Chlamydia trachomatis and Mycoplasma genitalium prevalence and associated factors among women presenting to a pregnancy termination and contraception clinic, 2009-2019.

BACKGROUND: Risk of pelvic inflammatory disease associated with Chlamydia trachomatis and Mycoplasma genitalium is increased after termination of pregnancy (TOP) and may be increased after insertion of intrauterine devices (IUDs). Screening prior to these procedures is recommended only for C. trachomatis. We examined C. trachomatis and M. genitalium prevalence and associated factors among women presenting to a pregnancy termination and contraception service over 10 years. METHODS: Retrospective analysis of clinical data collected from 17 573 women aged 15-45 years in 2009-2019 and for 266 M. genitalium positive women tested for macrolide resistance-associated mutations in 2016-2019. RESULTS: C. trachomatis and M. genitalium prevalence was 3.7% and 3.4%, respectively. In multivariable analyses, shared risk factors were younger age (p<0.001, for both C. trachomatis and M. genitalium), socioeconomic disadvantage (p=0.045 and p=0.008, respectively) and coinfection (p<0.001, for both sexually transmitted infections), with 10.1% of C. trachomatis positive women also positive for M. genitalium. Additional risk factors were earlier year of visit (p=0.001) for C. trachomatis and for M. genitalium residing outside a major city (p=0.013). The proportion of M. genitalium infections tested between 2016 and 2019 with macrolide resistance-associated mutations was 32.7%. CONCLUSIONS: Given the high level of antimicrobial resistance and the prevalence of coinfection, testing C. trachomatis positive women for M. genitalium could be considered in this setting to prevent further spread of resistant infections. Further research is required into the causal link between M. genitalium and pelvic inflammatory disease in women undergoing TOP and IUD insertion.

Gardnerella vaginalis Clade Distribution Is Associated With Behavioral Practices and Nugent Score in Women Who Have Sex With Women.

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.

Sexual practices have a significant impact on the vaginal microbiota of women who have sex with women.

Women-who-have-sex-with-women (WSW) are at increased risk of bacterial vaginosis (BV). We investigated the impact of practices and past BV on the vaginal microbiota within a two-year longitudinal cohort of Australian WSW. Self-collected vaginal swabs were used to characterise the vaginal microbiota using 16S-rRNA gene sequencing. Hierarchical clustering defined community state types (CSTs). Bacterial diversity was calculated using the Shannon diversity index and instability of the vaginal microbiota was assessed by change of CST and Bray-Curtis dissimilarity. Sex with a new partner increased the bacterial diversity (adjusted-coefficient = 0.41, 95%CI: 0.21,0.60, p < 0.001) and instability of the vaginal microbiota, in terms of both change of CST (adjusted-odds-ratio = 2.65, 95%CI: 1.34,5.22, p = 0.005) and increased Bray-Curtis dissimilarity (adjusted-coefficient = 0.21, 95%CI: 0.11,0.31, p < 0.001). Women reporting sex with a new partner were more likely than women reporting no new partner to have a vaginal microbiota characterised by Gardnerella vaginalis (adjusted-relative-risk-ratio[aRRR] = 3.45, 95%CI: 1.42,8.41, p = 0.006) or anaerobic BV-associated bacteria (aRRR = 3.62, 95%CI: 1.43,9.14, p = 0.007) relative to a Lactobacillus crispatus dominated microbiota. Sex with a new partner altered the vaginal microbiota of WSW by increasing the diversity and abundance of BV-associated bacteria. These findings highlight the influence of practices on the development of a non-optimal vaginal microbiota and provide microbiological support for the sexual exchange of bacteria between women.

The influence of sexual activity on the vaginal microbiota and Gardnerella vaginalis clade diversity in young women.

