool against Ae. albopictus based on considerations thoroughly discussed in the manuscript.}, } @article {pmid27770026, year = {2016}, author = {Lin, Y and Chen, ZX and Oliver, B and Harbison, ST}, title = {Microenvironmental Gene Expression Plasticity Among Individual Drosophila melanogaster.}, journal = {G3 (Bethesda, Md.)}, volume = {6}, number = {12}, pages = {4197-4210}, pmid = {27770026}, issn = {2160-1836}, mesh = {Animals ; Cluster Analysis ; Drosophila melanogaster/*genetics ; *Environment ; Female ; Gene Expression Profiling ; *Gene Expression Regulation ; *Gene-Environment Interaction ; Genetic Variation ; Genotype ; High-Throughput Nucleotide Sequencing ; Male ; Sex Characteristics ; Sex Factors ; Transcriptome ; }, abstract = {Differences in phenotype among genetically identical individuals exposed to the same environmental condition are often noted in genetic studies. Despite this commonplace observation, little is known about the causes of this variability, which has been termed microenvironmental plasticity. One possibility is that stochastic or technical sources of variance produce these differences. A second possibility is that this variation has a genetic component. We have explored gene expression robustness in the transcriptomes of 730 individual Drosophila melanogaster of 16 fixed genotypes, nine of which are infected with Wolbachia Three replicates of flies were grown, controlling for food, day/night cycles, humidity, temperature, sex, mating status, social exposure, and circadian timing of RNA extraction. Despite the use of inbred genotypes, and carefully controlled experimental conditions, thousands of genes were differentially expressed, revealing a unique and dynamic transcriptional signature for each individual fly. We found that 23% of the transcriptome was differentially expressed among individuals, and that the variability in gene expression among individuals is influenced by genotype. This transcriptional variation originated from specific gene pathways, suggesting a plastic response to the microenvironment; but there was also evidence of gene expression differences due to stochastic fluctuations. These observations reveal previously unappreciated genetic sources of variability in gene expression among individuals, which has implications for complex trait genetics and precision medicine.}, } @article {pmid27742209, year = {2016}, author = {Wu, CH and Zong, Q and Du, AL and Zhang, W and Yao, HC and Yu, XQ and Wang, YF}, title = {Knockdown of Dynamitin in testes significantly decreased male fertility in Drosophila melanogaster.}, journal = {Developmental biology}, volume = {420}, number = {1}, pages = {79-89}, doi = {10.1016/j.ydbio.2016.10.007}, pmid = {27742209}, issn = {1095-564X}, mesh = {Animals ; Cell Nucleus/metabolism ; Drosophila Proteins/*metabolism ; Drosophila melanogaster/*physiology ; Dyneins/metabolism ; Fertility ; Gene Expression Regulation ; *Gene Knockdown Techniques ; Intermediate Filament Proteins/*metabolism ; Male ; Mitochondria/metabolism ; Spermatids/metabolism ; Spermatogenesis ; Testis/*physiology ; Tubulin/metabolism ; }, abstract = {Dynamitin (Dmn) is a major component of dynactin, a multiprotein complex playing important roles in a variety of intracellular motile events. We previously found that Wolbachia bacterial infection resulted in a reduction of Dmn protein. As Wolbachia may modify sperm in male hosts, we speculate that Dmn may have a function in male fertility. Here we used nosGal4 to drive Dmn knock down in testes of Drosophila melanogaster to investigate the functions of Dmn in spermatogenesis. We found that knockdown of Dmn in testes dramatically decreased male fertility, overexpression of Dmn in Wolbachia-infected males significantly rescued male fertility, indicating an important role of Dmn in inducing male fertility defects following Wolbachia infection. Some scattered immature sperm with late canoe-shaped head distributed in the end of Dmn knockdown testis and only about half mature sperm were observed in the Dmn knockdown testis relative to those in the control. Transmission electron microscopy (TEM) exhibited fused spermatids in cysts and abnormal mitochondrial derivatives. Immunofluorescence staining showed significantly less abundance of tubulin around the nucleus of spermatid and scattered F-actin cones to different extents in the individualization complex (IC) during spermiogenesis in Dmn knockdown testes, which may disrupt the nuclear condensation and sperm individualization. Since dynein-dynactin complex has been shown to mediate transport of many cellular components, including mRNAs and organelles, these results suggest that Dmn may play an important role in Drosophila spermiogenesis by affecting transport of many important cytoplasmic materials.}, } @article {pmid27702765, year = {2016}, author = {Fromont, C and Riegler, M and Cook, JM}, title = {Phylogeographic analyses of bacterial endosymbionts in fig homotomids (Hemiptera: Psylloidea) reveal codiversification of both primary and secondary endosymbionts.}, journal = {FEMS microbiology ecology}, volume = {92}, number = {12}, pages = {}, doi = {10.1093/femsec/fiw205}, pmid = {27702765}, issn = {1574-6941}, mesh = {Animals ; Base Composition/genetics ; Biological Evolution ; DNA, Mitochondrial/genetics ; Enterobacteriaceae/classification/genetics/isolation & purification ; Evolution, Molecular ; Ficus ; Halomonadaceae/genetics/*isolation & purification ; Hemiptera/*microbiology ; Phylogeny ; Phylogeography ; Symbiosis/*genetics ; Wolbachia/genetics/*isolation & purification ; }, abstract = {While obligate primary (P-) endosymbionts usually cospeciate with their insect hosts, less is known about codiversification of secondary (S-) endosymbionts that are generally considered facultative. Typically, insects of the superfamily Psylloidea harbour one P- (Carsonella) and at least one S-endosymbiont, thought to compensate for Carsonella genome reduction. Most codiversification studies have used phylogenies of psyllids and their endosymbionts across and within host families or genera, but few have explored patterns within species. We focussed on P- and S-endosymbionts of three Mycopsylla (Homotomidae) species to explore whether they have congruent phylogenies and within-species geographic structures. The P-endosymbiont Carsonella, a S-endosymbiont and Wolbachia all had 100% prevalence, while Arsenophonus was only found in one species at low prevalence. Congruent phylogenies of Mycopsylla and P-endosymbionts across populations and species support strict cospeciation. S-endosymbiont phylogenies were also congruent across host species but low genetic variation in the S-endosymbiont was not correlated with host phylogeography, possibly due to a shorter evolutionary association. Between species, Wolbachia and Mycopsylla phylogenies were incongruent, probably due to horizontal transmission events. Our study is the first to explore endosymbionts of Mycopsylla and further supports the codivergence of Psylloidea hosts and P-endosymbionts, with obligate host interactions for both P- and S-endosymbionts.}, } @article {pmid27768689, year = {2016}, author = {Sikulu-Lord, MT and Milali, MP and Henry, M and Wirtz, RA and Hugo, LE and Dowell, FE and Devine, GJ}, title = {Near-Infrared Spectroscopy, a Rapid Method for Predicting the Age of Male and Female Wild-Type and Wolbachia Infected Aedes aegypti.}, journal = {PLoS neglected tropical diseases}, volume = {10}, number = {10}, pages = {e0005040}, pmid = {27768689}, issn = {1935-2735}, mesh = {Aedes/*growth & development/*microbiology ; Animals ; Female ; Insect Control ; Insect Vectors/*growth & development/*microbiology ; Male ; Pest Control, Biological ; Spectroscopy, Near-Infrared/*methods ; Wolbachia/*physiology ; }, abstract = {Estimating the age distribution of mosquito populations is crucial for assessing their capacity to transmit disease and for evaluating the efficacy of available vector control programs. This study reports on the capacity of the near-infrared spectroscopy (NIRS) technique to rapidly predict the ages of the principal dengue and Zika vector, Aedes aegypti. The age of wild-type males and females, and males and females infected with wMel and wMelPop strains of Wolbachia pipientis were characterized using this method. Calibrations were developed using spectra collected from their heads and thoraces using partial least squares (PLS) regression. A highly significant correlation was found between the true and predicted ages of mosquitoes. The coefficients of determination for wild-type females and males across all age groups were R2 = 0.84 and 0.78, respectively. The coefficients of determination for the age of wMel and wMelPop infected females were 0.71 and 0.80, respectively (P< 0.001 in both instances). The age of wild-type female Ae. aegypti could be identified as < or ≥ 8 days old with an accuracy of 91% (N = 501), whereas female Ae. aegypti infected with wMel and wMelPop were differentiated into the two age groups with an accuracy of 83% (N = 284) and 78% (N = 229), respectively. Our results also indicate NIRS can distinguish between young and old male wild-type, wMel and wMelPop infected Ae. aegypti with accuracies of 87% (N = 253), 83% (N = 277) and 78% (N = 234), respectively. We have demonstrated the potential of NIRS as a predictor of the age of female and male wild-type and Wolbachia infected Ae. aegypti mosquitoes under laboratory conditions. After field validation, the tool has the potential to offer a cheap and rapid alternative for surveillance of dengue and Zika vector control programs.