Figures
Abstract
Background
To determine prevalence and incidence of bacterial vaginosis (BV) and risk factors in young sexually-active Australian women.
Methods
1093 women aged 16–25 years were recruited from primary-care clinics. Participants completed 3-monthly questionnaires and self-collected vaginal smears 6-monthly for 12-months. The primary endpoint was a Nugent Score = 7–10 (BV) and the secondary endpoint was a NS = 4–10 (abnormal flora [AF]). BV and AF prevalence estimates and 95% confidence intervals (95%CI) were derived, and adjusted odds ratios (AOR) calculated to explore epidemiological associations with prevalent BV and AF. Proportional-hazards regression models were used to examine factors associated with incident BV and AF.
Results
At baseline 129 women had BV [11.8% (95%CI: 9.4–14.2)] and 188 AF (17.2%; 15.1–19.5). Prevalent BV was associated with having a recent female partner [AOR = 2.1; 1.0–4.4] and lack of tertiary-education [AOR = 1.9; 1.2–3.0]; use of an oestrogen-containing contraceptive (OCC) was associated with reduced risk [AOR = 0.6; 0.4–0.9]. Prevalent AF was associated with the same factors, and additionally with >5 male partners (MSP) in 12-months [AOR = 1.8; 1.2–2.5)], and detection of C.trachomatis or M.genitalium [AOR = 2.1; 1.0–4.5]. There were 82 cases of incident BV (9.4%;7.7–11.7/100 person-years) and 129 with incident AF (14.8%; 12.5–17.6/100 person-years). Incident BV and AF were associated with a new MSP [adjusted rate ratio (ARR) = 1.5; 1.1–2.2 and ARR = 1.5; 1.1–2.0], respectively. OCC-use was associated with reduced risk of incident AF [ARR = 0.7; 0.5–1.0].
Conclusion
This paper presents BV and AF prevalence and incidence estimates from a large prospective cohort of young Australian women predominantly recruited from primary-care clinics. These data support the concept that sexual activity is strongly associated with the development of BV and AF and that use of an OCC is associated with reduced risk.
Citation: Bradshaw CS, Walker J, Fairley CK, Chen MY, Tabrizi SN, Donovan B, et al. (2013) Prevalent and Incident Bacterial Vaginosis Are Associated with Sexual and Contraceptive Behaviours in Young Australian Women. PLoS ONE 8(3): e57688. https://doi.org/10.1371/journal.pone.0057688
Editor: Jacques Ravel, Institute for Genome Sciences, University of Maryland School of Medicine, United States of America
Received: December 10, 2012; Accepted: January 24, 2013; Published: March 5, 2013
Copyright: © 2013 Bradshaw et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Funding: This work was supported by the Commonwealth of Australia, as part of a National Chlamydia Pilot Program and the Australian National Health and Research Council (research grant number 509144). CSB, and JSH were supported by research fellowships issued by the Australian National Health and Medical Research Council (fellowship numbers 456164 and 566576 respectively). MP was supported by a Primary Health Care Research Evaluation and Development Mid-Career Fellowship, Department of Health and Aging. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Competing interests: The authors have declared that no competing interests exist.
Introduction
Bacterial vaginosis (BV) is the most common cause of abnormal vaginal discharge in women of reproductive age. However, internationally, most BV prevalence and incidence data are derived from testing women attending sexual/reproductive services, rather than from community-based screening studies. The widely cited 2001-4 U.S. National Health and Nutrition Survey (NHANES) showed 29% of 14–49 year old women had BV, with higher prevalence among African-American (51.6%) compared to Hispanic (32.1%) and Caucasian (23.2%) women [1]. While a study of asymptomatic women attending general practices in the United Kingdom, and a study of Australian university students, reported notably lower prevalence estimates of 9% and 5% respectively [2], [3]. Some of the highest community-based estimates have been reported from rural sub-Saharan Africa, with a systematic review of women attending antenatal facilities finding pooled BV prevalence estimates of 51% in East/Southern Africa and 38% in West/Central Africa [4].
BV incidence studies have generally been conducted in high prevalence populations, with a high proportion of participants from STI clinics or from disadvantaged backgrounds. These studies are likely to provide higher BV incidence estimates compared with community-based samples and have tended to report rates in excess of 20/100 woman-years [5], [6], [7], [8], [9], [10], [11], [12]. In contrast, a prospective study of 17–21 year old Australian university students reported a low incidence rate of 2.2/100 woman-years in sexually-active participants, and no incident BV in sexually-inactive women [13].
