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Bibliography on: Biofilm

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ESP: PubMed Auto Bibliography 05 Jun 2020 at 01:34 Created: 


Wikipedia: Biofilm A biofilm is any group of microorganisms in which cells stick to each other and often also to a surface. These adherent cells become embedded within a slimy extracellular matrix that is composed of extracellular polymeric substances (EPS). The EPS components are produced by the cells within the biofilm and are typically a polymeric conglomeration of extracellular DNA, proteins, and polysaccharides. Because they have three-dimensional structure and represent a community lifestyle for microorganisms, biofilms are frequently described metaphorically as cities for microbes. Biofilms may form on living or non-living surfaces and can be prevalent in natural, industrial and hospital settings. The microbial cells growing in a biofilm are physiologically distinct from planktonic cells of the same organism, which, by contrast, are single-cells that may float or swim in a liquid medium. Biofilms can be present on the teeth of most animals as dental plaque, where they may cause tooth decay and gum disease. Microbes form a biofilm in response to many factors, which may include cellular recognition of specific or non-specific attachment sites on a surface, nutritional cues, or in some cases, by exposure of planktonic cells to sub-inhibitory concentrations of antibiotics. When a cell switches to the biofilm mode of growth, it undergoes a phenotypic shift in behavior in which large suites of genes are differentially regulated.

Created with PubMed® Query: biofilm[title] NOT pmcbook NOT ispreviousversion

Citations The Papers (from PubMed®)

RevDate: 2020-06-04

Pourhajibagher M, Miri-Moosavi RS, Chiniforush N, et al (2020)

Anti-biofilm activity of Chlorella-mediated light activated disinfection: Ex vivo inhibition of intracanal mature Enterococcus faecalis biofilms via application of natural product.

Photodiagnosis and photodynamic therapy pii:S1572-1000(20)30207-6 [Epub ahead of print].

BACKGROUND: This study aimed to evaluate the effects of light-activated disinfection (LAD) as a strategy for optimizing root canal disinfection using Chlorella (Chlo) as a natural photosensitizer against Enterococcus faecalis biofilms ex vivo.

MATERIALS AND METHODS: The physical and chemical stability and absorption spectra of Chlo were determined. The mature microbial biofilm of E. faecalis was formed in root canals of 70 freshly extracted single-rooted premolars. After determining the minimum inhibitory concentration (MIC) of Chlo using the agar dilution testing, E. faecalis biofilms were exposed in the following groups (n = 10): 1- Chlo at a concentration of 2 × MIC, 1- Chlo at a concentration of 4 × MIC, 3- Diode laser, 4- 2 × MIC dose of Chlo-mediated LAD, 5- 4 × MIC dose of Chlo-mediated LAD, 6- 0.2% chlorohexidine (CHX), and 7- control group (E. faecalis biofilms without exposure to any photosensitizer and light source). The quantitative and qualitative evaluations of E. faecalis biofilms were done using counts of colony forming units (CFUs) and scanning electron microscope (SEM) and fluorescence microscope analysis, respectively.

RESULTS: According to the results, the MIC of Chlo was 125 μg/mL, which inhibited the growth of E. faecalis. To evaluate the anti-biofilm effects of Chlo, the 2× and 4 × MICs of Chlo (250 and 500 μg/mL, respectively) were used in the current study. The 4 × MIC dose (500 μg/mL) of Chlo-mediated LAD was significantly more effective compared to other groups (P < 0.05), while the lowest percentage of dead cells was detected in the diode laser irradiation group. In addition, there was no significant difference in the log10 CFU of E. faecalis between the biofilm treated with 500 μg/mL Chlo-mediated LAD (1.27 ± 0.05) versus 0.2% CHX (1.10 ± 0.06) (P < 0.05). As Moreover, SEM and fluorescence microscope images of the microbial biofilms showed that the highest percentage of dead bacteria was found in the 500 μg/mL Chlo-mediated LAD group.

CONCLUSIONS: The results of this study suggest that Chlo-mediated LAD can be used as an adjuvant therapy to eliminate the E. faecalis biofilms in the root canal system.

RevDate: 2020-06-04

Thaarup IC, T Bjarnsholt (2020)

Current in vitro biofilm-infected chronic wound models for developing new treatment possibilities.

Advances in wound care [Epub ahead of print].

SIGNIFICANCE: The prevalence of chronic wounds is increasing worldwide. The most recent estimates suggest that up to 2% of the population in the industrialized countries are affected.1 Recent Advances: During the past few decades, bacterial biofilms have been elucidated as one of the primary reasons why chronic wounds fail to heal.2,3 Critical Issues: There is a lack of direct causation and evidence of the role that biofilms play in persistent wounds, which complicates research on new treatment options, since it is still unknown which factors dominate. For this reason, several different in vitro wound models have been created, that mimic the biofilm infections observed in chronic wounds and other chronic infections. These different models are, amongst other purposes, used to test a variety of wound care products. However, chronic wounds are highly complex, and several different factors must be taken into consideration along with the infection, including physiochemical and human-supplemented factors. Furthermore, the limitations of using in vitro models, such as the lack of a responsive immune system should always be given due consideration.

FUTURE DIRECTIONS: Present understandings of all the elements and interactions that take place within chronic wounds are incomplete. As our insight of in vivo chronic wounds continues to expand, so too must the in vitro models used to mimic these infections evolve and adapt to new knowledge.

RevDate: 2020-06-04

Weigelt M, McNamara SA, Sanchez DP, et al (2020)

Evidence-Based Review of Anti-Biofilm Agents for Wound Care.

Advances in wound care [Epub ahead of print].

Significance Biofilms in vivo are small densely packed aggregations of microbes that are highly resistant to host immune responses and treatment. Biofilms attach to each other and to nearby surfaces. They are difficult to study and identify in a clinical setting as their quantification necessitates the use of advanced microscopy techniques such as confocal scanning laser microscopy (CLSM). Nonetheless it is likely that biofilms contribute to the pathophysiology of chronic skin wounds. Reducing, removing or preventing biofilms is thus a logical approach to help clinicians heal chronic wounds. Recent Advances Wound care products have demonstrated varying degrees of efficacy in destroying biofilms in in vitro and pre-clinical models as well as in some clinical studies. Critical Issues Controlled studies exploring the beneficial role of biofilm eradication and its relationship to healing in patients with chronic wounds are limited. This review aims to discuss the mode of action and clinical significance of currently available anti-biofilm products including surfactants, dressings and others, with a focus on levels of evidence for efficacy in disrupting biofilms and ability to improve wound healing outcomes. Future Directions Few available products have good evidence to support anti-biofilm activity and wound healing benefits. Novel therapeutic strategies are on the horizon. More high-quality clinical studies are needed. The development of non-invasive techniques to quantify biofilms will facilitate increased ease of research about biofilms in wounds and how to combat them.

RevDate: 2020-06-04

Pezzoni M, Pizarro RA, CS Costa (2020)

Role of quorum sensing in UVA-induced biofilm formation in Pseudomonas aeruginosa.

Microbiology (Reading, England) [Epub ahead of print].

Pseudomonas aeruginosa, a versatile bacterium present in terrestrial and aquatic environments and a relevant opportunistic human pathogen, is largely known for the production of robust biofilms. The unique properties of these structures complicate biofilm eradication, because they make the biofilms very resistant to diverse antibacterial agents. Biofilm development and establishment is a complex process regulated by multiple regulatory genetic systems, among them is quorum sensing (QS), a mechanism employed by bacteria to regulate gene transcription in response to population density. In addition, environmental factors such as UVA radiation (400-315 nm) have been linked to biofilm formation. In this work, we further investigate the mechanism underlying the induction of biofilm formation by UVA, analysing the role of QS in this phenomenon. We demonstrate that UVA induces key genes of the Las and Rhl QS systems at the transcriptional level. We also report that pelA and pslA genes, which are essential for biofilm formation and whose transcription depends in part on QS, are significantly induced under UVA exposure. Finally, the results demonstrate that in a relA strain (impaired for ppGpp production), the UVA treatment does not induce biofilm formation or QS genes, suggesting that the increase of biofilm formation due to exposure to UVA in P. aeruginosa could rely on a ppGpp-dependent QS induction.

RevDate: 2020-06-04

Zhang DX, Li Y, Yang XQ, et al (2020)

In vitro Antibiotic Susceptibility, Virulence Genes Distribution and Biofilm Production of Staphylococcus aureus Isolates from Bovine Mastitis in the Liaoning Province of China.

Infection and drug resistance, 13:1365-1375 pii:247765.

Purpose: The aim of this study was to identify the subtype, characterize the antimicrobial resistance, determine the virulence gene distribution, and analyze the biofilm production of Staphylococcus aureus isolates from bovine mastitis milk samples in the Liaoning Province of China.

Materials and Methods: In total, 56 Staph. aureus isolates were collected and identified in this study; the isolates were divided into different spa types based on the sequence of the polymorphic X region of the spa gene. Additionally, antimicrobial susceptibility was investigated using the broth microdilution method, and 18 virulence genes were detected using PCR. Biofilm formation was measured by spectrophotometry with crystal violet staining and observed using confocal laser scanning microscopy.

Results: There were 12.12% (56/462) milk samples that were positive for Staph. aureus. These isolates were nonsusceptible to sulfamethoxazole (100%), penicillin (76.9%), daptomycin (76.79%), clindamycin (69.64%), and oxacillin (60.71%); however, the majority of the isolates (80.4%) were susceptible to amoxicillin/clavulanate. The predominant virulence genes encoded the cytotoxins, hla (94.64%) and hlb (89.29%), and the adhesion factors clfA (89.29%), clfB (89.29%), and fnbB (80.36%). Comparatively, virulence genes related to other adhesion factors such as cna (8.93%) and enterotoxins, such as seg (26.79%), sea (16.07%), seb (7.14%), and sec (7.14%) were detected at relatively lower rates. The following eight spa types were identified: t267 (35.84%), t730 (22.64%), t518 (15.09%), t1190 (11.32%), t1456 (9.43%), t224 (1.88%), t9129 (1.88%), and t177 (1.88%). The highest biofilm production was observed for t267. Staph. aureus exhibited various patterns of biofilm formation, with the biofilm often being associated with a tower-shaped structure or a thicker biofilm.

Conclusion: Our results indicated that Staph. aureus isolates from dairy cows with mastitis in the Liaoning Province of China were non-susceptible to sulfamethoxazole, penicillin, daptomycin, oxacillin, and clindamycin. Additionally, the most prevalent subtype was t267, which displayed resistance to multiple antimicrobial agents and harbored several virulence genes, including clfA, clfB, fnbB, hla, and hlb.

RevDate: 2020-06-04

Bernegossi J, Fontana CR, Caiaffa KS, et al (2020)

Inhibitory Effect of a KSL-W Peptide-Loaded Poloxamer 407-Based Microemulsions for Buccal Delivery on Fusobacterium nucleatum Biofilm.

Journal of biomedical nanotechnology, 16(3):390-397.

KSL-W peptide has demonstrated antibacterial and antifungal activity and inhibitory effects against oral biofilm. This study aimed to check out the effect of chlorhexidine (CLX) or KSL-W peptide-loaded poloxamer 407-based microemulsions for buccal delivery on Fusobacterium nucleatum (F. nucleatum) biofilm. The formulation (F) containing 10% copolymer poloxamer 407 dispersion (1%), 40% oleic acid and 50% PPG-5-CETETH-20 was characterized by polarized light microscopy (PLM), small-angle X-ray scattering (SAXS), rheology, bioadhesive and syringeability; and in the treatment of a biofilm produced by F. nucleatum. The darkfield images obtained by PLM and the SAXS curves with an extended peak indicated that the system was characteristic of microemulsions. In a continuous analysis, microemulsions exhibited Newtonian behavior. In frequency, the oscillatory analysis profile presented predominantly viscous behavior. Bioadhesive force detected in the analysis of F (7.4 ± 1.81 mN˙ s) and syringeability (17.83± 5.97 N · mm) being adequate values for buccal administration. After 4 h, KSL-W-loaded F shown over 20% higher effectiveness than chlorhexidine-loaded microemulsions. In conclusion, the KSL-W-loaded microemulsions showed a considerable reduction in F. nucleatum biofilm formation and presented promising structural properties for buccal drug delivery.

RevDate: 2020-06-04

Engel AS, Kranz HT, Schneider M, et al (2020)

Biofilm formation on different dental restorative materials in the oral cavity.

BMC oral health, 20(1):162 pii:10.1186/s12903-020-01147-x.

BACKGROUND: Bacterial biofilms adhere to all tissues and surfaces in the oral cavity. Oral biofilms are responsible for the decay of human dental structures and the inflammatory degeneration of the alveolar bone. Moreover, oral biofilms on artificial materials influence the lifespan of dental prostheses and restoratives.

METHODS: To investigate in vivo oral biofilm formation and growth, five different dental restorative materials were analyzed and compared to human enamel. The roughness of the materials and the human enamel control probe were measured at the start of the study. The dental restorative materials and the human enamel control probe were placed in dental splints and worn for 3 h, 24 h and 72 h.

RESULTS: Scanning electron microscopy (SEM) revealed major differences between oral biofilm formation and growth on the materials compared to those on human enamel. Microbiological analyses showed that bacterial strains differed between the materials. Significant differences were observed in the roughness of the dental materials.

CONCLUSIONS: It can be concluded that material roughness affects biofilm formation on dental surfaces and restoratives, but other factors, such as surface charge, surface energy and material composition, may also have an influence.

RevDate: 2020-06-04

Ohsumi T, Takenaka S, Sakaue Y, et al (2020)

Adjunct use of mouth rinses with a sonic toothbrush accelerates the detachment of a Streptococcus mutans biofilm: an in vitro study.

BMC oral health, 20(1):161 pii:10.1186/s12903-020-01144-0.

BACKGROUND: The aim of this in vitro study was to examine the possible enhancement of the biofilm peeling effect of a sonic toothbrush following the use of an antimicrobial mouth rinse.

METHODS: The biofilm at a noncontact site in the interdental area was treated by sound wave convection with the test solution or by immersion in the solution. The biofilm peeling effect was evaluated by determining the bacterial counts and performing morphological observations. A Streptococcus mutans biofilm was allowed to develop on composite resin discs by cultivation with stirring at 50 rpm for 72 h. The specimens were then placed in recesses located between plastic teeth and divided into an immersion group and a combination group. The immersion group was treated with phosphate buffer, chlorhexidine digluconate Peridex™ (CHX) mouth rinse or Listerine® Fresh Mint (EO) mouth rinse. The combination group was treated with CHX or EO and a sonic toothbrush.

RESULTS: The biofilm thickness was reduced by approximately one-half compared with the control group. The combination treatment produced a 1 log reduction in the number of bacteria compared to the EO immersion treatment. No significant difference was observed in the biofilm peeling effect of the immersion group compared to the control group.

CONCLUSIONS: The combined use of a sonic toothbrush and a mouth rinse enhanced the peeling of the biofilm that proliferates in places that are difficult to reach using mechanical stress.

RevDate: 2020-06-04

Ionescu AC, Cazzaniga G, Ottobelli M, et al (2020)

Substituted Nano-Hydroxyapatite Toothpastes Reduce Biofilm Formation on Enamel and Resin-Based Composite Surfaces.

Journal of functional biomaterials, 11(2): pii:jfb11020036.

Background: Toothpastes containing nano-hydroxyapatite (n-HAp) substituted with metal ions provide calcium and phosphate ions to dental hard tissues, reducing demineralization, and promoting remineralization. Few data are available about the effect of these bioactive compounds on oral microbiota. Methods: This in vitro study evaluated the influence of two commercially-available substituted n-HAp-based toothpastes (α: Zn-carbonate substituted n-HAp; β: F, Mg, Sr-carbonate substituted n-HAp) on early colonization (EC, 12 h) and biofilm formation (BF, 24 h) by oral microbiota. Controls were brushed with distilled water. Artificial oral microcosm and Streptococcus mutans biofilms were developed using human enamel and a resin-based composite (RBC) as adherence surfaces. Two test setups, a shaking multiwell plate and a modified drip-flow reactor (MDFR), were used to simulate clinical conditions during the night (low salivary flow and clearance) and daytime, respectively. Energy-dispersive X-ray spectrometry (EDS) was used to evaluate specimens' surfaces after toothpaste treatment. Fluoride release from β toothpaste was evaluated. Viable adherent biomass was quantified by MTT assay, and biofilms' morphology was highlighted using confocal microscopy. Results: EDS showed the presence of remnants from the tested toothpastes on both adherence surfaces. β toothpaste showed significantly lower EC and BF compared to control using the artificial oral microcosm model, while α toothpaste showed lower EC and BF compared to control, but higher EC and BF compared to β toothpaste. The effect shown by β toothpaste was, to a minimal extent, due to fluoride release. Interestingly, this result was seen on both adherence surfaces, meaning that the tested toothpastes significantly influenced EC and BF even on RBC surfaces. Furthermore, the effect of toothpaste treatments was higher after 12 h than 24 h, suggesting that toothbrushing twice a day is more effective than brushing once. Conclusions: The efficacy of these treatments in reducing microbial colonization of RBC surfaces may represent a promising possibility in the prevention of secondary caries.

RevDate: 2020-06-04

Kostoglou D, Protopappas I, E Giaouris (2020)

Common Plant-Derived Terpenoids Present Increased Anti-Biofilm Potential against Staphylococcus Bacteria Compared to a Quaternary Ammonium Biocide.

Foods (Basel, Switzerland), 9(6): pii:foods9060697.

The antimicrobial actions of three common plant-derived terpenoids (i.e., carvacrol, thymol and eugenol) were compared to those of a typical quaternary ammonium biocide (i.e., benzalkonium chloride; BAC), against both planktonic and biofilm cells of two widespread Staphylococcus species (i.e., S. aureus and S. epidermidis). The minimum inhibitory and bactericidal concentrations (MICs, MBCs) of each compound against the planktonic cells of each species were initially determined, together with their minimum biofilm eradication concentrations (MBECs). Various concentrations of each compound were subsequently applied, for 6 min, against each type of cell, and survivors were enumerated by agar plating to calculate log reductions and determine the resistance coefficients (Rc) for each compound, as anti-biofilm effectiveness indicators. Sessile communities were always more resistant than planktonic ones, depending on the biocide and species. Although lower BAC concentrations were always needed to kill a specified population of either cell type compared to the terpenoids, for the latter, the required increases in their concentrations, to be equally effective against the biofilm cells with respect to the planktonic ones, were not as intense as those observed in the case of BAC, presenting thus significantly lower Rc. This indicates their significant anti-biofilm potential and advocate for their further promising use as anti-biofilm agents.

RevDate: 2020-06-03

Hülsen T, Sander EM, Jensen PD, et al (2020)

Application of purple phototrophic bacteria in a biofilm photobioreactor for single cell protein production: Biofilm vs suspended growth.

Water research, 181:115909 pii:S0043-1354(20)30446-2 [Epub ahead of print].

Single cell protein (SCP), has been proposed as alternative to effectively upgrade and recycle organics and nutrients from wastewater. Biomass recovery is a critical issue, and recovery as a biofilm is effective in comparison with sedimentation of suspended biomass. This study aims to determine the applicability of purple phototrophic bacteria (PPB) biofilm on infra-red irradiated, submerged surfaces for the treatment of pre-settled red meat processing wastewater, and SCP generation. PPB removed up to 66% of COD and 42% of TN and TP during batch operation with total areal productivities between 15 and 20 gVS m-2 d-1 achieved. More than 60% of the total biomass grew attached (as biofilm) with the remainder being suspended. The biofilm can be harvested at around 160 gTS L-1 with high protein (>96 g L-1) and low ash contents (>4.0% compared to >30% in the wastewater). The compositions of attached and suspended biomass differed significantly, where the suspended fraction resembled the wastewater composition (e.g. in terms of inert components). The PPB community was similar in the suspended and biofilm fractions while the biofilm had higher relative abundance of PPB representatives (57% vs 43%). A consistent product composition is highly relevant for the manufacturer and ultimately determines the value as feed, feed additive, or supplement.

RevDate: 2020-06-03

Meng J, Xu J, J Chen (2020)

The role of osmoregulated periplasmic glucans in the biofilm antibiotic resistance of Yersinia enterocolitica.

Microbial pathogenesis pii:S0882-4010(20)30650-1 [Epub ahead of print].

The formation of biofilms by bacteria is of great significance because it involves many physiological changes that serve to protect the cells from various stresses. One of the best-known biofilm-specific properties of bacteria is that bacteria that grow in biofilms are generally more resistant to antibiotics than their planktonic counterparts. In a previous study, osmoregulated periplasmic glucans (OPGs), catalyzed by the opgGH operon, were identified and found to function in Rcs signalling in Yersinia enterocolitica. In this study, the possible contribution of OPGs to antimicrobial resistance of Y. enterocolitica biofilms were investigated, and the results showed that OPGs, especially when overexpressed, conferred a high level of biofilm resistance to two different classes of antibiotics onto Y. enterocolitica. Subsequent analysis revealed that OPGs regulated the biofilm architecture in Y. enterocolitica by promoting the bacteria to form large cell aggregates. Moreover, the opgGH genes in biofilms showed higher expression than in planktonic cultures. OPGs were required to induce the expression of genes related to flagella, extracellular polysaccharide, and c-di-GMP biosynthesis in Y. enterocolitica biofilms and this effect was more significant when OPGs were overproduced. The current investigation showed an extension in the biological role of OPGs in Y. enterocolitica and provided a strong theoretical basis to further study this resistance mechanism at the molecular level to identify new drug targets or disinfectants for the treatment of infections caused by Y. enterocolitica within biofilms.