}, } @article {pmid27756651, year = {2016}, author = {Wiwatanaratanabutr, I and Grandjean, F}, title = {Impacts of temperature and crowding on sex ratio, fecundity and Wolbachia infection intensity in the copepod, Mesocyclops thermocyclopoides.}, journal = {Journal of invertebrate pathology}, volume = {141}, number = {}, pages = {18-23}, doi = {10.1016/j.jip.2016.10.003}, pmid = {27756651}, issn = {1096-0805}, mesh = {Animals ; Copepoda/*microbiology ; Female ; Fertility ; Gram-Negative Bacterial Infections/*veterinary ; Hot Temperature ; Male ; Real-Time Polymerase Chain Reaction ; Sex Ratio ; *Wolbachia ; }, abstract = {Wolbachia are a group of intracellular bacteria that cause reproductive alterations in arthropods. Here, we describe the effects of two environmental factors (crowding and temperature) on phenotypic expression of feminization, the host's fecundity and Wolbachia infection intensity among life cycle stages in the naturally Wolbachia-infected copepod, Mesocyclops thermocyclopoides. The copepod was first found to be co-infected with Wolbachia A- and B-supergroups Wolbachia strains based on wsp primers. The relative Wolbachia infection intensity within individuals was determined using quantitative real-time PCR and was significantly higher in the B-supergroup than in the A-supergroup. Experimental results of temperature effect on bacterial density in each developmental stage revealed a significant decrease in Wolbachia infection intensity following exposure to high temperature (37°C) in both sexes and implied that Wolbachia might survive in room temperature (25°C) better than in high temperature. Experimental results of crowding effects on Wolbachia infection intensity suggested a negative correlation between copepod nauplii and Wolbachia infection intensity. No effect of rearing temperature on the sex ratio was reported although the fecundity was significantly decreased by high temperature. The results showed that Wolbachia infection intensity to be correlated with crowding conditions and was decreased following exposure of elevated temperature.}, } @article {pmid27760540, year = {2016}, author = {Jiggins, FM}, title = {Open questions: how does Wolbachia do what it does?.}, journal = {BMC biology}, volume = {14}, number = {1}, pages = {92}, pmid = {27760540}, issn = {1741-7007}, mesh = {Aedes/microbiology ; Animals ; Female ; *Host-Pathogen Interactions ; Lepidoptera/microbiology ; Male ; Sex Ratio ; Symbiosis ; Wolbachia/*physiology ; }, abstract = {A common symbiont of insects, the bacterium Wolbachia has been implicated in phenomena as diverse as sex determination, pathogen defence and speciation and is being used in public health programs to prevent mosquitoes transmitting disease. Despite decades of research, we know remarkably little about how it exerts its effects.}, } @article {pmid27760142, year = {2016}, author = {Etebari, K and Asad, S and Zhang, G and Asgari, S}, title = {Identification of Aedes aegypti Long Intergenic Non-coding RNAs and Their Association with Wolbachia and Dengue Virus Infection.}, journal = {PLoS neglected tropical diseases}, volume = {10}, number = {10}, pages = {e0005069}, pmid = {27760142}, issn = {1935-2735}, mesh = {Aedes/*genetics/microbiology/virology ; Animals ; Dengue Virus/*physiology ; Genome, Insect ; *Host-Pathogen Interactions/genetics ; Mosquito Vectors/*genetics/microbiology/virology ; Pest Control, Biological ; RNA Interference ; RNA, Long Noncoding/*genetics/isolation & purification/metabolism ; Transcriptome ; Virus Replication ; Wolbachia/*physiology ; }, abstract = {Long intergenic non-coding RNAs (lincRNAs) are appearing as an important class of regulatory RNAs with a variety of biological functions. The aim of this study was to identify the lincRNA profile in the dengue vector Aedes aegypti and evaluate their potential role in host-pathogen interaction. The majority of previous RNA-Seq transcriptome studies in Ae. aegypti have focused on the expression pattern of annotated protein coding genes under different biological conditions. Here, we used 35 publically available RNA-Seq datasets with relatively high depth to screen the Ae. aegypti genome for lincRNA discovery. This led to the identification of 3,482 putative lincRNAs. These lincRNA genes displayed a slightly lower GC content and shorter transcript lengths compared to protein-encoding genes. Ae. aegypti lincRNAs also demonstrate low evolutionary sequence conservation even among closely related species such as Culex quinquefasciatus and Anopheles gambiae. We examined their expression in dengue virus serotype 2 (DENV-2) and Wolbachia infected and non-infected adult mosquitoes and Aa20 cells. The results revealed that DENV-2 infection increased the abundance of a number of host lincRNAs, from which some suppress viral replication in mosquito cells. RNAi-mediated silencing of lincRNA_1317 led to enhancement in viral replication, which possibly indicates its potential involvement in the host anti-viral defense. A number of lincRNAs were also differentially expressed in Wolbachia-infected mosquitoes. The results will facilitate future studies to unravel the function of lncRNAs in insects and may prove to be beneficial in developing new ways to control vectors or inhibit replication of viruses in them.}, } @article {pmid27734242, year = {2016}, author = {Zhang, X and Tang, S and Cheke, RA and Zhu, H}, title = {Modeling the Effects of Augmentation Strategies on the Control of Dengue Fever With an Impulsive Differential Equation.}, journal = {Bulletin of mathematical biology}, volume = {78}, number = {10}, pages = {1968-2010}, doi = {10.1007/s11538-016-0208-7}, pmid = {27734242}, issn = {1522-9602}, support = {//CIHR/Canada ; }, mesh = {Aedes/microbiology/virology ; Animals ; *Biological Control Agents ; Computer Simulation ; Dengue/*prevention & control/transmission ; Female ; Humans ; Male ; Mathematical Concepts ; Models, Biological ; Mosquito Vectors/microbiology/virology ; Sex Ratio ; Wolbachia/physiology ; }, abstract = {Dengue fever has rapidly become the world's most common vector-borne viral disease. Use of endosymbiotic Wolbachia is an innovative technology to prevent vector mosquitoes from reproducing and so break the cycle of dengue transmission. However, strategies such as population eradication and replacement will only succeed if appropriate augmentations with Wolbachia-infected mosquitoes that take account of a variety of factors are carried out. Here, we describe the spread of Wolbachia in mosquito populations using an impulsive differential system with four state variables, incorporating the effects of cytoplasmic incompatibility and the augmentation of Wolbachia-infected mosquitoes with different sex ratios. We then evaluated (a) how each parameter value contributes to the success of population replacement; (b) how different release quantities of infected mosquitoes with different sex ratios affect the success of population suppression or replacement; and (c) how the success of these two strategies can be realized to block the transmission of dengue fever. Analysis of the system's stability, bifurcations and sensitivity reveals the existence of forward and backward bifurcations, multiple attractors and the contribution of each parameter to the success of the strategies. The results indicate that the initial density of mosquitoes, the quantities of mosquitoes released in augmentations and their sex ratios have impacts on whether or not the strategies of population suppression or replacement can be achieved. Therefore, successful strategies rely on selecting suitable strains of Wolbachia and carefully designing the mosquito augmentation program.}, } @article {pmid27752109, year = {2016}, author = {Tamarozzi, F and Turner, JD and Pionnier, N and Midgley, A and Guimaraes, AF and Johnston, KL and Edwards, SW and Taylor, MJ}, title = {Wolbachia endosymbionts induce neutrophil extracellular trap formation in human onchocerciasis.}, journal = {Scientific reports}, volume = {6}, number = {}, pages = {35559}, pmid = {27752109}, issn = {2045-2322}, support = {MR/L018756/1/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Animals ; Cell-Derived Microparticles/metabolism ; Doxycycline/pharmacology/therapeutic use ; Extracellular Traps/drug effects/*metabolism ; Female ; Fluorescent Antibody Technique ; Humans ; Lipopeptides/metabolism ; Mice ; Mice, Inbred C57BL ; Microspheres ; Neutrophil Infiltration/drug effects ; Neutrophils/drug effects/*metabolism ; Onchocerciasis/drug therapy/*microbiology/pathology ; *Symbiosis/drug effects ; Toll-Like Receptor 6/metabolism ; Wolbachia/drug effects/*physiology ; }, abstract = {The endosymbiotic bacteria, Wolbachia, induce neutrophilic responses to the human helminth pathogen Onchocerca volvulus. The formation of Neutrophil Extracellular Traps (NETs), has been implicated in anti-microbial defence, but has not been identified in human helminth infection. Here, we demonstrate NETs formation in human onchocerciasis. Extracellular NETs and neutrophils were visualised around O. volvulus in nodules excised from untreated patients but not in nodules from patients treated with the anti-Wolbachia drug, doxycycline. Whole Wolbachia or microspheres coated with a synthetic Wolbachia lipopeptide (WoLP) of the major nematode Wolbachia TLR2/6 ligand, peptidoglycan associated lipoprotein, induced NETosis in human neutrophils in vitro. TLR6 dependency of Wolbachia and WoLP NETosis was demonstrated using purified neutrophils from TLR6 deficient mice. Thus, we demonstrate for the first time that NETosis occurs during natural human helminth infection and demonstrate a mechanism of NETosis induction via Wolbachia endobacteria and direct ligation of Wolbachia lipoprotein by neutrophil TLR2/6.