OBJECTIVES: To examine the influence of sexual activity on the composition and consistency of the vaginal microbiota over time, and distribution of Gardnerella vaginalis clades in young women. METHODS: Fifty-two participants from a university cohort were selected. Vaginal swabs were self-collected every 3-months for up to 12 months with 184 specimens analysed. The vaginal microbiota was characterised using Roche 454 V3/4 region 16S rRNA sequencing, and G.vaginalis clade typing by qPCR. RESULTS: A Lactobacillus crispatus dominated vaginal microbiota was associated with Caucasian ethnicity (adjusted relative risk ratio[ARRR] = 7.28, 95%CI:1.37,38.57,p = 0.020). An L.iners (ARRR = 17.51, 95%CI:2.18,140.33,p = 0.007) or G.vaginalis (ARRR = 14.03, 95%CI:1.22,160.69, p = 0.034) dominated microbiota was associated with engaging in penile-vaginal sex. Microbiota dominated by L.crispatus, L.iners or other lactobacilli exhibited greater longitudinal consistency of the bacterial communities present compared to ones dominated by heterogeneous non-lactobacilli (p<0.030); sexual activity did not influence consistency. Women who developed BV were more likely to have clade GV4 compared to those reporting no sex/practiced non-coital activities (OR = 11.82, 95%CI:1.87,74.82,p = 0.009). Specimens were more likely to contain multiple G.vaginalis clades rather than a single clade if women engaged in penile-vaginal sex (RRR = 9.55, 95%CI:1.33,68.38,p = 0.025) or were diagnosed with BV (RRR = 31.5, 95%CI:1.69,586.87,p = 0.021). CONCLUSIONS: Sexual activity and ethnicity influenced the composition of the vaginal microbiota of these young, relatively sexually inexperienced women. Women had consistent vaginal microbiota over time if lactobacilli were the dominant spp. present. Penile-vaginal sex did not alter the consistency of microbial communities but increased G.vaginalis clade diversity in young women with and without BV, suggesting sexual transmission of commensal and potentially pathogenic clades.

Incident bacterial vaginosis (BV) in women who have sex with women is associated with behaviors that suggest sexual transmission of BV.

BACKGROUND: Female same-sex partnerships provide a unique opportunity to study the pathogenesis and transmissibility of bacterial vaginosis (BV) because it can be diagnosed in both members of the partnership. We conducted a nationwide community-enrolled cohort study of women who have sex with women, including women coenrolled with their regular female sexual partner (FSP), to investigate the BV incidence rate and factors associated with incident BV. METHODS: Women who have sex with women, without prevalent BV in a cross-sectional study, were enrolled in a 24-month cohort study involving 3-monthly questionnaires and self-collected vaginal swabs that were scored by the Nugent method. We assessed the BV incidence rate per 100 woman-years (WY) and used univariate and multivariable Cox regression analysis to establish factors associated with BV acquisition. RESULTS: Two hundred ninety-eight participants were enrolled in the cohort; 122 were coenrolled with their regular FSP. There were 51 incident cases of BV (rate, 9.75/100 WY; 95% confidence interval [CI], 7.41-12.83). Incident BV was associated with exposure to a new sexual partner (adjusted hazard ratio [AHR], 2.51; 95% CI, 1.30-4.82), a partner with BV symptoms (AHR, 3.99; 95% CI, 1.39-11.45), receptive oral sex (AHR, 3.52; 95% CI, 1.41-8.79), and onset of BV symptoms (AHR, 2.80; 95% CI, 1.39-5.61). Women coenrolled with their BV-negative partner had a greatly reduced risk of incident BV (AHR, 0.26; 95% CI, .11-.61), and high concordance of Nugent category (74%), which was predominantly normal vaginal flora throughout follow-up. CONCLUSIONS: These data highlight the strong influence of sexual relationships and behaviors on BV acquisition and the vaginal microbiota. They provide epidemiological evidence to support exchange of vaginal bacterial species between women and the concept that BV is sexually transmitted.

Factors associated with participation and attrition in a longitudinal study of bacterial vaginosis in Australian women who have sex with women.