RevDate: 2020-06-03

Earl C, Arnaouteli S, Bamford N, et al (2020)

The majority of the matrix protein TapA is dispensable for Bacillus subtilis colony biofilm architecture.

Molecular microbiology [Epub ahead of print].

Biofilm formation is a co-operative behaviour where microbial cells become embedded in an extracellular matrix. This biomolecular matrix helps manifest the beneficial or detrimental outcome mediated by the collective of cells. Bacillus subtilis is an important bacterium for understanding the principles of biofilm formation. The protein components of the B. subtilis matrix include the secreted proteins BslA, which forms a hydrophobic coat over the biofilm, and TasA, which forms protease-resistant fibres needed for structuring. TapA is a secreted protein also needed for biofilm formation and helps in vivo TasA-fibre formation but is dispensable for in vitro TasA-fibre assembly. We show that TapA is subjected to proteolytic cleavage in the colony biofilm and that only the first 57 amino acids of the 253-amino acid protein are required for colony biofilm architecture. Through the construction of a strain which lacks all eight extracellular proteases, we show that proteolytic cleavage by these enzymes is not a prerequisite for TapA function. It remains unknown why TapA is synthesized at 253 amino acids when the first 57 are sufficient for colony biofilm structuring; the findings do not exclude the core conserved region of TapA having a second role beyond structuring the B. subtilis colony biofilm.

RevDate: 2020-06-03

Bin C, Al-Dhabi NA, Esmail GA, et al (2020)

Potential effect of Allium sativum bulb for the treatment of biofilm forming clinical pathogens recovered from periodontal and dental caries.

Saudi journal of biological sciences, 27(6):1428-1434.

Biofilm producing clinical bacterial isolates were isolated from periodontal and dental caries samples and identified as, Lactobacillus acidophilus, Streptococcus sanguis, S. salivarius, S. mutansand Staphylococcus aureus. Among the identified bacterial species, S. aureus and S. mutansshowed strong biofilm producing capacity. The other isolated bacteria, Streptococcus sanguis, S. salivarius showed moderate biofilm formation. These pathogens were subjected for the production of extracellular polysaccharides (EPS) in nutrient broth medium and the strain S. aureus synthesized more amounts of EPS (610 ± 11.2 µg/ml) than S. sanguis (480 ± 5.8 µg/ml).EPS production was found to be less in S. salivarius (52 ± 3.8 µg/ml).The solvent extract of A. sativum bulb showed the phytochemicals such as, carbohydrate, total protein, alkaloids, saponins, flavonoids, tannins and sterioids. The solvent extract of A. sativum bulb showed wide ranges of activity against the selected dental pathogens. The difference in antibacterial activity of the solvent extract revealed differences in solubility of phytochemicals in organic solvents. Ethanol extract was highly active againstS. aureus (25 ± 2 mm). The Minimum Inhibitory Concentration (MIC) of crude garlic bulb varied widely and this clearly showed that bacteria exhibits different level of susceptibility to secondary metabolites. MIC value ranged between 20 ± 2 mg/ml and 120 ± 6 mg/ml and Minimum Bactericidal Concentration (MBC) value ranged from 60 ± 5 mg/l to 215 ± 7 mg/ml. To conclude, A. sativum bulb can be effectively used to treat periodontal and dental caries infections.

RevDate: 2020-06-03

Warraich AA, Mohammed AR, Perrie Y, et al (2020)

Evaluation of anti-biofilm activity of acidic amino acids and synergy with ciprofloxacin on Staphylococcus aureus biofilms.

Scientific reports, 10(1):9021 pii:10.1038/s41598-020-66082-x.

Acidic amino acids, aspartic acid (Asp) and glutamic acid (Glu) can enhance the solubility of many poorly soluble drugs including ciprofloxacin (Cip). One of the mechanisms of resistance within a biofilm is retardation of drug diffusion due to poor penetration across the matrix. To overcome this challenge, this work set to investigate novel counter ion approach with acidic amino acids, which we hypothesised will disrupt the biofilm matrix as well as simultaneously improve drug effectiveness. The anti-biofilm activity of D-Asp and D-Glu was studied on Staphylococcus aureus biofilms. Synergistic effect of combining D-amino acids with Cip was also investigated as a strategy to overcome anti-microbial resistance in these biofilms. Interestingly at equimolar combinations, D-Asp and D-Glu were able to significantly disperse (at 20 mM and 40 mM) established biofilms and inhibit (at 10 mM, 20 mM and 40 mM) new biofilm formation in the absence of an antibiotic. Moreover, our study confirmed L-amino acids also exhibit anti-biofilm activity. The synergistic effect of acidic amino acids with Cip was observed at lower concentration ranges (<40 mM amino acids and <90.54 µM, respectively), which resulted in 96.89% (inhibition) and 97.60% (dispersal) reduction in CFU with exposure to 40 mM amino acids. Confocal imaging indicated that the amino acids disrupt the honeycomb-like extracellular DNA (eDNA) meshwork whilst also preventing its formation.

RevDate: 2020-06-03

Baig U, Ansari MA, Gondal MA, et al (2020)

Single step production of high-purity copper oxide-titanium dioxide nanocomposites and their effective antibacterial and anti-biofilm activity against drug-resistant bacteria.

Materials science & engineering. C, Materials for biological applications, 113:110992.

In the present research work, copper oxide-titanium dioxide nanocomposites were synthesized for the first time using advanced pulsed laser ablation in liquid (PLAL) technique for disinfection of drug-resistant pathogenic waterborne biofilm-producing bacterial strains. For this, a series of copper oxide-titanium dioxide nanocomposites were synthesized by varying the composition of copper oxide (5%, 10%, and 20%) with titanium dioxide. The pure titanium dioxide and copper oxide-titanium dioxide nanocomposites were characterized by advanced instrumental techniques. XRD, TEM, FE-SEM, EDX, elemental mapping and XPS analysis results consistently revealed the successful formation of copper oxide-titanium dioxide nanocomposites using PLAL technique. The antibacterial and antibiofilm activities of pure titanium dioxide and copper oxide-titanium dioxide nanocomposites were investigated against biofilm-producing strains of Methicillin-resistant Staphylococcus aureus and Pseudomonas aeruginosa by various methods. Our results revealed that the PLAL synthesized copper oxide-titanium dioxide nanocomposites showed enhanced anti-biofilm and antibacterial activity compared to pure titanium dioxide in a dose-dependent manner against targeted pathogens. Furthermore, the effects of pure titanium dioxide and copper oxide-titanium dioxide nanocomposites on bacterial morphology, biofilm formation, aggregation and their colonization by targeted pathogens were also examined using scanning electron microscopy. Microscopic images clearly showed that the cell envelope of almost all the cells were rumples, rough, had irregularities and abnormal appearance with the major damage being characterized by the formation of "pits". Many depressions and indentations were also seen in their cell envelope and the original shape of Pseudomonas aeruginosa cells changed from normal rod to swollen, large and elongated which indicates the loss of membrane integrity and damage of cell wall and membrane. The findings suggested that PLAL synthesized copper oxide-titanium dioxide nanocomposites have good potential for removal of biofilm or killing of pathogenic bacteria in water distribution network and for wastewater treatment, hospital and environmental applications. In addition, cytotoxic activity of pure TiO2 and PLAL synthesized copper oxide-titanium dioxide nanocomposites against normal and healthy cells (HEK-293) and cancerous cells (HCT-116) were also evaluated by MTT assay. The MTT assay results showed no cytotoxic effects on HEK-293 cells, which suggest TiO2 and PLAL synthesized copper oxide-titanium dioxide nanocomposites are non-toxic to the normal cells.

RevDate: 2020-06-03

Awasthi A, Sharma P, Jangir L, et al (2020)

Dose dependent enhanced antibacterial effects and reduced biofilm activity against Bacillus subtilis in presence of ZnO nanoparticles.

Materials science & engineering. C, Materials for biological applications, 113:111021.

Bacterial biofilms are self-produced matrix of sticky extracellular polymeric substances. They result in fouling in the food industry, water treatment plants, and possess significant environmental and industrial impacts. Nanoparticles have shown immense potential and have been effective in combating bacterial biofilm, which is the common cause of drug resistance development, biofouling in water treatment plants and the food industry. Hence, in order to explore the same, Zinc oxide nanoparticles have been synthesized by chemical synthesis method and their action against Bacillus subtilis biofilm formation was evaluated in this study by crystal violet and ROS assay. The dose-dependent reduction in biofilm biomass and density was observed as a result of nanoparticle exposure. There was considerable reduction in biofilm formation after treatment with ZnO nanoparticles. Change in surface morphology of the Bacillus subtilis cells was observed which could be due to oxidative stress induced by ZnO nanoparticles. The oxidative stress was estimated by measurement of catalase activity that also showed dose-dependent decrease.

RevDate: 2020-06-03

Liu MJ, Zhang YL, XY Wan (2020)

[Research progress on methicillin-resistant Staphylococcus aureus biofilm].

Zhonghua nei ke za zhi, 59(6):473-476.

RevDate: 2020-06-03

Cai H, Wang Y, Wu K, et al (2020)

Enhanced Hydrophilic and Electrophilic Properties of Polyvinyl Chloride (PVC) Biofilm Carrier.

Polymers, 12(6): pii:polym12061240.

Polyvinyl chloride (PVC) biofilm carrier is used as a carrier for bacterial adsorption in wastewater treatment. The hydrophilicity and electrophilicity of its surface play an important role in the adsorption of bacteria. The PVC biofilm carrier was prepared by extruder, and its surface properties were investigated. In order to improve the hydrophilicity and electrophilic properties of the PVC biofilm carrier, polyvinyl alcohol (PVA) and cationic polyacrylamide (cPAM) were incorporated into polyvinyl chloride (PVC) by blending. Besides, the surface area of the PVC biofilm carrier was increased by azodicarbonamide modified with 10% by weight of zinc oxide (mAC). The surface contact angle of PVC applied by PVA and cPAM at 5 wt %, 15 wt % was 81.6°, which was 18.0% lower than pure PVC. It shows the significant improvement of the hydrophilicity of PVC. The zeta potential of pure PVC was -9.59 mV, while the modified PVC was 14.6 mV, which proves that the surface charge of PVC changed from negative to positive. Positive charge is more conducive to the adsorption of bacteria. It is obvious from the scanning electron microscope (SEM) images that holes appeared on the surface of the PVC biofilm carrier after adding mAC, which indicates the increase of PVC surface area.

RevDate: 2020-06-02

Rzhepishevska O, Hakobyan S, Ruhal R, et al (2013)

The surface charge of anti-bacterial coatings alters motility and biofilm architecture.

Biomaterials science, 1(6):589-602.

Bacterial biofilms affect many areas of human activity including food processing, transportation, public infrastructure, and most importantly healthcare. This study addresses the prevention of biofilms and shows that the surface charge of an abiotic substrate influences bacterial motility as well as the morphology and physiology of the biofilm. Grafting-from polymerisation was used to create polymer brush surfaces with different characteristics, and the development of Pseudomonas aeruginosa biofilms was followed using confocal microscopy. Interestingly, two types of biofilms developed on these surfaces: mushroom structures with high levels of cyclic diguanylate (c-di-GMP) were found on negatively charged poly (3-sulphopropylmethacrylate) (SPM) and zwitterionic poly (2-(methacryloyloxy)ethyl)dimethyl-3-sulphoproyl) ammonium hydroxide) (MEDSAH), while flat biofilms developed on glass, positively charged poly (2-(methacryloyloxy)-ethyl trimethyl ammonium chloride) (METAC), protein-repellent poly oligo(ethylene glycol methyl ether methacrylate) (POEGMA) and hydrophobic polymethylmethacrylate (PMMA). The results show that of all the surfaces studied, overall the negatively charged polymer brushes were most efficient in reducing bacterial adhesion and biofilm formation. However, the increased level of regulatory c-di-GMP in mushroom structures suggests that bacteria are capable of a quick physiological response when exposed to surfaces with varying physicochemical characteristics enabling some bacterial colonization also on negatively charged surfaces.

RevDate: 2020-06-02

Xiao B, Zou Z, Bhandari J, et al (2020)

Exposure to diode laser (810nm) affects the bacterial adherence and biofilm formation in a E. faecalis biofilm model.

Photodiagnosis and photodynamic therapy pii:S1572-1000(20)30125-3 [Epub ahead of print].

BACKGROUND: A large number of studies have shown that diode laser can effectively killEnterococcus faecalis (E. faecalis). However, to our knowledge there has been little information regarding high-level analysis of sterilization mechanism on E. faecalis biofilm models after laser irradiation. This study provides emphasis on the E. faecalis viability and exopolysaccharide content after laser irradiation. It also aims to examine whether diode laser affects the biofilm formation and adherence of E. faecalis biofilm at the level of gene expression.

METHODS: E. faecalis (ATCC 29212) was inoculated for biofilm formation. After growing for 48 h, the biofilms were submitted to the following treatments, twice daily (n = 6): (i) Diode laser with energy densities as 28.7 J/cm2; (ii) 1% sodium hypochlorite (NaOCl) was used as a positive control; (iii) 0.9% physiological saline (NaCl) was used as a negative control. After 120 h of biofilm growth, bactericidal activity of diode laser was evaluated by using plate count method and scanning electron microscopy (SEM). The distribution of extracellular polysaccharide was assessed by Confocal laser scanning microscopy (CLSM). Anthrone method was used to quantify the amount of water-soluble exopolysaccharide (WSE) and water-insoluble exopolysaccharides (WIE) in E. faecalis. Real-time quantitative polymerase chain reaction (RT-qPCR) was employed to assess transcription of genes (gelE, ace and esp) related to formation and adherence of E. faecalis biofilm.

RESULTS: Compared with NaCl, diode laser significantly destabilizedE. faecalis biofilm, which showing a decrease in the number of bacteria and inhibiting the adherence of biofilm (p < 0.05). From Anthrone method and CLSM, the study found that diode laser lowered the concentration of WSE and WIE in biofilms (p < 0.05). In addition, transcription of gene gelE, ace and esp were also reduced after irradiation of diode laser (p < 0.05).

CONCLUSION: By inhibiting the synthesis of WIE and WSE, diode laser may reduce the formation of E. faecalis biofilm. The expression of specific genes (gelE, ace and esp) involved in bacterial adherence and biofilm formation were down regulated under the exposure to diode laser (810 nm), which in turn is expected to decrease the pathogenicity of E. faecalis.

RevDate: 2020-06-02

Qin H, Liu Y, Cao X, et al (2020)

RpoS is a pleiotropic regulator of motility, biofilm formation, exoenzymes, siderophore and prodigiosin production, and trade-off during prolonged stationary phase in Serratia marcescens.

PloS one, 15(6):e0232549 pii:PONE-D-20-03043.

Prodigiosin is an important secondary metabolite produced by Serratia marcescens. It can help strains resist stresses from other microorganisms and environmental factors to achieve self-preservation. Prodigiosin is also a promising secondary metabolite due to its pharmacological characteristics. However, pigmentless S. marcescens mutants always emerge after prolonged starvation, which might be a way for the bacteria to adapt to starvation conditions, but it could be a major problem in the industrial application of S. marcescens. To identify the molecular mechanisms of loss of prodigiosin production, two mutants were isolated after 16 days of prolonged incubation of wild-type (WT) S. marcescens 1912768R; one mutant (named 1912768WR) exhibited reduced production of prodigiosin, and a second mutant (named 1912768W) was totally defective. Comparative genomic analysis revealed that the two mutants had either mutations or deletions in rpoS. Knockout of rpoS in S. marcescens 1912768R had pleiotropic effects. Complementation of rpoS in the ΔrpoS mutant further confirmed that RpoS was a positive regulator of prodigiosin production and that its regulatory role in prodigiosin biosynthesis was opposite that in Serratia sp. ATCC 39006, which had a different type of pig cluster; further, rpoS from Serratia sp. ATCC 39006 and other strains complemented the prodigiosin defect of the ΔrpoS mutant, suggesting that the pig promoters are more important than the genes in the regulation of prodigiosin production. Deletion of rpoS strongly impaired the resistance of S. marcescens to stresses but increased membrane permeability for nutritional competence; competition assays in rich and minimum media showed that the ΔrpoS mutant outcompeted its isogenic WT strain. All these data support the idea that RpoS is pleiotropic and that the loss of prodigiosin biosynthesis in S. marcescens 1912768R during prolonged incubation is due to a mutation in rpoS, which appears to be a self-preservation and nutritional competence (SPANC) trade-off.

RevDate: 2020-06-02

Zhang J, Chen C, Chen J, et al (2020)

Dual Mode of Anti-biofilm Action of G3 against Streptococcus mutans.

ACS applied materials & interfaces [Epub ahead of print].

Oral biofilms, formed by multiple microorganisms and their extracellular polymeric substances (EPS), seriously affect people's life. Emergence of the resistance of biofilms to conventional antibiotics and their side effects to the oral cavity have posed a great challenge in the treatment of dental diseases. Recently, antimicrobial peptides (AMPs) have been recognized as a promising alternative to conventional antibiotics due to their broad antibacterial spectrum, high antibacterial activity, and specific mechanism. However, the research of their anti-biofilm behaviors is still in its infancy and the underlying mechanism remains unclear. In this study, we investigated the anti-biofilm activities of a designed helical peptide (G3) against Streptococcus mutans (S. mutans), one of the primary causative pathogens of caries. The results indicated that G3 inhibited S. mutans biofilm formation by interfering with different stages of biofilm development. At the initial stage, G3 inhibited the bacterial adhesion by decreasing the bacterial surface charges, hydrophobicity, membrane integrity, and adhesion-related genes transcription. At the later stage, G3 interacted with extracellular DNA (eDNA) to destabilize the 3D architecture of mature biofilms and thus dispersed them. The high activity of G3 against S. mutans biofilms, along with its specific modes of action, endows it great application potential in preventing and treating dental plaque diseases.

RevDate: 2020-06-02

Al Marjania MF, Kouhsari E, Ali FS, et al (2020)

Evaluation of type II toxin-antitoxin systems, antibiotic resistance profiles, and biofilm quorum sensing genes in Acinetobacter baumannii isolates in Iraq.

Infectious disorders drug targets pii:IDDT-EPUB-106916 [Epub ahead of print].

BACKGROUND: Bacterial Toxin-Antitoxin (TAs) systems are extensive two-component elements in the bacterial genome, which involved in many key biological functions including growth arrest, survival, biofilm formation, plasmid maintenance, defense against phages, persistence and virulence.

AIM: This study aimed to assess the molecular determinants involved in TAs, biofilm quorum sensing, and antibiotic resistance profiles in Acinetobacter baumannii isolated from Baghdad`s hospitals in Iraq.

METHODS: A total of 127 A. baumannii isolates were collected from 2160 different clinical samples. The antimicrobial susceptibility test was performed using disk diffusion test. All isolates were characterized for molecular determinants involved in TAs and biofilm formation using the well-known PCR-based sequencing assay.

RESULTS: A high multi-drug resistant (MDR) (96.06%; 122/127) and imipenem resistance (84.25%; 107/127) rates were observed from A.baumannii isolates. Results showed the presence of rhlIR gene in three isolates (2.36%), and lasIR gene appeared in two isolates (1.57%) isolates, whilst, mazEF, ccdAB, and relBE genes have not detected among isolates.

CONCLUSION: A high MDR and imipenem resistance rates within a low prevalence of rhlIR, and lasIR genes could be found in clinical A. baumannii isolates from some of Iraqi hospitals.

RevDate: 2020-06-02

Labine M, DePledge L, Feirer N, et al (2020)

Enzymatic and Mutational Analysis of the PruA Pteridine Reductase Required for Pterin-Dependent Control of Biofilm Formation in Agrobacterium tumefaciens.

Journal of bacteriology pii:JB.00098-20 [Epub ahead of print].

Pterins are ubiquitous biomolecules with diverse functions including roles as cofactors, pigments, and redox mediators. Recently, a novel pterin-dependent signaling pathway that controls biofilm formation was identified in the plant pathogen, Agrobacterium tumefaciens A key player in this pathway is a pteridine reductase termed PruA, where its enzymatic activity has been shown to control surface attachment and limit biofilm formation. Here, we biochemically characterize PruA to investigate the catalytic properties and substrate specificity of this pteridine reductase. PruA demonstrates maximal catalytic efficiency with dihydrobiopterin and comparable activities with the stereoisomers dihydromonapterin and dihydroneopterin. Since A. tumefaciens does not synthesize or utilize biopterins, the likely physiological substrate is dihydromonapterin or dihydroneopterin, or both. Notably, PruA does not exhibit pteridine reductase activity with dihydrofolate or fully oxidized pterins. Site-directed mutagenesis studies of a conserved tyrosine residue, the key component of a putative catalytic triad, indicate that this tyrosine is not directly involved in PruA catalysis but may be important for substrate or cofactor binding. Additionally, mutagenesis of the arginine residue in the N-terminal TGX3RXG motif significantly reduces the catalytic efficiency of PruA, supporting its proposed role in pterin binding and catalysis. Finally, we report the enzymatic characterization of PruA homologs from Pseudomonas aeruginosa and Brucella abortus, thus expanding the roles and potential significance of pteridine reductases in diverse bacteria.Importance Biofilms are complex multicellular communities that are formed by diverse bacteria. In the plant pathogen, Agrobacterium tumefaciens, the transition from a free-living motile state to a non-motile biofilm state is governed by a novel signaling pathway involving small molecules called pterins. The involvement of pterins in biofilm formation is unexpected and prompts many questions about the molecular details of this pathway. This work biochemically characterizes the PruA pteridine reductase involved in the signaling pathway to reveal its enzymatic properties and substrate preference, thus providing important insight into pterin biosynthesis and its role in A. tumefaciens biofilm control. Additionally, the enzymatic characteristics of related pteridine reductases from mammalian pathogens are examined to uncover potential roles of these enzymes in other bacteria.

