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Tania Crucitti - One of the best experts on this subject based on the ideXlab platform.
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contraceptive rings promote vaginal lactobacilli in a high bacterial vaginosis prevalence population a randomised open label longitudinal study in rwandan women
PLOS ONE, 2018Co-Authors: Tania Crucitti, Liselotte Hardy, Lambert Mwambarangwe, Irith De Baetselier, Janneke Van De Wijgert, Stephen Agaba, Jozefien Buyze, Evelyne Kestelyn, Thérèse Delvaux, Vicky JespersAbstract:Background Hormonal contraception has been associated with a reduced risk of vaginal dysbiosis, which in turn has been associated with reduced prevalence of sexually transmitted infections (STIs), including HIV. Vaginal rings are used or developed as delivery systems for contraceptive hormones and antimicrobial drugs for STI and HIV prevention or treatment. We hypothesized that a contraceptive vaginal ring (CVR) containing oestrogen enhances a lactobacilli-dominated vaginal microbial community despite biomass accumulation on the CVR’s surface. Methods We enrolled 120 women for 12 weeks in an open-label NuvaRing® study at Rinda Ubuzima, Kigali, Rwanda. Vaginal and ring microbiota were assessed at baseline and each ring removal visit by Gram stain Nugent scoring (vaginal only), quantitative PCR for Lactobacillus species, Gardnerella vaginalis and Atopobium vaginae, and fluorescent in situ hybridization to visualize cell-adherent bacteria. Ring biomass was measured by crystal violet staining. Results Bacterial vaginosis (BV) prevalence was 48% at baseline. The mean Nugent Score decreased significantly with ring use. The presence and mean log10 concentrations of Lactobacillus species in vaginal secretions increased significantly whereas those of G. vaginalis and presence of A. vaginae decreased significantly. Biomass accumulated on the CVRs with a species composition mirroring the vaginal microbiota. This ring biomass composition and optical density after crystal violet staining did not change significantly over time. Conclusions NuvaRing® promoted lactobacilli-dominated vaginal microbial communities in a population with high baseline BV prevalence despite the fact that biomass accumulated on the rings.
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Scanning electron microscope observation of the biomass accumulated on contraceptive vaginal rings used for three weeks.
2018Co-Authors: Tania Crucitti, Liselotte Hardy, Lambert Mwambarangwe, Irith De Baetselier, Janneke Van De Wijgert, Stephen Agaba, Jozefien Buyze, Evelyne Kestelyn, Thérèse Delvaux, Vicky JespersAbstract:Pictures 1a-3a: The contraceptive vaginal ring was used by a woman with a Nugent Score of 10 and with presence of Lactobacillus iners, Gardnerella vaginalis and Atopobium vaginae as measured by qPCR. A vaginal biofilm consisting of Gardnerella vaginalis and Atopobium vaginae was identified using fluorescence in situ hybridization. On the ring Lactobacillus species, Gardnerella vaginalis and Atopobium vaginae were detected using qPCR, the crystal violet optical density was 3.7555 Pictures 1b-3b: Biomass on a contraceptive vaginal ring after use by a woman with a Nugent Score of 0 and presence of Lactobacillus iners only. No biofilm was visualized using fluorescence in situ hybridization. On the ring Lactobacillus species was detected using qPCR, the crystal violet optical density was 3.7225.
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association of vaginal dysbiosis and biofilm with contraceptive vaginal ring biomass in african women
PLOS ONE, 2017Co-Authors: Liselotte Hardy, Vicky Jespers, Lambert Mwambarangwe, Viateur Musengamana, Irith De Baetselier, Janneke Van De Wijgert, Jozefien Buyze, Tania CrucittiAbstract:We investigated the presence, density and bacterial composition of contraceptive vaginal ring biomass and its association with the vaginal microbiome. Of 415 rings worn by 120 Rwandese women for three weeks, the biomass density was assessed with crystal violet and the bacterial composition of biomass eluates was assessed with quantitative polymerase chain reaction (qPCR). The biomass was visualised after fluorescence in situ hybridisation (FISH) and with scanning electron microscopy (SEM). The vaginal microbiome was assessed with Nugent scoring and vaginal biofilm was visualised after FISH. All vaginal rings were covered with biomass (mean optical density (OD) of 3.36; standard deviation (SD) 0.64). Lactobacilli were present on 93% of the rings, Gardnerella vaginalis on 57%, and Atopobium vaginae on 37%. The ring biomass density was associated with the concentration of A. vaginae (OD +0.03; 95% confidence interval (CI) 0.01–0.05 for one log increase; p = 0.002) and of G. vaginalis (OD +0.03; (95% CI 0.01–0.05; p = 0.013). The density also correlated with Nugent Score: rings worn by women with a BV Nugent Score (mean OD +0.26), and intermediate Score (mean OD +0.09) had a denser biomass compared to rings worn by participants with a normal Score (p = 0.002). Furthermore, presence of vaginal biofilm containing G. vaginalis (p = 0.001) and A. vaginae (p = 0.005) correlated with a denser ring biomass (mean OD +0.24 and +0.22 respectively). With SEM we observed either a loose network of elongated bacteria or a dense biofilm. We found a correlation between vaginal dysbiosis and the density and composition of the ring biomass, and further research is needed to determine if these relationships are causal. As multipurpose vaginal rings to prevent pregnancy, HIV, and other sexually transmitted diseases are being developed, the potential impact of ring biomass on the vaginal microbiota and the release of active pharmaceutical ingredients should be researched in depth.
