Normal Nugent Score Research Articles

Impact of testosterone use on the vaginal microbiota of transgender men, including susceptibility to bacterial vaginosis: study protocol for a prospective, observational study

IntroductionThe effect of testosterone (T) therapy on the vaginal microbiota of transgender men (TGM) is not well characterised, although one cross-sectional study comparing the vaginal microbiota of cisgender women to...

Vaginal Secretion Epithelium Count as a Prognostic Indicator of High Abundance of Ureaplasmas in Women with a Normal Nugent Score.

Genital tract ureaplasma infections are associated with numerous complications, ranging from inflammation, through infertility, to problematic pregnancy. In the course of ureaplasma infection, the risk of human papillomavirus infection increases. Diagnostic tests for urea-plasma infections are not always carried out, especially in women with the normal Nugent test results. The study attempts to check whether it is possible to find a prognostic indicator that could suggest a high abundance of ureaplasmas (≥ 104 CFU/ml) at the stage of the initial examination of vaginal discharge. Such a prognostic factor could qualify women for further tests to detect infections with these atypical bacteria. Six hundred twenty-seven white women with a score of 0–3 on the Nugent scale were tested, including 322 patients with a high abundance of ureaplasmas (≥ 104 CFU/ml) and 305 who tested negative for these bacteria. Ureaplasma infections were detected statistically significant in women who had few or no epithelial cells in the genital swab specimens compared to the results obtained for women with numerous or very numerous epithelial cells (p < 0.001). The risk of the high density of ureaplasmas was 38.7% higher with fewer or no epithelial cells than with high numbers. In patients aged 18–40 years with few or no epithelial cells, a high density of ureaplasmas (≥ 104 CFU/ml) was observed significantly more frequently (p = 0.003). Determining the number of epithelial cells in Gram-stained slides may be the prognostic indicator of ureaplasma infection. Testing for genital ureaplasma infection should be considered, especially in women of childbearing age (18–40 years), even if the Nugent test value is normal and pH ≤ 4.6.

Protozoan-Viral-Bacterial Co-Infections Alter Galectin Levels and Associated Immunity Mediators in the Female Genital Tract.

Co-infections with sexually transmittable pathogens are common and more likely in women with disturbed vaginal bacteriome. Among those pathogens, the protozoan parasite Trichomonas vaginalis (TV) is most common after accounting for the highly persistent DNA viruses human papillomavirus (HPV) and genital herpes. The parasitic infection often concurs with the dysbiotic syndrome diagnosed as bacterial vaginosis (BV) and both are associated with risks of superimposed viral infections. Yet, the mechanisms of microbial synergisms in evading host immunity remain elusive. We present clinical and experimental evidence for a new role of galectins, glycan-sensing family of proteins, in mixed infections. We assessed participants of the HIV Epidemiology Research Study (HERS) at each of their incident TV visits (223 case visits) matched to controls who remained TV-negative throughout the study. Matching criteria included age, race, BV (by Nugent score), HIV status, hysterectomy, and contraceptive use. Non-matched variables included BV status at 6 months before the matched visit, and variables examined at baseline, within 6 months of and/or at the matched visit e.g. HSV-2, HPV, and relevant laboratory and socio-demographic parameters. Conditional logistic regression models using generalized estimating equations calculated odds ratios (OR) for incident TV occurrence with each log10 unit higher cervicovaginal concentration of galectins and cytokines. Incident TV was associated with higher levels of galectin-1, galectin-9, IL-1β and chemokines (ORs 1.53 to 2.91, p <0.001). Galectin-9, IL-1β and chemokines were up and galectin-3 down in TV cases with BV or intermediate Nugent versus normal Nugent scores (p <0.001). Galectin-9, IL-1β and chemokines were up in TV-HIV and down in TV-HPV co-infections. In-vitro, TV synergized with its endosymbiont Trichomonasvirus (TVV) and BV bacteria to upregulate galectin-1, galectin-9, and inflammatory cytokines. The BV-bacterium Prevotella bivia alone and together with TV downregulated galectin-3 and synergistically upregulated galectin-1, galectin-9 and IL-1β, mirroring the clinical findings of mixed TV–BV infections. P. bivia also downregulated TVV+TV-induced anti-viral response e.g. IP-10 and RANTES, providing a mechanism for conducing viral persistence in TV-BV co-infections. Collectively, the experimental and clinical data suggest that galectin-mediated immunity may be dysregulated and exploited by viral–protozoan–bacterial synergisms exacerbating inflammatory complications from dysbiosis and sexually transmitted infections.