OBJECTIVE: A number of social and sexual risk factors for bacterial vaginosis (BV) have been described. It is important to understand whether these factors are associated with non-participation or attrition of participants from longitudinal studies in order to examine potential for recruitment or attrition bias. We describe factors associated with participation and attrition in a 24-month prospective cohort study, investigating incident BV among Australian women who have sex with women. STUDY DESIGN AND SETTING: Participants negative for prevalent BV were offered enrolment in a longitudinal cohort study. Participants self-collected vaginal samples and completed questionnaires 3-monthly to endpoint (BV-positive/BV-negative by 24 months). Factors associated with participation in the cohort study were examined by logistic regression and factors associated with attrition from the cohort were examined by Cox regression. RESULTS: The cross-sectional study recruited 457 women. 334 BV-negative women were eligible for the cohort and 298 (89%, 95%CI 85, 92) enrolled. Lower educational levels (aOR 2.72, 95%CI 1.09, 6.83), smoking (aOR 2.44, 95%CI 1.13, 5.27), past BV symptoms (aOR 3.42, 95%CI 1.16, 10.10) and prior genital warts (aOR 2.71, 95%CI 1.14, 6.46) were associated with non-participation; a partner co-enrolling increased participation (aOR 3.73, 95%CI 1.43, 9.70). 248 participants (83%, 95%CI 78, 87) were retained to study endpoint (BV-negative at 24 months or BV-positive at any stage). Attrition was associated being <30 yrs (aHR 2.15, 95%CI 1.13, 4.10) and a male partner at enrolment (aHR 6.12, 95%CI 1.99, 18.82). CONCLUSION: We achieved high participation and retention levels in a prospective cohort study and report factors influencing participation and retention of participants over a 24-month study period, which will assist in the design and implementation of future cohort studies in sexual health and disease.

The influence of behaviors and relationships on the vaginal microbiota of women and their female partners: the WOW Health Study.

BACKGROUND: A community-based study of women who have sex with women (WSW) was performed to determine the burden of bacterial vaginosis (BV), and behavioral factors influencing the vaginal microbiota of women and their female sexual partners (FSPs), as measured by Nugent score (NS). METHODS: In a cross-sectional study of 18-55-year-old WSW recruited nationally, participants completed questionnaires and self-collected vaginal swab samples weekly on 3 occasions. BV was defined as an NS of 7-10. Factors associated with BV, stability of NS category, and concordance of these categories in co-enrolled couples were examined with multivariable logistic regression analysis. RESULTS: A total of 458 participants were recruited; 192 were co-enrolled with their FSP (96 couples). BV was detected in 125 women (27%; 95% confidence interval [CI], 23%-32%). BV was associated with ≥4 lifetime FSPs (adjusted odds ratio [AOR], 1.9; 95% CI, 1.2-3.1), an FSP with BV symptoms (AOR, 2.9; 1.0-8.2) and smoking, with ≥30 cigarettes per week showing greatest odds (AOR, 2.7; 1.5-5.0). Of 428 women returning ≥2 swab samples, 375 (88%) had a stable NS category across all samples, predominantly reflecting normal flora. Co-enrolled WSW were less likely to have BV (31% vs 23%; P = .07), and the majority (70%) were concordant for NS category (κ = 0.47; P ≤ .01), with most concordant for normal flora. Concordant NS category was associated with a relationship of >6 months (AOR, 4.7; 95% CI, 1.4-16.4) and frequent sexual contact (more than once per month; AOR, 2.7; 1.0-7.1). CONCLUSIONS: BV is associated with key behaviors and smoking practices in WSW, but longer-duration, sexually active WSW partnerships support a stable favorable vaginal microbiota.

The potential of metatranscriptomics for identifying screening targets for bacterial vaginosis.