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the presence of the putative gardnerella vaginalis sialidase a gene in vaginal specimens is associated with bacterial vaginosis biofilm
PLOS ONE, 2017Co-Authors: Liselotte Hardy, Vicky Jespers, Lambert Mwambarangwe, Viateur Musengamana, Mario Vaneechoutte, Jozefien Buyze, Magelien Van Den Bulck, Tania CrucittiAbstract:Bacterial vaginosis (BV) is a difficult-to-treat recurrent condition in which health-associated lactobacilli are outnumbered by other anaerobic bacteria, such as Gardnerella vaginalis. Certain genotypes of G. vaginalis can produce sialidase, while others cannot. Sialidase is known to facilitate the destruction of the protective mucus layer on the vaginal epithelium by hydrolysis of sialic acid on the glycans of mucous membranes. This process possibly facilitates adhesion of bacterial cells on the epithelium since it has been linked with the development of biofilm in other pathogenic conditions. Although it has not been demonstrated yet, it is probable that G. vaginalis benefits from this mechanism by attaching to the vaginal epithelium to initiate biofilm development. In this study, using vaginal specimens of 120 women enrolled in the Ring Plus study, we assessed the association between the putative G. vaginalis sialidase A gene by quantitative polymerase chain reaction (qPCR), the diagnosis of BV according to Nugent Score, and the occurrence of a BV-associated biofilm dominated by G. vaginalis by fluorescence in situ hybridisation (FISH). We detected the putative sialidase A gene in 75% of the G. vaginalis-positive vaginal specimens and found a strong association (p<0.001) between the presence of a G. vaginalis biofilm, the diagnosis of BV according to Nugent and the detection of high loads of the G. vaginalis sialidase A gene in the vaginal specimens. These results could redefine diagnosis of BV, and in addition might guide research for new treatment.
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a fruitful alliance the synergy between atopobium vaginae and gardnerella vaginalis in bacterial vaginosis associated biofilm
Sexually Transmitted Infections, 2016Co-Authors: Liselotte Hardy, Vicky Jespers, Lambert Mwambarangwe, Viateur Musengamana, Mario Vaneechoutte, Said Abdellati, Irith De Baetselier, Janneke Van De Wijgert, Tania CrucittiAbstract:Objectives Bacterial vaginosis (BV) is characterised by a change in the microbial composition of the vagina. The BV-associated organisms outnumber the health-associated Lactobacillus species and form a polymicrobial biofilm on the vaginal epithelium, possibly explaining the difficulties with antibiotic treatment. A better understanding of vaginal biofilm with emphasis on Atopobium vaginae and Gardnerella vaginalis may contribute to a better diagnosis and treatment of BV. Methods To this purpose, we evaluated the association between the presence of both bacteria by fluorescence in situ hybridisation (FISH) and BV by Nugent scoring in 463 vaginal slides of 120 participants participating in a clinical trial in Rwanda. Results A bacterial biofilm was detected in half of the samples using a universal bacterial probe. The biofilm contained A. vaginae in 54.1% and G. vaginalis in 82.0% of the samples. A. vaginae was accompanied by G. vaginalis in 99.5% of samples. The odds of having a Nugent Score above 4 were increased for samples with dispersed G. vaginalis and/or A. vaginae present (OR 4.5; CI 2 to 10.3). The probability of having a high Nugent Score was even higher when a combination of adherent G. vaginalis and dispersed A. vaginae was visualised (OR 75.6; CI 13.3 to 429.5) and highest when both bacteria were part of the biofilm (OR 119; CI 39.9 to 360.8). Conclusions Our study, although not comprehensive at studying the polymicrobial biofilm in BV, provided a strong indication towards the importance of A. vaginae and the symbiosis of A. vaginae and G. vaginalis in this biofilm. Trial registration number NCT01796613.
Liselotte Hardy - One of the best experts on this subject based on the ideXlab platform.
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contraceptive rings promote vaginal lactobacilli in a high bacterial vaginosis prevalence population a randomised open label longitudinal study in rwandan women
PLOS ONE, 2018Co-Authors: Tania Crucitti, Liselotte Hardy, Lambert Mwambarangwe, Irith De Baetselier, Janneke Van De Wijgert, Stephen Agaba, Jozefien Buyze, Evelyne Kestelyn, Thérèse Delvaux, Vicky JespersAbstract:Background Hormonal contraception has been associated with a reduced risk of vaginal dysbiosis, which in turn has been associated with reduced prevalence of sexually transmitted infections (STIs), including HIV. Vaginal rings are used or developed as delivery systems for contraceptive hormones and antimicrobial drugs for STI and HIV prevention or treatment. We hypothesized that a contraceptive vaginal ring (CVR) containing oestrogen enhances a lactobacilli-dominated vaginal microbial community despite biomass accumulation on the CVR’s surface. Methods We enrolled 120 women for 12 weeks in an open-label NuvaRing® study at Rinda Ubuzima, Kigali, Rwanda. Vaginal and ring microbiota were assessed at baseline and each ring removal visit by Gram stain Nugent scoring (vaginal only), quantitative PCR for Lactobacillus species, Gardnerella vaginalis and Atopobium vaginae, and fluorescent in situ hybridization to visualize cell-adherent bacteria. Ring biomass was measured by crystal violet staining. Results Bacterial vaginosis (BV) prevalence was 48% at baseline. The mean Nugent Score decreased significantly with ring use. The presence and mean log10 concentrations of Lactobacillus species in vaginal secretions increased significantly whereas those of G. vaginalis and presence of A. vaginae decreased significantly. Biomass accumulated on the CVRs with a species composition mirroring the vaginal microbiota. This ring biomass composition and optical density after crystal violet staining did not change significantly over time. Conclusions NuvaRing® promoted lactobacilli-dominated vaginal microbial communities in a population with high baseline BV prevalence despite the fact that biomass accumulated on the rings.
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Scanning electron microscope observation of the biomass accumulated on contraceptive vaginal rings used for three weeks.