Risk and protective factors associated with BV chronicity among women in Rakai, Uganda

ObjectivesTo assess risk and protective factors associated with bacterial vaginosis (BV) chronicity ascertained by Nugent score criteria.MethodsA longitudinal cohort study included 255 sexually experienced, postmenarcheal women who provided weekly self-collected...

Sexually Transmitted Infections Among Kenyan Adolescent Girls and Young Women With Limited Sexual Experience.

Objectives: Globally, the highest rates of sexually transmitted infections (STIs) are among the 15-24 age group. Studying adolescent girls and young women (AGYW) pre-sexual debut could identify risk factors for STI acquisition.Methods: We recruited a prospective cohort of low-risk AGYW aged 16-20 in Kenya. Participants were HIV and HSV-2 seronegative and reported no history of sexual intercourse or reported sex with one partner. Participants underwent genital exams, nucleic acid testing of vaginal swabs for Neisseria gonorrhea (NG), Chlamydia trachomatis (CT), Trichomonas vaginalis (TV), and vaginal gram stains for vaginal dysbiosis by Nugent score. STI correlates were described using χ2 test and t-test.Results: We enrolled 400 AGYW, of which 322 (80.5%) reported never having had sex, while 78 (19.5%) reported prior sex with 1 partner. Among the 78 participants reporting prior sex, 20 (25.6%) reported contraception use in the last 3 months, with 60% using only emergency contraceptive pills. Despite self-reported history, of 373 subjects who underwent STI testing, 49 subjects (13.1%) tested positive for STIs, with 41 CT, 5 GC, and 3 TV cases. Of these 49 subjects, 33 (67.3%) reported no prior sexual intercourse. Bacterial vaginosis was rare and 90% of subjects had a normal Nugent score (0–3).Conclusions: Upon baseline evaluation of a cohort of low risk AGYW, we found high numbers of STIs, especially CT, which is not routinely screened for in Kenyan settings. Interventions to address STIs and unintended pregnancy should target girls pre-sexual debut, including those who do not self-identify as at risk.

When normal isn't normal: heterogeneity in dominant lactobacillus species among women having a nugent score of 0-3

To describe differences in prevalence, concentration, and relative abundance of five vaginal Lactobacillus species across the spectrum of normal Nugent scores (0-3).

Vaginal bacteriome of Nigerian women in health and disease: A study with 16S rRNA metagenomics

Introduction: The argument on what bacteria make up healthy vagina and bacterial vaginosis (BV) remain unresolved. Black women most often are placed in grade IV vaginal communities as lacking Lactobacillus-dominated microbes. We sought to determine the vaginal microbiota compositions of healthy and those with BV using 16S rRNA metagenomics methods. Materials and Methods: Twenty-eight women provided vaginal swabs for Nugent scoring. Fifteen had BV (Nugent score 7–10), whereas 13 were normal (Nugent score 0–3). DNA was extracted and 16S rRNA V4 region amplified using custom bar-coded primers prior to sequencing with MiSeq platform. Sequence reads were imported into Illumina BaseSpace Metagenomics pipeline for 16S rRNA recognition. Distribution of taxonomic categories at different levels of resolution was done using Greengenes databases. Manhattan principal component analysis was used for similarity clustering. Results: Non-BV subjects were colonized by 12 taxonomic phyla that represent 182 genera and 357 species. Overall, 23 phyla representing 388 genera and 805 species were identified in BV subjects. Firmicutes represented 95% of the sequence reads in non-BV subjects with Lactobacillus-dominated genera and Lactobacillus crispatus–dominated species, followed by Proteobacteria (3.78%), Actinobacteria (0.74%), and Bacteriodetes (0.05%). In BV subjects, Firmicutes represented 59% of the classified sequence reads, followed by Bacteroidetes (19%), Actinobacteria (15.8%), Fusobacteria (4.08%), Proteobacteria (1.48%), and Tenericutes (1.25%). Conclusion: Non-BV healthy Black African, Nigerian women had Lactobacillus genera as the predominant microbiota, contrary to published reports. The study shows that BV subjects had varying proportions of diverse bacteria similar to studies from other parts of the world.