}, } @article {pmid27748992, year = {2017}, author = {Hunter, MS and Asiimwe, P and Himler, AG and Kelly, SE}, title = {Host nuclear genotype influences phenotype of a conditional mutualist symbiont.}, journal = {Journal of evolutionary biology}, volume = {30}, number = {1}, pages = {141-149}, doi = {10.1111/jeb.12993}, pmid = {27748992}, issn = {1420-9101}, mesh = {Animals ; Female ; Genetic Variation ; *Genotype ; *Hemiptera ; *Phenotype ; *Rickettsia ; *Symbiosis ; }, abstract = {Arthropods commonly carry maternally inherited intracellular bacterial symbionts that may profoundly influence host biology and evolution. The intracellular symbiont Rickettsia sp. nr. bellii swept rapidly into populations of the sweetpotato whitefly Bemisia tabaci in the south-western USA. Previous laboratory experiments showed female-bias and fitness benefits were associated with Rickettsia infection, potentially explaining the high frequencies of infection observed in field populations, but the effects varied with whitefly genetic line. Here, we explored whether host extranuclear or nuclear genes influenced the variation in the Rickettsia-host phenotype in two genetic lines of the whitefly host, each with Rickettsia-infected and uninfected sublines. Introgression between the Rickettsia-infected subline of one genetic line and the Rickettsia-uninfected subline of the other was used to create two new sublines, each with the maternally inherited extranuclear genetic lineages of one line (Rickettsia, two other symbionts and the mitochondria) and the nuclear genotype of the other. Performance assays comparing the original and new lines showed that in addition to Rickettsia, the interaction of Rickettsia infection with host nuclear genotype influenced development time and the sex ratio of the progeny, whereas the extranuclear genotype did not. Host nuclear genotype, but not extranuclear genotype, also influenced the titre of Rickettsia. Our results support the hypothesis that differences in host nuclear genotype alone may explain considerable within-population variation in host-symbiont phenotype and may contribute to the observed variation in Rickettsia-whitefly interactions worldwide.}, } @article {pmid27738153, year = {2016}, author = {Servick, K}, title = {Winged warriors.}, journal = {Science (New York, N.Y.)}, volume = {354}, number = {6309}, pages = {164-167}, doi = {10.1126/science.354.6309.164}, pmid = {27738153}, issn = {1095-9203}, mesh = {Aedes/*genetics/*microbiology ; Animals ; Animals, Genetically Modified/*genetics ; Brazil ; Chikungunya Fever/prevention & control ; Communicable Disease Control/*methods ; Dengue/prevention & control ; Female ; Humans ; Male ; Mosquito Control/*methods ; Wolbachia ; Zika Virus Infection/prevention & control ; }, } @article {pmid27734894, year = {2016}, author = {Brown, AM and Wasala, SK and Howe, DK and Peetz, AB and Zasada, IA and Denver, DR}, title = {Genomic evidence for plant-parasitic nematodes as the earliest Wolbachia hosts.}, journal = {Scientific reports}, volume = {6}, number = {}, pages = {34955}, pmid = {27734894}, issn = {2045-2322}, mesh = {Animals ; Biological Evolution ; DNA, Bacterial/genetics ; Female ; Gene Library ; *Genome, Bacterial ; Genomics ; Homeostasis ; In Situ Hybridization, Fluorescence ; Iron/metabolism ; Likelihood Functions ; Male ; Nematoda/*microbiology ; Phenotype ; Phylogeny ; Plants/*parasitology ; *Symbiosis ; Wolbachia/*physiology ; }, abstract = {Wolbachia, one of the most widespread endosymbionts, is a target for biological control of mosquito-borne diseases (malaria and dengue virus), and antibiotic elimination of infectious filarial nematodes. We sequenced and analyzed the genome of a new Wolbachia strain (wPpe) in the plant-parasitic nematode Pratylenchus penetrans. Phylogenomic analyses placed wPpe as the earliest diverging Wolbachia, suggesting two evolutionary invasions into nematodes. The next branches comprised strains in sap-feeding insects, suggesting Wolbachia may have first evolved as a nutritional mutualist. Genome size, protein content, %GC, and repetitive DNA allied wPpe with mutualistic Wolbachia, whereas gene repertoire analyses placed it between parasite (A, B) and mutualist (C, D, F) groups. Conservation of iron metabolism genes across Wolbachia suggests iron homeostasis as a potential factor in its success. This study enhances our understanding of this globally pandemic endosymbiont, highlighting genetic patterns associated with host changes. Combined with future work on this strain, these genomic data could help provide potential new targets for plant-parasitic nematode control.}, } @article {pmid27731327, year = {2017}, author = {Kemp, DJ and Thomson, FE and Edwards, W and Iturbe-Ormaetxe, I}, title = {Incomplete offspring sex bias in Australian populations of the butterfly Eurema hecabe.}, journal = {Heredity}, volume = {118}, number = {3}, pages = {284-292}, pmid = {27731327}, issn = {1365-2540}, mesh = {Animals ; Australia ; Butterflies/*genetics/microbiology ; Female ; Male ; Pedigree ; *Sex Ratio ; Wolbachia ; }, abstract = {Theory predicts unified sex ratios for most organisms, yet biases may be engendered by selfish genetic elements such as endosymbionts that kill or feminize individuals with male genotypes. Although rare, feminization is established for Wolbachia-infected Eurema butterflies. This paradigm is presently confined to islands in the southern Japanese archipelago, where feminized phenotypes produce viable all-daughter broods. Here, we characterize sex bias for E. hecabe in continental Australia. Starting with 186 wild-caught females, we reared >6000 F1-F3 progeny in pedigree designs that incorporated selective antibiotic treatments. F1 generations expressed a consistent bias across 2 years and populations that was driven by an ~5% incidence of broods comprising ⩾80% daughters. Females from biased lineages continued to overproduce daughters over two generations of outcrossing to wild males. Treatment with antibiotics of differential strength influenced sex ratio only in biased lineages by inducing an equivalent incomplete degree of son overproduction. Brood sex ratios were nevertheless highly variable within lineages and across generations. Intriguingly, the cytogenetic signature of female karyotype was uniformly absent, even among phenotypic females in unbiased lineages. Molecular evidence supported the existence of a single Wolbachia strain at high prevalence, yet this was not clearly linked to brood sex bias. In sum, we establish an inherited, experimentally reversible tendency for incomplete offspring bias. Key features of our findings clearly depart from the Japanese feminization paradigm and highlight the potential for more subtle degrees of sex distortion in arthropods.}, } @article {pmid27727237, year = {2016}, author = {Bordenstein, SR and Bordenstein, SR}, title = {Eukaryotic association module in phage WO genomes from Wolbachia.}, journal = {Nature communications}, volume = {7}, number = {}, pages = {13155}, pmid = {27727237}, issn = {2041-1723}, support = {R01 GM085163/GM/NIGMS NIH HHS/United States ; R21 HD086833/HD/NICHD NIH HHS/United States ; }, mesh = {Ankyrin Repeat ; Bacteriophages/*genetics ; Conserved Sequence ; Eukaryota/*metabolism ; Furin/metabolism ; Gene Transfer, Horizontal ; Genes, Viral ; *Genome, Viral ; Models, Biological ; Phylogeny ; Spider Venoms/chemistry ; Tetratricopeptide Repeat ; Viral Proteins/metabolism ; Wolbachia/*virology ; }, abstract = {Viruses are trifurcated into eukaryotic, archaeal and bacterial categories. This domain-specific ecology underscores why eukaryotic viruses typically co-opt eukaryotic genes and bacteriophages commonly harbour bacterial genes. However, the presence of bacteriophages in obligate intracellular bacteria of eukaryotes may promote DNA transfers between eukaryotes and bacteriophages. Here we report a metagenomic analysis of purified bacteriophage WO particles of Wolbachia and uncover a eukaryotic association module in the complete WO genome. It harbours predicted domains, such as the black widow latrotoxin C-terminal domain, that are uninterrupted in bacteriophage genomes, enriched with eukaryotic protease cleavage sites and combined with additional domains to forge one of the largest bacteriophage genes to date (14,256 bp). To the best of our knowledge, these eukaryotic-like domains have never before been reported in packaged bacteriophages and their phylogeny, distribution and sequence diversity imply lateral transfers between bacteriophage/prophage and animal genomes. Finally, the WO genome sequences and identification of attachment sites will potentially advance genetic manipulation of Wolbachia.