2018Co-Authors: Tania Crucitti, Liselotte Hardy, Lambert Mwambarangwe, Irith De Baetselier, Janneke Van De Wijgert, Stephen Agaba, Jozefien Buyze, Evelyne Kestelyn, Thérèse Delvaux, Vicky JespersAbstract:Pictures 1a-3a: The contraceptive vaginal ring was used by a woman with a Nugent Score of 10 and with presence of Lactobacillus iners, Gardnerella vaginalis and Atopobium vaginae as measured by qPCR. A vaginal biofilm consisting of Gardnerella vaginalis and Atopobium vaginae was identified using fluorescence in situ hybridization. On the ring Lactobacillus species, Gardnerella vaginalis and Atopobium vaginae were detected using qPCR, the crystal violet optical density was 3.7555 Pictures 1b-3b: Biomass on a contraceptive vaginal ring after use by a woman with a Nugent Score of 0 and presence of Lactobacillus iners only. No biofilm was visualized using fluorescence in situ hybridization. On the ring Lactobacillus species was detected using qPCR, the crystal violet optical density was 3.7225.
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association of vaginal dysbiosis and biofilm with contraceptive vaginal ring biomass in african women
PLOS ONE, 2017Co-Authors: Liselotte Hardy, Vicky Jespers, Lambert Mwambarangwe, Viateur Musengamana, Irith De Baetselier, Janneke Van De Wijgert, Jozefien Buyze, Tania CrucittiAbstract:We investigated the presence, density and bacterial composition of contraceptive vaginal ring biomass and its association with the vaginal microbiome. Of 415 rings worn by 120 Rwandese women for three weeks, the biomass density was assessed with crystal violet and the bacterial composition of biomass eluates was assessed with quantitative polymerase chain reaction (qPCR). The biomass was visualised after fluorescence in situ hybridisation (FISH) and with scanning electron microscopy (SEM). The vaginal microbiome was assessed with Nugent scoring and vaginal biofilm was visualised after FISH. All vaginal rings were covered with biomass (mean optical density (OD) of 3.36; standard deviation (SD) 0.64). Lactobacilli were present on 93% of the rings, Gardnerella vaginalis on 57%, and Atopobium vaginae on 37%. The ring biomass density was associated with the concentration of A. vaginae (OD +0.03; 95% confidence interval (CI) 0.01–0.05 for one log increase; p = 0.002) and of G. vaginalis (OD +0.03; (95% CI 0.01–0.05; p = 0.013). The density also correlated with Nugent Score: rings worn by women with a BV Nugent Score (mean OD +0.26), and intermediate Score (mean OD +0.09) had a denser biomass compared to rings worn by participants with a normal Score (p = 0.002). Furthermore, presence of vaginal biofilm containing G. vaginalis (p = 0.001) and A. vaginae (p = 0.005) correlated with a denser ring biomass (mean OD +0.24 and +0.22 respectively). With SEM we observed either a loose network of elongated bacteria or a dense biofilm. We found a correlation between vaginal dysbiosis and the density and composition of the ring biomass, and further research is needed to determine if these relationships are causal. As multipurpose vaginal rings to prevent pregnancy, HIV, and other sexually transmitted diseases are being developed, the potential impact of ring biomass on the vaginal microbiota and the release of active pharmaceutical ingredients should be researched in depth.
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the presence of the putative gardnerella vaginalis sialidase a gene in vaginal specimens is associated with bacterial vaginosis biofilm
PLOS ONE, 2017Co-Authors: Liselotte Hardy, Vicky Jespers, Lambert Mwambarangwe, Viateur Musengamana, Mario Vaneechoutte, Jozefien Buyze, Magelien Van Den Bulck, Tania CrucittiAbstract:Bacterial vaginosis (BV) is a difficult-to-treat recurrent condition in which health-associated lactobacilli are outnumbered by other anaerobic bacteria, such as Gardnerella vaginalis. Certain genotypes of G. vaginalis can produce sialidase, while others cannot. Sialidase is known to facilitate the destruction of the protective mucus layer on the vaginal epithelium by hydrolysis of sialic acid on the glycans of mucous membranes. This process possibly facilitates adhesion of bacterial cells on the epithelium since it has been linked with the development of biofilm in other pathogenic conditions. Although it has not been demonstrated yet, it is probable that G. vaginalis benefits from this mechanism by attaching to the vaginal epithelium to initiate biofilm development. In this study, using vaginal specimens of 120 women enrolled in the Ring Plus study, we assessed the association between the putative G. vaginalis sialidase A gene by quantitative polymerase chain reaction (qPCR), the diagnosis of BV according to Nugent Score, and the occurrence of a BV-associated biofilm dominated by G. vaginalis by fluorescence in situ hybridisation (FISH). We detected the putative sialidase A gene in 75% of the G. vaginalis-positive vaginal specimens and found a strong association (p<0.001) between the presence of a G. vaginalis biofilm, the diagnosis of BV according to Nugent and the detection of high loads of the G. vaginalis sialidase A gene in the vaginal specimens. These results could redefine diagnosis of BV, and in addition might guide research for new treatment.