RevDate: 2020-06-02

Fiamengo TE, Runcan EE, Premanandan C, et al (2020)

Evaluation of Biofilm Production by Escherichia coli Isolated From Clinical Cases of Canine Pyometra.

Topics in companion animal medicine, 39:100429.

Many Escherichia coli (E. coli) strains produce biofilm that confers antimicrobial resistance. However, studies of biofilm production by E. coli from canine pyometra are lacking. Objectives were to elucidate the role of biofilm production by E. coli in pyometra by: (1) assessing the ability of E. coli to produce biofilm in vitro, and (2) confirming biofilm in situ. Endometrial biopsies were obtained from bitches with pyometra and preserved for microscopic analysis (n = 25). An endometrial swab was submitted for aerobic culture. Samples with confirmed E. coli were evaluated further for biofilm production in vitro and in vivo. Seventy percent of cases (16/23) resulted in pure growth of 1 or 2 E. coli strains, totaling 20 isolates. Fifteen isolates (15/20, 75%) had higher optical densities then negative controls (P < .05). On histopathology, all tissues exhibited endometrial inflammation and mucus was located within endometrial glands and occasionally overlying epithelium on 14 slides (14/16, 88%). Bacteria was noted in 50% of slides (8/16). During FISH acellular debris within the uterine lumen consistent with biofilm was noted on 94% of samples (15/16) and E coli was positively identified on all samples (15/15). Areas suggestive of the presence of biofilm were observed on all samples on scanning electron microscopy; but, bacteria consistent with E. coli were only visualized in 9 samples (9/16, 56%). In conclusion, we demonstrated that relevant strains of E. coli produce biofilm in vitro and in vivo, which may be considered in the development of new pyometra treatments aimed at disrupting these E. coli biofilm.

RevDate: 2020-06-01

Liu C, Yan B, Duan J, et al (2020)

Biofilm inhibition effect of an ivermectin/silyl acrylate copolymer coating and the colonization dynamics.

The Science of the total environment, 736:139599 pii:S0048-9697(20)33119-3 [Epub ahead of print].

Ivermectin is now being used as a substitute for toxic organic biocide in marine antifouling coatings due to its environmentally friendly nature and the efficacy against parasites. However, the release performance of ivermectin from the hydrolyzed acrylic-based resin into the seawater is not clear. Moreover, the efficiency and mechanism of ivermectin in preventing biofilm or slime formation have not been fully investigated. In this study, a coating was developed by mixing ivermectin with an acrylic-based resin, silyl acrylate copolymer, and a 45-day in situ antifouling test was conducted in the Yellow Sea. Direct observation and confocal microscope investigation indicated that the polymer coating with ivermectin was effective against biofilm formation. High-throughput sequencing analysis showed that ivermectin can selectively inhibit the adhesion of microorganisms. Abundances of Gammaproteobacteria and Alphaproteobacteria decreased significantly with the increased concentration of ivermectin. As for the eukaryote community, species of Stolidobranchia and unidentified_Bacillariophyceae were proved to be sensitive to ivermectin. Therefore, the ivermectin/silyl acrylate copolymer coating is a promising substitute for marine antifouling material.

RevDate: 2020-06-01

Jeronimo LP, Choi MR, Yeon SH, et al (2020)

Effects of povidone-iodine composite on the elimination of bacterial biofilm.

International forum of allergy & rhinology [Epub ahead of print].

BACKGROUND: Povidone-iodine (PVP-I) is well known as an antiseptic and exhibits extensive activity against various pathogens. However, due to its uniquely unpleasant nature, it cannot be used locally to deactivate various sinonasal pathogens. Therefore, we developed a PVP-I composite that blocks the unpleasant odor of PVP-I for use as a local antiseptic in the sinonasal cavity and evaluated its effect on bacterial biofilm's formation and elimination in in vivo and in vitro models.

METHODS: MTT, lactate dehydrogenase, and live/dead staining assay were performed to examine the cellular toxicity of PVP-I composites on the primary human nasal epithelial and RPMI 2650 cells. Crystal violet assay was performed to quantify bacterial biofilm after treating with various agents, including PVP-I and antibiotics. Hematoxylin-and-eosin staining, live/dead staining assay, and scanning electron microscopy were conducted to evaluate the effect of PVP-I on biofilm formation in a mice biofilm model.

RESULTS: It was observed that the PVP-I composite did not have any significant toxic effect on the nasal epithelial cells. Furthermore, the PVP-I composite effectively inhibited the formation of bacterial biomass within a dose-dependent manner after 48 hours of incubation with Pseudomonas aeruginosa and Staphylococcus aureus. In mice, it effectively eliminated biofilm from the mucosa of the nasal cavity and maxillary sinus at the tested concentrations.

CONCLUSION: The results of this study indicate that the PVP-I composite is a promising compound that could be used locally to prevent the formation of biofilms and to eliminate them from the sinonasal cavity.

RevDate: 2020-06-01

Shakerimoghaddam A, Razavi D, Rahvar F, et al (2020)

Evaluate the effect of zinc oxide and silver nanoparticles (ZnO-Ag NPs) on biofilm and icaA gene expression in methicillin-resistant Staphylococcus aureus (MRSA) isolated from burn wound infection.

Journal of burn care & research : official publication of the American Burn Association pii:5849936 [Epub ahead of print].

Methicillin-resistant Staphylococcus aureus is the cause of nosocomial and community-acquired infections. This study aimed to evaluate the effect of zinc oxide and silver nanoparticles (ZnO-Ag NPs) on biofilms formation and icaA gene expression in methicillin-resistant S. aureus (MRSA).

RevDate: 2020-06-01

Nguyen BVG, Nagakubo T, Toyofuku M, et al (2020)

Synergy between Sophorolipid Biosurfactant and SDS Increases the Efficiency of P. aeruginosa Biofilm Disruption.

Langmuir : the ACS journal of surfaces and colloids [Epub ahead of print].

Biofilms are communities of bacteria encased in self-secreted extracellular polymeric substances (EPS) that adhere stubbornly to submerged surfaces. Once established, these communities can cause serious chronic illnesses in medical settings, while they can promote corrosion and biofouling in industrial settings. Due to the difficulty of their removal, strongly oxidizing chemicals and detergents can be used to degrade and remove biofilms by killing the cells and degrading the matrix; however, the choice of compounds is limited in delicate environments due to the potential damage they may cause. In the case of detergents, most are synthesized from nonrenewable petrochemicals that have a degree of aquatic toxicity. There is a growing need to identify and characterize alternatives to synthetic surfactants. Biosurfactants, which are surfactants produced by microorganisms, are a promising alternative since they can be synthesized from renewable resources, have low environmental toxicity, and have been shown to have higher degrees of specificity in the mechanism of action. Sophorolipids are a class of glycolipid surfactants produced by yeast that have demonstrated great promise due to large yields from renewable feedstocks and for antimicrobial properties; however, the effect of the application of sophorolipids to Gram-negative bacterial biofilms has not been well studied. We investigate the antibiofilm properties of sophorolipids by demonstrating its ability to cause the catastrophic disruption of Pseudomonas aeruginosa PAO1 biofilms in microfluidic channels. We show that while sophorolipids inflict little damage to the bacteria, they weaken the EPS biofilm matrix, leading to surface-detachment and breakup of the biofilm. Furthermore, we find that sophorolipids act cooperatively with the widely used surfactant, sodium dodecyl sulfate. When combined, concentrations ∼100-fold lower than the minimum effective concentration, when used independently, recover potency. Biosurfactants are typically expensive to produce, thus our work demonstrates a means to improve efficacy while simultaneously reducing both cost and the amount of environmentally harmful substances used.

RevDate: 2020-06-01

Qu Y, Li Y, Cameron DR, et al (2020)

Hyperosmotic Infusion and Oxidized Surfaces Are Essential for Biofilm Formation of Staphylococcus capitis From the Neonatal Intensive Care Unit.

Frontiers in microbiology, 11:920.

Staphylococcus capitis is an opportunistic pathogen often implicated in bloodstream infections in the neonatal intensive care unit (NICU). This is assisted by its ability to form biofilms on indwelling central venous catheters (CVC), which are highly resistant to antibiotics and the immune system. We sought to understand the fundamentals of biofilm formation by S. capitis in the NICU, using seventeen clinical isolates including the endemic NRCS-A clone and assessing nine commercial and two modified polystyrene surfaces. S. capitis clinical isolates from the NICU initiated biofilm formation only in response to hyperosmotic conditions, followed by a developmental progression driven by icaADBC expression to establish mature biofilms, with polysaccharide being their major extracellular polymer substance (EPS) matrix component. Physicochemical features of the biomaterial surface, and in particular the level of the element oxygen present on the surface, significantly influenced biofilm development of S. capitis. A lack of highly oxidized carbon species on the surface prevented the immobilization of S. capitis EPS and the formation of mature biofilms. This information provides guidance in regard to the preparation of hyperosmolar total parenteral nutrition and the engineering of CVC surfaces that can minimize the risk of catheter-related bloodstream infections caused by S. capitis in the NICU.

RevDate: 2020-06-01

Adnan M, Patel M, Deshpande S, et al (2020)

Effect of Adiantum philippense Extract on Biofilm Formation, Adhesion With Its Antibacterial Activities Against Foodborne Pathogens, and Characterization of Bioactive Metabolites: An in vitro-in silico Approach.

Frontiers in microbiology, 11:823.

Adiantum philippense (A. philippense), an ethnomedicinally important fern, has become an interesting herb in the search for novel bioactive metabolites, which can also be used as therapeutic agents. Primarily, in this study, A. philippense crude extract was screened for its phytochemical constituents, antagonistic potential, and effect on bacterial adhesion and biofilm formation against common food pathogens. Phytochemical profiling of A. philippense was carried out by using High Resolution-Liquid Chromatography and Mass Spectroscopy (HR-LCMS) followed by antibacterial activity via agar cup/well diffusion, broth microdilution susceptibility methods, and growth curve analysis. Antibiofilm potency and efficacy were assessed on the development, formation, and texture of biofilms through light microscopy, fluorescent microscopy, scanning electron microscopy, and the assessment of exopolysaccharide production. Correspondingly, a checkerboard test was performed to evaluate the combinatorial effect of A. philippense and chloramphenicol. Lastly, molecular docking studies of identified phytochemicals with adhesin proteins of tested food pathogens, which helps the bacteria in surface attachment and leads to biofilm formation, were assessed. A. philippense crude extract was found to be active against all tested food pathogens, displaying the rapid time-dependent kinetics of bacterial killing. A. philippense crude extract also impedes the biofilm matrix by reducing the total content of exopolysaccharide, and, likewise, the microscopic images revealed a great extent of disruption in the architecture of biofilms. A synergy was observed between A. philippense crude extract and chloramphenicol for E. coli, S. aureus, and P. aeruginosa, whereas an additive effect was observed for S. flexneri. Various bioactive phytochemicals were categorized from A. philippense crude extract using HR-LCMS. The molecular docking of these identified phytochemicals was interrelated with the active site residues of adhesin proteins, IcsA, Sortase A, OprD, EspA, and FimH from S. flexneri, S. aureus, P. aeruginosa, and E. coli, respectively. Thus, our findings represent the bioactivity and potency of A. philippense crude extract against food pathogens not only in their planktonic forms but also against/in biofilms for the first time. We have also correlated these findings with the possible mechanism of biofilm inhibition via targeting adhesin proteins, which could be explored further to design new bioactive compounds against biofilm producing foodborne bacterial pathogens.

RevDate: 2020-06-01

Su T, He J, Li N, et al (2020)

A Rational Designed PslG With Normal Biofilm Hydrolysis and Enhanced Resistance to Trypsin-Like Protease Digestion.

Frontiers in microbiology, 11:760.

A glycosyl hydrolase produced by Pseudomonas aeruginosa, PslG, has become a promising candidate for biofilm treatment because of its ability to inhibit and disperse biofilms by disrupting exopolysaccharide matrix at nanomolar concentrations. However, as a protein, PslG used for treatment may be degraded by the ubiquitous proteases (of which trypsin-like serine proteases are a major group) secreted by human cells. This would lead to an insufficient effective concentration of PslG. Here, based on the result of liquid chromatography-tandem mass spectrometry (LC-MS/MS) and structural analysis, we generate a PslG mutant (K286A/K433S) with greatly enhanced trypsin resistance. This measure raises IC50 (the concentration of trypsin that can degrade 50% of protein in 30 min at 37°C) from 0.028 mg mL-1 of the wild-type PslG to 0.283 mg mL-1 of PslG K286A/K433S . In addition, biofilm inhibition assay shows that PslG K286A/K433S is much more efficient than wild-type PslG in the presence of trypsin. This indicates that PslG K286A/K433S is a better biofilm inhibitor than wild-type PslG in clinical use where trypsin-like proteases widely exist.

RevDate: 2020-06-01

Priya A, SK Pandian (2020)

Piperine Impedes Biofilm Formation and Hyphal Morphogenesis of Candida albicans.

Frontiers in microbiology, 11:756.

Candida albicans is the primary etiological agent associated with the pathogenesis of candidiasis. Unrestricted growth of C. albicans in the oral cavity may lead to oral candidiasis, which can progress to systemic infections in worst scenarios. Biofilm of C. albicans encompasses yeast and hyphal forms, where hyphal formation and yeast to hyphal morphological transitions are contemplated as the key virulence elements. Current clinical repercussions necessitate the identification of therapeutic agent that can limit the biofilm formation and escalating the susceptibility of C. albicans to immune system and conventional antifungals. In the present study, a plant-derived alkaloid molecule, piperine, was investigated for the antibiofilm and antihyphal activities against C. albicans. Piperine demonstrated a concentration-dependent antibiofilm activity without exerting negative impact on growth and metabolic activity. Inhibition in the hyphal development was witnessed through confocal laser-scanning microscopy and scanning electron microscopy. Interestingly, piperine displayed a tremendous potential to inhibit the virulence-associated colony morphologies, such as filamentation and wrinkling. Furthermore, piperine regulated morphological transitions between yeast and hyphal forms by inhibiting hyphal extension and swapping hyphal phase to yeast forms yet under filamentation-inducing circumstances. Remarkably, piperine-challenged C. albicans exhibited low potential for spontaneous antibiofilm resistance development. In addition, piperine effectively reduced in vivo colonization and prolonged survival of C. albicans-infected Caenorhabditis elegans, thereby expounding the distinct antivirulent potential. Transcriptomic analysis revealed piperine significantly downregulating the expression of several biofilm related and hyphal-specific genes (ALS3, HWP1, EFG1, CPH1, etc.). Furthermore, no acute toxicity was observed in the HBECs and nematodes exposed to piperine. Altogether, results from this study reveals the potential of piperine to inhibit biofilm and hyphal morphogenesis, and its in vivo efficacy and innocuous nature to HBECs suggests that piperine may be considered as a potential candidate for the treatment of biofilm-associated C. albicans infection, especially for oral candidiasis.

RevDate: 2020-06-01

Jeon DM, An JS, Lim BS, et al (2020)

Orthodontic bonding procedures significantly influence biofilm composition.

Progress in orthodontics, 21(1):14 pii:10.1186/s40510-020-00314-8.

BACKGROUND: Because changes in surface properties affect bacterial adhesion, orthodontic bonding procedures may significantly influence biofilm formation and composition around orthodontic appliances. However, most studies used a mono-species biofilm model under static conditions, which does not simulate the intraoral environment and complex interactions of oral microflora because the oral cavity is a diverse and changeable environment. In this study, a multi-species biofilm model was used under dynamic culture conditions to assess the effects of the orthodontic bonding procedure on biofilm formation and compositional changes in two main oral pathogens, Streptococcus mutans and Porphyromonas gingivalis.

METHODS: Four specimens were prepared with bovine incisors and bonding adhesive: untreated enamel surface (BI), enamel surface etched with 37% phosphoric acid (ET), primed enamel surface after etching (PR), and adhesive surface (AD). Surface roughness (SR), surface wettability (SW), and surface texture were evaluated. A multi-species biofilm was developed on each surface and adhesion amounts of Streptococcus mutans, Porphyromonas gingivalis, and total bacteria were analyzed at day 1 and day 4 using real-time polymerase chain reaction. After determining the differences in biofilm formation, SR, and SW between the four surfaces, relationships between bacteria levels and surface properties were analyzed.

RESULTS: The order of SR was AD < PR < BI < ET, as BI and ET showed more irregular surface texture than PR and AD. For SW, ET had the greatest value followed by PR, BI, and AD. S. mutans and P. gingivalis showed greater adhesion to BI and ET with rougher and more wettable surfaces than to AD with smoother and less wettable surfaces. The adhesion of total bacteria and S. mutans significantly increased over time, but the amount of P. gingivalis decreased. The adhesion amounts of all bacteria were positively correlated with SR and SW, irrespective of incubation time.

CONCLUSIONS: Within the limitations of this study, changes in SR and SW associated with orthodontic bonding had significant effects on biofilm formation and composition of S. mutans and P. gingivalis.

RevDate: 2020-05-31

Liang J, Liang D, Liang Y, et al (2020)

Effects of a derivative of reutericin 6 and gassericin A on the biofilm of Streptococcus mutans in vitro and caries prevention in vivo.

Odontology pii:10.1007/s10266-020-00529-5 [Epub ahead of print].

It is known that Streptococcus mutans (S. mutans) is the leading cariogenic pathogen. Recently, an increasing number of antimicrobial peptides (AMPs) have been brought into consideration as anti-caries agents. Here, we designed and synthesized an AMP derived from reutericin 6 and/or gassericin A, named LN-7, and explored its effect on biofilm of S. mutans UA159 in vitro and development of dental caries in vivo. Antibacterial assays showed that LN-7 was more active against S. mutans (3.2 μM) than many peptide-based agents, capable of killing other types of Streptococci in oral cavity. In addition, LN-7 presented fast killing kinetics, with more than 97% S. mutans killed within 5 min. The mechanism of the antimicrobial activity mainly lies on the disruption of bacterial membrane. Effects of LN-7 on the biofilm formation and the viability of preformed biofilm were quantified by crystal violet staining, which showed that LN-7 could effectively inhibit the biofilm accumulation of S. mutans. Moreover, the biofilm of S. mutans treated with LN-7 displayed notable changes in bacterial viability and morphology, observed by confocal laser scanning microscopy and scanning electron microscopy. In addition, topical oral treatment with LN-7 could suppress the development of dental caries in vivo, reducing the occurrence of severe dental lesion in a rodent model. These results reveal a new peptide-based agent as a topical treatment for dental caries, opening the door to clinical studies to explore its potential for caries prevention.

RevDate: 2020-05-31

Eroshenko DV, Polyudova TV, AA Pyankova (2020)

VapBC and MazEF toxin/antitoxin systems in the regulation of biofilm formation and antibiotic tolerance in nontuberculous mycobacteria.

International journal of mycobacteriology, 9(2):156-166.

Background: Mycobacterium smegmatis and other nontuberculous mycobacteria (NTM) are widely distributed in the environment, but a significant increase of NTM infections has taken place in the last few decades. The objective of this study was to determine the role of toxin-antitoxin (TA) vapBC and mazEF systems that act as effectors of persistence in the stress response of NTM.

Methods: The growth ability and the biofilm formation of NTM were evaluated by conventional methods. Bacterial cell viability was determined using MTT staining, agar plating, or the method of limiting dilutions. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of antibiotics were estimated using broth and agar dilution methods.

Results: Despite a comparable growth dynamics and biofilm formation on solid/liquid interface with the wild type, a M. smegmatis vapBC, mazEF, and vapBC × mazEF deletion mutant produced more abundant pellicle and were more susceptible to heat shock. Significant differences were also found in the resistance wild type of NTM to isoniazid and ciprofloxacin reflected by higher MBC/MIC ratios. The proposed method of cultivation of agar blocks without visible growth after MIC determination into a liquid medium allows us to detect transition of all wild type of NTM strains to a dormant state in the presence of subMICs of isoniazid and ciprofloxacin while all deletion mutants failed to form dormant cells.

Conclusion: Our data suggest that both vapBC and mazEF TA systems putatively involved in the heat and antibiotic stress response of NTM via their key role in transition to the dormant state.

RevDate: 2020-05-31

Wagner EM, Pracser N, Thalguter S, et al (2020)

Identification of biofilm hotspots in a meat processing environment: Detection of spoilage bacteria in multi-species biofilms.

