Voltage-dependent cationic channels formed by a cytolytic toxin produced by Gardnerella vaginalis.
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A cytolytic toxin produced by G. vaginalis was incorporated in artificial membranes and giant liposomes. The toxin formed ionic channels when incorporated in lipid bilayers. The electrical properties of such channels were studied. Current records revealed a unitary conductance of 126 pS (in symmetrical 150 mM KCl). The open state probability of the cytolysin formed channels was a function of the applied membrane potential. The permeability ratio of cations to anions was estimated to be 6.5. (+info)
An altered immunity hypothesis for the development of symptomatic bacterial vaginosis.
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The hypothesis is advanced that the transition from a Lactobacillus-dominated vaginal microflora to a microflora characteristic of bacterial vaginosis (BV), as well as development of the adverse consequences of BV in some women but not in others, are due to alterations in innate immunity. A microbial-induced inhibition of Toll-like receptor expression and/or activity may block induction of proinflammatory immunity and lead to the proliferation of atypical vaginal bacteria. A lack of 70-kDa heat-shock protein production and release in response to abnormal flora would compound this failure to activate antimicrobial immune responses. A deficit in vaginal mannose-binding lectin concentrations would further decrease the capacity for microbial killing and increase the likelihood of bacterial migration from the vagina to the upper genital tract. (+info)
Effect of biofilm phenotype on resistance of Gardnerella vaginalis to hydrogen peroxide and lactic acid.
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OBJECTIVE: Bacterial vaginosis is the most common vaginal disorder worldwide. Certain lactobacilli produce H2O2 and lactic acid, which normally suppress growth of anaerobes; however, in bacterial vaginosis, Gardnerella vaginalis and other anaerobes proliferate, and the number of lactobacilli decreases. G. vaginalis colonizes the vaginal epithelium as a biofilm, which likely plays a role in colonization and relapsing infection. STUDY DESIGN: We developed an in vitro model for G. vaginalis biofilm formation and compared susceptibilities of biofilms vs planktonic cultures to H2O2 and lactic acid. The structure and composition of the biofilm matrix were studied in order to design a method for biofilm dissolution. RESULTS: Biofilms tolerated 5-fold and 4-8 fold higher concentrations of H2O2 and lactic acid (respectively) than planktonic cultures. Proteolytic dissolution of biofilms reduced sensitivity to H2O2 and lactic acid. CONCLUSION: Increased tolerance to H2O2 and lactic acid suggests that biofilm formation contributes to the survival of G. vaginalis in the presence of lactobacilli. (+info)
The susceptibility of organisms associated with bacterial vaginosis to spermicidal compounds, in vitro.
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OBJECTIVES: Bacterial vaginosis (BV) is a prevalent vaginal infection that is now regarded as a risk factor in more serious pelvic and obstetric complications. Spermicides are known to have antimicrobial activity against other sexually transmitted diseases and the aim of this study was to test whether the causative organisms of BV were also susceptible to spermicides, in vitro. DESIGN: Minimal Inhibitory Concentrations of five spermicidal compounds were determined for the organisms associated with BV, in an agar dilution technique. LOCATION: The Department of Experimental and Clinical Microbiology, University of Sheffield Medical School, UK. SPERMICIDES AND ORGANISMS: Nonoxynol-9, Nonoxynol-11, Docusate sodium, Benzalkonium chloride and Menfegol were tested against 20 strains each of Gardnerella vaginalis, Bacteroides and Mobiluncus organisms, isolated from patients with BV who attended the Department of Genitourinary Medicine, the Royal Hallamshire Hospital, Sheffield. MAIN OUTCOME MEASURES: The susceptibility of BV-associated organisms to spermicidal compounds, in vitro. RESULTS: G vaginalis, Mobiluncus spp, Bacteroides bivius and Bacteroides disiens were all susceptible to the five spermicides tested, with MICs ranging between less than or equal to 19 and 5000 mg/l (0.0019%-0.5%). CONCLUSION: The concentrations of spermicides incorporated in contraceptive preparations are usually between 3% and 8%, which are far in excess of the MICs found for BV organisms. Their usage could exert a significant antimicrobial effect and be a useful prophylactic in preventing the infection. (+info)
Isolation of the Bacillus subtilis antimicrobial peptide subtilosin from the dairy product-derived Bacillus amyloliquefaciens.