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a fruitful alliance the synergy between atopobium vaginae and gardnerella vaginalis in bacterial vaginosis associated biofilm
Sexually Transmitted Infections, 2016Co-Authors: Liselotte Hardy, Vicky Jespers, Lambert Mwambarangwe, Viateur Musengamana, Mario Vaneechoutte, Said Abdellati, Irith De Baetselier, Janneke Van De Wijgert, Tania CrucittiAbstract:Objectives Bacterial vaginosis (BV) is characterised by a change in the microbial composition of the vagina. The BV-associated organisms outnumber the health-associated Lactobacillus species and form a polymicrobial biofilm on the vaginal epithelium, possibly explaining the difficulties with antibiotic treatment. A better understanding of vaginal biofilm with emphasis on Atopobium vaginae and Gardnerella vaginalis may contribute to a better diagnosis and treatment of BV. Methods To this purpose, we evaluated the association between the presence of both bacteria by fluorescence in situ hybridisation (FISH) and BV by Nugent scoring in 463 vaginal slides of 120 participants participating in a clinical trial in Rwanda. Results A bacterial biofilm was detected in half of the samples using a universal bacterial probe. The biofilm contained A. vaginae in 54.1% and G. vaginalis in 82.0% of the samples. A. vaginae was accompanied by G. vaginalis in 99.5% of samples. The odds of having a Nugent Score above 4 were increased for samples with dispersed G. vaginalis and/or A. vaginae present (OR 4.5; CI 2 to 10.3). The probability of having a high Nugent Score was even higher when a combination of adherent G. vaginalis and dispersed A. vaginae was visualised (OR 75.6; CI 13.3 to 429.5) and highest when both bacteria were part of the biofilm (OR 119; CI 39.9 to 360.8). Conclusions Our study, although not comprehensive at studying the polymicrobial biofilm in BV, provided a strong indication towards the importance of A. vaginae and the symbiosis of A. vaginae and G. vaginalis in this biofilm. Trial registration number NCT01796613.
Suzanne M Garland - One of the best experts on this subject based on the ideXlab platform.
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gardnerella vaginalis clade distribution is associated with behavioral practices and Nugent Score in women who have sex with women
The Journal of Infectious Diseases, 2019Co-Authors: Erica Plummer, Lenka A Vodstrcil, Gerald L Murray, Christopher K Fairley, Jennifer Danielewski, Suzanne M Garland, Eric P F Chow, Dieter M BulachAbstract:BACKGROUND: Gardnerella vaginalis is detected in women with and without bacterial vaginosis (BV). Identification of 4 G. vaginalis clades raised the possibility that pathogenic and commensal clades exist. We investigated the association of behavioral practices and Nugent Score with G. vaginalis clade distribution in women who have sex with women (WSW). METHODS: Longitudinal self-collected vaginal specimens were analyzed using established G. vaginalis species-specific and clade-typing polymerase chain reaction assays. Logistic regression assessed factors associated with detection of G. vaginalis clades, and multinomial regression assessed factors associated with number of clades. RESULTS: Clades 1, 2, and 3 and multiclade communities (<2 clades) were associated with Nugent-BV. Clade 1 (odds ratio [OR], 3.36; 95% confidence interval [CI], 1.65-6.84) and multiclade communities (relative risk ratio [RRR], 9.51; 95% CI, 4.36-20.73) were also associated with Lactobacillus-deficient vaginal microbiota. Clade 4 was neither associated with Nugent-BV nor Lactobacillus-deficient microbiota (OR, 1.49; 95% CI, 0.67-3.33). Specific clades were associated with differing behavioral practices. Clade 1 was associated with increasing number of recent sexual partners and smoking, whereas clade 2 was associated with penile-vaginal sex and sharing of sex toys with female partners. CONCLUSIONS: Our results suggest that G. vaginalis clades have varying levels of pathogenicity in WSW, with acquisition occurring through sexual activity. These findings suggest that partner treatment may be an appropriate strategy to improve BV cure.
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p364 gardnerella vaginalis clade distribution is associated with behaviours and Nugent Score in women who have sex with women
Sexually Transmitted Infections, 2019Co-Authors: Erica Plummer, Lenka A Vodstrcil, Gerald L Murray, Christopher K Fairley, Jennifer Danielewski, Suzanne M Garland, Eric P F Chow, Dieter M Bulach, Katherine A Fethers, Jane S HockingAbstract:Background Gardnerella vaginalis (GV) can be detected in women with BV and without BV. Identification of four genetically distinct GV-clades (defined using clade-specific genetic markers) led to the hypothesis that there may be both pathogenic and commensal GV-clades. We conducted a study investigating the distribution and behavioural associations of GV-clades in a cohort of women-who-have-sex-with-women (WSW). Methods Women self-collected vaginal swabs and completed behavioural questionnaires 3-monthly for 24 months or until incident BV(Nugent Score[NS]=7–10). qPCR assays were used to detect GV and the four GV-clades. Multinomial logistic regression assessed factors associated with number of GV-clades. Generalized estimating equations population-averaged models assessed factors associated with each GV-clade. Models accounted for repeated measures. Results 369 specimens from 101 women were analysed. GV was detected in 181 specimens, and most GV-positive specimens had multiple clades present (n=119/181, 66%). Detection of multiple GV-clades was associated with smoking (adjusted relative risk ratio [RRR]:2.52; 95%CI:1.25,5.07), increased lifetime female sex partners (FSP; adjRRR:2.43; 95%CI:1.09,5.38), and a NS=4-6 (intermediate microbiota) or NS=7-10 (Nugent BV) relative to no clades. GV4 was the most prevalent clade (n=136/369; 37%; 95% CI: 32,42%), followed by GV1 (n=116/369; 31%; 95% CI: 27,36%) and GV2 (n=76/369; 21%; 95% CI: 17,25%). GV3 was uncommon (n=17/369; 5%; 95% CI:3,7%). GV1 was associated with a NS=7–10 (adjusted odds ratio[AOR]:3.87; 95%CI:1.75,8.56), smoking (AOR:2.74; 95%CI:1.28,5.87) and report of any sexual partners (AOR:3.41; 95%CI:1.18,9.86). GV2 was associated with NS=4–6 (AOR:3.28; 95%CI:1.00,10.77), sharing of sex-toys (AOR:2.30; 95%CI:1.05,5.04) and recent male sex partners (AOR:6.58; 95%CI:2.02,21.40). GV4 presence was associated with increased lifetime FSPs (AOR:3.17; 95%CI:1.25,5.07). Conclusion GV1 and presence of multiple GV-clades was associated with Nugent BV in WSW, whereas GV2 was associated with intermediate microbiota. Individual GV-clades were associated with a range of differing sexual behaviours in adjusted analyses. These associations are of uncertain importance, but do provide support for exchange of GV-clades between sexual partners. Disclosure No significant relationships.