International journal of food microbiology, 328:108668 pii:S0168-1605(20)30162-8 [Epub ahead of print].

Biofilms are comprised of microorganisms embedded in a self-produced matrix that normally adhere to a surface. In the food processing environment they are suggested to be a source of contamination leading to food spoilage or the transmission of food-borne pathogens. To date, research has mainly focused on the presence of (biofilm-forming) bacteria within food processing environments, without measuring the associated biofilm matrix components. Here, we assessed the presence of biofilms within a meat processing environment, processing pork, poultry and beef, by the detection of microorganisms and at least two biofilm matrix components. Sampling included 47 food contact surfaces and 61 non-food contact surfaces from eleven rooms within an Austrian meat processing plant, either during operation or after cleaning and disinfection. The 108 samples were analysed for the presence of microorganisms by cultivation and targeted quantitative real-time PCR based on 16S rRNA. Furthermore, the presence of the major matrix components carbohydrates, extracellular DNA and proteins was evaluated. Overall, we identified ten biofilm hotspots, among them seven of which were sampled during operation and three after cleaning and disinfection. Five biofilms were detected on food contact surfaces (cutters and associated equipment and a screw conveyor) and five on non-food contact surfaces (drains and water hoses) resulting in 9.3 % of the sites being classified as biofilm positive. From these biofilm positive samples, we cultivated bacteria of 29 different genera. The most prevalent bacteria belonged to the genera Brochothrix (present in 80 % of biofilms), Pseudomonas and Psychrobacter (isolated from 70 % biofilms). From each biofilm we isolated bacteria from four to twelve different genera, indicating the presence of multi-species biofilms. This work ultimately determined the presence of multi-species biofilms within the meat processing environment, thereby identifying various sources of potential contamination. Especially the identification of biofilms in water hoses and associated parts highlights the need of a frequent monitoring at these sites. The knowledge gained about the presence and composition of biofilms (i.e. chemical and microbiological) will help to prevent and reduce biofilm formation within food processing environments.

RevDate: 2020-05-31

Wang J, Liu Q, Dong D, et al (2020)

In-situ monitoring of the unstable bacterial adhesion process during wastewater biofilm formation: A comprehensive study.

Environment international, 140:105722 pii:S0160-4120(20)30641-3 [Epub ahead of print].

The initial bacterial adhesion phase is a pivotal and unstable step in the formation of biofilms. The initiation of biofilm formation is an unstable process caused by the reversible adhesion of bacteria, which is always time-consuming and yet to be elucidated. In this study, impedance-based real time cell analysis (RTCA) was employed to comprehensively investigate the initial bacterial adhesion process. Results showed that the time required for the unstable adhesion process was significantly (p < 0.05) reduced by increasing the initial concentration of bacteria, which is mainly attributed to the large deposition rate of bacteria at high concentrations. In addition, the unstable adhesion process is also regulated by shear stress, derived in this work from orbital shaking. Shear stress improves the reversibility of unstable bacterial attachment. Furthermore, attachment characteristics during the unstable phase vary between different species of bacteria (Sphingomonas rubra, Nakamurella multipartita and mixed bacteria). The S. rubra strain and mixed culture were more prone to adhere to the substratum surface during the unstable process, which was attributed to the smaller xDLVO energy barrier and motility of species in comparison with N. multipartita. Meanwhile, the molecular composition of extracellular polymeric substances (EPS) in the initial attachment phase presented a significant difference in expressed proteins, indicating the important role of proteins in EPS that strengthen bacterial adhesion. Overall, these findings suggest that during the biofilm reactor start-up process, seed sludge conditions, including the bacterial concentration, composition and hydraulics, need to be carefully considered.

RevDate: 2020-05-31

Huyan J, Tian Z, Zhang Y, et al (2020)

Dynamics of class 1 integrons in aerobic biofilm reactors spiked with antibiotics.

Environment international, 140:105816 pii:S0160-4120(20)31771-2 [Epub ahead of print].

Class 1 integrons are strongly associated with the dissemination of antibiotic resistance in bacteria. However, little is known about whether the presence of antibiotics affects the abundance of integrons and antibiotic resistance genes during biological wastewater treatment. To explore the roles of class 1 integrons in spreading antibiotic resistance genes in environmental compartments, the dynamics of integrons were followed in biofilm reactors treating synthetic wastewater respectively spiked with streptomycin (STM) and oxytetracycline (OTC). The relative abundance of the integron-integrase gene (intI1) increased 12 or 29-fold respectively when treated with STM or OTC, under incrementally increasing dosage regimes from 0 to 50 mg L-1. Significant increases in intI1 abundance initially occurred at an antibiotic dose of 0.1 mg L-1. At the beginning of the experiment, 51% to 64% of integrons carried no gene cassettes. In STM and OTC spiked systems, there was a significant increase in the proportion of integrons that contained resistance gene cassettes, particularly at intermediate and higher antibiotic concentrations. Gene cassettes encoding resistance to aminoglycosides, trimethoprim, beta-lactam, erythromycin, and quaternary ammonium compounds were all detected in the treated systems. Three tetracycline resistance genes (tetA, tetC, tetG) were significantly correlated with the abundance of intI1 (p < 0.01), despite no tet resistance being present as a gene cassette. Genome sequencing of isolates showed synteny between the tet resistance genes and intI1, mediated through linkage to transposable elements including Tn3, IS26 and ISCR3. Class 1 integrons appeared to be under positive selection in the presence of antibiotics, and might have actively acquired new gene cassettes during the experiment.

RevDate: 2020-05-31

Zhang Z, Liu Y, Lu M, et al (2020)

Rhodiola rosea extract inhibits the biofilm formation and the expression of virulence genes of cariogenic oral pathogen Streptococcus mutans.

Archives of oral biology, 116:104762 pii:S0003-9969(20)30140-0 [Epub ahead of print].

OBJECTIVE: The present study aimed to evaluate the effect of Rhodiola rosea extract (RE) on Streptococcus mutans biofilm formation and the relevant mechanism of its action.

METHODS: The effect of RE on the biofilm formation and extracellular polysaccharides (EPS) synthesis of S. mutans was assessed by confocal laser scanning microscopy (CLSM), crystal violet staining and CFU counting method. Scanning electron microscopy (SEM) was applied to observe the surface morphology of S. mutans biofilms formed on glass coverslips and dental enamel. To study the relevant mechanism, quantitative real time PCR (qRT-PCR) and zymogram assay were applied to measure the expression of virulence genes and the enzymatic activity of glucosyltransferases (Gtfs) under the treatment of RE. The CCK-8 assay was also performed on macrophages (RAWs) and human oral keratinocytes (HOKs) in order to evaluate its biocompatibility.

RESULTS: As a result, RE inhibited the biofilm formation and EPS synthesis of S. mutans. RE also suppressed the expression of gtf genes and quorum sensing (QS) system as well as the enzymatic activity of Gtf proteins. Moreover, RE exhibited a good biocompatibility to human cells.

CONCLUSIONS: This study provides the evidence for RE as a novel anti-biofilm agent for clinical use.

RevDate: 2020-05-30

Xiong F, Zhao X, Wen D, et al (2020)

Effects of N-acyl-homoserine lactones-based quorum sensing on biofilm formation, sludge characteristics, and bacterial community during the start-up of bioaugmented reactors.

The Science of the total environment, 735:139449 pii:S0048-9697(20)32966-1 [Epub ahead of print].

Bioaugmentation is an effective technology for treating wastewater containing recalcitrant organic pollutants. However, it is restricted by several technical problems, including the difficult colonization and survival of the inoculated bacteria, and the time-consuming start-up process. Considering the important roles of quorum sensing (QS) in regulating microbial behaviors, this study investigated the effects of N-acyl-homoserine lactones (AHLs)-based manipulation on the start-up of biofilm reactors bioaugmented with a pyridine-degrading strain Paracoccus sp. BW001. The results showed that, in the presence of two specific exogenous AHLs (C6-HSL and 3OC6-HSL), the biofilm formation process on carriers was significantly accelerated, producing thick and structured biofilms. The protein and polysaccharide contents of the extracellular polymeric substances (EPS) and soluble microbial products (SMP) in sludge were also elevated, possibly due to the increased abundance of several EPS-producing bacterial genera. Specifically, the stability and complexity of protein structures were improved. Besides the reactor running time, the AHL-manipulation was proved to be the main factor that drove the shift of bacterial community structures in the reactors. The addition of exogenous AHLs significantly increased the succession rate of bacterial communities and decreased the bacterial alpha diversity. Most importantly, the final proportions of the inoculated strain BW001 were elevated by nearly 100% in both sludge and biofilm communities via the AHL-manipulation. These findings strongly elucidated that AHL-based QS was deeply involved in biofilm formation, sludge characteristics, and microbial community construction in bioaugmented reactors, providing a promising start-up strategy for bioaugmentation technology.

RevDate: 2020-05-30

Gieroba B, Krysa M, Wojtowicz K, et al (2020)

The FT-IR and Raman Spectroscopies as Tools for Biofilm Characterization Created by Cariogenic Streptococci.

International journal of molecular sciences, 21(11): pii:ijms21113811.

Fourier transform infrared (FT-IR) and Raman spectroscopy and mapping were applied to the analysis of biofilms produced by bacteria of the genus Streptococcus. Bacterial biofilm, also called dental plaque, is the main cause of periodontal disease and tooth decay. It consists of a complex microbial community embedded in an extracellular matrix composed of highly hydrated extracellular polymeric substances and is a combination of salivary and bacterial proteins, lipids, polysaccharides, nucleic acids, and inorganic ions. This study confirms the value of Raman and FT-IR spectroscopies in biology, medicine, and pharmacy as effective tools for bacterial product characterization.

RevDate: 2020-05-30

Bossù M, Selan L, Artini M, et al (2020)

Characterization of Scardovia wiggsiae Biofilm by Original Scanning Electron Microscopy Protocol.

Microorganisms, 8(6): pii:microorganisms8060807.

Early childhood caries (ECC) is a severe manifestation of carious pathology with rapid and disruptive progression. The ECC microbiota includes a wide variety of bacterial species, among which is an anaerobic newly named species, Scardovia wiggsiae, a previously unidentified Bifidobacterium. Our aim was to provide the first ultrastructural characterization of S. wiggsiae and its biofilm by scanning electron microscopy (SEM) using a protocol that faithfully preserved the biofilm architecture and allowed an investigation at very high magnifications (order of nanometers) and with the appropriate resolution. To accomplish this task, we analyzed Streptococcusmutans' biofilm by conventional SEM and VP-SEM protocols, in addition, we developed an original procedure, named OsO4-RR-TA-IL, which avoids dehydration, drying and sputter coating. This innovative protocol allowed high-resolution and high-magnification imaging (from 10000× to 35000×) in high-vacuum and high-voltage conditions. After comparing three methods, we chose OsO4-RR-TA-IL to investigate S.wiggsiae. It appeared as a fusiform elongated bacterium, without surface specialization, arranged in clusters and submerged in a rich biofilm matrix, which showed a well-developed micro-canalicular system. Our results provide the basis for the development of innovative strategies to quantify the effects of different treatments, in order to establish the best option to counteract ECC in pediatric patients.

RevDate: 2020-05-29

Elsayed MM, Elgohary FA, Zakaria AI, et al (2020)

Novel eradication methods for Staphylococcus aureus biofilm in poultry farms and abattoirs using disinfectants loaded onto silver and copper nanoparticles.

Environmental science and pollution research international pii:10.1007/s11356-020-09340-9 [Epub ahead of print].

Recent developments in the nanotechnology field have created opportunities to design new biomaterials for Staphylococcus aureus biofilm eradication. These biomaterials including disinfectant-loaded nanoparticles could overcome the limitations of conventional disinfectants. The objective of this study was to assess the biocidal activity of five commercial disinfectants (DC&R®, VirkonS®, TH4++, Tek-Trol, and peracetic acid) alone and as with silver and copper nanocomposites on S. aureus biofilm at different concentrations and exposure times. Consequently, 227 samples were collected from two broiler farms, two-layer farms, and three abattoirs at El-Dakahlia Province, Egypt, during summer 2018. The samples were collected from birds as well as the surrounding environment. S. aureus strains were isolated and biofilm producers were phenotypically evaluated by Congo red agar (CRA) test. Besides, 4 biofilm-associated genes including bap, fnbA, cna, and ebps were genotypically detected by PCR technology. Out of 227 collected samples, 141 (62.1%) strains were identified as S. aureus, while 127 strains (90.1%) were S. aureus biofilm producers for all examined samples except for hand swabs of abattoir workers. The prevalence of fnbA and bap genes was 79.5% (101/127) and 20.5% (26/127), respectively but, no strains harbored cna or ebps genes. Tested nanocomposites were prepared using an aqueous solution of metal salts such as copper sulfate and silver nitrate and added to the same amount of disinfectant solution. The obtained nanocomposites were characterized by transmission electron microscopy (TEM) and zeta potential which showed spherical and elongated particles and with a surface charge of disinfectants-silver and copper nanocomposites-of 2.92 and 3.43 mV, respectively. Complete eradication of S. aureus biofilm was observed after treatment with disinfectants loaded onto silver (AgNPs) and copper (CuNPs) nanoparticles in varying concentrations as well as at different exposure times in comparing to disinfectants alone. Our results exhibited the potential applications of disinfectant nanocomposites in complete eradication of S. aureus biofilm in farms and abattoirs without developing of disinfectant resistant bacteria.

RevDate: 2020-05-29

Sen CK, Mathew-Steiner SS, Das A, et al (2020)

Electroceutical Management of Bacterial Biofilm and Surgical Infection.

Antioxidants & redox signaling [Epub ahead of print].

SIGNIFICANCE: In the host-microbe microenvironment, bioelectrical factors influence microbial, host as well as host-microbe interactions. This article discusses relevant mechanistic underpinnings of this novel paradigm. It also addresses how such knowledge may be leveraged to develop novel electroceutical solutions to manage biofilms in the context of surgical infection. Recent Advances: Systematic review and meta-analysis of several hundred wound studies reported a 78.2% prevalence of biofilms in chronic wounds. Biofilm infection is a major cause of delayed wound healing. In the host-microbe microenvironment, bioelectrical factors influence microbial, host as well as host-microbe interactions.

CRITICAL ISSUES: Rapid biological responses are driven by electrical signals generated by minute currents of ions moving across cell membranes. Bacterial life, growth and function relies on a bioelectrical milieu which when perturbed impairs their ability to form biofilm, a major threat to healthcare. Viral stability depends on electrostatic forces. Weak electrical field strength, otherwise safe for humans, can achieve such benefit. In the host, electric field enhanced keratinocyte migration, bolstered immune defenses, improved mitochondrial function and demonstrated multiple other effects consistent with supporting wound healing. Deeper mechanistic understanding of electrical principles will inform the design of next generation electroceuticals.

FUTURE DIRECTIONS: This is an opportune moment in time as there is a surge of interest in electroceuticals in medicine. Projected to reach $35.5 billion by 2025, electroceuticals are becoming a cynosure in the global market. WHO reports that more than 50% of surgical site infections can be antibiotic resistant. Electroceuticals offer a serious alternative.

RevDate: 2020-05-29

Noumi E, Merghni A, Alreshidi M, et al (2020)

Phenotypic and Genotypic Characterization with MALDI-TOF-MS Based Identification of Staphylococcus spp. Isolated from Mobile Phones with their Antibiotic Susceptibility, Biofilm Formation, and Adhesion Properties.

International journal of environmental research and public health, 17(11): pii:ijerph17113761.

Cell phones, smartphones, and tablets are extensively used in social and professional life, so they are frequently exposed to bacteria. The main goal of the present work was to isolate and characterize Staphylococci strains from students' cell phone mobiles. Subsequently, 24 Staphylococci strains were tested against a wide range of antibiotics, for the distribution of some virulence-related genes and their ability to form biofilm. Staphylococcus spp. were cultured from all studied devices on chromogenic medium and identified using the matrix-assisted laser desorption/ionization (MALDI), time-of-flight (TOF) mass spectrometry (MS) technique (MALDI-TOF-MS). The results obtained showed that S.aureus was the dominant species (19 strains, 79.1%), followed by S.warneri (3 strains, 12.5%), and S.haemolyticus (2 strains, 8.3%). Isolated strains showed high percentages of hydrolytic enzymes production, resistance to many tested antibiotics, and 37.5% expressed the mecA gene. The tested strains were highly adhesive to polystyrene and glass and expressed implicated icaA (62.5%) and icaD (66.6%) genes. All Staphylococcus spp. strains tested were found to possess proteases and the α-hemolysin gene. Our results highlighted the importance of mobile phones as a great source of Staphylococcus spp., and these species were found to be resistant to many antibiotics with multiple antibiotic resistance (MAR) index ranging from (0.444) to (0.812). Most of the studied strains are able to form biofilm and expressed many virulence genes. Phylogenetic analysis based on the phenotypic and genetic characters highlighted the phenotypic and genetic heterogeneity of the S.aureus population studied. Further analyses are needed to elucidate the human health risks associated with the identified Staphylococci strains.

RevDate: 2020-05-29

Abdel-Aziz MM, Emam TM, MM Raafat (2020)

Hindering of Cariogenic Streptococcus mutans Biofilm by Fatty Acid Array Derived from an Endophytic Arthrographis kalrae Strain.

Biomolecules, 10(5): pii:biom10050811.

Streptococcus mutans has been considered as the major etiological agent of dental caries, mostly due to its arsenal of virulence factors, including strong biofilm formation, exopolysaccharides production, and high acid production. Here, we present the antivirulence activity of fatty acids derived from the endophytic fungus Arthrographis kalrae isolated from Coriandrum sativum against Streptococcus mutans. The chemical composition of the fatty acids was analyzed by gas chromatography-mass spectrometry GC-MS and revealed nine compounds representing 99.6% of fatty acids, where unsaturated and saturated fatty acids formed 93.8% and 5.8 % respectively. Oleic and linoleic acids were the major unsaturated fatty acids. Noteworthy, the fatty acids at the concentration of 31.3 mg L-1 completely inhibited Streptococcus mutans biofilm, and water insoluble extracellular polysaccharide production in both polystyrene plates, and tooth model assay using saliva-coated hydroxyapatite discs. Inhibition of biofilm correlated significantly and positively with the inhibition of water insoluble extracellular polysaccharide (R=1, p <0.0001). Furthermore, Arthrographis kalrae fatty acids at a concentration of 7.8 mg L-1 exhibited acidogenesis-mitigation activity. They did not show bactericidal activity against Streptococcus mutans and cytotoxic activity against human oral fibroblast cells at the concentration used. On the other hand, saliva-coated hydroxyapatite discs treated with sub-minimum biofilm inhibitory concentration of fatty acids showed disturbed biofilm architecture with a few unequally distributed clumped matrices using fluorescence microscopy. Our findings revealed that the intracellular fatty acid arrays derived from endophytic Arthrographis kalrae could contribute to the biofilm-preventing alternatives, specifically Streptococcus mutans biofilms.

RevDate: 2020-05-29

Swolana D, Kępa M, Idzik D, et al (2020)

The Antibacterial Effect of Silver Nanoparticles on Staphylococcus epidermidis Strains with Different Biofilm-Forming Ability.

Nanomaterials (Basel, Switzerland), 10(5): pii:nano10051010.

Among many infectious diseases, infections caused by pathogens of Staphylococcus species exert a substantial influence upon human health, mainly due to their continuous presence on human skin and mucous membranes. For that reason, an intensive search for new, effective anistaphyloccocal agents can currently be observed worldwide. In recent years, there has been growing interest in nanoparticles, as compounds with potential antibacterial effect. The antibacterial activity of silver containing substances has been well recognized, but thoughtful studies focused on the effect of silver nanoparticles on bacterial biofilm are scarce. The aim of this study was to assess the influence of silver nanoparticles (AgNPs) with particle sizes in the range between 10 and 100 nm, and a concentration range from 1 to 10 µg/mL, upon Staphylococcus epidermidis strains with different biofilm-forming abilities (BFAs). The studies revealed the highest level of antimicrobial activity for AgNPs in relation to S. epidermidis strains with BFA, and what is more, the observed effect was proportional to the increasing particles' size, and strains not forming biofilm were more susceptible to silver nanoparticles with the smallest examined size, which was 10 nm.

RevDate: 2020-05-29

Staszczyk M, Jurczak A, Magacz M, et al (2020)

Effect of Polyols and Selected Dental Materials on the Ability to Create a Cariogenic Biofilm-On Children Caries-Associated Streptococcus Mutans Isolates.

International journal of environmental research and public health, 17(10): pii:ijerph17103720.

Secondary caries is a disease associated with the formation of biofilm on the border of the tooth and dental filling. Its development is strongly influenced by the dietary sweet foods and the type of dental material. The aim of the study was to assess the effect of sweeteners on the ability of clinical Streptococcus mutans strains to form biofilm on dental materials. Strains were isolated from plaque samples from 40 pediatric patients from the 3-6 ICADS II group. The ability to form biofilm was tested on composite and glass ionomer dental materials used for milk teeth filling in the presence of sucrose, xylitol, sorbitol, and erythritol. The bacterial film mass after 12, 24, 48, and 72 h and the number of bacterial colonies significantly decreased (p < 0.01) compared to the initial value for 5% erythritol and sorbitol on examined materials. A greater inhibitory effect was noted for glass ionomers compared to composites. Sucrose and xylitol supported biofilm formation, while erythritol had the best inhibitory effect. The use of fluoride-releasing glass ionomers exerted an effect synergistic to erythritol, i.e., inhibited plaque formation and the amount of cariogenic S. mutans. Selection of proper type of dental material together with replacing sucrose with polyols can significantly decrease risk of secondary caries development. Erithritol in combination with glass ionomer seems to be the most effective in secondary caries prevention.