}, } @article {pmid27726483, year = {2017}, author = {Leite, GL and de Paulo, PD and Zanuncio, JC and Tavares, WS and Alvarenga, AC and Dourado, LR and Bispo, EP and Soares, MA}, title = {Herbicide toxicity, selectivity and hormesis of nicosulfuron on 10 Trichogrammatidae (Hymenoptera) species parasitizing Anagasta (= Ephestia) kuehniella (Lepidoptera: Pyralidae) eggs.}, journal = {Journal of environmental science and health. Part. B, Pesticides, food contaminants, and agricultural wastes}, volume = {52}, number = {1}, pages = {70-76}, doi = {10.1080/03601234.2016.1229476}, pmid = {27726483}, issn = {1532-4109}, mesh = {Animals ; Female ; Herbicides/*toxicity ; Hormesis/*drug effects ; Hymenoptera/*drug effects ; Lepidoptera/drug effects/*parasitology ; Male ; Moths ; Ovum/drug effects/parasitology ; Pest Control, Biological ; Pyridines/*toxicity ; Sex Ratio ; Sulfonylurea Compounds/*toxicity ; }, abstract = {Selective agrochemicals including herbicides that do not affect non-target organisms such as natural enemies are important in the integrated pest management (IPM) programs. The aim of this study was to evaluate the herbicide toxicity, selectivity and hormesis of nicosulfuron, recommended for the corn Zea mays L. (Poaceae) crop, on 10 Trichogrammatidae (Hymenoptera) species. A female of each Trichogramma spp. or Trichogrammatoidea annulata De Santis, 1972 was individually placed in plastic test tubes (no choice) with a cardboard containing 45 flour moth Anagasta (= Ephestia) kuehniella Zeller, 1879 (Lepidoptera: Pyralidae) eggs. Parasitism by these natural enemies was allowed for 48 h and the cardboards were sprayed with the herbicide nicosulfuron at 1.50 L.ha[-1], along with the control (only distilled water). Nicosulfuron reduced the emergence rate of Trichogramma bruni Nagaraja, 1983 females, but increased that of Trichogramma pretiosum Riley, 1879, Trichogramma acacioi Brun, Moraes and Smith, 1984 and T. annulata females. Conversely, this herbicide increased the emergence rate of Trichogramma brasiliensis Ashmead, 1904, T. bruni, Trichogramma galloi Zucchi, 1988 and Trichogramma soaresi Nagaraja, 1983 males and decreased those of T. acacioi, Trichogramma atopovilia Oatman and Platner, 1983 and T. pretiosum males. In addition, nicosulfuron reduced the sex ratio of T. galloi, Trichogramma bennetti Nagaraja and Nagarkatti, 1973 and T. pretiosum and increased that of T. acacioi, T. bruni, T. annulata, Trichogramma demoraesi Nagaraja, 1983, T. soaresi and T. brasiliensis. The herbicide nicosulfuron was "harmless" (class 1, <30% reduction) for females and the sex ratio of all Trichogrammatidae species based on the International Organization for Biological Control (IOBC) classification. The possible hormesis effect of nicosulfuron on Trichogrammatidae species and on the bacterium Wolbachia sp. (Rickettsiales: Rickettsiaceae) was also discussed.}, } @article {pmid27721807, year = {2016}, author = {Dittmer, J and van Opstal, EJ and Shropshire, JD and Bordenstein, SR and Hurst, GD and Brucker, RM}, title = {Disentangling a Holobiont - Recent Advances and Perspectives in Nasonia Wasps.}, journal = {Frontiers in microbiology}, volume = {7}, number = {}, pages = {1478}, pmid = {27721807}, issn = {1664-302X}, abstract = {The parasitoid wasp genus Nasonia (Hymenoptera: Chalcidoidea) is a well-established model organism for insect development, evolutionary genetics, speciation, and symbiosis. The host-microbiota assemblage which constitutes the Nasonia holobiont (a host together with all of its associated microbes) consists of viruses, two heritable bacterial symbionts and a bacterial community dominated in abundance by a few taxa in the gut. In the wild, all four Nasonia species are systematically infected with the obligate intracellular bacterium Wolbachia and can additionally be co-infected with Arsenophonus nasoniae. These two reproductive parasites have different transmission modes and host manipulations (cytoplasmic incompatibility vs. male-killing, respectively). Pioneering studies on Wolbachia in Nasonia demonstrated that closely related Nasonia species harbor multiple and mutually incompatible Wolbachia strains, resulting in strong symbiont-mediated reproductive barriers that evolved early in the speciation process. Moreover, research on host-symbiont interactions and speciation has recently broadened from its historical focus on heritable symbionts to the entire microbial community. In this context, each Nasonia species hosts a distinguishable community of gut bacteria that experiences a temporal succession during host development and members of this bacterial community cause strong hybrid lethality during larval development. In this review, we present the Nasonia species complex as a model system to experimentally investigate questions regarding: (i) the impact of different microbes, including (but not limited to) heritable endosymbionts, on the extended phenotype of the holobiont, (ii) the establishment and regulation of a species-specific microbiota, (iii) the role of the microbiota in speciation, and (iv) the resilience and adaptability of the microbiota in wild populations subjected to different environmental pressures. We discuss the potential for easy microbiota manipulations in Nasonia as a promising experimental approach to address these fundamental aspects.}, } @article {pmid27721806, year = {2016}, author = {Bouchon, D and Zimmer, M and Dittmer, J}, title = {The Terrestrial Isopod Microbiome: An All-in-One Toolbox for Animal-Microbe Interactions of Ecological Relevance.}, journal = {Frontiers in microbiology}, volume = {7}, number = {}, pages = {1472}, pmid = {27721806}, issn = {1664-302X}, abstract = {Bacterial symbionts represent essential drivers of arthropod ecology and evolution, influencing host traits such as nutrition, reproduction, immunity, and speciation. However, the majority of work on arthropod microbiota has been conducted in insects and more studies in non-model species across different ecological niches will be needed to complete our understanding of host-microbiota interactions. In this review, we present terrestrial isopod crustaceans as an emerging model organism to investigate symbiotic associations with potential relevance to ecosystem functioning. Terrestrial isopods comprise a group of crustaceans that have evolved a terrestrial lifestyle and represent keystone species in terrestrial ecosystems, contributing to the decomposition of organic matter and regulating the microbial food web. Since their nutrition is based on plant detritus, it has long been suspected that bacterial symbionts located in the digestive tissues might play an important role in host nutrition via the provisioning of digestive enzymes, thereby enabling the utilization of recalcitrant food compounds (e.g., cellulose or lignins). If this were the case, then (i) the acquisition of these bacteria might have been an important evolutionary prerequisite for the colonization of land by isopods, and (ii) these bacterial symbionts would directly mediate the role of their hosts in ecosystem functioning. Several bacterial symbionts have indeed been discovered in the midgut caeca of terrestrial isopods and some of them might be specific to this group of animals (i.e., Candidatus Hepatoplasma crinochetorum, Candidatus Hepatincola porcellionum, and Rhabdochlamydia porcellionis), while others are well-known intracellular pathogens (Rickettsiella spp.) or reproductive parasites (Wolbachia sp.). Moreover, a recent investigation of the microbiota in Armadillidium vulgare has revealed that this species harbors a highly diverse bacterial community which varies between host populations, suggesting an important share of environmental microbes in the host-associated microbiota. In this review, we synthesize our current knowledge on the terrestrial isopod microbiome and identify future directions to (i) fully understand the functional roles of particular bacteria (both intracellular or intestinal symbionts and environmental gut passengers), and (ii) whether and how the host-associated microbiota could influence the performance of terrestrial isopods as keystone species in soil ecosystems.}, } @article {pmid27713560, year = {2016}, author = {Kumar, N and Lin, M and Zhao, X and Ott, S and Santana-Cruz, I and Daugherty, S and Rikihisa, Y and Sadzewicz, L and Tallon, LJ and Fraser, CM and Dunning Hotopp, JC}, title = {Efficient Enrichment of Bacterial mRNA from Host-Bacteria Total RNA Samples.