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AIMS: To purify and characterize an antimicrobial protein (bacteriocin) isolated from the dairy product-derived Bacillus amyloliquefaciens. METHODS AND RESULTS: An unknown bacterial species cultured from the Yogu Farm probiotic dairy beverage was identified through 16S ribosomal RNA analysis as B. amyloliquefaciens, a phylogenetically close relative of Bacillus subtilis. The cell-free supernatant (CFS) of overnight cultures was active against Listeria monocytogenes and also against clinical isolates of Gardnerella vaginalis and Streptococcus agalactiae. At the same time, several isolates of vaginal probiotic Lactobacilli were resistant to the CFS. The nature of the compound causing inhibitory activity was confirmed as proteinaceous by enzymatic digestion. The protein was isolated using ammonium sulfate precipitation, and further purified via column chromatography. PCR analysis was conducted to determine relatedness to other bacteriocins produced by Bacillus spp. CONCLUSION: The antimicrobial protein isolated from B. amyloliquefaciens was shown to be subtilosin, a bacteriocin previously reported as produced only by B. subtilis. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report of intra-species horizontal gene transfer for subtilosin and the first fully characterized bacteriocin isolated from B. amyloliquefaciens. Finally, this is the first report on subtilosin's activity against bacterial vaginosis-associated pathogens. (+info)
Gardnerella vaginalis bacteremia in a previously healthy man: case report and characterization of the isolate.
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Gardnerella vaginalis in women causes vaginitis or infections in other sites, such as the urinary tract, but is an infrequent cause of bacteremia. Bacteremia in men is very rare and is typically associated with immunocompromised states. Here we describe G. vaginalis bacteremia in a previously healthy man with renal calculi and urosepsis. (+info)
Quantitative determination by real-time PCR of four vaginal Lactobacillus species, Gardnerella vaginalis and Atopobium vaginae indicates an inverse relationship between L. gasseri and L. iners.
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BACKGROUND: Most studies of the vaginal microflora have been based on culture or on qualitative molecular techniques. Here we applied existing real-time PCR formats for Lactobacillus crispatus, L. gasseri and Gardnerella vaginalis and developed new formats for Atopobium vaginae, L. iners and L. jensenii to obtain a quantitative non culture-based determination of these species in 71 vaginal samples from 32 pregnant and 28 non-pregnant women aged between 18 and 45 years. RESULTS: The 71 vaginal microflora samples of these women were categorized, using the Ison and Hay criteria, as refined by Verhelst et al. (2005), as follows: grade Ia: 8 samples, grade Iab: 10, grade Ib: 13, grade I-like: 10, grade II: 11, grade III: 12 and grade IV: 7.L. crispatus was found in all but 5 samples and was the most frequent Lactobacillus species detected. A significantly lower concentration of L. crispatus was found in grades II (p < 0.0001) and III (p = 0.002) compared to grade I. L. jensenii was found in all grades but showed higher concentration in grade Iab than in grade Ia (p = 0.024). A. vaginae and G. vaginalis were present in high concentrations in grade III, with log10 median concentrations (log10 MC), respectively of 9.0 and 9.2 cells/ml. Twenty (38.5%) of the 52 G. vaginalis positive samples were also positive for A. vaginae. In grade II we found almost no L. iners (log10 MC: 0/ml) but a high concentration of L. gasseri (log10 MC: 8.7/ml). By contrast, in grade III we found a high concentration of L. iners (log10 MC: 8.3/ml) and a low concentration of L. gasseri (log10 MC: 0/ml). These results show a negative association between L. gasseri and L. iners (r = -0.397, p = 0.001) and between L. gasseri and A. vaginae (r = -0.408, p < 0.0001). CONCLUSION: In our study we found a clear negative association between L. iners and L. gasseri and between A. vaginae and L. gasseri. Our results do not provide support for the generally held proposition that grade II is an intermediate stage between grades I and III, because L. gasseri, abundant in grade II is not predominant in grade III, whereas L. iners, abundant in grade III is present only in low numbers in grade II samples. (+info)
Vaginal IL-8 levels are positively associated with Candida albicans and inversely with lactobacilli in HIV-infected women.
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