RevDate: 2020-05-29

Iseppi R, Di Cerbo A, Aloisi P, et al (2020)

In Vitro Activity of Essential Oils Against Planktonic and Biofilm Cells of Extended-Spectrum β-Lactamase (ESBL)/Carbapenamase-Producing Gram-Negative Bacteria Involved in Human Nosocomial Infections.

Antibiotics (Basel, Switzerland), 9(5): pii:antibiotics9050272.

The aim of this study was to analyze the antibacterial activity of four essential oils (EOs), Melaleuca alternifolia, Eucalyptus globulus, Mentha piperita, and Thymus vulgaris, in preventing the development and spread of extended-spectrum β-lactamase (ESBL)-producing Escherichia coli and Klebsiella pneumoniae, metallo-beta-lactamase (MBL)-producing Pseudomonas aeruginosa and carbapenemase (KPC)-producing Klebsiella pneumoniae. A total of 60 strains were obtained from the stock collection from the Microbiology Laboratory of Hesperia Hospital, Modena, Italy. Twenty ESBL-producing E. coli, 5 K. pneumoniae, 13 KPC-producing K. pneumoniae, and 20 MBL-producing P. aeruginosa were cultured and reconfirmed as ESBL and carbapenamase producers. Polymerase chain reaction was used for the detection of genes responsible for antibiotic resistance (ESBL and KPC/MBL). Antibacterial activity of the EOs was determined using the agar disk diffusion assay, and minimal inhibitory concentrations (MICs) were also evaluated. Lastly, adhesion capability and biofilm formation on polystyrene and glass surfaces were studied in 24 randomly selected strains. M. alternifolia and T. vulgaris EOs showed the best antibacterial activity against all tested strains and, as revealed by agar disk diffusion assay, M. alternifolia was the most effective, even at low concentrations. This effect was also confirmed by MICs, with values ranging from 0.5 to 16 µg/mL and from 1 to 16 µg/mL, for M. alternifolia and T. vulgaris EOs, respectively. The EOs' antibacterial activity compared to antibiotics confirmed M. alternifolia EO as the best antibacterial agent. T. vulgaris EO also showed a good antibacterial activity with MICs lower than both reference antibiotics. Lastly, a significant anti-biofilm activity was observed for the two EOs (*P < 0.05 and **P < 0.01 for M. alternifolia and T. vulgaris EOs, respectively). A good antibacterial and anti-biofilm activity of M. alternifolia and T. vulgaris EOs against all selected strains was observed, thus demonstrating a future possible use of these EOs to treat infections caused by ESBL/carbapenemase-producing strains, even in association with antibiotics.

RevDate: 2020-05-28

de Assis Graciano Dos Santos F, Leite-Andrade MC, de Sousa Brandão I, et al (2020)

Anti-biofilm effect by the combined action of fluconazole and acetylsalicylic acid against species of Candida parapsilosis complex.

Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases pii:S1567-1348(20)30209-4 [Epub ahead of print].

The Candida parapsilosis complex has been associated with highly refractory infections mainly due to the presence of biofilms. High glucose levels enable the development of this virulence factor which can aggravate the clinical condition of patients with diabetes mellitus, those using parenteral nutrition, with invasive medical device, including others. Combined antifungal therapy, such as azole and cyclooxygenase inhibitors, may be an alternative in such infections since they modulate prostaglandin production favoring the adhesion and development of biofilms. Thus, the present study aimed to evaluate the influence of glucose supplementation in the formation and detection of Candida parapsilosis complex biofilms and to treat them using fluconazole and a cyclooxygenase inhibitor in combination. Protein spectra evaluation allowed the differentiation between species from the complex (score > 2) in our studies. All isolates were able to form active biofilms at different glucose concentrations. In addition, a significant reduction in biofilm formation was observed when fluconazole and acetylsalicylic acid were combined. The ultrastructural analysis presented typical biofilm characteristics by species from the complex. These data support new combined therapies for the treatment of fungal infections, especially with those which are resistant and therapeutic failure is associated with virulence factors.

RevDate: 2020-05-28

Frassinetti S, Falleni A, R Del Carratore (2020)

Effect of itraconazole on Staphylococcus aureus biofilm and extracellular vesicles formation.

Microbial pathogenesis pii:S0882-4010(20)30633-1 [Epub ahead of print].

Staphylococcus aureus is a leading cause of a wide range of clinical chronic infections mainly due to the establishment of a biofilm. Biofilm, a population of bacteria within a self-produced matrix of extracellular polymeric substance, decreases the susceptibility to antibiotics, immune defenses and contributes to antimicrobial resistance. To date antibiotic combination has been considered a strategy to combat S. aureus infection, but this approach does not solves the main pharmacokinetic problem caused by biofilms, consisting in insufficient drug penetration within the structure. Therefore, new antimicrobial agents that could overcome this resistance need to be discovered. Fighting staphylococcal resistance and biofilm formation is an important goal of the pharmaceutical research. Some fungicide has been observed to have antibacterial effect anyway their use as antibiotics on S.aureus has been poorly studied. The aim of this work was to investigate the effect of the fungicide itraconazole (IT) on S. aureus biofilm formation and explore by SEM the morphological alteration after treatment. A strong biofilm disaggregation and morphologically different extracellular vesicles (EV) production were observed starting from sublethal IT doses. This suggests that IT resistance phenomena on the part of S. aureus are more difficult to establish respect other antibiotics. The adjuvant properties of IT could be used to combat bacterial biofilm and/or to improve antibiotic treatment. Moreover because the production of EV represents a secretory pathway involved in intercellular communication shared to mammalian cells, fungi, and bacteria, our study is important to increase information that can be generalized to higher organisms.

RevDate: 2020-05-28

Okaro U, George S, Valdes S, et al (2020)

A non-coding RNA controls transcription of a gene encoding a DNA binding protein that modulates biofilm development in Bartonella henselae.

Microbial pathogenesis pii:S0882-4010(20)30638-0 [Epub ahead of print].

Bartonella henselae (Bh) is a Gram-negative zoonotic bacterium that can grow as large aggregates and form biofilms in vitro dependent upon the adhesin BadA. Previously, we reported that the Houston-1 strain of Bh has a family of nine small, highly-expressed intergenic transcripts called Bartonellaregulatory transcripts, Brt1-9. Each of the Brts bears a stem and loop structure on the 3' end followed by a gene encoding a DNA binding protein called the Transcriptional regulatory proteins, Trp1-9. RNA-seq analysis of laboratory-grown bacteria revealed the trps were poorly transcribed suggesting that the 3' stem and loop on the Brts results in transcript termination upstream of the trp genes under these conditions. Here we demonstrate that transcription of brt1 continues into trp1 when Bh is grown in a biofilm. Deletion of brt1, or just the 3' terminus of brt1 (containing the stem and loop structure), resulted in increased transcription of both trp1 and badA and increased biofilm formation. Trp1 was shown to directly bind the putative badA promoter region as demonstrated by an electrophoretic mobility shift assay (EMSA). Our data suggest that the 3' end of brt1 responds to a stimulus generated by growth of Bh in an in vitro biofilm to allow increased trp1 transcription. We further show that transcription of trp1 increases under conditions consistent with the mammalian host but is not highly expressed in the cat flea vector until the bacterium is excreted into the flea feces. Based on these data, we hypothesize that the 3' end of Brt1 functions to control trp1 transcription and Trp1 in turn results in increased badA expression and enhanced biofilm formation.

RevDate: 2020-05-28

Kim HR, Shin DS, Jang HI, et al (2020)

Anti-biofilm and anti-virulence effects of zerumbone against Acinetobacter baumannii.

Microbiology (Reading, England) [Epub ahead of print].

Acinetobacter baumannii is a multidrug-resistant opportunistic pathogen that affects patients with a compromised immune system and is becoming increasingly important as a hospital-derived infection. This pathogen is difficult to treat owing to its intrinsic multidrug resistance and ability to form antimicrobial-tolerant biofilms. In the present study, we aimed to assess the potential use of zerumbone as a novel anti-biofilm and/or anti-virulence agent against A. baumannii. The results showed that zerumbone at sub-inhibitory doses decreased biofilm formation and disrupted established A. baumannii biofilms. The zerumbone-induced decrease in biofilm formation was dose-dependent based on the results of microtitre plate biofilm assays and confocal laser scanning microscopy. In addition, our data validated the anti-virulence efficacy of zerumbone, wherein it significantly interfered with the motility of A. baumannii. To support these phenotypic results, transcriptional analysis revealed that zerumbone downregulated the expression of biofilm- and virulence-associated genes (adeA, adeB, adeC and bap) in A. baumannii. Overall, our findings suggested that zerumbone might be a promising bioactive agent for the treatment of biofilm- and virulence-related infections caused by multidrug-resistant A. baumannii.

RevDate: 2020-05-28

Badar W, MA Ullah Khan (2020)

Analytical study of biosynthesised silver nanoparticles against multi-drug resistant biofilm-forming pathogens.

IET nanobiotechnology, 14(4):331-340.

The emergence of the huge number of multi-drug resistant (MDR) bacteria requires an alternative to the drugs. Silver nanoparticles (AgNPs) are a strong candidate for this due to their bactericidal properties, which can be better concluded by understanding their morphology and chemistry. The study hypothesised that AgNPs synthesised using leaves of Syzygium cumini can be used to treat locally emerging MDRs forming biofilms on indwelling medical devices. Synthesised particles were characterised by methods like UV-visible spectroscopy, X-ray powder diffraction, scanning electron microscopy, transmission electron microscopy, and Zetasizer. Fourier transform infrared spectroscopy, and high-performance liquid chromatography were used to predict phytochemicals present in the leaves. The shape of particles is revealed to be relatively spherical, with average size to be around 10-100 nm. Phenolic compounds are attributed to the formation of nanoparticles, stability analysis shows particles to be stable, and zeta potential determined the surface charge to be -20.1 mV. Biosynthesised particles are found to possess efficient antibacterial activity MDR bacteria developing biofilms in medical devices; hence, it is concluded that S. cumini based NPs can be used to develop a layer on implant-related medical devices. Toxicity evaluation against A594 cancer cells portrays AgNPs to be potential tumour reduction agents in a concentration-dependent manner.

RevDate: 2020-05-28

Cusimano MG, Ardizzone F, Nasillo G, et al (2020)

Biogenic iron-silver nanoparticles inhibit bacterial biofilm formation due to Ag+ release as determined by a novel phycoerythrin-based assay.

Applied microbiology and biotechnology pii:10.1007/s00253-020-10686-w [Epub ahead of print].

Silver nanoparticles (Ag-NPs) can be considered as a cost-effective alternative to antibiotics. In the presence of Fe(III)-citrate and Ag+, Klebsiella oxytoca DSM 29614 produces biogenic Ag-NPs embedded in its peculiar exopolysaccharide (EPS). K. oxytoca DSM 29614 was cultivated in a defined growth medium-containing citrate (as sole carbon source) and supplemented with Ag+ and either low or high Fe(III) concentration. As inferred from elemental analysis, transmission and scanning electron microscopy, Fourier transform infrared spectrometry and dynamic light scattering, Ag-EPS NPs were produced in both conditions and contained also Fe. The production yield of high-Fe/Ag-EPS NPs was 12 times higher than the production yield of low-Fe/Ag-EPS NPs, confirming the stimulatory effect of iron. However, relative Ag content and Ag+ ion release were higher in low-Fe/Ag-EPS NPs than in high-Fe/Ag-EPS NPs, as revealed by emission-excitation spectra by luminescent spectrometry using a novel ad hoc established phycoerythrin fluorescence-based assay. Interestingly, high and low-Fe/Ag-EPS NPs showed different and growth medium-dependent minimal inhibitory concentrations against Staphylococcus aureus ATCC 29213 and Pseudomonas aeruginosa ATCC 15442. In addition, low-Fe/Ag-EPS NPs exert inhibition of staphylococcal and pseudomonal biofilm formation, while high-Fe/Ag-EPS NPs inhibits staphylococcal biofilm formation only. Altogether, these results, highlighting the different capability of Ag+ release, support the idea that Fe/Ag-EPS NPs produced by K. oxytoca DSM 29614 can be considered as promising candidates in the development of specific antibacterial and anti-biofilm agents.Key points • Klebsiella oxytoca DSM 29614 produces bimetal nanoparticles containing Fe and Ag.• Fe concentration in growth medium affects nanoparticle yield and composition.• Phycoerythrin fluorescence-based assay was developed to determine Ag+release.• Antimicrobial efficacy of bimetal nanoparticle parallels Ag+ions release.

RevDate: 2020-05-28

Lima JY, Moreira C, Nunes Freitas PN, et al (2020)

Structuring biofilm communities living in pesticide contaminated water.

Heliyon, 6(5):e03996 pii:e03996.

The wide use of pesticides in agriculture expose microbiota to stressful conditions that require the development of survival strategies. The bacterial response to many pollutants has not been elucidated in detail, as well as the evolutionary processes that occur to build adapted communities. The purpose of this study was to evaluate the bacterial population structure and adaptation strategies in planktonic and biofilm communities in limited environments, as tanks containing water used for washing herbicide containers. This biodiversity, with high percentage of nonculturable microorganisms, was characterized based on habitat and abiotic parameters using molecular and bioinformatics tools. According to water and wastewater standards, the physicochemical conditions of the tank water were inadequate for survival of the identified bacteria, which had to develop survival strategies in this hostile environment. The biodiversity decreased in the transition from planktonic to biofilm samples, indicating a possible association between genetic drift and selection of individuals that survive under stressful conditions, such as heating in water and the presence of chlorine, fluorine and agrochemicals over a six-month period. The abundance of Enterobacter, Acinetobacter and Pseudomonas in biofilms from water tanks was linked to essential processes, deduced from the genes attributed to these taxonomic units, and related to biofilm formation, structure and membrane transport, quorum sensing and xenobiotic degradation. These characteristics were randomly combined and fixed in the biofilm community. Thus, communities of biofilm bacteria obtained under these environmental conditions serve as interesting models for studying herbicide biodegradation kinetics and the prospects of consortia suitable for use in bioremediation in reservoirs containing herbicide-contaminated wastewater, as biofilters containing biofilm communities capable of degrading herbicides.

RevDate: 2020-05-28

Zhang J, Brown J, Scurr D, et al (2020)

Cryo-OrbiSIMS for 3D molecular imaging of a bacterial biofilm in its native state.

Analytical chemistry [Epub ahead of print].

Secondary ion mass spectrometry (SIMS) is gaining popularity for molecular imaging in the life-sciences since it is label-free and allows imaging in two and three dimensions. The recent introduction of the OrbiSIMS has significantly improved the utility for biological imaging through combining sub-cellular spatial resolution with high-performance Orbitrap mass spectrometry. SIMS instruments operate in high-vacuum and samples are typically analysed in a freeze-dried state. Consequently, the molecular and structural information may not be well-preserved. We report a method for molecular imaging of biological materials, preserved in a native state, by using an OrbiSIMS instrument, equipped with cryogenic sample handling, and a high-pressure freezing protocol compatible with mass spectrometry. The performance is demonstrated by imaging a challenging sample (>90% water) of a mature Pseudomonas aeruginosa biofilm in its native state. The 3D distribution of quorum sensing signaling molecules, nucleobases and bacterial membrane molecules are revealed with high spatial-resolution and high mass-resolution. We discover that analysis in the frozen-hydrated state yields a 10,000 fold increase in signal intensity for polar molecules, such as amino acid, which has important implications for SIMS imaging of metabolites and pharmaceuticals.

RevDate: 2020-05-27

Hillman KM, RC Sims (2020)

Struvite formation associated with the microalgae biofilm matrix of a rotating algal biofilm reactor (RABR) during nutrient removal from municipal wastewater.

Water science and technology : a journal of the International Association on Water Pollution Research, 81(4):644-655.

Struvite was observed within the microalgae biofilm matrix of an outdoor, pilot-scale rotating algal biofilm reactor (RABR) designed to remove nitrogen and phosphorus from municipal anaerobic digester filtrate. The bottom layer of cells (2.5-month growth) and two top layers of cells (1-week and 2.5-month growth) were evaluated on east- and west-facing sides of the RABR. Sun orientation and shading effects of upper biofilm layers impacted the species composition and microalgae content of the bottom biofilm layers. Struvite formed within the microalgae biofilm matrix, and a higher struvite content appeared to be correlated with a higher microalgae content. The highest struvite content (expressed as %wt. of total solids) was observed in the east- and west-facing bottom layers of growth and west-facing 1-week growth (5.0%, 4.3%, and 4.1%, respectively). The lowest struvite content was observed in east- and west-facing 2.5-month growth and east-facing 1-week growth (1.1%, 1.5%, and 1.1%, respectively). Despite RABR influent component ion molar ratios with potential for various magnesium and calcium precipitates, microalgae biofilm provided pH and nucleation sites favorable to struvite precipitation. This evaluation is the first in the refereed literature the authors are aware of that reports on the association of struvite formation in the presence of a microalgae biofilm.

RevDate: 2020-05-27

Badal D, Jayarani AV, Kollaran MA, et al (2020)

Pseudomonas aeruginosa biofilm formation on endotracheal tubes requires multiple two-component systems.

Journal of medical microbiology [Epub ahead of print].

Introduction. Indwelling medical devices such as endotracheal tubes (ETTs), urinary catheters, vascular access devices, tracheostomies and feeding tubes are often associated with hospital-acquired infections. Bacterial biofilm formed on the ETTs in intubated patients is a significant risk factor associated with ventilator-associated pneumonia. Pseudomonas aeruginosa is one of the four frequently encountered bacteria responsible for causing pneumonia, and the biofilm formation on ETTs. However, understanding of biofilm formation on ETT and interventions to prevent biofilm remains lagging. The ability to sense and adapt to external cues contributes to their success. Thus, the biofilm formation is likely to be influenced by the two-component systems (TCSs) that are composed of a membrane-associated sensor kinase and an intracellular response regulator.Aim. This study aims to establish an in vitro method to analyse the P. aeruginosa biofilm formation on ETTs, and identify the TCSs that contribute to this process.Methodology. In total, 112 P. aeruginosa PA14 TCS mutants were tested for their ability to form biofilm on ETTs, their effect on quorum sensing (QS) and motility.Results. Out of 112 TCS mutants studied, 56 had altered biofilm biomass on ETTs. Although the biofilm formation on ETTs is QS-dependent, none of the 56 loci controlled quorum signal. Of these, 18 novel TCSs specific to ETT biofilm were identified, namely, AauS, AgtS, ColR, CopS, CprR, NasT, KdpD, ParS, PmrB, PprA, PvrS, RcsC, PA14_11120, PA14_32580, PA14_45880, PA14_49420, PA14_52240, PA14_70790. The set of 56 included the GacS network, TCS proteins involved in fimbriae synthesis, TCS proteins involved in antimicrobial peptide resistance, and surface-sensing. Additionally, several of the TCS-encoding genes involved in biofilm formation on ETTs were found to be linked to flagellum-dependent swimming motility.Conclusions. Our study established an in vitro method for studying P. aeruginosa biofilm formation on the ETT surfaces. We also identified novel ETT-specific TCSs that could serve as targets to prevent biofilm formation on indwelling devices frequently used in clinical settings.

RevDate: 2020-05-27

Parducho KR, Beadell B, Ybarra TK, et al (2020)

The Antimicrobial Peptide Human Beta-Defensin 2 Inhibits Biofilm Production of Pseudomonas aeruginosa Without Compromising Metabolic Activity.

Frontiers in immunology, 11:805.

Biofilm production is a key virulence factor that facilitates bacterial colonization on host surfaces and is regulated by complex pathways, including quorum sensing, that also control pigment production, among others. To limit colonization, epithelial cells, as part of the first line of defense, utilize a variety of antimicrobial peptides (AMPs) including defensins. Pore formation is the best investigated mechanism for the bactericidal activity of AMPs. Considering the induction of human beta-defensin 2 (HBD2) secretion to the epithelial surface in response to bacteria and the importance of biofilm in microbial infection, we hypothesized that HBD2 has biofilm inhibitory activity. We assessed the viability and biofilm formation of a pyorubin-producing Pseudomonas aeruginosa strain in the presence and absence of HBD2 in comparison to the highly bactericidal HBD3. At nanomolar concentrations, HBD2 - independent of its chiral state - significantly reduced biofilm formation but not metabolic activity, unlike HBD3, which reduced biofilm and metabolic activity to the same degree. A similar discrepancy between biofilm inhibition and maintenance of metabolic activity was also observed in HBD2 treated Acinetobacter baumannii, another Gram-negative bacterium. There was no evidence for HBD2 interference with the regulation of biofilm production. The expression of biofilm-related genes and the extracellular accumulation of pyorubin pigment, another quorum sensing controlled product, did not differ significantly between HBD2 treated and control bacteria, and in silico modeling did not support direct binding of HBD2 to quorum sensing molecules. However, alterations in the outer membrane protein profile accompanied by surface topology changes, documented by atomic force microscopy, was observed after HBD2 treatment. This suggests that HBD2 induces structural changes that interfere with the transport of biofilm precursors into the extracellular space. Taken together, these data support a novel mechanism of biofilm inhibition by nanomolar concentrations of HBD2 that is independent of biofilm regulatory pathways.