128. Sexually Transmitted Infections Among Adolescent Girls in Thika, Kenya

BackgroundGlobally, the rates of sexually transmitted infections (STIs) are highest among 15- to 24-year-old girls, especially in Sub-Saharan African countries where the adolescent sexual health is poor. Recruiting girls presexual debut could identify risk factors for STI acquisition.MethodsWe recruited a prospective cohort of “low-risk” adolescent girls aged 16–20 in Kenya. To be eligible, girls were HIV and HSV-2 seronegative and reported no history of sexual intercourse or reported having sex with only one partner. Demographic data were collected, and girls had nucleic acid testing of vaginal swabs for Neisseria gonorrhea, Chlamydia trachomatis, and Trichomonas vaginalis, and vaginal gram stains for bacterial vaginosis (BV).ResultsWe enrolled 400 girls, with a median age of 18.6 years. In this cohort, 322 (80.5%) girls reported never having had sex, while 78 (19.5%) reported prior sex with 1 partner. Of those reporting prior sex, only 20 (25.6%) reported contraception use in the last 3 months, with 60% using only emergency contraceptive pills. The median age of sexual partners was 22 (IQR 19–25). Of the 373 participants with an STI swab result, 49 participants (13.1%) tested positive for STIs at entry into the study, with 41 chlamydia, 5 gonorrhea, and 3 trichomonas cases. Of these 49 participants, 33 (67.3%) had denied prior sexual intercourse. Testing positive for STIs was, however, significantly different among those reporting prior sexual intercourse vs. reporting never having had sex, 21.1% vs. 11.1% (P = 0.02). BV was rare (5.6%) in the cohort, with 90% of participants with a normal Nugent score of 0–3.ConclusionIn the initial testing of a sexually inexperienced cohort of girls, we found unexpectedly high numbers of prevalent STIs, especially chlamydia which is not routinely screened for in Kenyan settings. Additionally, lack of sexual activity appeared overreported. BV was rare, with much lower prevalence than in adult women in Africa. Our data suggest that prior to initiation of sexual activity, most girls in this Kenyan cohort have vaginal microflora that is dominated by Lactobacillus. Interventions to address STIs, including pre-exposure prophylaxis for HIV, should be targeted at girls at a young age, presexual debut, and in nonmedical settings where girls can be reached who do not selfidentify as at risk for STI.Disclosures All authors: No reported disclosures.

Study on the effects of an oral lactobacilli and lactoferrin complex in women with intermediate vaginal microbiota.

In the vagina of healthy reproductive-aged women, several microbial species maintain a finely tuned mutualistic relationship with the host providing the first-line of defense against the colonization by opportunistic pathogens, which are the leading cause of dysbiosis or vaginal infections (bacterial vaginosis, vulvovaginal candidiasis, and aerobic vaginitis). The use of probiotic lactobacilli to prevent vaginal infections has a good rationale, and an excellent safety record, but so far only a few strains have been clinically proven to be effective, particularly to prevent BV. The aim of the clinical trial was to evaluate the changes in Nugent score in women with intermediate vaginal microbiota treated with oral Lactobacillus acidophilus GLA-14 and Lactobacillus rhamnosus HN001 mixture, in combination with bovine lactoferrin RCX™ (Respecta®) or placebo, for 15days. Vaginal swabs were collected from each woman at baseline and at the end of probiotic treatment and analyzed by RT-PCR. Both symptoms of abnormal vaginal micorbiota and adverse effects were assessed throughout the study. The results showed that oral intake of lactobacilli/lactoferrin mixture led to significant vaginal colonization by L. acidophilus GLA-14 and L. rhamnosus HN001 showing that both strains can colonize vagina following oral ingestion. The effect of such colonization is correlated with the restoration of normal Nugent score (values 0-3) and an improvement of symptoms of abnormal vaginal micorbiota including itching and discharge. Oral consumption of lactobacilli/lactoferrin complex corroborates the effectiveness of using lactobacilli for supporting vaginal health and provides a rational basis for future studies on vaginal infections.

Treatment for Symptomatic Bacterial Vaginosis: ARandomized Controlled Trial.