}, journal = {Scientific reports}, volume = {6}, number = {}, pages = {34850}, pmid = {27713560}, issn = {2045-2322}, support = {DP2 OD007372/OD/NIH HHS/United States ; HHSN272200900009C/AI/NIAID NIH HHS/United States ; R01 AI047885/AI/NIAID NIH HHS/United States ; U19 AI110820/AI/NIAID NIH HHS/United States ; }, mesh = {Animals ; Brugia malayi/microbiology ; Drosophila/microbiology ; Ehrlichia chaffeensis/genetics ; *Genetic Techniques ; High-Throughput Nucleotide Sequencing/methods ; Host-Pathogen Interactions/*genetics ; Microfluidic Analytical Techniques ; Poly A/chemistry ; RNA, Bacterial/chemistry/*isolation & purification ; RNA, Messenger/chemistry/*isolation & purification ; RNA, Ribosomal/chemistry ; Reverse Transcriptase Polymerase Chain Reaction/methods ; Symbiosis ; Wolbachia/genetics/pathogenicity ; Wolves/microbiology ; }, abstract = {Despite numerous advances in genomics and bioinformatics, technological hurdles remain to examine host-microbe transcriptomics. Sometimes the transcriptome of either or both can be ascertained merely by generating more sequencing reads. However, many cases exist where bacterial mRNA needs to be enriched further to enable cost-effective sequencing of the pathogen or endosymbiont. While a suitable method is commercially available for mammalian samples of this type, development of such methods has languished for invertebrate samples. Furthermore, a common method across multiple taxa would facilitate comparisons between bacteria in invertebrate vectors and their vertebrate hosts. Here, a method is described to concurrently remove polyadenylated transcripts, prokaryotic rRNA, and eukaryotic rRNA, including those with low amounts of starting material (e.g. 100 ng). In a Wolbachia-Drosophila system, this bacterial mRNA enrichment yielded a 3-fold increase in Wolbachia mRNA abundance and a concomitant 3.3-fold increase in the percentage of transcripts detected. More specifically, 70% of the genome could be recovered by transcriptome sequencing compared to 21% in the total RNA. Sequencing of similar bacterial mRNA-enriched samples generated from Ehrlichia-infected canine cells covers 93% of the Ehrlichia genome, suggesting ubiquitous transcription across the entire Ehrlichia chaffeensis genome. This technique can potentially be used to enrich bacterial mRNA in many studies of host-microbe interactions.}, } @article {pmid27708633, year = {2016}, author = {Chen, S and Zhao, J and Joshi, D and Xi, Z and Norman, B and Walker, ED}, title = {Persistent Infection by Wolbachia wAlbB Has No Effect on Composition of the Gut Microbiota in Adult Female Anopheles stephensi.}, journal = {Frontiers in microbiology}, volume = {7}, number = {}, pages = {1485}, pmid = {27708633}, issn = {1664-302X}, support = {R37 AI021884/AI/NIAID NIH HHS/United States ; }, abstract = {The bacteria in the midgut of Anopheles stephensi adult females from laboratory colonies were studied by sequencing the V4 region of 16S rRNA genes, with respect to three experimental factors: stable or cured Wolbachia infection; sugar or blood diet; and age. Proteobacteria and Bacteroidetes dominated the community [>90% of operational taxonomic units (OTUs)]; most taxa were in the classes Flavobacteriia, Gammaproteobacteria, and Alphaproteobacteria, and were assigned to Elizabethkingia (46.9%), Asaia (6.4%) and Pseudomonas (6.0%), or unclassified Enterobacteriaceae (37.2%). Bacterial communities were similar between Wolbachia-cured and Wolbachia-infected mosquito lines, indicating that the gut microbiota were not dysregulated in the presence of Wolbachia. The proportion of Enterobacteriaceae was higher in mosquitoes fed a blood meal compared to those provided a sugar meal. Collectively, the bacterial community had a similar structure in older Wolbachia-infected mosquitoes 8 days after the blood meal, as in younger Wolbachia-infected mosquitoes before a blood meal, except that older mosquitoes had a higher proportion of Enterobacteriaceae and lower proportion of Elizabethkingia. Consistent presence of certain predominant bacteria (Elizabethkingia, Asaia, Pseudomonas, and Enterobacteriaceae) suggests they would be useful for paratransgenesis to control malaria infection, particularly when coupled to a Wolbachia-based intervention strategy.}, } @article {pmid27707863, year = {2016}, author = {van Nouhuys, S and Kohonen, M and Duplouy, A}, title = {Wolbachia increases the susceptibility of a parasitoid wasp to hyperparasitism.}, journal = {The Journal of experimental biology}, volume = {219}, number = {Pt 19}, pages = {2984-2990}, doi = {10.1242/jeb.140699}, pmid = {27707863}, issn = {1477-9145}, mesh = {Animals ; Butterflies/parasitology ; Disease Resistance ; Estonia ; Finland ; Host-Pathogen Interactions ; Larva/parasitology ; Parasites/*microbiology ; Species Specificity ; Virulence ; Wasps/*microbiology/pathogenicity ; Wolbachia/*physiology ; }, abstract = {The success of maternally transmitted endosymbiotic bacteria, such as Wolbachia, is directly linked to their host reproduction but in direct conflict with other parasites that kill the host before it reaches reproductive maturity. Therefore, symbionts that have evolved strategies to increase their host's ability to evade lethal parasites may have high penetrance, while detrimental symbionts would be selected against, leading to lower penetrance or extinction from the host population. In a natural population of the parasitoid wasp Hyposoter horticola in the Åland Islands (Finland), the Wolbachia strain wHho persists at an intermediate prevalence (∼50%). Additionally, there is a negative correlation between the prevalence of Wolbachia and a hyperparasitoid wasp, Mesochorus cf. stigmaticus, in the landscape. Using a manipulative field experiment, we addressed the persistence of Wolbachia at this intermediate level, and tested whether the observed negative correlation could be due to Wolbachia inducing either susceptibility or resistance to parasitism. We show that infection with Wolbachia does not influence the ability of the wasp to parasitize its butterfly host, Melitaea cinxia, but that hyperparasitism of the wasp increases in the presence of wHho. Consequently, the symbiont is detrimental, and in order to persist in the host population, must also have a positive effect on fitness that outweighs the costly burden of susceptibility to widespread parasitism.}, } @article {pmid27706105, year = {2016}, author = {Benelli, G and Jeffries, CL and Walker, T}, title = {Biological Control of Mosquito Vectors: Past, Present, and Future.}, journal = {Insects}, volume = {7}, number = {4}, pages = {}, pmid = {27706105}, issn = {2075-4450}, support = {//Wellcome Trust/United Kingdom ; 101285//Wellcome Trust/United Kingdom ; }, abstract = {Mosquitoes represent the major arthropod vectors of human disease worldwide transmitting malaria, lymphatic filariasis, and arboviruses such as dengue virus and Zika virus. Unfortunately, no treatment (in the form of vaccines or drugs) is available for most of these diseases andvectorcontrolisstillthemainformofprevention. Thelimitationsoftraditionalinsecticide-based strategies, particularly the development of insecticide resistance, have resulted in significant efforts to develop alternative eco-friendly methods. Biocontrol strategies aim to be sustainable and target a range of different mosquito species to reduce the current reliance on insecticide-based mosquito control. In thisreview, weoutline non-insecticide basedstrategiesthat havebeenimplemented orare currently being tested. We also highlight the use of mosquito behavioural knowledge that can be exploited for control strategies.}, } @article {pmid27703857, year = {2016}, author = {García-Vázquez, D and Ribera, I}, title = {The origin of widespread species in a poor dispersing lineage (diving beetle genus Deronectes).}, journal = {PeerJ}, volume = {4}, number = {}, pages = {e2514}, pmid = {27703857}, issn = {2167-8359}, abstract = {In most lineages, most species have restricted geographic ranges, with only few reaching widespread distributions. How these widespread species reached their current ranges is an intriguing biogeographic and evolutionary question, especially in groups known to be poor dispersers. We reconstructed the biogeographic and temporal origin of the widespread species in a lineage with particularly poor dispersal capabilities, the diving beetle genus Deronectes (Dytiscidae). Most of the ca. 60 described species of Deronectes have narrow ranges in the Mediterranean area, with only four species with widespread European distributions. We sequenced four mitochondrial and two nuclear genes of 297 specimens of 109 different populations covering the entire distribution of the four lineages of Deronectes, including widespread species. Using Bayesian probabilities with an a priori evolutionary rate, we performed (1) a global phylogeny/phylogeography to estimate the relationships of the main lineages within each group and root them, and (2) demographic analyses of the best population coalescent model for each species group, including a reconstruction of the geographical history estimated from the distribution of the sampled localities. We also selected 56 specimens to test for the presence of Wolbachia, a maternally transmitted parasite that can alter the patterns of mtDNA variability. All species of the four studied groups originated in the southern Mediterranean peninsulas and were estimated to be of Pleistocene origin. In three of the four widespread species, the central and northern European populations were nested within those in the northern areas of the Anatolian, Balkan and Iberian peninsulas respectively, suggesting a range expansion at the edge of the southern refugia. In the Mediterranean peninsulas the widespread European species were replaced by vicariant taxa of recent origin. The fourth species (D. moestus) was proven to be a composite of unrecognised lineages with more restricted distributions around the Western and Central Mediterranean. The analysis of Wolbachia showed a high prevalence of infection among Deronectes, especially in the D. aubei group, where all sequenced populations were infected with the only exception of the Cantabrian Mountains, the westernmost area of distribution of the lineage. In this group there was a phylogenetic incongruence between the mitochondrial and the nuclear sequence, although no clear pattern links this discordance to the Wolbachia infection. Our results suggest that, in different glacial cycles, populations that happened to be at the edge of the newly deglaciated areas took advantage of the optimal ecological conditions to expand their ranges to central and northern Europe. Once this favourable ecological window ended populations become isolated, resulting in the presence of closely related but distinct species in the Mediterranean peninsulas.