BACKGROUND: The ribosomal RNA content of a sample collected from a woman with bacterial vaginosis (BV) was analysed to determine the active microbial community, and to identify potential targets for further screening. METHODOLOGY/PRINCIPAL FINDINGS: The sample from the BV patient underwent total RNA extraction, followed by physical subtraction of human rRNA and whole transcriptome amplification. The metatranscriptome was sequenced using Roche 454 titanium chemistry. The bioinformatics pipeline MG-RAST and desktop DNA analysis platforms were utilised to analyse results. Bacteria of the genus Prevotella (predominately P. amnii) constituted 36% of the 16S rRNA reads, followed by Megasphaera (19%), Leptotrichia/Sneathia (8%) and Fusobacterium (8%). Comparison of the abundances of several bacteria to quantitative PCR (qPCR) screening of extracted DNA revealed comparable relative abundances. This suggests a correlation between what was present and transcriptionally active in this sample: however distinct differences were seen when compared to the microbiome determined by 16S rRNA gene amplicon sequencing. To assess the presence of P. amnii in a larger pool of samples, 90 sexually active women were screened using qPCR. This bacterium was found to be strongly associated with BV (P<0.001, OR 23.3 (95%CI:2.9-190.7)) among the 90 women. CONCLUSIONS/SIGNIFICANCE: This study highlighted the potential of metatranscriptomics as a tool for characterising metabolically active microbiota and identifying targets for further screening. Prevotella amnii was chosen as an example target, being the most metabolically active species present in the single patient with BV, and was found to be detected at a high concentration by qPCR in 31% of cohort with BV, with an association with both oral and penile-vaginal sex.

Bacterial vaginosis (BV) candidate bacteria: associations with BV and behavioural practices in sexually-experienced and inexperienced women.

BACKGROUND: In recent years several new fastidious bacteria have been identified that display a high specificity for BV; however no previous studies have comprehensively assessed the behavioural risk associations of these bacterial vaginosis-candidate organisms (BV-COs). METHODS: We examined the associations between 8 key previously described BV-COs and BV status established by Nugent's score (NS). We also examined the sexual practices associated with each BV-CO. We incorporated 2 study populations: 193 from a sexually-inexperienced university population and 146 from a highly sexually-active clinic population. Detailed behavioural data was collected by questionnaire and vaginal smears were scored by the Nugent method. Stored samples were tested by quantitative PCR assays for the 8 BV-COs: Atopobium vaginae, Gardnerella vaginalis, Leptotrichia spp., Megasphaera type I, Sneathia spp., and the Clostridia-like bacteria BVAB1, BVAB2 and BVAB3. Associations between BV-COs and BV and behaviours were examined by univariate and multivariable analyses. RESULTS: On univariate analysis, all BV-COs were more common in BV compared to normal flora. However, only Megasphaera type I, BVAB2, A. vaginae and G. vaginalis were significantly independently associated with BV by multivariable analysis. Six of the eight BV-COs (Megasphaera type I, BVAB2, BVAB3, Sneathia, Leptotrichia and G. vaginalis) were rare or absent in sexually-unexposed women, and demonstrated increasing odds of detection with increasing levels of sexual activity and/or numbers of lifetime sexual partners. Only G. vaginalis and A. vaginae were commonly detected in sexually-unexposed women. Megasphaera type I was independently associated with women-who-have-sex-with women (WSW) and lifetime sexual partner numbers, while unprotected penile-vaginal-sex was associated with BVAB2 detection by multivariate analysis. CONCLUSIONS: Four of eight key BV-COs were significantly associated with BV after adjusting for the presence of other BV-COs. The majority of BV-COs were absent or rare in sexually-unexposed women, and associated with increasing sexual exposure, suggesting potential sexual transmission of BV-COs.

Low incidence of bacterial vaginosis in cohort of young Australian women.

Cohort study of incident bacterial vaginosis (BV) in Australian students. BV only occurred in vaginally sexually active women (BV incidence = 2.2 [95% confidence interval, 0.8-4.9]/100 woman-years), and no incident BV occurred in women who did not engage in vaginal sex during the cohort period (0 [95% confidence interval, 0-3.2]/100 woman-years), P = 0.10. In university students, incident BV was uncommon and absent in sexually abstinent women.

Early sexual experiences and risk factors for bacterial vaginosis.