To compare the efficacy of multiple doses of vaginal clindamycin with a single oral dose of secnidazole for the treatment of bacterial vaginosis. Double-blinded randomized controlled trial. Shifa Foundation Community Health Center, from March 2012 till February 2015. After obtaining written informed consent, a pelvic examination was performed for the confirmation of symptoms of milky white vaginal discharge on speculum examination, positive Amine test and presence of clue cells on microscopy. Pregnant women, known diabetes or any immunocompromised condition, were excluded. Blinding of the patient, doctor, and the pharmacist was done. Study cohort was then divided into two groups, Group Areceived medicine pack Awhich contained active clindamycin and placebo oral preparation, whereas group B was given pack B which contained active 2-gm secnidazole with placebo vaginal cream. Primary outcome and therapeutic success were defined by correction of two out of three (normal Nugent score, negative Amine test, and no milky white discharge) on day 15. At 15th day of treatment, 96.6% participants in vaginal clindamycin group (Group A), recovered from the bacterial vaginosis; whereas, (group B) 23% patients were cured in oral secnidazole group. Multiple doses of vaginal clindamycin are superior to single dose of oral secnidazole for the treatment of bacterial vaginosis.

A longitudinal analysis of the vaginal microbiota and vaginal immune mediators in women from sub-Saharan Africa

In cross-sectional studies increased vaginal bacterial diversity has been associated with vaginal inflammation which can be detrimental for health. We describe longitudinal changes at 5 visits over 8 weeks in vaginal microbiota and immune mediators in African women. Women (N = 40) with a normal Nugent score at all visits had a stable lactobacilli dominated microbiota with prevailing Lactobacillus iners. Presence of prostate-specific antigen (proxy for recent sex) and being amenorrhoeic (due to progestin-injectable use), but not recent vaginal cleansing, were significantly associated with microbiota diversity and inflammation (controlled for menstrual cycle and other confounders). Women (N = 40) with incident bacterial vaginosis (Nugent 7–10) had significantly lower concentrations of lactobacilli and higher concentrations of Gardnerella vaginalis, Atopobium vaginae, and Prevotella bivia, at the incident visit and when concentrations of proinflammatory cytokines (IL-1β, IL-12p70) were increased and IP-10 and elafin were decreased. A higher ‘composite-qPCR vaginal-health-score’ was directly associated with decreased concentrations of proinflammatory cytokines (IL-1α, IL-8, IL-12(p70)) and increased IP-10. This longitudinal study confirms the inflammatory nature of vaginal dysbiosis and its association with recent vaginal sex and progestin-injectable use. A potential role for proinflammatory mediators and IP-10 in combination with the vaginal-health-score as predictive biomarkers for vaginal dysbiosis merits further investigation.

Effect of probiotics on vaginal health in pregnancy. EFFPRO, a randomized controlled trial

Preterm delivery is a leading cause of neonatal morbidity and death. It often results from chorioamnionitis, which is a complication of bacterial vaginosis. Probiotics are effective in the treatment of bacterial vaginosis in women who were not pregnant; studies in pregnant woman are missing. The purpose of this study was to evaluate whether an oral probiotic food supplement supports the maintenance or restoration of a normal vaginal microbiota during pregnancy. We conducted a randomized, placebo-controlled, triple-blind, parallel group trial. Oral Lactobacillus rhamnosus GR-1and L reuteri RC-14 (109 colony-forming units) or placebo were administered for 8 weeks to women with <12 completed weeks of pregnancy. Participants were enrolled at Tuebingen University Hospital and 10 recruiting gynecologic practices. Vaginal swabs were taken before and after intervention and analyzed according to the Nugent scoring system. Telephone interviews were performed before and after intervention and after delivery. Primary outcome was the proportion of swabs with normal Nugent score (<4) after intervention, compared by Fisher's exact test in an intention-to-treat analysis. Three hundred twenty pregnant women were enrolled. Vaginal swabs were analyzed from 290 women before and 271 women after intervention. The proportion of normal vaginal microbiota decreased from 82.6 to 77.8% in the treatment group and from 79.1 to 74.3% in the placebo group, with no significant difference across groups after intervention (P=.297). Oral probiotics may be suitable for implementation in antenatal care but, as administered here, had no effect on vaginal health during mid gestation. Other application routes or probiotic preparations may be more effective in supporting vaginal microbiota during pregnancy.