}, } @article {pmid27693536, year = {2016}, author = {Kuesel, AC}, title = {Research for new drugs for elimination of onchocerciasis in Africa.}, journal = {International journal for parasitology. Drugs and drug resistance}, volume = {6}, number = {3}, pages = {272-286}, pmid = {27693536}, issn = {2211-3207}, support = {001/WHO_/World Health Organization/International ; }, mesh = {Africa/epidemiology ; Animals ; *Disease Eradication ; Drug Approval ; Drug Discovery/*trends ; Drug Therapy/*methods ; Filaricides/*isolation & purification/*therapeutic use ; Onchocerciasis/*epidemiology/*prevention & control ; }, abstract = {Onchocerciasis is a parasitic, vector borne disease caused by the filarial nematode Onchocerca volvulus. More than 99% of the population at risk of infection live in Africa. Onchocerciasis control was initiated in West Africa in 1974 with vector control, later complemented by ivermectin mass drug administration and in the other African endemic countries in 1995 with annual community directed treatment with ivermectin (CDTI.) This has significantly reduced infection prevalence. Together with proof-of-concept for onchocerciasis elimination with annual CDTI from foci in Senegal and Mali, this has resulted in targeting onchocerciasis elimination in selected African countries by 2020 and in 80% of African countries by 2025. The challenges for meeting these targets include the number of endemic countries where conflict has delayed or interrupted control programmes, cross-border foci, potential emergence of parasite strains with low susceptibility to ivermectin and co-endemicity of loiasis, another parasitic vector borne disease, which slows down or prohibits CDTI implementation. Some of these challenges could be addressed with new drugs or drug combinations with a higher effect on Onchocerca volvulus than ivermectin. This paper reviews the path from discovery of new compounds to their qualification for large scale use and the support regulatory authorities provide for development of drugs for neglected tropical diseases. The status of research for new drugs or treatment regimens for onchocerciasis along the path to regulatory approval and qualification for large scale use is reviewed. This research includes new regimens and combinations of ivermectin and albendazole, antibiotics targeting the O. volvulus endosymbiont Wolbachia, flubendazole, moxidectin and emodepside and discovery of new compounds.}, } @article {pmid27692043, year = {2016}, author = {Schnettler, E and Sreenu, VB and Mottram, T and McFarlane, M}, title = {Wolbachia restricts insect-specific flavivirus infection in Aedes aegypti cells.}, journal = {The Journal of general virology}, volume = {97}, number = {11}, pages = {3024-3029}, pmid = {27692043}, issn = {1465-2099}, support = {MC_UU_12014/8/MRC_/Medical Research Council/United Kingdom ; MC_UU_12014/1/MRC_/Medical Research Council/United Kingdom ; G0801822/MRC_/Medical Research Council/United Kingdom ; MC_UP_A550_1031/MRC_/Medical Research Council/United Kingdom ; MC_UU_12014/12/MRC_/Medical Research Council/United Kingdom ; }, mesh = {Aedes/*microbiology/*virology ; Animals ; Drosophila/microbiology ; Flavivirus/*physiology ; Insect Vectors/microbiology/virology ; Species Specificity ; Wolbachia/*physiology ; }, abstract = {Mosquito-borne viruses are known to cause disease in humans and livestock and are often difficult to control due to the lack of specific antivirals and vaccines. The Wolbachia endosymbiont has been widely studied for its ability to restrict positive-strand RNA virus infection in mosquitoes, although little is known about the precise antiviral mechanism. In recent years, a variety of insect-specific viruses have been discovered in mosquitoes and an interaction with mosquito-borne viruses has been reported for some of them; however, nothing is known about the effect of Wolbachia on insect-specific virus infection in mosquitoes. Here, we show that transinfection of the Drosophila-derived wMelPop Wolbachia strain into Aedes aegypti-derived cells resulted in inhibition and even clearance of the persistent cell-fusing agent flavivirus infection in these cells. This broadens the antiviral activity of Wolbachia from acute infections to persistent infections and from arboviruses to mosquito-specific viruses. In contrast, no effect on the Phasi Charoen-like bunyavirus persistent infection in these cells was observed, suggesting a difference in Wolbachia inhibition between positive- and negative-strand RNA viruses.}, } @article {pmid27684942, year = {2016}, author = {Faria, VG and Martins, NE and Magalhães, S and Paulo, TF and Nolte, V and Schlötterer, C and Sucena, É and Teixeira, L}, title = {Drosophila Adaptation to Viral Infection through Defensive Symbiont Evolution.}, journal = {PLoS genetics}, volume = {12}, number = {9}, pages = {e1006297}, pmid = {27684942}, issn = {1553-7404}, support = {P 27630/FWF_/Austrian Science Fund FWF/Austria ; }, abstract = {Microbial symbionts can modulate host interactions with biotic and abiotic factors. Such interactions may affect the evolutionary trajectories of both host and symbiont. Wolbachia protects Drosophila melanogaster against several viral infections and the strength of the protection varies between variants of this endosymbiont. Since Wolbachia is maternally transmitted, its fitness depends on the fitness of its host. Therefore, Wolbachia populations may be under selection when Drosophila is subjected to viral infection. Here we show that in D. melanogaster populations selected for increased survival upon infection with Drosophila C virus there is a strong selection coefficient for specific Wolbachia variants, leading to their fixation. Flies carrying these selected Wolbachia variants have higher survival and fertility upon viral infection when compared to flies with the other variants. These findings demonstrate how the interaction of a host with pathogens shapes the genetic composition of symbiont populations. Furthermore, host adaptation can result from the evolution of its symbionts, with host and symbiont functioning as a single evolutionary unit.}, } @article {pmid27681542, year = {2016}, author = {Mohanty, I and Rath, A and Mahapatra, N and Hazra, RK}, title = {Wolbachia: A biological control strategy against arboviral diseases.}, journal = {Journal of vector borne diseases}, volume = {53}, number = {3}, pages = {199-207}, pmid = {27681542}, issn = {0972-9062}, mesh = {Animals ; Antibiosis ; Arbovirus Infections/epidemiology/*prevention & control ; Arboviruses/*growth & development ; Culicidae/*microbiology ; Disease Transmission, Infectious/*prevention & control ; Humans ; Mosquito Control/*methods ; Wolbachia/*growth & development ; }, abstract = {Vector-borne diseases particularly those transmitted by mosquitoes like Dengue are among the leading causes of mortality and morbidity in human population. There are no effective vaccines or treatment against dengue fever till date and the control methods are limited. So, new approaches are urgently in need to reverse these trends. Vector control is currently the primary intervention tool. Strategies that reduce or block pathogen transmission by mosquitoes have been proposed as a means of augmenting current control measures to reduce the growing burden of vector-borne diseases. Wolbachia an endosymbiont of arthropod vectors is being explored as a novel ecofriendly control strategy. Studies in Drosophila have shown that Wolbachia can confer resistance to diverse RNA viruses and protect flies from virus-induced mortality. This review was focused on biology of the Wolbachia and its implication as a control measure for arboviral diseases mainly Dengue and Chikungunya.}, } @article {pmid27680689, year = {2017}, author = {Gunadi, A and Bansal, R and Finer, JJ and Michel, A}, title = {Establishment of in vitro soybean aphids, Aphis glycines (Hemiptera: Aphididae): a tool to facilitate studies of aphid symbionts, plant-insect interactions and insecticide efficacy.}, journal = {Pest management science}, volume = {73}, number = {6}, pages = {1229-1235}, doi = {10.1002/ps.4448}, pmid = {27680689}, issn = {1526-4998}, mesh = {Animals ; Aphids/drug effects/microbiology/*physiology ; Buchnera ; Enterobacteriaceae ; Insecticides/*pharmacology ; Neonicotinoids/pharmacology ; Nitro Compounds/pharmacology ; Oxazines/pharmacology ; Glycine max/*physiology ; Symbiosis ; Thiamethoxam ; Thiazoles/pharmacology ; Wolbachia ; ortho-Aminobenzoates/pharmacology ; }, abstract = {BACKGROUND: Studies on plant-insect interactions of the soybean aphid, Aphis glycines (Matsumura), can be influenced by environmental fluctuations, status of the host plant and variability in microbial populations. Maintenance of aphids on in vitro-grown plants minimizes environmental fluctuations, provides uniform host materials and permits the selective elimination of aphid-associated microbes for more standardized controls in aphid research.