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Heat map depicting vaginal bacterial communities analysed in this study.
2017Co-Authors: Lenka A Vodstrcil, Christopher K Fairley, Suzanne M Garland, Eric P F Chow, Katherine A Fethers, Jane S Hocking, Jimmy Twin, Matthew G. Law, Erica L. Plummer, Sepehr N. TabriziAbstract:Heat map of proportions of bacterial taxa identified from 184 specimens from 52 participants. Values are expressed as arcsine transformed relative abundances. Nugent Score categories and sexual activity categories for each specimen are indicated. Community state type (CST) groupings shown based on previous literature [4, 6, 16], where CST I aligns with specimens dominated by Lactobacillus crispatus, CST II with specimens dominated by L. gasseri, CST III with specimens dominated by L. iners, CST IV with specimens dominated by G. vaginalis and CST V with specimens dominated by L. jensenii. CST VI represents specimens dominated by other heterogeneous non-lactobacilli (predominantly Atopobium vaginae and Prevotella spp.) and CST VII specimens dominated by Bifidobacterium breve.
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recurrence of bacterial vaginosis is significantly associated with posttreatment sexual activities and hormonal contraceptive use
Clinical Infectious Diseases, 2013Co-Authors: Lenka A Vodstrcil, Suzanne M Garland, Jane S Hocking, Catriona S Bradshaw, Anna N Morton, Matthew Law, Marie Pirotta, Deborah De Guingand, Christopher K FairleyAbstract:Background. Bacterial vaginosis (BV) recurrence following recommended therapies is common yet whether recurrence is due to persistent infection or re-infection is unknown. Our aim was to determine behaviours associated with BV recurrence in women enrolled in a randomized-controlled trial.Methods. Symptomatic 18-50 year old females with BV (>/=3Amsels criteria and Nugent Score (NS)=4-10) were enrolled in a 3-arm randomised double-blind placebo-controlled trial at Melbourne Sexual Health Centre Australia 2009-10. 450 participants received 7-days of oral metronidazole were equally randomised to: vaginal-clindamycin a lactobacillus vaginal-probiotic or vaginal-placebo and completed a questionnaire. At 123 & 6-months participants self-collected vaginal smears and completed questionnaires. Primary endpoint was NS=7-10. Cox regression was used to estimate hazard ratios (HR) for risk of BV recurrence associated with baseline and longitudinal characteristics allowing for repeated measures from participants and stratifying for treatment.Results. 404 (90%) women who provided post-randomization data were included in analyses. Cumulative 6-month BV recurrence was 28% (95%CI 24-33%) and not associated with treatment. After adjustment for frequency of sex and age BV recurrence was associated with having the same pre/post-treatment regular sexual partner(RSP) (Adjusted HR=1.9;95%CI 1.2-3.0) inconsistent condom use (AHR=1.9;1.0-3.3) and being born outside Australia (AHR=1.5;1.0-2.1); use of an oestrogen-containing contraceptive was protective (AHR=0.5;0.3-0.8).Conclusions. BV recurrence was significantly increased by remaining with the same pre/post-treatment RSP and inconsistent condom use and halved with use of oestrogen-containing contraceptives. Behavioural and contraceptive practices appear to play a significant role in modifying the effectiveness of antibiotic therapies in the treatment of BV. These findings have implications for clinical practice.
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the association of atopobium vaginae and gardnerella vaginalis with bacterial vaginosis and recurrence after oral metronidazole therapy
The Journal of Infectious Diseases, 2006Co-Authors: Catriona S Bradshaw, Christopher K Fairley, Suzanne M Garland, Sepehr N. Tabrizi, Anna N Morton, E RudlandAbstract:Background. We investigated associations between Atopobium vaginae and bacterial vaginosis (BV) and the role that A. vaginae plays in recurrent BV after oral metronidazole therapy. Methods. Women with abnormal vaginal discharge or odor were enrolled in a cross-sectional study (n p ); the proportion of those infected with Gardnerella vaginalis and A. vaginae was determined by polymerase 358 chain reaction. Women with BV (Nugent Score [NS] 7–10 or 4–6 with 3 Amsel criteria; ) were treated n p 139 with oral metronidazole (400 mg twice a day for 7 days) and examined at 1, 3, 6, and 12 months or until they reached an NS of 7–10 and recurrence of A. vaginae and G. vaginalis infection was established. Results. A. vaginae and G. vaginalis were highly sensitive for BV—96% (95% confidence interval [CI], 91%– 98%) and 99% (95% CI, 97%–100%), respectively. However, A. vaginalis was more specific for BV (77% [95% CI, 71%–82%]) than was G. vaginalis (35% [95% CI, 29%–42%]). G. vaginalis was detected in 100% and A. vaginae in 75% of women with recurrent BV; higher organism loads were present in women with recurrent BV. A. vaginae was rarely detected without G. vaginalis, and women in whom both organisms were detected had higher rates of recurrent BV (83%) than women infected with G. vaginalis only (38%) ( ). P ! .001 Conclusions. Infection with A. vaginae is more specific for BV than infection with G. vaginalis. The higher recurrence rates in women in whom both A. vaginae and G. vaginalis were detected suggest that A. vaginae makes a significant contribution to BV. However, its etiological role remains unclear.
Vicky Jespers - One of the best experts on this subject based on the ideXlab platform.