BACKGROUND: We have undertaken a cross-sectional study that investigates the association between bacterial vaginosis (BV) and sexual practices in sexually experienced and inexperienced women. METHODS: Participants were 17-21-year-old females who attend Melbourne University, Australia. Study kits that contained an information and consent form, questionnaire, swab, and slide were distributed. Information regarding demographic characteristics and a broad range of sexual practices were collected. Gram-stained, self-collected vaginal smears were scored with the Nugent method. Associations between BV and behaviors were examined by univariate and multivariate analysis. RESULTS: BV was diagnosed in 25 (4.7%) of 528 women (95% confidence interval [CI], 3.1%-6.9%). Importantly, BV was not detected in women (n = 83) without a history of coital or noncoital sexual contact (0%; 95% CI, 0%-4.3%). BV was detected in 3 (3.8%) of 78 women (95% CI, 0.8%-10.8%) with noncoital sexual experience only and in 22 (6.0%) of 367 women (95% CI, 3.8%-8.9%) who reported penile-vaginal sex. BV was associated with a history of any genital contact with a sexual partner (P=.02). BV was strongly associated with >3 penile-vaginal sex partners in the prior year (adjusted odds ratio, 7.1; 95% CI, 2.7-18.4) by multivariable analysis. CONCLUSIONS: This study shows a strong association between BV and penile-vaginal sex with multiple partners but found no BV in sexually inexperienced women, once a history of noncoital sexual practices was elicited. Our findings indicate that BV is not present in truly sexually inexperienced women.

Sexual risk factors and bacterial vaginosis: a systematic review and meta-analysis.

We performed a systematic review and meta-analysis of the association between sexual risk factors and bacterial vaginosis (BV). Forty-three studies reported new or multiple sexual partners and condom use relative to prevalent, incident, or recurrent BV. The summary estimate of the relative risk for the association between BV new or multiple male partners was 1.6 (95% confidence interval, 1.5-1.8), between BV and any female partners was 2.0 (95% confidence interval, 1.7-2.3), and between BV and condom use was 0.8 (95% confidence interval, 0.8-0.9). This review is the first to summarize available observational data for BV. It shows that BV is significantly associated with sexual contact with new and multiple male and female partners and that decreasing the number of unprotected sexual encounters may reduce incident and recurrent infection. Investigation of sexual transmission of BV is limited by the absence of a clear microbiological etiology; however, we have shown that the epidemiological profile of BV is similar to that of established sexually transmitted infections.

"Let's not talk about sex": reconsidering the public health approach to sexually transmissible infections in remote Indigenous populations in Australia.

Sexually transmissible infections (STIs) are hyperendemic in some remote Indigenous populations in Australia. Screening programs have had some success in reducing the prevalence of STIs in specific populations, but there has been little overall improvement in the past 10 years. We question the usefulness of current practice and urge consideration of a new and radical approach. Instead of a "screen, treat and contact trace" strategy, we suggest adopting the same approach as currently accepted for trachoma control: populations reaching a threshold prevalence for a set of marker STIs (identified through sentinel surveillance) should be offered a treatment program aimed at the entire sexually active population. We also recommend a parallel program of health promotion and "life skills" education and outline the arguments for such a departure from currently accepted public health policy.

Genital ulcer disease in central Australia: predictors of testing and outcomes.

OBJECTIVES: To identify factors associated with the uptake and outcome of testing for infectious causes of genital ulcer disease (GUD) in central Australia. METHODS: Prospective investigation of GUD cases in central Australia from February 2002 to August 2003. Data were collected from primary health care clinics in remote indigenous communities of Central Australia. RESULTS: During the study period, 134 cases of GUD were reported and investigated. Of these cases, 71 (53%) were in women (age range 14-75, median 28) and 63 (47%) in men (14-63, median 28). Testing for syphilis was undertaken for 111 (82.8%) cases, 75 (56%) were tested for herpes simplex virus infection, and 82 (61.2%) for donovanosis. Testing for at least one of the three sexually transmissible pathogens of interest was undertaken in 128 (95.5%) cases, while 99 (73.9%) were tested for two pathogens and 41 (30.6%) for all three agents. Of subjects tested, 19.8% had new syphilis infection, 51% had herpes simplex virus infection and 7% had donovanosis. In 19 of 41 (46.3%) subjects fully investigated no cause for genital ulceration was found. CONCLUSION: This study provides the first quantitative description of GUD diagnosis in central Australia. Logistic constraints limited the systematic application of diagnostic tests. Current treatment protocols may need to be reassessed in light of the higher than expected detection of genital herpes as a cause of GUD.