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

CONCLUSIONS: The molecular detection of canine vector-borne disease (CVBD) agents, some of which are zoonotic, reinforces the need to implement efficient prophylactic measures, such as insect repellents and macrocyclic lactones (including compliance to administration), in the geographical areas where these agents are distributed, with the view to prevent infection and disease among mammalian hosts including humans.}, } @article {pmid27159727, year = {2017}, author = {Gaikwad, S and Warudkar, A and Shouche, Y}, title = {Efficacy of DNA barcoding for the species identification of spiders from Western Ghats of India.}, journal = {Mitochondrial DNA. Part A, DNA mapping, sequencing, and analysis}, volume = {28}, number = {5}, pages = {638-644}, doi = {10.3109/24701394.2016.1166219}, pmid = {27159727}, issn = {2470-1408}, mesh = {Animals ; DNA Barcoding, Taxonomic/*methods ; Electron Transport Complex IV/genetics ; Genetic Variation ; India ; Phylogeny ; Sequence Analysis, DNA/*methods ; Species Specificity ; Spiders/*classification/enzymology/genetics ; }, abstract = {DNA barcoding has emerged as an additional tool for taxonomy and as an aid to taxonomic impediments. Due to their extensive morphological variation, spiders are taxonomically challenging. Therefore, all over the world, attempts are being made to DNA barcode species of spiders. Till now no attempts were made to DNA barcode Indian spiders despite their rich diversity. We have generated DNA barcodes for 60 species (n = 112) of spiders for the first time from India. Although only 17 species were correctly identified at the species level, DNA barcoding correctly discriminated 99% of the species studied here. We have also found high intraspecies nucleotide divergence in Plexippus paykulli suggesting cryptic diversity that needs to be studied in detail. Our study also showed non-specific amplification of the Cytochrome Oxidase I (COI) gene of endosymbiont bacteria Wolbachia. However, these cases are very rare and could be resolved by the use of modified or group specific primers.}, } @article {pmid27159401, year = {2016}, author = {, }, title = {Correction: Wolbachia Blocks Viral Genome Replication Early in Infection without a Transcriptional Response by the Endosymbiont or Host Small RNA Pathways.}, journal = {PLoS pathogens}, volume = {12}, number = {5}, pages = {e1005639}, pmid = {27159401}, issn = {1553-7374}, abstract = {[This corrects the article DOI: 10.1371/journal.ppat.1005536.].}, } @article {pmid27156023, year = {2016}, author = {Dutra, HL and Rocha, MN and Dias, FB and Mansur, SB and Caragata, EP and Moreira, LA}, title = {Wolbachia Blocks Currently Circulating Zika Virus Isolates in Brazilian Aedes aegypti Mosquitoes.}, journal = {Cell host & microbe}, volume = {19}, number = {6}, pages = {771-774}, pmid = {27156023}, issn = {1934-6069}, mesh = {Aedes/*microbiology/*virology ; Animals ; Antibiosis ; Brazil ; Disease Transmission, Infectious/prevention & control ; Female ; Insect Vectors/microbiology/virology ; Male ; Saliva/microbiology/virology ; Symbiosis ; Wolbachia/*physiology ; Zika Virus/isolation & purification/*physiology ; Zika Virus Infection/microbiology/prevention & control/transmission/virology ; }, abstract = {The recent association of Zika virus with cases of microcephaly has sparked a global health crisis and highlighted the need for mechanisms to combat the Zika vector, Aedes aegypti mosquitoes. Wolbachia pipientis, a bacterial endosymbiont of insect, has recently garnered attention as a mechanism for arbovirus control. Here we report that Aedes aegypti harboring Wolbachia are highly resistant to infection with two currently circulating Zika virus isolates from the recent Brazilian epidemic. Wolbachia-harboring mosquitoes displayed lower viral prevalence and intensity and decreased disseminated infection and, critically, did not carry infectious virus in the saliva, suggesting that viral transmission was blocked. Our data indicate that the use of Wolbachia-harboring mosquitoes could represent an effective mechanism to reduce Zika virus transmission and should be included as part of Zika control strategies.}, } @article {pmid27151484, year = {2016}, author = {Suroowan, S and Mahomoodally, F and Ragoo, L}, title = {Management and Treatment of Dengue and Chikungunya - Natural Products to the Rescue.}, journal = {Combinatorial chemistry & high throughput screening}, volume = {19}, number = {7}, pages = {554-564}, doi = {10.2174/1386207319666160506123401}, pmid = {27151484}, issn = {1875-5402}, mesh = {Aedes/virology ; Animals ; Anti-Inflammatory Agents, Non-Steroidal/therapeutic use ; Antiviral Agents/*chemistry/*pharmacology ; Biological Products/*chemistry ; Chikungunya Fever/*drug therapy/epidemiology/etiology/transmission ; Dengue/*drug therapy/epidemiology/transmission ; Humans ; Mosquito Control ; Mosquito Vectors/virology ; Neglected Diseases/drug therapy ; Plant Extracts/chemistry/pharmacology ; }, abstract = {Neglected tropical diseases (NTDs) flourish mostly in impoverished developing nations of the world. It is estimated that NTDs plague up to 1 billion people every year thereby inducing a massive economic and health burden worldwide. Following explosive outbreaks mostly in Asia, Latin America, Europe and the Indian Ocean, two common NTDs namely, Chikungunya and Dengue both transmitted by an infected mosquito vector principally Aedes aegypti have emerged as a major public health threat. Given the limitations of conventional medicine in specifically targeting the Chikungunya and Dengue virus (CHIKV and DENV), natural products present an interesting avenue to explore in the quest of developing novel anti; mosquito, CHIKV and DENV agents. In this endeavor, a number of plant extracts, isolated phytochemicals, essential oils and seaweeds have shown promising larvicidal and insecticidal activity against some mosquito vectors as well as anti CHIKV and DENV activity invitro. Other natural products that have depicted good potential against these diseases include; the symbiotic bacterial genus Wolbachia which can largely reduce the life span and infectivity of mosquito vectors and the marine Cyanobacterium Trichodesmium erythraeum which has shown anti- CHIKV activity at minimal cytotoxic level. The impetus of modern drug discovery approaches such as high throughput screening, drug repositioning, synthesis and computer-aided drug design will undeniably enhance the process of developing more stable lead molecules from natural products which have shown promising antiviral activity in-vitro.}, } @article {pmid27144810, year = {2016}, author = {Cattel, J and Martinez, J and Jiggins, F and Mouton, L and Gibert, P}, title = {Wolbachia-mediated protection against viruses in the invasive pest Drosophila suzukii.}, journal = {Insect molecular biology}, volume = {25}, number = {5}, pages = {595-603}, doi = {10.1111/imb.12245}, pmid = {27144810}, issn = {1365-2583}, mesh = {Animals ; Dicistroviridae/*physiology ; Drosophila/*microbiology/virology ; Insect Viruses/*physiology ; Introduced Species ; Nodaviridae/*physiology ; Symbiosis ; Wolbachia/*physiology ; }, abstract = {The maternally inherited bacterium Wolbachia is well known for spreading in natural populations by manipulating the reproduction of its arthropod hosts, but can also have mutualist effects that increase host fitness. In mosquitoes and Drosophila some Wolbachia strains can lead to an increase in survival of virus-infected insects, and in most cases this is associated with reduced accumulation of the virus in host tissues. We investigated if the Wolbachia strain wSuz, which naturally infects Drosophila suzukii, is able to confer protection against Drosophila C virus and Flock House virus in different host genetic backgrounds. We found that this strain can increase host survival upon infection with these two viruses. In some cases this effect was associated with lower viral titres, suggesting that it confers resistance to the viruses rather than allowing the flies to tolerate infection. Our results indicate that, in D. suzukii, the antiviral protection provided by Wolbachia is not correlated to its density as found in other Drosophila species. This study demonstrates a phenotypic effect induced by wSuz on its native host which could explain its maintenance in natural populations of D. suzukii.