RevDate: 2020-05-27

Aqawi M, Gallily R, Sionov RV, et al (2020)

Cannabigerol Prevents Quorum Sensing and Biofilm Formation of Vibrio harveyi.

Frontiers in microbiology, 11:858.

Cannabigerol (CBG) is a non-psychoactive cannabinoid naturally present in trace amounts in the Cannabis plant. So far, CBG has been shown to exert diverse activities in eukaryotes. However, much less is known about its effects on prokaryotes. In this study, we investigated the potential role of CBG as an anti-biofilm and anti-quorum sensing agent against Vibrio harveyi. Quorum sensing (QS) is a cell-to-cell communication system among bacteria that involves small signaling molecules called autoinducers, enabling bacteria to sense the surrounding environment. The autoinducers cause alterations in gene expression and induce bioluminescence, pigment production, motility and biofilm formation. The effect of CBG was tested on V. harveyi grown under planktonic and biofilm conditions. CBG reduced the QS-regulated bioluminescence and biofilm formation of V. harveyi at concentrations not affecting the planktonic bacterial growth. CBG also reduced the motility of V. harveyi in a dose-dependent manner. We further observed that CBG increased LuxO expression and activity, with a concomitant 80% downregulation of the LuxR gene. Exogenous addition of autoinducers could not overcome the QS-inhibitory effect of CBG, suggesting that CBG interferes with the transmission of the autoinducer signals. In conclusion, our study shows that CBG is a potential anti-biofilm agent via inhibition of the QS cascade.

RevDate: 2020-05-27

Silva AV, Edel M, Gescher J, et al (2020)

Exploring the Effects of bolA in Biofilm Formation and Current Generation by Shewanella oneidensis MR-1.

Frontiers in microbiology, 11:815.

Microbial electrochemical technologies (METs) have emerged in recent years as a promising alternative green source of energy, with microbes consuming organic matter to produce energy or valuable byproducts. It is the ability of performing extracellular electron transfer that allows these microbes to exchange electrons with an electrode in these systems. The low levels of current achieved have been the limiting factor for the large-scale application of METs. Shewanella oneidensis MR-1 is one of the most studied electroactive organisms regarding extracellular electron transfer, and it has been shown that biofilm formation is a key factor for current generation. The transcription factor bolA has been identified as a central player in biofilm formation in other organisms, with its overexpression leading to increased biofilm. In this work we explore the effect of this gene in biofilm formation and current production by S. oneidensis MR-1. Our results demonstrate that an increased biofilm formation and consequent current generation was achieved by the overexpression of this gene. This information is crucial to optimize electroactive organisms toward their practical application in METs.

RevDate: 2020-05-27

Akinpelu S, Ajayi A, Smith SI, et al (2020)

Efflux pump activity, biofilm formation and antibiotic resistance profile of Klebsiella spp. isolated from clinical samples at Lagos University Teaching Hospital.

BMC research notes, 13(1):258 pii:10.1186/s13104-020-05105-2.

OBJECTIVE: Nosocomial and community acquired multidrug resistant Klebsiella infections are wide spread resulting in high morbidity and mortality due to limited number of antibiotics treatment options. This study investigated efflux pump activity, biofilm forming potential and antibiotic susceptibility profile of Klebsiella spp. isolated from clinical samples in a tertiary hospital in Lagos Nigeria. Eighteen clinical Klebsiella spp. isolated from urine, blood and sputum were subjected to antibiotic susceptibility testing using the disc diffusion method. Efflux pump activity was evaluated by the ethidium bromide cartwheel method and biofilm forming ability was determined by the tissue culture plate technique.

RESULTS: All 18 (100%) Klebsiella isolates were resistant to cefuroxime, cefixime, amoxicillin - clavulanate, ampicillin + cloxacillin, cefotaxime, and imipenem. Seventeen (94.4%) were resistant to ofloxacin while sixteen (88.9%) were resistance to nalidixic acid, Gentamicin and levofloxacin. All Klebsiella isolates possessed active efflux pump with the ability to form biofilm. However, their biofilm forming capabilities varied as 4 (22.2%) were strong, 3 (16.7%) were moderate and 11 (61.1%) were weak biofilm formers. Findings in this study reveal multiple factors at play in mediating the high level of antibiotic resistance observed in Klebsiella isolates. Hence a multifaceted approach is advocated in managing the infections caused by the pathogen.

RevDate: 2020-05-27

Teul J, Deja S, Celińska-Janowicz K, et al (2020)

LC-QTOF-MS and 1H NMR Metabolomics Verifies Potential Use of Greater Omentum for Klebsiella pneumoniae Biofilm Eradication in Rats.

Pathogens (Basel, Switzerland), 9(5): pii:pathogens9050399.

Bacterial wound infections are a common problem associated with surgical interventions. In particular, biofilm-forming bacteria are hard to eradicate, and alternative methods of treatment based on covering wounds with vascularized flaps of tissue are being developed. The greater omentum is a complex organ covering the intestines in the abdomen, which support wound recovery following surgical procedures and exhibit natural antimicrobial activity that could improve biofilm eradication. We investigated changes in rats' metabolome following Klebsiella pneumoniae infections, as well as the greater omentum's ability for Klebsiella pneumoniae biofilm eradication. Rats received either sterile implants or implants covered with Klebsiella pneumoniae biofilm (placed in the peritoneum or greater omentum). Metabolic profiles were monitored at days 0, 2, and 5 after surgery using combined proton nuclear magnetic resonance (1H NMR) and high performance liquid chromatography quadrupole time-of-flight tandem mass spectrometry (LC-QTOF‑MS) measurements of urine samples followed by chemometric analysis. Obtained results indicated that grafting of the sterile implant to the greater omentum did not cause major disturbances in rats' metabolism, whereas the sterile implant located in the peritoneum triggered metabolic perturbations related to tricarboxylic acid (TCA) cycle, as well as choline, tryptophan, and hippurate metabolism. Presence of implants colonized with Klebsiella pneumoniae biofilm resulted in similar levels of metabolic perturbations in both locations. Our findings confirmed that surgical procedures utilizing the greater omentum may have a practical use in wound healing and tissue regeneration in the future.

RevDate: 2020-05-27

Ferriol-González C, P Domingo-Calap (2020)

Phages for Biofilm Removal.

Antibiotics (Basel, Switzerland), 9(5): pii:antibiotics9050268.

Biofilms are clusters of bacteria that live in association with surfaces. Their main characteristic is that the bacteria inside the biofilms are attached to other bacterial cells and to the surface by an extracellular polymeric matrix. Biofilms are capable of adhering to a wide variety of surfaces, both biotic and abiotic, including human tissues, medical devices, and other materials. On these surfaces, biofilms represent a major threat causing infectious diseases and economic losses. In addition, current antibiotics and common disinfectants have shown limited ability to remove biofilms adequately, and phage-based treatments are proposed as promising alternatives for biofilm eradication. This review analyzes the main advantages and challenges that phages can offer for the elimination of biofilms, as well as the most important factors to be taken into account in order to design effective phage-based treatments.

RevDate: 2020-05-26

Sapata DM, Ramos AL, Sábio S, et al (2020)

Evaluation of biofilm accumulation on and deactivation force of orthodontic Ni-Ti archwires before and after exposure to an oral medium: A prospective clinical study.

Journal of dental research, dental clinics, dental prospects, 14(1):41-47.

Background. This in vitro study aimed to evaluate biofilm accumulation on and deactivation force of orthodontic nickeltitanium (NiTi) archwires before and after exposure to an oral medium. Methods. Four commercial brands of orthodontic NiTi 0.016" archwires were examined before and after exposure to the oral medium for 4 weeks. Six archwire segments, 30 mm in length, from each manufacturer were tested in a device with four selfligating brackets, channel 0.022", adapted to a universal test machine to evaluate the deactivation force between 0.5 and 3 mm of deflection. The presence of biofilm on the archwire surfaces was evaluated by scanning electron microscopy, before and after exposure to the oral medium. The Wilcoxon and kappa tests were applied to the biofilm scores, three-way ANOVA for repeated measures (Bonferroni post-test), and linear regression between biofilm and deactivation force. Results. The exposure to the oral medium promoted moderate to severe presence of debris on the archwire surfaces and caused a reduction in deactivation force for the Ormco and GAC brands, while maintaining them with adequate force levels. The MORELLI and ORTHOMETRIC archwires underwent no significant reduction in deactivation force; moreover, these maintained elevated levels of force after exposure to the oral medium. The Spearman test indicated a low correlation between biofilm accumulation and deflection force for the Morelli (R2=0.132 and P=0.683) and Orthometric (R2=0.308 and P=0.330) brands. On the other hand, the GAC (R=0.767 and P=0.004) and ORMCO (R=0.725 and P=0.008) brands exhibited statistically significant correlation between these variables. Conclusion. Exposure to the oral medium for one month might give rise to significant changes in the dissipation of forces of orthodontic NiTi archwires, resulting from biofilm accumulation.

RevDate: 2020-05-26

Kart D, AS Kuştimur (2019)

Investigation of Gelatinase Gene Expression and Growth of Enterococcus faecalis Clinical Isolates in Biofilm Models.

Turkish journal of pharmaceutical sciences, 16(3):356-361.

Objectives: Enterococcus faecalis is the major reason for biofilm-related infections and it also interacts with Staphylococcus aureus in biofilms. Gelatinase (gelE) enzyme is an important virulence factor of E. faecalis for biofilm formation. This study aimed to compare the biofilm producing E. faecalis isolates from urine and urinary catheters. The influence of S. aureus on the growth of E. faecalis biofilm cells was also investigated in a dual biofilm model in vitro. Another aim was to evaluate E. faecalis gelE gene expression during biofilm formation.

Materials and Methods: Firstly, crystal violet staining was used to measure the total biofilm biomass of the isolates. Secondly, plate counting was performed to determine the biofilm formation ability of E. faecalis isolates and the effect of S. aureus on E. faecalis biofilm formation. Finally, the gelE expression profile of the isolates was assessed by quantitative real time-polymerase chain reaction.

Results: According to crystal violet staining and plate counting, all E. faecalis isolates were biofilm producers and the number of E. faecalis sessile cells increased in the presence of S. aureus. Among the 21 E. faecalis isolates, ten expressed high levels of the gelE gene, while eight of them had low expression profiles (p<0.05).

Conclusion: When they grow together, S. aureus may give some advantages to E. faecalis such as increasing sessile cell growth. The expression of the gelE gene was not affected by E. faecalis biofilm formation of the isolates collected from the patients with urinary tract infections.

RevDate: 2020-05-26

Zakaria BS, BR Dhar (2020)

Changes in syntrophic microbial communities, EPS matrix, and gene-expression patterns in biofilm anode in response to silver nanoparticles exposure.

The Science of the total environment, 734:139395 pii:S0048-9697(20)32912-0 [Epub ahead of print].

Understanding the toxic effect of silver nanoparticles (AgNPs) on various biological wastewater treatment systems is of significant interest to researchers. In recent years, microbial electrochemical technologies have opened up new opportunities for bioenergy and chemicals production from organic wastewater. However, the effects of AgNPs on microbial electrochemical systems are yet to be understood fully. Notably, no studies have investigated the impact of AgNPs on a microbial electrochemical system fed with a complex fermentable substrate. Here, we investigated the impact of AgNPs (50 mg/L) exposure to a biofilm anode in a microbial electrolysis cell (MEC) fed with glucose. The volumetric current density was 29 ± 2.0 A/m3 before the AgNPs exposure, which decreased to 20 ± 2.2 A/m3 after AgNPs exposure. The biofilms produced more extracellular polymeric substances (EPS) to cope with the AgNPs exposure, while carbohydrate to protein ratio in EPS considerably increased from 0.4 to 0.7. Scanning electron microscope (SEM) imaging also confirmed the marked excretion of EPS, forming a thick layer covering the anode biofilms after AgNPs injection. Transmission electron microscope (TEM) imaging showed that AgNPs still penetrated some microbial cells, which could explain the deterioration of MEC performance after AgNPs exposure. The relative expression level of the quorum signalling gene (LuxR) increased by 30%. Microbial community analyses suggested that various fermentative bacterial species (e.g., Bacteroides, Synergistaceae_vadinCA02, Dysgonomonas, etc.) were susceptible to AgNPs toxicity, which led to the disruption of their syntrophic partnership with electroactive bacteria. The abundance of some specific electroactive bacteria (e.g., Geobacter species) also decreased. Moreover, decreased relative expressions of various extracellular electron transfer associated genes (omcB, omcC, omcE, omcZ, omcS, and pilA) were observed. However, the members of family Enterobacteriaceae, known to perform a dual function of fermentation and anodic respiration, became dominant after biofilm anode exposed to AgNPs. Thus, EPS extraction provided partial protection against AgNPs exposure.

RevDate: 2020-05-26

Baattrup-Pedersen A, Graeber D, Kallestrup H, et al (2020)

Effects of low flow and co-occurring stressors on structural and functional characteristics of the benthic biofilm in small streams.

The Science of the total environment, 733:139331 pii:S0048-9697(20)32848-5 [Epub ahead of print].

Low flow and co-occurring stress is a more and more frequent phenomenon these years in small agricultural streams as a consequence of climate change. In the present study we explored short and longer term structural responses of the stream benthic algae community and biofilm metabolism to multiple stress in small streams applying a semi-experimental approach. We hypothesized that i) a reduction in flow in combination with secondary stress (nutrients and sediments) have immediate effects on the benthic algae community in terms of biomass (chlorophyll a, biovolume), taxonomic and trait (lifeform and size distribution) compositions as well as on metabolism (GPP and CR), and ii) that changes in the benthic algae community persist due to altered environmental settings but that functional redundancy among benthic algae species provides a high level of resilience in metabolism (GPP and CR). Overall, we found that stress imposed by nutrients was less pronounced than stress imposed by fine sediments under low flow, and that nutrient enrichment to some extent mitigated effects of fine sediments. Fine sediment deposition mediated a decline in the fraction of erect algae and/or algae with mucilage stalks but this did not happen under co-occurring stress from both sediments and nutrients. Additionally, fine sediment deposition mediated a decline in GPP of the biofilm, but again this did not happen under co-occurring stress from nutrients. We conclude that 1) the benthic algae community and biofilm metabolism displayed similar resilience to stress imposed by low flow and co-occurring stress from nutrients and sediments on a short and longer time scale and 2) as structure-function adaptations may occur at several trophic levels in the biofilm, more research is needed to explore mechanisms underlying mitigating effects of nutrients in response to sediment deposition under low flow.

RevDate: 2020-05-26

Badi S, Salah Abbassi M, Snoussi M, et al (2020)

High rates of antibiotic resistance and biofilm production in Escherichia coli isolates from food products of animal and vegetable origins in Tunisia: a real threat to human health.

International journal of environmental health research [Epub ahead of print].

The aim of this study was to compare the antibiotic susceptibility of eighty Escherichia coli isolates from vegetables and food products of animal origin in Tunisia, and to study their genes encoding antibiotic resistance and in vitro biofilm forming capacity. Antimicrobial susceptibilities were determined, as well as PCR investigation of genes associated with antibiotic resistance. Biofilm formation was tested using four different methods: the microtiter plate-, MTT-staining-, XTT-staining-, and the Congo Red Agar assays. High antibiotic resistance rates were observed for amoxicillin (68.7%), amoxicillin/clavulanic acid (73.7%), gentamicin (68.7%), kanamycin (66.2%), nalidixic acid (36.2%), streptomycin (68.7%) and tetracycline (35%). The majority of isolates was multidrug resistant and biofilm producer. MTT testing showed that vegetables isolates were significantly higher biofilm producers compared to foods of animal origins. This study showed that E. coli isolates from food products were reservoirs of genes encoding antibiotic-resistance and have a high propensity to produce biofilm.

RevDate: 2020-05-26

Li M, Shi D, Li Y, et al (2020)

Recombination of T4-like Phages and Its Activity against Pathogenic Escherichia coli in Planktonic and Biofilm Forms.

Virologica Sinica pii:10.1007/s12250-020-00233-2 [Epub ahead of print].

The increasing emergence of multi-drug resistant Escherichia coli (E. coli) has become a global concern, primarily due to the limitation of antimicrobial treatment options. Phage therapy has been considered as a promising alternative for treating infections caused by multi-drug resistant E. coli. However, the application of phages as a promising antimicrobial agent is limited by their narrow host range and specificity. In this research, a recombinant T4-like phage, named WGqlae, has been obtained by changing the receptor specificity determinant region of gene 37, using a homologous recombination platform of T4-like phages established by our laboratory previously. The engineered phage WGqlae can lyse four additional hosts, comparing to its parental phages WG01 and QL01. WGqlae showed similar characteristics, including thermo and pH stability, optimal multiplicity of infection and one-step growth curve, to the donor phage QL01. In addition, sequencing results showed that gene 37 of recombinant phage WGqlae had genetically stable even after 20 generations. In planktonic test, phage WGqlae had significant antimicrobial effects on E. coli DE192 and DE205B. The optical density at 600 nm (OD600) of E. coli in phage WGqlae treating group was significantly lower than that of the control group (P < 0.01). Besides, phage WGqlae demonstrated an obvious inhibitory effect on the biofilm formation and the clearance of mature biofilms. Our study suggested that engineered phages may be promising candidates for future phage therapy applications against pathogenic E. coli in planktonic and biofilm forms.

RevDate: 2020-05-26

Vishwakarma J, S V L (2020)

Unraveling the anti-biofilm potential of green algal sulfated polysaccharides against Salmonella enterica and Vibrio harveyi.

Applied microbiology and biotechnology pii:10.1007/s00253-020-10653-5 [Epub ahead of print].

One of the main reasons for the bacterial resistance to antibiotics is caused by biofilm formation of microbial pathogens during bacterial infections. Salmonella enterica and Vibrio harveyi are known to form biofilms and represent a major health concern worldwide, causing human infections responsible for morbidity and mortality. The current study aims to investigate the effect of purified sulfated polysaccharides (SPs) from Chlamydomonas reinhardtii (Cr) on planktonic and biofilm growth of these bacteria. The effect of Cr-SPs on bacterial planktonic growth was assessed by using the agar well diffusion method, which showed clear zones ranging from 13 to 26 mm in diameter from 0.5 to 8 mg/mL of Cr-SPs against both the bacteria. Time-kill activity and reduction in clonogenic propagation further help to understand the anti-microbial potential of Cr-SPs. The minimum inhibitory concentration of Cr-SPs against S. enterica and V. harveyi was as low as 440 μg/mL and 490 μg/mL respectively. Cr-SPs inhibited bacterial cell attachment up to 34.65-100% at 0.5-8 mg/mL in S. enterica and V. harveyi respectively. Cr-SPs also showed 2-fold decrease in the cell surface hydrophobicity, indicating their potential to prevent bacterial adherence. Interestingly, Cr-SPs efficiently eradicated the preformed biofilms. Increased reduction in total extracellular polysaccharide (EPS) and extracellular DNA (eDNA) content in a dose-dependent manner demonstrates Cr-SPs ability to interact and destroy the bacterial EPS layer. SEM analysis showed that Cr-SPs effectively distorted preformed biofilms and also induced morphological changes. Furthermore, Cr-SPs also showed anti-quorum-sensing potential by reducing bacterial urease and protease activities. These results indicate the potential of Cr-SPs as an anti-biofilm agent and will help to develop them as alternative therapeutics against biofilm-forming bacterial infections. KEY POINTS: • Cr-SPs not only inhibited biofilm formation but also eradicated preformed biofilms. • Cr-SPs altered bacterial cell surface hydrophobicity preventing biofilm formation. • Cr-SPs efficiently degraded eDNA of the EPS layer disrupting mature biofilms. • Cr-SPs reduced activity of quorum-sensing-mediated enzymes like protease and urease.

RevDate: 2020-05-25

Zhang L, Liang E, Cheng Y, et al (2020)

Is combined medication with natural medicine a promising therapy for bacterial biofilm infection?.

Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie, 128:110184 pii:S0753-3322(20)30376-0 [Epub ahead of print].

Bacterial biofilms widely exist in nature and seriously threaten global public health. Biofilms always cause persistent infection and seriously aggravate the occurrence of antibiotic resistance, which makes the treatment of bacterial infection difficult. Current conventional therapies, such as antibiotics, bacteriophages and quorum sensing inhibitors, are widely used to combat biofilms. However, these therapies are inadequate for the safe and effective treatment of biofilms. Antibiotics often produce resistance in treated bacteria, and antibacterial peptides are easily decomposed by proteases, so their efficacy is reduced. These results indicate that the treatment of biofilms needs further improvement. Increasing evidence has shown that natural medicine therapies have significant inhibitory effects on biofilms. This review summarized and analyzed the efficacy characteristics and corresponding mechanisms of conventional and natural medicine therapies combatting biofilms. By comparison, the advantages and disadvantages of those therapies have been classified and interpreted, so we have inferred that combined medication with natural medicines will be a more effective strategy against biofilms. This review lays a promising foundation for the development of antibiofilm agents and provides novel thinking for the treatment of bacterial biofilm infections.