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contraceptive rings promote vaginal lactobacilli in a high bacterial vaginosis prevalence population a randomised open label longitudinal study in rwandan women
PLOS ONE, 2018Co-Authors: Tania Crucitti, Liselotte Hardy, Lambert Mwambarangwe, Irith De Baetselier, Janneke Van De Wijgert, Stephen Agaba, Jozefien Buyze, Evelyne Kestelyn, Thérèse Delvaux, Vicky JespersAbstract:Background Hormonal contraception has been associated with a reduced risk of vaginal dysbiosis, which in turn has been associated with reduced prevalence of sexually transmitted infections (STIs), including HIV. Vaginal rings are used or developed as delivery systems for contraceptive hormones and antimicrobial drugs for STI and HIV prevention or treatment. We hypothesized that a contraceptive vaginal ring (CVR) containing oestrogen enhances a lactobacilli-dominated vaginal microbial community despite biomass accumulation on the CVR’s surface. Methods We enrolled 120 women for 12 weeks in an open-label NuvaRing® study at Rinda Ubuzima, Kigali, Rwanda. Vaginal and ring microbiota were assessed at baseline and each ring removal visit by Gram stain Nugent scoring (vaginal only), quantitative PCR for Lactobacillus species, Gardnerella vaginalis and Atopobium vaginae, and fluorescent in situ hybridization to visualize cell-adherent bacteria. Ring biomass was measured by crystal violet staining. Results Bacterial vaginosis (BV) prevalence was 48% at baseline. The mean Nugent Score decreased significantly with ring use. The presence and mean log10 concentrations of Lactobacillus species in vaginal secretions increased significantly whereas those of G. vaginalis and presence of A. vaginae decreased significantly. Biomass accumulated on the CVRs with a species composition mirroring the vaginal microbiota. This ring biomass composition and optical density after crystal violet staining did not change significantly over time. Conclusions NuvaRing® promoted lactobacilli-dominated vaginal microbial communities in a population with high baseline BV prevalence despite the fact that biomass accumulated on the rings.
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Scanning electron microscope observation of the biomass accumulated on contraceptive vaginal rings used for three weeks.
2018Co-Authors: Tania Crucitti, Liselotte Hardy, Lambert Mwambarangwe, Irith De Baetselier, Janneke Van De Wijgert, Stephen Agaba, Jozefien Buyze, Evelyne Kestelyn, Thérèse Delvaux, Vicky JespersAbstract:Pictures 1a-3a: The contraceptive vaginal ring was used by a woman with a Nugent Score of 10 and with presence of Lactobacillus iners, Gardnerella vaginalis and Atopobium vaginae as measured by qPCR. A vaginal biofilm consisting of Gardnerella vaginalis and Atopobium vaginae was identified using fluorescence in situ hybridization. On the ring Lactobacillus species, Gardnerella vaginalis and Atopobium vaginae were detected using qPCR, the crystal violet optical density was 3.7555 Pictures 1b-3b: Biomass on a contraceptive vaginal ring after use by a woman with a Nugent Score of 0 and presence of Lactobacillus iners only. No biofilm was visualized using fluorescence in situ hybridization. On the ring Lactobacillus species was detected using qPCR, the crystal violet optical density was 3.7225.
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association of vaginal dysbiosis and biofilm with contraceptive vaginal ring biomass in african women
PLOS ONE, 2017Co-Authors: Liselotte Hardy, Vicky Jespers, Lambert Mwambarangwe, Viateur Musengamana, Irith De Baetselier, Janneke Van De Wijgert, Jozefien Buyze, Tania CrucittiAbstract:We investigated the presence, density and bacterial composition of contraceptive vaginal ring biomass and its association with the vaginal microbiome. Of 415 rings worn by 120 Rwandese women for three weeks, the biomass density was assessed with crystal violet and the bacterial composition of biomass eluates was assessed with quantitative polymerase chain reaction (qPCR). The biomass was visualised after fluorescence in situ hybridisation (FISH) and with scanning electron microscopy (SEM). The vaginal microbiome was assessed with Nugent scoring and vaginal biofilm was visualised after FISH. All vaginal rings were covered with biomass (mean optical density (OD) of 3.36; standard deviation (SD) 0.64). Lactobacilli were present on 93% of the rings, Gardnerella vaginalis on 57%, and Atopobium vaginae on 37%. The ring biomass density was associated with the concentration of A. vaginae (OD +0.03; 95% confidence interval (CI) 0.01–0.05 for one log increase; p = 0.002) and of G. vaginalis (OD +0.03; (95% CI 0.01–0.05; p = 0.013). The density also correlated with Nugent Score: rings worn by women with a BV Nugent Score (mean OD +0.26), and intermediate Score (mean OD +0.09) had a denser biomass compared to rings worn by participants with a normal Score (p = 0.002). Furthermore, presence of vaginal biofilm containing G. vaginalis (p = 0.001) and A. vaginae (p = 0.005) correlated with a denser ring biomass (mean OD +0.24 and +0.22 respectively). With SEM we observed either a loose network of elongated bacteria or a dense biofilm. We found a correlation between vaginal dysbiosis and the density and composition of the ring biomass, and further research is needed to determine if these relationships are causal. As multipurpose vaginal rings to prevent pregnancy, HIV, and other sexually transmitted diseases are being developed, the potential impact of ring biomass on the vaginal microbiota and the release of active pharmaceutical ingredients should be researched in depth.