}, } @article {pmid27132518, year = {2016}, author = {Khoo, JJ and Chen, F and Kho, KL and Ahmad Shanizza, AI and Lim, FS and Tan, KK and Chang, LY and AbuBakar, S}, title = {Bacterial community in Haemaphysalis ticks of domesticated animals from the Orang Asli communities in Malaysia.}, journal = {Ticks and tick-borne diseases}, volume = {7}, number = {5}, pages = {929-937}, doi = {10.1016/j.ttbdis.2016.04.013}, pmid = {27132518}, issn = {1877-9603}, mesh = {Anaplasma/genetics/isolation & purification ; Animals ; *Animals, Domestic ; Bacillus/genetics/isolation & purification ; Bacteria/*isolation & purification ; Cats/parasitology ; Chickens/parasitology ; Coxiella/genetics/isolation & purification ; DNA, Bacterial/genetics ; Dogs/parasitology ; Ehrlichia/genetics/isolation & purification ; Ixodidae/*microbiology ; Malaysia/epidemiology ; *Microbiota ; Mycobacterium/genetics/isolation & purification ; Phylogeny ; Polymerase Chain Reaction ; Pseudomonas/genetics/isolation & purification ; RNA, Ribosomal, 16S/genetics ; Rickettsia/genetics/isolation & purification ; Sphingomonas/genetics/isolation & purification ; Tick Infestations/epidemiology/*veterinary ; }, abstract = {Ticks are vectors in the transmission of many important infectious diseases in human and animals. Ticks can be readily found in the semi-forested areas such as the settlements of the indigenous people in Malaysia, the Orang Asli. There is still minimal information available on the bacterial agents associated with ticks found in Malaysia. We performed a survey of the bacterial communities associated with ticks collected from domestic animals found in two Orang Asli villages in Malaysia. We collected 62 ticks, microscopically and molecularly identified as related to Haemaphysalis wellingtoni, Haemaphysalis hystricis and Haemaphysalis bispinosa. Bacterial 16s rRNA hypervariable region (V6) amplicon libraries prepared from the tick samples were sequenced on the Ion Torrent PGM platform. We detected a total of 392 possible bacterial genera after pooling and sequencing 20 samples, indicating a diverse bacterial community profile. Dominant taxa include the potential tick endosymbiont, Coxiella. Other dominant taxa include the tick-associated pathogen, Rickettsia, and environmental bacteria such as Bacillus, Mycobacterium, Sphingomonas and Pseudomonas. Other known tick-associated bacteria were also detected, including Anaplasma, Ehrlichia, Rickettsiella and Wolbachia, albeit at very low abundance. Specific PCR was performed on selected samples to identify Rickettsia and Coxiella. Sequence of Rickettsia felis, which causes spotted fever in human and cats, was identified in one sample. Coxiella endosymbionts were detected in three samples. This study provides the baseline knowledge of the microbiome of ticks in Malaysia, focusing on tick-associated bacteria affecting the Orang Asli communities. The role of the herein found Coxiella and Rickettsia in tick physiology or disease transmission merits further investigation.}, } @article {pmid27126029, year = {2016}, author = {Dobson, SL and Bordenstein, SR and Rose, RI}, title = {Wolbachia mosquito control: Regulated.}, journal = {Science (New York, N.Y.)}, volume = {352}, number = {6285}, pages = {526-527}, doi = {10.1126/science.352.6285.526-b}, pmid = {27126029}, issn = {1095-9203}, mesh = {Animals ; Culicidae/*microbiology ; Dengue/*prevention & control ; Mosquito Control/*methods ; *Wolbachia ; }, } @article {pmid27126028, year = {2016}, author = {O'Neill, SL}, title = {Wolbachia mosquito control: Tested.}, journal = {Science (New York, N.Y.)}, volume = {352}, number = {6285}, pages = {526}, doi = {10.1126/science.352.6285.526-a}, pmid = {27126028}, issn = {1095-9203}, mesh = {Animals ; Culicidae/*microbiology ; Dengue/*prevention & control ; Mosquito Control/*methods ; *Wolbachia ; }, } @article {pmid27124663, year = {2016}, author = {Aliota, MT and Walker, EC and Uribe Yepes, A and Velez, ID and Christensen, BM and Osorio, JE}, title = {The wMel Strain of Wolbachia Reduces Transmission of Chikungunya Virus in Aedes aegypti.}, journal = {PLoS neglected tropical diseases}, volume = {10}, number = {4}, pages = {e0004677}, pmid = {27124663}, issn = {1935-2735}, support = {R21 AI117413/AI/NIAID NIH HHS/United States ; #AI117413-01/AI/NIAID NIH HHS/United States ; }, mesh = {Aedes/*microbiology/physiology/*virology ; Animals ; *Antibiosis ; Chikungunya Fever/prevention & control/*transmission ; Chikungunya virus/*isolation & purification ; Disease Transmission, Infectious/*prevention & control ; Insect Vectors ; Saliva/virology ; Survival Analysis ; Wolbachia/*growth & development/physiology ; }, abstract = {BACKGROUND: New approaches to preventing chikungunya virus (CHIKV) are needed because current methods are limited to controlling mosquito populations, and they have not prevented the invasion of this virus into new locales, nor have they been sufficient to control the virus upon arrival. A promising candidate for arbovirus control and prevention relies on the introduction of the intracellular bacterium Wolbachia into Aedes aegypti mosquitoes. This primarily has been proposed as a tool to control dengue virus (DENV) transmission; however, evidence suggests Wolbachia infections confer protection for Ae. aegypti against CHIKV. Although this approach holds much promise for limiting virus transmission, at present our understanding of the ability of CHIKV to infect, disseminate, and be transmitted by wMel-infected Ae. aegypti currently being used at Wolbachia release sites is limited.

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

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

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

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

CONCLUSIONS: The risk analysis was designed to incorporate the interacting complexity of hazards that may affect the release of the technology into the environment. The risk analysis was a small, but important, implementation phase in the success of this innovative research introducing a new technology to combat dengue transmission in the environment.}, } @article {pmid27035832, year = {2016}, author = {Hunter, CM and Huang, W and Mackay, TF and Singh, ND}, title = {The Genetic Architecture of Natural Variation in Recombination Rate in Drosophila melanogaster.}, journal = {PLoS genetics}, volume = {12}, number = {4}, pages = {e1005951}, pmid = {27035832}, issn = {1553-7404}, support = {R01 GM045146/GM/NIGMS NIH HHS/United States ; R01 GM45146/GM/NIGMS NIH HHS/United States ; }, mesh = {Animals ; Drosophila melanogaster/*genetics/microbiology ; Female ; Genome-Wide Association Study ; Male ; *Recombination, Genetic ; Wolbachia/isolation & purification ; }, abstract = {Meiotic recombination ensures proper chromosome segregation in many sexually reproducing organisms. Despite this crucial function, rates of recombination are highly variable within and between taxa, and the genetic basis of this variation remains poorly understood. Here, we exploit natural variation in the inbred, sequenced lines of the Drosophila melanogaster Genetic Reference Panel (DGRP) to map