RevDate: 2020-05-25

Zhou X, Ahmad JI, van der Hoek JP, et al (2020)

Thermal energy recovery from chlorinated drinking water distribution systems: Effect on chlorine and microbial water and biofilm characteristics.

Environmental research, 187:109655 pii:S0013-9351(20)30548-X [Epub ahead of print].

Thermal energy recovery from drinking water has a high potential in the application of sustainable building and industrial cooling. However, drinking water and biofilm microbial qualities should be concerned because the elevated water temperature after cold recovery may influence the microbial activities in water and biofilm phases in drinking water distribution systems (DWDSs). In this study, the effect of cold recovery on microbial qualities was investigated in a chlorinated DWDS. The chlorine decay was slight (1.1%-15.5%) due to a short contact time (~60 s) and was not affected by the cold recovery (p > 0.05). The concentrations of cellular ATP and intact cell numbers in the bulk water were partially inactivated by the residual chlorine, with the removal rates of 10.1%-16.2% and 22.4%-29.4%, respectively. The chlorine inactivation was probably promoted by heat exchangers but was not further enhanced by higher temperatures. The higher water temperature (25 °C) enhanced the growth of biofilm biomass on pipelines. Principle coordination analysis (PCoA) showed that the biofilms on the stainless steel plates of HEs and the plastic pipe inner surfaces had totally different community compositions. Elevated temperatures favored the growth of Pseudomonas spp. and Legionella spp. in the biofilm after cold recovery. The community functional predictions revealed more abundances of five human diseases (e.g. Staphylococcis aureus infection) and beta-lactam resistance pathways in the biofilms at higher temperature. Compared with a previous study with a non-chlorinated DWDS, chlorine dramatically reduced the biofilm biomass growth but raised the relative abundances of the chlorine-resistant genera (i.e. Pseudomonas and Sphingomonas) in bacterial communities.

RevDate: 2020-05-25

Tu C, Chen T, Zhou Q, et al (2020)

Biofilm formation and its influences on the properties of microplastics as affected by exposure time and depth in the seawater.

The Science of the total environment, 734:139237 pii:S0048-9697(20)32754-6 [Epub ahead of print].

The effects of microbial colonization and biofilm formation on microplastics in the marine and coastal environments have aroused global concern recently. However, the simultaneous influences of exposure time and depth on biofilm formation, and subsequently on the properties variations of microplastics is less studied. In this study, polyethylene (PE) film was exposed at three depths (2 m, 6 m, and 12 m) for three time periods (30 days, 75 days, and 135 days) in the coastal seawater of Yellow Sea, China. The results show that the total amount of biofilms markedly increased with exposure time, but decreased with water depth. Typical morphologies and compositions of biofilms such as coccus-, rod-, disc-shaped bacteria and filaments, as well as a dense layer of extracellular polymeric substances were observed on the surfaces of the PE microplastics. Biofilm formation could decrease the hydrophobicity of PE microplastics, and increase the abundances of hydrophilic C-O and CO groups on the surface of PE. Alphaproteobacteria, Gammaproteobacteria and Bacteroidia were identified as the core microbiome of the PE associated biofilms, while the dominant bacteria families vary from the early to the late phases of the biofilm formation. Our results indicate that microplastics associated biofilms could affect the environmental processes and fates of microplastics in the marine and coastal environment.

RevDate: 2020-05-25

Hujslová M, Gryndlerová H, Bystrianský L, et al (2020)

Biofilm and planktonic microbial communities in highly acidic soil (pH < 3) in the Soos National Nature Reserve, Czech Republic.

Extremophiles : life under extreme conditions pii:10.1007/s00792-020-01177-x [Epub ahead of print].

Biofilm formation is a typical life strategy used by microorganisms populating acidic water systems. The same strategy might be used by microbes in highly acidic soils that are, however, neglected in this regard. In the present study, the microbial community in such highly acidic soil in the Soos National Nature Reserve (Czech Republic) has been investigated using high-throughput DNA sequencing and the organisms associated with biofilm life mode and those preferring planktonic life were distinguished using the biofilm trap technique. Our data show the differences between biofilm and planktonic microbiota fraction, although the majority of the organisms were capable of using both life modes. The by far most abundant prokaryotic genus was Acidiphilium and fungi were identified among the most abundant eukaryotic elements in biofilm formations. On the other hand, small flagellates from diverse taxonomical groups predominated in plankton. The application of cellulose amendment as well as the depth of sampling significantly influenced the composition of the detected microbial community.

RevDate: 2020-05-24

Lundström T, Lingström P, Wattle O, et al (2020)

Equine saliva components during mastication, and in vivo pH changes in the oral biofilm of sound and carious tooth surfaces after sucrose exposure.

Acta veterinaria Scandinavica, 62(1):21 pii:10.1186/s13028-020-00518-2.

BACKGROUND: The role of saliva composition and dietary sugar in development of infundibular caries in equine cheek teeth is not fully understood. This study analysed electrolyte and urea concentrations in saliva in relation to different forage and measured pH changes after sucrose application in vivo in sound and carious cheek teeth.

RESULTS: Forage type had no effect on the equine saliva electrolyte concentrations, which varied considerably both intra- and inter-individually. Chewing resulted in increased values for all electrolytes except bicarbonate. Compared with stimulated human saliva, horse saliva after mastication, contained higher amounts of potassium, calcium and bicarbonate, and less phosphate. The in vivo pH measurements showed a lower resting pH and a more pronounced pH drop after sucrose application in carious teeth compared to sound teeth.

CONCLUSIONS: No large differences were found between the composition of equine saliva and human saliva. A more pronounced acidogenicity was found for the carious than sound teeth. Thus, the caries process in equine cheek teeth seems to follow the same pattern as in human teeth, caused by acid production by oral microorganisms after sugar consumption.

RevDate: 2020-05-23

Yang Z, Wang Z, Lei M, et al (2020)

Effects of Spo0A on Clostridium acetobutylicum with an emphasis on biofilm formation.

World journal of microbiology & biotechnology, 36(6):80 pii:10.1007/s11274-020-02859-6.

Clostridium acetobutylicum is a well-known strain for biofuel production. In previous work, it was found that this strain formed biofilm readily during fermentation processes. Biofilm formation could protect cells and enhance productivities under environmental stresses in our previous work. To explore the molecular mechanism of biofilm formation, Spo0A of C. acetobutylicum was selected to investigate its influences on biofilm formation and other physiological performances. When spo0A gene was disrupted, the spo0A mutant could hardly form biofilm. The aggregation and adhesion abilities of the spo0A mutant as well as its swarming motility were dramatically reduced compared to those of wild type strain. Sporulation was also negatively influenced by spo0A disruption, and solvent production was almost undetectable in the spo0A mutant fermentation. Furthermore, proteomic differences between wild type strain and the spo0A mutant were consistent with physiological performances. This is the first study confirming a genetic clue to C. acetobutylicum biofilm and will be valuable for biofilm optimization through genetic engineering in the future.

RevDate: 2020-05-23

Song Y, Sun M, Feng L, et al (2020)

Lactobacillus plantarum-12 Exopolysaccharides Have Anti-Biofilm Activity Against Shigella flexneri.

Applied and environmental microbiology pii:AEM.00694-20 [Epub ahead of print].

In the developing countries, Shigella flexneri is the most common enteric pathogen causing bacillary dysentery. And the biofilm formation of S. flexneri can cause the emergence of antibiotic-resistant strains and serious threat to food safety and human health. In this study, the effects of Lactobacillus plantarum-12 exopolysaccharides (L-EPS) and S. flexneri exopolysaccharides (S-EPS) on the S. flexneri CMCC51574 biofilm formation were investigated. The results showed that L-EPS could decrease the polysaccharides production in the extracellular polymeric matrix of S. flexneri, and inhibit the biofilm formation of S. flexneri L-EPS could decrease the minimum biofilm elimination concentration (MBEC) of the antibiotics against S. flexneri biofilm and inhibit S. flexneri adhesion and invasion to the HT-29 cell monolayers, which might be ascribed to the S. flexneri biofilm disturbance by L-EPS. While the S-EPS exhibited the exactly opposite effects compared to L-EPS. The monosaccharide composition analysis showed that L-EPS was composed of mannose, glucuronic acid, galactosamine, glucose, galactose and xylose, with the molar ratio of 32.26: 0.99: 1.79: 5.63: 0.05: 4.07, and the S-EPS was composed of mannose, glucuronic acid, galactosamine, glucose, galactose, with the molar ratio of 25.43: 2.28: 7.13: 5.35. The L-EPS was separated to the neutral polysaccharide L-EPS 1-1 and the acidic polysaccharide L-EPS 2-1 by the ion exchange chromatography and gel chromatography. The L-EPS 2-1 exerted higher anti-biofilm activity than the L-EPS 1-1. The anti-biofilm activity of L-EPS might be associated with its structure.Importance:S. flexneri is a wide-spread food-borne pathogen causing food contamination and responsible for food poisoning outbreaks through various foods in developing countries. The biofilm formation of S. flexneri was not only difficult to be eliminated, but also increased the drug-resistant of the strain. In the present study, it was demonstrated that the L-EPS secreted by the Lactobacillus plantrum-12 could inhibit the biofilm formation, adhesion and invasion to HT-29 cells of S. flexneri And the L-EPS could also decrease the minimum biofilm elimination concentration (MBEC) of the antibiotics against S. flexneri biofilm. Therefore, the L-EPS was a bioactive macromolecule with the potential ability against the infections of the S. flexneri.

RevDate: 2020-05-23

Khan F, Yu H, YM Kim (2020)

Bactericidal Activity of Usnic Acid-Chitosan Nanoparticles against Persister Cells of Biofilm-Forming Pathogenic Bacteria.

Marine drugs, 18(5): pii:md18050270.

The present study aimed to prepare usnic acid (UA)-loaded chitosan (CS) nanoparticles (UA-CS NPs) and evaluate its antibacterial activity against biofilm-forming pathogenic bacteria. UA-CS NPs were prepared through simple ionic gelification of UA with CS, and further characterized using Fourier transform infrared spectroscopy, X-ray diffraction, and field-emission transmission electron microscopy. The UA-CS NPs presented a loading capacity (LC) of 5.2%, encapsulation efficiency (EE) of 24%, and a spherical shape and rough surface. The maximum release of UA was higher in pH 1.2 buffer solution as compared to that in pH 6.8 and 7.4 buffer solution. The average size and zeta potential of the UA-CS NPs was 311.5 ± 49.9 nm in diameter and +27.3 ± 0.8 mV, respectively. The newly prepared UA-CS NPs exhibited antibacterial activity against persister cells obtained from the stationary phase in batch culture, mature biofilms, and antibiotic-induced gram-positive and gram-negative pathogenic bacteria. Exposure of sub-inhibitory concentrations of UA-CS NPs to the bacterial cells resulted in a change in morphology. The present study suggests an alternative method for the application of UA into nanoparticles. Furthermore, the anti-persister activity of UA-CS NPs may be another possible strategy for the treatment of infections caused by biofilm-forming pathogenic bacteria.

RevDate: 2020-05-23

Sun X, Chen B, Xia B, et al (2020)

Impact of mariculture-derived microplastics on bacterial biofilm formation and their potential threat to mariculture: A case in situ study on the Sungo Bay, China.

Environmental pollution (Barking, Essex : 1987), 262:114336.

Microplastics (MPs) pollution in the marine environment has attracted considerable global attention. However, the colonization of microorganisms on mariculture-derived MPs and their effects on mariculture remain poorly understood. In this study, the MPs (fishing nets, foams and floats) and a natural substrate, within size ranges (1-4 mm), were then incubated for 21 days in Sungo Bay (China), and the composition and diversity of bacterial communities attached on all substrates were investigated. Results showed that bacterial communities on MPs mainly originated from their surrounding seawater and sediment, with an average contribution on total MPs adherent population of 47.91% and 37.33%, respectively. Principle coordinate analysis showed that community similarity between MPs and surrounding seawater decreased with exposure time. In addition, lower average bacterial community diversity and higher relative abundances of bacteria from the genera Vibrio, Pseudoalteromonas and Alteromonas on MPs than those in their surrounding seawater and sediments indicated that MPs might enrich potential pathogens and bacteria related with carbohydrate metabolism. They are responsible for the significant differences in KEGG Orthology pathways (infectious disease and carbohydrate metabolism) between MPs and seawater. The KO pathway (Infectious Diseases) associated with MPs was also significantly higher than those with feathers in the nearshore area. MPs might be vectors for enrichment of potentially pathogenic Vibrio, and enhance the ecological risk of MPs to mariculture industry.

RevDate: 2020-05-22

Liu W, Lu H, Chu X, et al (2020)

Tea polyphenols inhibits biofilm formation, attenuates the quorum sensing-controlled virulence and enhances resistance to Klebsiella pneumoniae infection in Caenorhabditis elegans model.

Microbial pathogenesis pii:S0882-4010(20)30632-X [Epub ahead of print].

Bacteria cells can communicate with each other via quorum sensing (QS) system. Various physiological characteristics including virulence factors and biofilm formation are controlled by QS. So interrupting the bacterial communication is an alternative strategy instead of antibiotics for control bacterial infection. The aim of this study was to investigate the effects of tea polyphenols (TPs) on quorum sensing and virulence factors of Klebsiella pneumoniae. In vitro study showed that the anti-QS activity of tea polyphenols against Chromobacterium violaceum in violacein production. At sub-MICs, TPs inhibited the motility, reduced protease and exopolysaccharide (EPS) production and also biofilm formation in K. pneumoniae. In addition, in vivo study showed that tea polyphenols at 200 μg/mL and 400 μg/mL increased the survival rate of Caenorhabditis elegans to 73.3% and 82.2% against K. pneumonia infection. Our findings suggest that tea polyphenols can act as an effective QS inhibitor and can serve as a novel anti-virulence agent for the management of bacterial pathogens.

RevDate: 2020-05-22

Brzezinka MS, Walczak M, Kalwasińska A, et al (2020)

Biofilm formation during biodegradation of polylactide, poly (3,4 hydroxybutyrate) and poly(ε-caprolactone) in activated sludge.

International journal of biological macromolecules pii:S0141-8130(20)33260-8 [Epub ahead of print].

Biodegradable materials, namely pure polylactide (PLA), poly (3,4-hydroxybutyrate) (PHB), poly(ε-caprolactone) (PCL) were investigated to assess their degradability by activated sludge. The study aimed at the isolation of biofilm-forming bacteria and the determination of their hydrolytic activity toward the PLA, PHB, and PCL with embedded PHMG derivatives. The biological oxygen demand and physical properties (tensile strength, water vapor permeability, surface structure) of materials indicated that PCL was the best biodegradable film. Aeromonas and Rhodococcus isolated from the polymers' surface during the process of decomposition showed the ability to form biofilms. The introduction of PHMG derivatives into PLA, PCL, and PHB films did not affect biofilm formation and hydrolase activity for most of the isolates. PHMG derivatives at the concentration of 1% disturbed the degradation process.

RevDate: 2020-05-22

Arutyunov AS, Tsareva TV, Kirakosyan LG, et al (2020)

[Features and significance of adhesion of bacteria and fungi of the oral cavity as the initial stage of the formation of a microbial biofilm on dental polymer materials].

Stomatologiia, 99(2):79-84.

OBJECTIVE: Characteristics of the adhesion of yeast fungi and oral bacteria of various types in vitro to samples of polymeric materials for fixed structures of dental prostheses, obtained using various technologies: adjective digital 3D printing and traditional methods.

MATERIAL AND METHODS: Conducted model experiments on the adhesion of bacterial (including the main periodontopic-pathogenic species - P. gingivalis, P. intermedia, etc.) and fungal pathogens (C. albicans) to standard samples of polymer materials NextDent C & B Micro Filled Hybrid («NextDent», Netherlands), Detax Freeprint temp UV («Detax», Germany), obtained by digital additive 3D printing technology, and Luxatemp Automix Plus («DMG», Germany) and Acrytemp («Zhermack», Italy) - by the traditional method as a control. Removal of adhering microbes from the material was carried out using an ultrasound machine (exposure time 10 minutes, power 60 kHz).

RESULTS: The dependence of the degree of microbial adhesion on the nature of the material and processing technology (3D printing, milling) was established. The materials of NextDent C & B Micro Filled Hybrid and Detax Freeprint temp UV showed high resistance to adhesion of clinical isolates of periodontal pathogenic bacteria and C. albicans fungi.

CONCLUSION: The lowest adhesion values for periodontopathogenic species and C. albicans fungi were detected when using samples of materials obtained by 3D printing: NextDent C & B Micro Filled Hybrid and Detax Freeprint temp UV compared to Luxatemp Automix Plus and Acrytemp polymers.

RevDate: 2020-05-22

Lee M, Ponraja G, McLeod K, et al (2020)

Breast Implant Illness: A Biofilm Hypothesis.

Plastic and reconstructive surgery. Global open, 8(4):e2755.

Background: "Breast implant illness" (BII) is a poorly defined cluster of nonspecific symptoms, attributed by patients as being caused by their breast implants. These symptoms can include joint pain, skin and hair changes, concentration, and fatigue. Many patients complaining of BII symptoms are dismissed as psychosomatic. There are currently over 10,000 peer-reviewed articles on breast implants, but at the time of commencing this study, only 2 articles discussed this entity. At the same time, mainstream media and social media are exploding with nonscientific discussion about BII.

Methods: We have prospectively followed 50 consecutive patients, self-referring for explantation due to BII. We analyzed their preoperative symptoms and followed up each patient with a Patient-Reported Outcome Questionnaire. All implants and capsules were, if possible, removed en bloc. Explanted implants were photographed. Implant shell and capsule sent for histology and microbiological culture.

Results: BII symptoms were not shown to correlate with any particular implant type, surface, or fill. There was no significant finding as to duration of implant or location of original surgery. Chronic infection was found in 36% of cases with Propionibacterium acnes the most common finding. Histologically, synoviocyte metaplasia was found in a significantly greater incidence than a matched cohort that had no BII symptoms (P = 0.0164). Eighty-four percent of patients reported partial or complete resolution of BII symptoms on Patient-Reported Outcome Questionnaire. None of the 50 patients would consider having breast implants again.

Conclusion: The authors believe BII to be a genuine entity worthy of further study. We have identified microbiological and histological abnormalities in a significant number of patients identifying as having BII. A large proportion of these patients have reported resolution or improvement of their symptoms in patient-reported outcomes. Improved microbiology culture techniques may identify a larger proportion of chronic infection, and further investigation of immune phenotypes and toxicology may also be warranted in this group.

RevDate: 2020-05-22

Tahrioui A, Duchesne R, Bouffartigues E, et al (2019)

Extracellular DNA release, quorum sensing, and PrrF1/F2 small RNAs are key players in Pseudomonas aeruginosa tobramycin-enhanced biofilm formation.

NPJ biofilms and microbiomes, 5(1):15 pii:10.1038/s41522-019-0088-3.

Biofilms are structured microbial communities that are the leading cause of numerous chronic infections which are difficult to eradicate. Within the lungs of individuals with cystic fibrosis (CF), Pseudomonas aeruginosa causes persistent biofilm infection that is commonly treated with aminoglycoside antibiotics such as tobramycin. However, sublethal concentrations of this aminoglycoside were previously shown to increase biofilm formation by P. aeruginosa, but the underlying adaptive mechanisms still remain elusive. Herein, we combined confocal laser scanning microscope analyses, proteomics profiling, gene expression assays and phenotypic studies to unravel P. aeruginosa potential adaptive mechanisms in response to tobramycin exposure during biofilm growth. Under this condition, we show that the modified biofilm architecture is related at least in part to increased extracellular DNA (eDNA) release, most likely as a result of biofilm cell death. Furthermore, the activity of quorum sensing (QS) systems was increased, leading to higher production of QS signaling molecules. We also demonstrate upon tobramycin exposure an increase in expression of the PrrF small regulatory RNAs, as well as expression of iron uptake systems. Remarkably, biofilm biovolumes and eDNA relative abundances in pqs and prrF mutant strains decrease in the presence of tobramycin. Overall, our findings offer experimental evidences for a potential adaptive mechanism linking PrrF sRNAs, QS signaling, biofilm cell death, eDNA release, and tobramycin-enhanced biofilm formation in P. aeruginosa. These specific adaptive mechanisms should be considered to improve treatment strategies against P. aeruginosa biofilm establishment in CF patients' lungs.

RevDate: 2020-05-22

Toledano-Osorio M, Osorio R, Aguilera FS, et al (2020)

Polymeric nanoparticles protect the resin-dentin bonded interface from cariogenic biofilm degradation.

Acta biomaterialia pii:S1742-7061(20)30263-4 [Epub ahead of print].