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the presence of the putative gardnerella vaginalis sialidase a gene in vaginal specimens is associated with bacterial vaginosis biofilm
PLOS ONE, 2017Co-Authors: Liselotte Hardy, Vicky Jespers, Lambert Mwambarangwe, Viateur Musengamana, Mario Vaneechoutte, Jozefien Buyze, Magelien Van Den Bulck, Tania CrucittiAbstract:Bacterial vaginosis (BV) is a difficult-to-treat recurrent condition in which health-associated lactobacilli are outnumbered by other anaerobic bacteria, such as Gardnerella vaginalis. Certain genotypes of G. vaginalis can produce sialidase, while others cannot. Sialidase is known to facilitate the destruction of the protective mucus layer on the vaginal epithelium by hydrolysis of sialic acid on the glycans of mucous membranes. This process possibly facilitates adhesion of bacterial cells on the epithelium since it has been linked with the development of biofilm in other pathogenic conditions. Although it has not been demonstrated yet, it is probable that G. vaginalis benefits from this mechanism by attaching to the vaginal epithelium to initiate biofilm development. In this study, using vaginal specimens of 120 women enrolled in the Ring Plus study, we assessed the association between the putative G. vaginalis sialidase A gene by quantitative polymerase chain reaction (qPCR), the diagnosis of BV according to Nugent Score, and the occurrence of a BV-associated biofilm dominated by G. vaginalis by fluorescence in situ hybridisation (FISH). We detected the putative sialidase A gene in 75% of the G. vaginalis-positive vaginal specimens and found a strong association (p<0.001) between the presence of a G. vaginalis biofilm, the diagnosis of BV according to Nugent and the detection of high loads of the G. vaginalis sialidase A gene in the vaginal specimens. These results could redefine diagnosis of BV, and in addition might guide research for new treatment.
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a fruitful alliance the synergy between atopobium vaginae and gardnerella vaginalis in bacterial vaginosis associated biofilm
Sexually Transmitted Infections, 2016Co-Authors: Liselotte Hardy, Vicky Jespers, Lambert Mwambarangwe, Viateur Musengamana, Mario Vaneechoutte, Said Abdellati, Irith De Baetselier, Janneke Van De Wijgert, Tania CrucittiAbstract:Objectives Bacterial vaginosis (BV) is characterised by a change in the microbial composition of the vagina. The BV-associated organisms outnumber the health-associated Lactobacillus species and form a polymicrobial biofilm on the vaginal epithelium, possibly explaining the difficulties with antibiotic treatment. A better understanding of vaginal biofilm with emphasis on Atopobium vaginae and Gardnerella vaginalis may contribute to a better diagnosis and treatment of BV. Methods To this purpose, we evaluated the association between the presence of both bacteria by fluorescence in situ hybridisation (FISH) and BV by Nugent scoring in 463 vaginal slides of 120 participants participating in a clinical trial in Rwanda. Results A bacterial biofilm was detected in half of the samples using a universal bacterial probe. The biofilm contained A. vaginae in 54.1% and G. vaginalis in 82.0% of the samples. A. vaginae was accompanied by G. vaginalis in 99.5% of samples. The odds of having a Nugent Score above 4 were increased for samples with dispersed G. vaginalis and/or A. vaginae present (OR 4.5; CI 2 to 10.3). The probability of having a high Nugent Score was even higher when a combination of adherent G. vaginalis and dispersed A. vaginae was visualised (OR 75.6; CI 13.3 to 429.5) and highest when both bacteria were part of the biofilm (OR 119; CI 39.9 to 360.8). Conclusions Our study, although not comprehensive at studying the polymicrobial biofilm in BV, provided a strong indication towards the importance of A. vaginae and the symbiosis of A. vaginae and G. vaginalis in this biofilm. Trial registration number NCT01796613.
Gregor Reid - One of the best experts on this subject based on the ideXlab platform.
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a study of the vaginal microbiome in healthy canadian women utilizing cpn60 based molecular profiling reveals distinct gardnerella subgroup community state types
PLOS ONE, 2015Co-Authors: Arianne Albert, John J Schellenberg, Janet E Hill, Gregor Reid, Bonnie Chaban, Emily C Wagner, Matthew G Links, Julie Van Schalkwyk, Sean M Hemmingsen, Deborah MoneyAbstract:The vaginal microbiota is important in women’s reproductive and overall health. However, the relationships between the structure, function and dynamics of this complex microbial community and health outcomes remain elusive. The objective of this study was to determine the phylogenetic range and abundance of prokaryotes in the vaginal microbiota of healthy, non-pregnant, ethnically diverse, reproductive-aged Canadian women. Socio-demographic, behavioural and clinical data were collected and vaginal swabs were analyzed from 310 women. Detailed profiles of their vaginal microbiomes were generated by pyrosequencing of the chaperonin-60 universal target. Six community state types (CST) were delineated by hierarchical clustering, including three Lactobacillus-dominated CST (L. crispatus, L. iners, L. jensenii), two Gardnerella-dominated (subgroups A and C) and an “intermediate” CST which included a small number of women with microbiomes dominated by seven other species or with no dominant species but minority populations of Streptococcus, Staphylococcus, Peptoniphilus, E. coli and various Proteobacteria in co-dominant communities. The striking correspondence between Nugent Score and deep sequencing CST continues to reinforce the basic premise provided by the simpler Gram stain method, while additional analyses reveal detailed cpn60-based phylogeny and estimated abundance in microbial communities from vaginal samples. Ethnicity was the only demographic or clinical characteristic predicting CST, with differences in Asian and White women (p = 0.05). In conclusion, this study confirms previous work describing four cpn60-based subgroups of Gardnerella, revealing previously undescribed CST. The data describe the range of bacterial communities seen in Canadian women presenting with no specific vaginal health concerns, and provides an important baseline for future investigations of clinically important cohorts.