A DNA tool for early detection of vaginal dysbiosis in African women

A next-generation diagnostic tool for bacterial vaginosis, consisting of quantitative and/or qualitative molecular criteria, has not yet been identified. The optimal diagnostic tool should not only diagnose bacterial vaginosis in diverse populations, but should also detect early signs of transition to dysbiosis.We evaluated a tool based on log10-transformed qPCR data for Lactobacillus crispatus, Lactobacillus iners, Lactobacillus jensenii, Lactobacillus gasseri, Lactobacillus vaginalis, Lactobacillus genus, Atopobium vaginae and Gardnerella vaginalis in vaginal specimens of 426 African women to detect dysbiosis and predict transition to dysbiosis.G. vaginalis (p = 0.204) and A. vaginae (p = 0.001) were more commonly present in women who evolved to an intermediate (Nugent 4–6) or bacterial vaginosis score (Nugent 7–10) compared to women who continued to have a normal Nugent score. The combination of G. vaginalis, A. vaginae and Lactobacillus genus counts performed best for diagnostic accuracy for bacterial vaginosis--sensitivity 93.4% and specificity 83.6%; and for predictive accuracy for bacterial vaginosis--sensitivity 79% and specificity 52%. L. crispatus combinations did not perform well.We conclude that a triple-G. vaginalis-A. vaginae-Lactobacillus genus-qPCR tool holds promise for research in sub-Saharan Africa or when developed as a next-generation clinical diagnostic modality for bacterial vaginosis, ideally engineered as a rapid assay.

The Contribution of Cervicovaginal Infections to the Immunomodulatory Effects of Hormonal Contraception.

ABSTRACTParticular types of hormonal contraceptives (HCs) and genital tract infections have been independently associated with risk of HIV-1 acquisition. We examined whether immunity in women using injectable depot medroxyprogesterone acetate (DMPA), combined oral contraceptives (COC), or no HCs differs by the presence of cervicovaginal infections. Immune mediators were quantified in cervical swabs from 832 HIV-uninfected reproductive-age Ugandans and Zimbabweans. Bacterial infections and HIV were diagnosed by PCR, genital herpes serostatus by enzyme-linked immunosorbent assay (ELISA), altered microflora by Nugent score, and Trichomonas vaginalis and Candida albicans infection by wet mount. Generalized linear models utilizing Box-Cox-Power transformation examined associations between levels of mediators, infection status, and HCs. In no-HC users, T. vaginalis was associated with broadest spectrum of aberrant immunity (higher interleukin 1β [IL-1β], IL-8, macrophage inflammatory protein 3α [MIP-3α], β-defensin 2 [BD2], and IL-1 receptor antigen [IL-1RA]). In women with a normal Nugent score and no genital infection, compared to the no-HC group, COC users showed higher levels of IL-1β, IL-6, IL-8, and IL-1RA, while DMPA users showed higher levels of RANTES and lower levels of BD2, both associated with HIV seroconversion. These effects of COC were blunted in the presence of gonorrhea, chlamydia, trichomoniasis, candidiasis, and an abnormal Nugent score; however, RANTES was increased among COC users with herpes, chlamydia, and abnormal Nugent scores. The effect of DMPA was exacerbated by lower levels of IL-1RA in gonorrhea, chlamydia, or herpes, SLPI in gonorrhea, and IL-1β, MIP-3α, and IL-1RA/IL1β ratio in trichomoniasis. Thus, the effects of HC on cervical immunity depend on the genital tract microenvironment, and a weakened mucosal barrier against HIV may be a combined resultant of genital tract infections and HC use.

Hydrogen Peroxide-Producing Lactobacilli Are Associated With Lower Levels of Vaginal Interleukin-1β, Independent of Bacterial Vaginosis.

The presence of hydrogen peroxide (H2O2)-producing lactobacilli in the vagina is associated with decreased rates of preterm birth and HIV acquisition. We hypothesize that this is due to immunomodulatory effects of these species. Concentrations of interleukin (IL)-1β, IL-6, IL-8, secretory leukocyte protease inhibitor, and human β-defensin 2 were quantified from vaginal swabs from 4 groups of women: women with and without bacterial vaginosis (BV) by Nugent score, further stratified by detection of H2O2-producing lactobacilli by semiquantitative culture. Ten quantitative polymerase chain reaction assays characterized the presence and quantity of select Lactobacillus and BV-associated species in each group. Levels of immune markers and bacteria were compared between the 4 groups using analysis of variance, Kruskal-Wallis, Mann-Whitney U, or χ tests. Swabs from 110 women from 4 groups were included: 26 had a normal Nugent score (BV-), and no H2O2-producing lactobacilli detected (H2O2-); 47 were BV-, H2O2+; 27 BV+, H2O2-; and 10 BV+, H2O2+. The groups were similar in age, marital status, and reproductive history, but not ethnicity: the BV-, H2O2- group had more white participants (P = 0.02). In women with and without BV, IL-1β was lower in the H2O2+ groups. Human β-defensin 2 was lowest in BV+ H2O2- women and highest in BV-, H2O2-. Secretory leukocyte protease inhibitor was lower in women with BV and did not differ by the presence of H2O2-producing lactobacilli. In regression analysis, higher quantities of Lactobacillus crispatus were associated with lower quantities of IL-1β. Detection and quantity of BV-associated species by quantitative polymerase chain reaction was significantly different between women with and without BV, but not between women with and without H2O2-producing lactobacilli within those groups. The presence of H2O2-producing lactobacilli is associated with lower levels of some vaginal proinflammatory cytokines, even in women with BV.