The objective was to assess doxycycline (Dox) and zinc (Zn) doped nanoparticles' (NPs) potential to protect the resin-dentin interface from cariogenic biofilm. Three groups of polymeric NPs were tested: unloaded, loaded with zinc and with doxycycline. NPs were applied after dentin etching. The disks were exposed to a cariogenic biofilm challenge in a Drip-Flow Reactor during 72 h and 7 d. Half of the specimens were not subjected to biofilm formation but stored 72 h and 7 d. LIVE/DEAD® viability assay, nano-dynamic mechanical assessment, Raman spectroscopy and field emission electron microscopy (FESEM) analysis were performed. The measured bacterial death rates, at 7 d were 46% for the control group, 51% for the undoped-NPs, 32% for Dox-NPs, and 87% for Zn-NPs; being total detected bacteria reduced five times in the Dox-NPs group. Zn-NPs treated samples reached, in general, the highest complex modulus values at the resin-dentin interface over time. Regarding the mineral content, Zn-NPs-treated dentin interfaces showed the highest mineralization degree associated to the phosphate peak and the relative mineral concentration. FESEM images after Zn-NPs application permitted to observe remineralization of the etched and non-resin infiltrated collagen layer, and bacteria were scarcely encountered. The combined antibacterial and remineralizing effects, when Zn-NPs were applied, reduced biofilm formation. Dox-NPs exerted an antibacterial role but did not remineralize the bonded interface. Undoped-NPs did not improve the properties of the interfaces. Application of Zn-doped NPs during the bonding procedure is encouraged. Statement of significance Application of Zn-doped nanoparticles on acid etched dentin reduced biofilm formation and viability at the resin-dentin interface due to both remineralization and antibacterial properties. Doxycycline-doped nanoparticles also diminished oral biofilm viability, but did not remineralize the resin-dentin interface.

RevDate: 2020-05-22

Rocca DM, Silvero C MJ, Aiassa V, et al (2020)

Rapid and effective phototreatment of biofilm infections using low doses of amoxicillin-coated gold nanoparticles.

Photodiagnosis and photodynamic therapy pii:S1572-1000(20)30165-4 [Epub ahead of print].

Bacterial biofilm are complex microbial communities covered by a matrix of extracellular polymeric substances, which develops when a community of microorganisms irreversibly adheres to a living or inert surface. This structure is considered an important virulence factor because it is difficult to eradicate and often responsible for treatment failures. This adherent community represents one of the greatest problems in public health due to the continued emergence of conventional antibiotic-therapy resistance. Photodynamic Antimicrobial Therapy (PACT) is a therapeutic alternative and promises to be an effective treatment against multiresistant bacteria biofilm, demonstrating a broad spectrum of action. This work demonstrates the reduction in biofilms of relevant clinical isolates (as Pseudomonas aeruginosa and Staphylococcus aureus) treated with PACT using low concentrations of amoxicillin-coated gold nanoparticles (amoxi@AuNP) as a photosensitizer. Moreover, the viability reduction of 60% in S. aureus biofilms and 70% in P. aeruginosa biofilms were obtained after three hours of irradiation with white light and amoxi@AuNP. Scanning electron microscopy analysis revealed that amoxi@AuNP could penetrate and cause damage to the biofilm matrix, and interact with bacteria cells. A strong biofilm production in P. aeruginosa was observed by confocal laser scanning microscopy using acridine orange as a probe, and a markedly decrease in live bacteria was appreciated when PACT was applied. The use of amoxi@AuNP for PACT allows the viability reduction of clinical Gram positive and Gram negative biofilms. This novel strategy needs shorter irradiation times and lower concentrations of nanoparticles than other reports described. This could be attributed to two major innovations: the selectivity for the bacterial wall given by the amoxicillin and the polydispersity of size and shapes with seems to contribute to the photo-antibacterial capacity.

RevDate: 2020-05-22

Hans S, Purkait D, Nandan S, et al (2020)

Rec A disruption unveils cross talk between DNA repair and membrane damage, efflux pump activity, biofilm formation in Mycobacterium smegmatis.

Microbial pathogenesis pii:S0882-4010(20)30628-8 [Epub ahead of print].

Tuberculosis (TB) caused by Mycobacterium tuberculosis (MTB) has emerged in recent decades as one of the leading causes of mortality worldwide. The burden of TB is alarmingly high, with one third affected global population as reported by WHO. Short-course treatment with an antibiotic is a powerful weapon to treat infection of susceptible MTB strain, however; MTB has developed resistance to anti-TB drugs, which is an escalating global health crisis. Thus there is urgent need to identify new drug targets. RecA is a 38 kilodalton protein required for the repair and maintenance of DNA and regulation of the SOS response. The objective of this study is to understand the effect of disruption of RecA gene (deletion mutant ΔdisA from previous study) in a surrogate model for MTB, Mycobacterium smegmatis. This study demonstrated that disruption of RecA causes enhanced susceptibility towards rifampicin and generation of ROS leading to lipid peroxidation and impaired membrane homeostasis as depicted by altered cell membrane permeability and efflux pump activity. Mass spectrometry based lipidomic analysis revealed decreased mycolic acid moieties, phosphatidylinositol mannosides (PIM), Phthiocerol dimycocerosate (DIM). Furthermore, biofilm formation was considerably reduced. Additionally, we have validated all the disrupted phenotypes by RT-PCR which showed a good correlation with the biochemical assays. Lastly, RecA mutant displayed reduced infectivity in Caenorhabditis elegans illustrating its vulnerability as antimycobacterial target. Together, present study establishes a link between DNA repair, drug efflux and biofilm formation and validates RecA as an effective drug target. Intricate studies are needed to further understand and exploit this therapeutic opportunity.

RevDate: 2020-05-21

Tang M, Wei X, Wan X, et al (2020)

The role and relationship with efflux pump of biofilm formation in Klebsiella pneumoniae.

Microbial pathogenesis pii:S0882-4010(20)30531-3 [Epub ahead of print].

This study aimed to identify the role and relationship with efflux pump of biofilm formation in Klebsiella pneumoniae. Sixty-one K. pneumoniae clinical isolates were collected between January and June of 2017 from the affiliated hospital of southwest medical university in Luzhou, China. The minimum inhibitory concentration (MIC) and minimum biofilm eradication concentration (MBEC) were determined using broth microdilution method. Crystal violet (CV) staining and confocal laser scanning microscope (CLSM) were used to monitor biofilm formation. Efflux pump expression was investigated qualitatively and quantitatively by polymerase chain reaction (PCR) and reverse transcriptase quantitative PCR (RT-qPCR). Crystal violet staining was performed to evaluate the effect of efflux pump inhibitor carbonyl cyanide m-chlorophenyl hydrazine (CCCP) on K. pneumoniae biofilms. Our results showed that crystal violet staining and CLSM had good consistency in biofilm detection. Biofilm formation was an independent biological behavior of the strain and measured at 24 h was reasonable. Biofilms up-regulated antimicrobial resistance and expression of efflux pump gene acrA, emrB, oqxA, and qacEΔ1 in K. pneumoniae. CCCP inhibited biofilms but dose-dependent effect was obvious. Altogether, our data demonstrates that biofilm formation, as well as its interaction with efflux pump, promotes antimicrobial resistance in K. pneumoniae.

RevDate: 2020-05-21

Yang D, JI Reyes-De-Corcuera (2020)

Continuous flow system for biofilm formation using controlled concentrations of Pseudomonas putida from chicken carcass and coupled to electrochemical impedance detection.

Biofouling [Epub ahead of print].

Most studies dealing with monitoring the dynamics of biofilm formation use microbial suspensions at high concentrations. These conditions do not always represent food or water distribution systems. A continuous flow system capable of controlling the concentration of the microbial suspension stream from 104 to 106 CFU ml-1 is reported. Pseudomonas putida biofilms formed using 100-fold, 1,000-fold or 10,000-fold diluted bacterial suspensions were monitored in-line by electrochemical impedance spectroscopy (EIS) and total plate counts. Randles equivalent circuit model and a modified Randles model with biofilm elements were used to fit the EIS data. In Randles equivalent circuit, the charge transfer resistance decreased as the biofilm formed. The log colony counts of the biofilm correlated to the charge transfer resistance. In the biofilm model, the biofilm resistance and the double layer capacitance decreased as the biofilm formed. The log colony counts of the biofilm correlated to the biofilm resistance.

RevDate: 2020-05-20

Dong Y, Li S, Zhao D, et al (2020)

IolR, a negative regulator of the myo-inositol metabolic pathway, inhibits cell autoaggregation and biofilm formation by downregulating RpmA in Aeromonas hydrophila.

NPJ biofilms and microbiomes, 6(1):22 pii:10.1038/s41522-020-0132-3.

Aeromonas hydrophila is the causative agent of motile Aeromonad septicemia in fish. Previous studies have shown that the myo-inositol metabolism is essential for the virulence of this bacterium. IolR is a transcription inhibitor that negatively regulates myo-inositol metabolic activity. While in the process of studying the inositol catabolism in A. hydrophila Chinese epidemic strain NJ-35, we incidentally found that ΔiolR mutant exhibited obvious autoaggregation and increased biofilm formation compared to the wild type. The role of surface proteins in A. hydrophila autoaggregation was confirmed by different degradation treatments. Furthermore, calcium promotes the formation of aggregates, which disappear in the presence of the calcium chelator EGTA. Transcriptome analysis, followed by targeted gene deletion, demonstrated that biofilm formation and autoaggregation caused by the inactivation of iolR was due to the increased transcription of a RTX-family adhesion gene, rmpA. Further, IolR was determined to directly regulate the transcription of rmpA. These results indicated that iolR is negatively involved in autoaggregation and biofilm formation in A. hydrophila, and this involvement was associated with its inhibition on the expression of rmpA.

RevDate: 2020-05-20

Luze H, Holzer J, Nischwitz SP, et al (2020)

The importance of in vivo biofilm models for clinical practice.

RevDate: 2020-05-20

Ma Q, Wang H, Chen ZN, et al (2020)

Removal of biofilm is essential for long-term ventilation tube retention.

World journal of clinical cases, 8(9):1592-1599.

BACKGROUND: Although long-term retention of a ventilation tube is required in many ear diseases, spontaneous removal of conventional ventilation tube is observed in patients within 3 to 12 mo. To address this issue, we aimed to determine a new method for long-term retention of the ventilation tube.

AIM: To explore the value of removing the biofilm for long-term retention of tympanostomy ventilation tubes.

METHODS: A case-control study design was used to evaluate the safety and effectiveness of long-term tube retention by directly removing the biofilm (via surgical exfoliation) in patients who underwent myringotomy with ventilation tube placement. The patients were randomly divided into two groups: Control group and treatment group. Patients in the treatment group underwent regular biofilm exfoliation surgery in the clinic, whereas those in the control group did not have their biofilm removed. Only conventional ventilation tubes were placed in this study. Outcome measures were tube position and patency. Tube retention time and any complications were documented.

RESULTS: Eight patients with biofilm removal and eight patients without biofilm removal as a control group were enrolled in the study. The tympanostomy tube retention time was significantly longer in the treatment group (43.5 ± 26.4 mo) than in the control group (9.5 ± 6.9 mo) (P = 0.003). More tympanostomy tubes were found to be patent and in correct position in the treatment group during the follow-up intervals than in the control group (P = 0.01).

CONCLUSION: Despite the use of short-term ventilation tubes, direct biofilm removal can be a well-tolerated and effective treatment for long-term tube retention of tympanostomy ventilation tubes in patients who underwent myringotomy.

RevDate: 2020-05-20

Soukarieh F, Liu R, Romero M, et al (2020)

Hit Identification of New Potent PqsR Antagonists as Inhibitors of Quorum Sensing in Planktonic and Biofilm Grown Pseudomonas aeruginosa.

Frontiers in chemistry, 8:204.

Current treatments for Pseudomonas aeruginosa infections are becoming less effective because of the increasing rates of multi-antibiotic resistance. Pharmacological targeting of virulence through inhibition of quorum sensing (QS) dependent virulence gene regulation has considerable therapeutic potential. In P. aeruginosa, the pqs QS system regulates the production of multiple virulence factors as well as biofilm maturation and is a promising approach for developing antimicrobial adjuvants for combatting drug resistance. In this work, we report the hit optimisation for a series of potent novel inhibitors of PqsR, a key regulator of the pqs system, bearing a 2-((5-methyl-5H-[1,2,4]triazino[5,6-b]indol-3-yl)thio) acetamide scaffold. The initial hit compound 7 (PAO1-L IC50 0.98 ± 0.02 μM, PA14 inactive at 10 μM) was obtained through a virtual screening campaign performed on the PqsR ligand binding domain using the University of Nottingham Managed Chemical Compound Collection. Hit optimisation gave compounds with enhanced potency against strains PAO1-L and PA14, evaluated using P. aeruginosa pqs-based QS bioreporter assays. Compound 40 (PAO1-L IC50 0.25 ± 0.12 μM, PA14 IC50 0.34 ± 0.03 μM) is one of the most potent PqsR antagonists reported showing significant inhibition of P. aeruginosa pyocyanin production and pqs system signaling in both planktonic cultures and biofilms. The co-crystal structure of 40 with the PqsR ligand binding domain revealed the specific binding interactions occurring between inhibitor and this key regulatory protein.

RevDate: 2020-05-20

Pijls BG, Sanders IMJG, Kujiper EJ, et al (2020)

Induction heating for eradicating Staphylococcus epidermidis from biofilm.

Bone & joint research, 9(4):192-199 pii:10.1302_2046-3758.94.BJR-2019-0274.R1.

Aims: Induction heating is a noninvasive, nonantibiotic treatment modality that can potentially be used to cause thermal damage to the bacterial biofilm on the metal implant surface. The purpose of this study was to determine the effectiveness of induction heating on killing Staphylococcus epidermidis from biofilm and to determine the possible synergistic effect of induction heating and antibiotics.

Methods: S. epidermidis biofilms were grown on titanium alloy (Ti6Al4V) coupons for 24 hours (young biofilm) and seven days (mature biofilm). These coupons with biofilm were heated to temperatures of 50°C, 55°C, 60°C, 65°C, 70°C, 80°C, and 90°C for 3.5 minutes and subsequently exposed to vancomycin and rifampicin at clinically relevant concentrations.

Results: For the young biofilm, total eradication was observed at 65°C or higher for 3.5 minutes followed by 24 hours of vancomycin 10 mg/l and rifampicin 1 mg/l. For the mature biofilm, total eradication was observed at 60°C for 3.5 minutes followed by 24 hours of vancomycin 10 mg/l and rifampicin 1 mg/l. Total eradication was also observed at 60°C for 3.5 minutes followed by 24 hours of vancomycin 1 mg/l and rifampicin 1 mg/l followed by another thermal shock of 60°C for 3.5 minutes (two thermal shocks).

Conclusion: Induction heating of Ti6Al4V coupons is effective in reducing bacterial load in vitro for S. epidermidis biofilms. Induction heating and antibiotics have a synergistic effect resulting in total eradication of the biofilm at 60°C or higher for clinically relevant concentrations of vancomycin and rifampicin.Cite this article:Bone Joint Res. 2020;9(4):192-199.

RevDate: 2020-05-20

Said MB, Saad MB, Bousselmi L, et al (2020)

Use of the catalytic complex TiO2/Red Cabbage Anthocyanins to reduce the biofilm formation by planktonic bacteria.

Environmental technology [Epub ahead of print].

The bacterial cells dwelling within the biofilm usually develop resistance against common disinfectants. In this current study, to improve the effectiveness of photocatalytic treatment, a natural sensitizer in combination with unsupported titanium dioxide nanoparticles (TiO2-NPs) was used to optimize the absorbance of NPs in the visible region and, to enhance the catalytic activity of the semiconductor. Different kinetic parameters were determined according to the first-order and the biphasic models to evaluate the ability of tested bacteria to form biofilm under different photocatalytic treatment conditions. As a result, the addition of red cabbage anthocyanins (RCA) as photosensitizer allows the enhancement of biocide activity of TiO2-NPs and the reduction of biofilm formation by tested bacteria.

RevDate: 2020-05-20

Souza C, Mota HF, Faria YV, et al (2020)

Resistance to Antiseptics and Disinfectants of Planktonic and Biofilm-Associated Forms of Corynebacterium striatum.

Microbial drug resistance (Larchmont, N.Y.) [Epub ahead of print].

Disinfection and antisepsis are of primary importance in controlling nosocomial infections and outbreaks by pathogens expressing multiple resistance to antimicrobial agents (multidrug-resistant [MDR]) used in therapy. Nowadays, infections related to health services (HAIs) due to MDR and multidrug-susceptible (MDS) Corynebacterium striatum should not be underestimated, including patients using invasive medical devices. The virulence potential of C. striatum needs further investigation. Currently, susceptibility profiles of planktonic and/or sessile forms of four C. striatum strains of different pulsed-field gel electrophoresis types were examined as biocides based on the manufacturer's recommendations: 2% glutaraldehyde (GA), 2% peracetic acid (PA), 1% potassium monopersulfate (Virkon®; VK), 1% sodium hypochlorite (SH), and 70% ethyl alcohol (ET). Time-kill assays using 2% bovine serum albumin (BSA) were performed for evaluation of influence of organic matter on biocides effects. Planktonic forms expressed GA resistance at different levels. C. striatum viability was observed until 2, 4, 20, and 30 min for MDR 2369/II, MDS 1954/IV, MDR 1987/I, and MDS 1961/III strains, respectively. In contrast to GA, the biocides PA, VK24h, SH, and ET had higher effective bacterial mortality. However, storage of VK (48 hr) reduced their biocide activities. Moreover, mature biofilms were produced on abiotic substrates, including steel surfaces. Post-treatment with GA (30 min), survival of sessile forms was ≥100% than planktonic forms of all C. striatum tested strains. Independent of biocides tested, BSA increased the survival of planktonic and sessile forms (p ≤ 0.005). Present data indicated that hospital staff should be aware of dissemination and eradication of HAIs by C. striatum presenting resistance to biocides, including high-level disinfectants, such as GA.


ESP Quick Facts

ESP Origins

In the early 1990's, Robert Robbins was a faculty member at Johns Hopkins, where he directed the informatics core of GDB — the human gene-mapping database of the international human genome project. To share papers with colleagues around the world, he set up a small paper-sharing section on his personal web page. This small project evolved into The Electronic Scholarly Publishing Project.

ESP Support

In 1995, Robbins became the VP/IT of the Fred Hutchinson Cancer Research Center in Seattle, WA. Soon after arriving in Seattle, Robbins secured funding, through the ELSI component of the US Human Genome Project, to create the original ESP.ORG web site, with the formal goal of providing free, world-wide access to the literature of classical genetics.

ESP Rationale

Although the methods of molecular biology can seem almost magical to the uninitiated, the original techniques of classical genetics are readily appreciated by one and all: cross individuals that differ in some inherited trait, collect all of the progeny, score their attributes, and propose mechanisms to explain the patterns of inheritance observed.

ESP Goal

In reading the early works of classical genetics, one is drawn, almost inexorably, into ever more complex models, until molecular explanations begin to seem both necessary and natural. At that point, the tools for understanding genome research are at hand. Assisting readers reach this point was the original goal of The Electronic Scholarly Publishing Project.

ESP Usage

Usage of the site grew rapidly and has remained high. Faculty began to use the site for their assigned readings. Other on-line publishers, ranging from The New York Times to Nature referenced ESP materials in their own publications. Nobel laureates (e.g., Joshua Lederberg) regularly used the site and even wrote to suggest changes and improvements.

ESP Content

When the site began, no journals were making their early content available in digital format. As a result, ESP was obliged to digitize classic literature before it could be made available. For many important papers — such as Mendel's original paper or the first genetic map — ESP had to produce entirely new typeset versions of the works, if they were to be available in a high-quality format.

ESP Help

Early support from the DOE component of the Human Genome Project was critically important for getting the ESP project on a firm foundation. Since that funding ended (nearly 20 years ago), the project has been operated as a purely volunteer effort. Anyone wishing to assist in these efforts should send an email to Robbins.

ESP Plans

With the development of methods for adding typeset side notes to PDF files, the ESP project now plans to add annotated versions of some classical papers to its holdings. We also plan to add new reference and pedagogical material. We have already started providing regularly updated, comprehensive bibliographies to the ESP.ORG site.


Order from Amazon

This is a must read book for anyone with an interest in invasion biology. The full title of the book lays out the author's premise — The New Wild: Why Invasive Species Will Be Nature's Salvation. Not only is species movement not bad for ecosystems, it is the way that ecosystems respond to perturbation — it is the way ecosystems heal. Even if you are one of those who is absolutely convinced that invasive species are actually "a blight, pollution, an epidemic, or a cancer on nature", you should read this book to clarify your own thinking. True scientific understanding never comes from just interacting with those with whom you already agree. R. Robbins

Electronic Scholarly Publishing
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Bellingham, WA 98226

E-mail: RJR8222 @

Papers in Classical Genetics

The ESP began as an effort to share a handful of key papers from the early days of classical genetics. Now the collection has grown to include hundreds of papers, in full-text format.

Digital Books

Along with papers on classical genetics, ESP offers a collection of full-text digital books, including many works by Darwin (and even a collection of poetry — Chicago Poems by Carl Sandburg).


ESP now offers a much improved and expanded collection of timelines, designed to give the user choice over subject matter and dates.


Biographical information about many key scientists.

Selected Bibliographies

Bibliographies on several topics of potential interest to the ESP community are now being automatically maintained and generated on the ESP site.

ESP Picks from Around the Web (updated 07 JUL 2018 )