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improved cure of bacterial vaginosis with single dose of tinidazole 2 g lactobacillus rhamnosus gr 1 and lactobacillus reuteri rc 14 a randomized double blind placebo controlled trial
Canadian Journal of Microbiology, 2009Co-Authors: Rafael Chacon Ruiz Martinez, Silvio Antonio Franceschini, Maristela Carbol Patta, Silvana Maria Quintana, Bruna Carrer Gomes, Elaine Cristina Pereira De Martinis, Gregor ReidAbstract:Bacterial vaginosis (BV) is the most prevalent vaginal infection worldwide and is characterized by depletion of the indigenous lactobacilli. Antimicrobial therapy is often ineffective. We hypothesized that probiotic Lactobacillus rham- nosus GR-1 and Lactobacillus reuteri RC-14 might provide an adjunct to antimicrobial treatment and improve cure rates. Sixty-four Brazilian women diagnosed with BV were randomly assigned to receive a single dose of tinidazole (2 g) sup- plemented with either 2 placebo capsules or 2 capsules containing L. rhamnosus GR-1 and L. reuteri RC-14 every morning for the following 4 weeks. At the end of treatment (day 28), the probiotic group had a significantly higher cure rate of BV (87.5%) than the placebo group (50.0%) (p = 0.001). In addition, according to the Gram-stain Nugent Score, more women were assessed with ''normal'' vaginal microbiota in the probiotic group (75.0% vs. 34.4% in the placebo group; p = 0.011). This study shows that probiotic lactobacilli can provide benefits to women being treated with antibiotics for an in- fectious condition.
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augmentation of antimicrobial metronidazole therapy of bacterial vaginosis with oral probiotic lactobacillus rhamnosus gr 1 and lactobacillus reuteri rc 14 randomized double blind placebo controlled trial
Microbes and Infection, 2006Co-Authors: Kingsley C Anukam, Gregor Reid, Emmanuel O Osazuwa, Ijeoma Ahonkhai, Michael Ngwu, Gibson I Osemene, Andrew A BruceAbstract:This study enrolled 125 premenopausal women diagnosed with bacterial vaginosis (BV) by presence of vaginal irritation, discharge and 'fishy' odor, and Nugent criteria and detection of sialidase enzyme. The subjects were treated with oral metronidazole (500 mg) twice daily from days 1 to 7, and randomized to receive oral Lactobacillus rhamnosus GR-1 (1 x 10(9)) and Lactobacillus reuteri RC-14 (1 x 10(9)) or placebo twice daily from days 1 to 30. Primary outcome was cure of BV as determined by normal Nugent Score, negative sialidase test and no symptoms or signs of BV at day 30. A total of 106 subjects returned for 30-day follow-up, of which 88% were cured in the antibiotic/probiotic group compared to 40% in the antibiotic/placebo group (p 10(5) CFU/ml) were recovered from the vagina of 96% probiotic-treated subjects compared to 53% controls at day 30. In summary, this study showed efficacious use of lactobacilli and antibiotic in the eradication of BV in black African women.
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evaluation of the bacterial vaginal flora of 20 postmenopausal women by direct Nugent Score and molecular polymerase chain reaction and denaturing gradient gel electrophoresis techniques
The Journal of Infectious Diseases, 2002Co-Authors: Jeremy P Burton, Gregor ReidAbstract:To monitor the bacterial vaginal microflora of postmenopausal women, several approaches were used: (1) Nugent scoring of Gram-stained smears from vaginal swabs; (2) testing for bacteria, using specific polymerase chain reaction (PCR) primers; (3) PCR-denaturing gradient gel electrophoresis (DGGE) analysis of total bacterial DNA, amplified using eubacterial and Lactobacillus-specific primers; and (4) sequence analysis of amplified DNA fragments of interest from denaturing gradient gels. Of 20 women studied, 70% had either intermediate-grade bacterial colonization or bacterial vaginosis (BV), as defined by Nugent criteria, at the first sampling point. Subjects with normal Nugent Scores had >/=1 species of Lactobacillus detected by sequencing of dominant DNA fragments from denaturing gradient gels generated using eubacterial primers. Samples from women with Nugent Scores that indicated BV had varied bacterial DGGE profiles, which emphasizes the diverse nature of such infections. This study indicates that asymptomatic BV appears to be much more common than is currently perceived; these findings may have implications for the health of postmenopausal women.
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evaluation of the bacterial vaginal flora of 20 postmenopausal women by direct Nugent Score and molecular polymerase chain reaction and denaturing gradient gel electrophoresis techniques
The Journal of Infectious Diseases, 2002Co-Authors: Jeremy P Burton, Gregor ReidAbstract:To monitor the bacterial vaginal microflora of postmenopausal women, several approaches were used: (1) Nugent scoring of Gram-stained smears from vaginal swabs; (2) testing for bacteria, using specific polymerase chain reaction (PCR) primers; (3) PCR‐denaturing gradient gel electrophoresis (DGGE) analysis of total bacterial DNA, amplified using eubacterial and Lactobacillus-specific primers; and (4) sequence analysis of amplified DNA fragments of interest from denaturing gradient gels. Of 20 women studied, 70% had either intermediate-grade bacterial colonization or bacterial vaginosis (BV), as defined by Nugent criteria, at the first sampling point. Subjects with normal Nugent Scores had 1 species of Lactobacillus detected by sequencing of dominant DNA fragments from denaturing gradient gels generated using eubacterial primers. Samples from women with Nugent Scores that indicated BV had varied bacterial DGGE profiles, which emphasizes the diverse nature of such infections. This study indicates that asymptomatic BV appears to be much more common than is currently perceived; these findings may have implications for the health of postmenopausal women. Urinary tract infections (UTIs) and vaginal infections, including yeast vaginitis and bacterial vaginosis (BV), collectively represent perhaps the most common affliction in women. Such infections occur with greater frequency after menopause, but, although there have been some bacteriological studies of the vaginal flora [1], these studies have not led to new therapeutic options. The single exception is the finding that the use of vaginal estriol can lead to a recovery of the normal lactobacilli flora and a reduced risk of infection [2]. The relationship between estrogen levels and bacterial colonization has long been implicated in the elevated incidence of BV [2‐4], but predisposing microbial factors have not yet been adequately explored. The correlation between a healthy vaginal tract and dominant colonization by members of the bacterial genus Lactobacillus has led people to believe that these commensal organisms play a major role in preventing certain types of vaginal infections. To prevent recurrences of urogenital infections, at