BioMiCo: a supervised Bayesian model for inference of microbial community structure.

BackgroundMicrobiome samples often represent mixtures of communities, where each community is composed of overlapping assemblages of species. Such mixtures are complex, the number of species is huge and abundance information for many species is often sparse. Classical methods have a limited value for identifying complex features within such data.ResultsHere, we describe a novel hierarchical model for Bayesian inference of microbial communities (BioMiCo). The model takes abundance data derived from environmental DNA, and models the composition of each sample by a two-level hierarchy of mixture distributions constrained by Dirichlet priors. BioMiCo is supervised, using known features for samples and appropriate prior constraints to overcome the challenges posed by many variables, sparse data, and large numbers of rare species. The model is trained on a portion of the data, where it learns how assemblages of species are mixed to form communities and how assemblages are related to the known features of each sample. Training yields a model that can predict the features of new samples. We used BioMiCo to build models for three serially sampled datasets and tested their predictive accuracy across different time points. The first model was trained to predict both body site (hand, mouth, and gut) and individual human host. It was able to reliably distinguish these features across different time points. The second was trained on vaginal microbiomes to predict both the Nugent score and individual human host. We found that women having normal and elevated Nugent scores had distinct microbiome structures that persisted over time, with additional structure within women having elevated scores. The third was trained for the purpose of assessing seasonal transitions in a coastal bacterial community. Application of this model to a high-resolution time series permitted us to track the rate and time of community succession and accurately predict known ecosystem-level events.ConclusionBioMiCo provides a framework for learning the structure of microbial communities and for making predictions based on microbial assemblages. By training on carefully chosen features (abiotic or biotic), BioMiCo can be used to understand and predict transitions between complex communities composed of hundreds of microbial species.Electronic supplementary materialThe online version of this article (doi:10.1186/s40168-015-0073-x) contains supplementary material, which is available to authorized users.

The significance of Lactobacillus crispatus and L. vaginalis for vaginal health and the negative effect of recent sex: a cross-sectional descriptive study across groups of African women.

BackgroundWomen in sub-Saharan Africa are vulnerable to acquiring HIV infection and reproductive tract infections. Bacterial vaginosis (BV), a disruption of the vaginal microbiota, has been shown to be strongly associated with HIV infection. Risk factors related to potentially protective or harmful microbiota species are not known.MethodsWe present cross-sectional quantitative polymerase chain reaction data of the Lactobacillus genus, five Lactobacillus species, and three BV-related bacteria (Gardnerella vaginalis, Atopobium vaginae, and Prevotella bivia) together with Escherichia coli and Candida albicans in 426 African women across different groups at risk for HIV. We selected a reference group of adult HIV-negative women at average risk for HIV acquisition and compared species variations in subgroups of adolescents, HIV-negative pregnant women, women engaging in traditional vaginal practices, sex workers and a group of HIV-positive women on combination antiretroviral therapy. We explored the associations between presence and quantity of the bacteria with BV by Nugent score, in relation to several factors of known or theoretical importance.ResultsThe presence of species across Kenyan, South African and Rwandan women was remarkably similar and few differences were seen between the two groups of reference women in Kenya and South Africa. The Rwandan sex workers and HIV-positive women had the highest G. vaginalis presence (p = 0.006). Pregnant women had a higher Lactobacillus genus mean log (7.01 genome equivalents (geq)/ml) compared to the reference women (6.08 geq/ml). L. vaginalis (43%) was second to L. iners (81.9%) highly present in women with a normal Nugent score. Recent sexual exposure negatively affected the presence of L. crispatus (<0.001), L. vaginalis (p = 0.001), and Lactobacillus genus (p < 0.001). Having more than one sexual partner in the last three months was associated with an increased prevalence of G. vaginalis (p = 0.044) and L. iners (p = 0.001).ConclusionsAlthough the composition of species across the studied African countries was similar, the presence of protective species i.e. L. crispatus and L. vaginalis in women with a normal Nugent score appeared lower compared to non-African studies. Furthermore, Lactobacillus species were negatively affected by sexual behavioural. Strategies to support protective Lactobacillus species are urgently needed.Trial registrationThe study is registered at the Trial Registration at the National Health Research Ethics Council South Africa with the number DOH2709103223.Electronic supplementary materialThe online version of this article (doi:10.1186/s12879-015-0825-z) contains supplementary material, which is available to authorized users.

Prevalence ofGardnerella vaginalisamong women with lactobacillus-predominant vaginal flora

To determine the prevalence of Gardnerella vaginalis in women with normal vaginal flora. Women without symptoms or signs of vaginal infection and five or fewer lifetime sexual partners were recruited for a longitudinal study of vaginal flora. Negative Amsel criteria and a Nugent score of 0-3 were required for enrolment. Vaginal specimens were self-collected daily for Gram stain and every 3 days for PCR for G vaginalis for 30 days. Women completed daily diaries recording sexual activity, symptoms and menses. Twenty women were recruited for the study with 19 completing all specimens and 1 lost to follow-up. During the 30-day study period, 13/19 (68.4%) of women had normal Nugent scores (0-3) whereas 6/19 (31.6%) of women had at least 2 days of Nugent scores in the intermediate range (p=0.09). Among the 19 women, 9 (47%) were negative for G vaginalis by PCR throughout the study period whereas 10 (53%) had at least one specimen that demonstrated the presence of G vaginalis by PCR. Of those women with intermediate flora on Gram stain during the course of the study 5/6 (83.3%) were positive for G vaginalis while 5/13 (38.5%) of those women with only normal Nugent scores were positive for G vaginalis. Thus, 61.5% of women with normal Nugent scores had no evidence of G vaginalis by serial PCR. Gardnerella may not be part of the normal flora in women with optimal vaginal health.

P1.032 Bacterial Species in the Vaginal Microbiome Correlated by Nugent Score: Cross-Sectional Data from the Microbicide Safety Biomarkers Study in Kenya, Rwanda, and South Africa

BackgroundStudies characterising the normal and disturbed vaginal microbiome in African women are scarce.MethodsWomen were recruited in three African countries and followed up over seven visits. Vaginal specimens were examined using...

Quantification of bacterial species of the vaginal microbiome in different groups of women, using nucleic acid amplification tests

BackgroundThe vaginal microbiome plays an important role in urogenital health. Quantitative real time Polymerase Chain Reaction (qPCR) assays for the most prevalent vaginal Lactobacillus species and bacterial vaginosis species G. vaginalis and A. vaginae exist, but qPCR information regarding variation over time is still very limited. We set up qPCR assays for a selection of seven species and defined the temporal variation over three menstrual cycles in a healthy Caucasian population with a normal Nugent score. We also explored differences in qPCR data between these healthy women and an ‘at risk’ clinic population of Caucasian, African and Asian women with and without bacterial vaginosis (BV), as defined by the Nugent score.ResultsTemporal stability of the Lactobacillus species counts was high with L. crispatus counts of 108 copies/mL and L. vaginalis counts of 106 copies/mL. We identified 2 types of ‘normal flora’ and one ‘BV type flora’ with latent class analysis on the combined data of all women. The first group was particularly common in women with a normal Nugent score and was characterized by a high frequency of L. crispatus, L. iners, L. jensenii, and L. vaginalis and a correspondingly low frequency of L. gasseri and A. vaginae. The second group was characterized by the predominance of L. gasseri and L. vaginalis and was found most commonly in healthy Caucasian women. The third group was commonest in women with a high Nugent score but was also seen in a subset of African and Asian women with a low Nugent score and was characterized by the absence of Lactobacillus species (except for L. iners) but the presence of G. vaginalis and A. vaginae.ConclusionsWe have shown that the quantification of specific bacteria by qPCR contributes to a better description of the non-BV vaginal microbiome, but we also demonstrated that differences in populations such as risk and ethnicity also have to be taken into account. We believe that our selection of indicator organisms represents a feasible strategy for the assessment of the vaginal microbiome and could be useful for monitoring the microbiome in safety trials of vaginal products.