While the aetiology of BV and whether it is sexually transmitted remains unclear, epidemiological studies show a consistent association between BV and sexual activity. Some of the strongest evidence comes from a meta-analysis of 43 studies which found prevalent and incident BV were associated with new or multiple male partnerships, report of female partners and inconsistent condom use [14]. Observational studies have, however, also reported associations between increased risk of BV and non-sexual behavioural practices such as smoking [15] and vaginal douching/cleaning practices [16], and a number of cross-sectional and longitudinal studies have shown a reduced risk of BV in women using hormonal contraceptives [6], [17], [18], [19], [20], [21]. Current antibiotic regimens for BV have limited effectiveness with up to 60% of women experiencing recurrence within 12 months [17]. Improving our understanding of how behavioural factors are associated with the development and recurrence of BV is integral to developing more effective approaches for the prevention and treatment of BV.
We present BV prevalence and incidence estimates, derived using the Nugent method, and examine the associated risk factors for the development of BV and abnormal flora (AF), in a large cohort study of young sexually-active women predominantly recruited from primary-care clinics. These are the first community-based estimates for the Australian population.
Methods
Recruitment, Clinical and Laboratory Methods
Women aged 16 to 25 years attending 29 primary-care services (general practice/family planning/sexual health services) in three Australian states, for any reason, were invited to participate in a cohort study to determine C.trachomatis incidence and re-infection rates (detailed methods described elsewhere) [22]. Participants were also invited to enroll in a sub-study to determine the prevalence and incidence of BV and Mycoplasma genitalium. Women were eligible if they were sexually-active, aged 16–25 years, not pregnant and contactable by post. Research assistants approached consecutive eligible women at clinic sites and obtained written informed-consent. Participants completed 3-monthly questionnaires on demographic, sexual and behavioural data, recent antibiotic and contraceptive use, and genital symptoms. They self-collected a vaginal swab and smeared the sample onto a glass slide in clinic at baseline, and then at home at 6 and 12 months. Samples and questionnaires were returned by post.
BV was defined as a Nugent score (NS) of 7–10, intermediate flora as a NS = 4–6, and normal flora as a NS = 0–3 [23]. Abnormal flora (AF) was defined as a NS = 4–10. All slides were scored by an experienced microbiologist. An independent senior microbiologist reread all NS 6 and 7 slides, had 50 problematic slides referred to her for a second opinion, and additionally randomly audited 10% of slides in each Nugent category. Concordance in the random audit was 92%; 9/112 slides were found to have a minor discrepancy defined as a difference of ≥2 in Nugent score that did not result in misclassification of BV, or normal versus intermediate flora. Use of self-collected samples for BV diagnosis has been shown to be comparable to clinician-collected samples [24]. Women with symptomatic BV were treated with metronidazole 400 mg orally twice daily or 2%-clindamycin cream vaginally for 7 nights.
Participants were tested for C.trachomatis and M.genitalium at baseline and during follow-up by PCR using the Cobas-TaqMan CT-assay (Roche Applied Science) and a PCR-assay targeting the 517 bp region of the 16S rRNA gene of M.genitalium, respectively [25]. Detailed laboratory methodology and results have been described elsewhere [22], [26].
Statistical Methods
Data were analysed using STATA 12.0 (StataCorp LP,College Station,USA). Assuming a design effect of 2 to take account of within-clinic correlation, a sample size of 1000 women was sufficient to generate 95% confidence intervals (CIs) of 7.2–12.8/100 person-years if the estimated incidence rate was 10/100 person-years.
BV and AF prevalence estimates and 95% CIs were calculated adjusting for within-clinic correlation. Adjusted odds ratios (AOR) and robust standard errors were calculated to explore epidemiological associations of variables with the primary outcome, prevalent BV (NS 7–10 versus NS 0–6), and secondary outcome, AF (NS 0–3 versus NS 4–10).
Women with a NS<7 at baseline were included in the cohort analysis. Incident BV was defined as the first occurrence of a NS = 7–10 in a participant with a previous NS<7; this led to a participant being censored from the incidence analysis. The association between risk factors and incident BV was investigated using a discrete-time version of the proportional-hazards regression model [27]. Variables were included in the model on the basis of the likelihood ratio test, or if they had been associated with BV in previous studies. Follow-up concluded 18 months after enrolment. The same statistical methods were applied to examination associations with incident AF.
Written informed consent was obtained from all participants. All eligible women were assessed for competency by research staff prior to being invited into the study. The research staff worked closely with the clinical staff to ensure only competent women were approached. All ethics committees approved the inclusion of participants over the age of 16 without parental or guardian consent. Ethics approval to conduct this study was obtained from ten Human Research Ethics Committees throughout Australia including: The Australian Capital Territory (ACT) Health Human Research Ethics Committee, ACT; The Alfred Health Ethics Committee, Melbourne; The University of Ballarat Ethics Committee, Ballarat, Victoria; Family Planning Victoria Human Research Ethics Committee, Victoria; The North Coast Area Health Service, Human Research Ethics Committee, New South Wales (NSW); The South Eastern Sydney Area Health Service, Human Research Ethics Committee - Northern Area, NSW; The University of Melbourne, Human Research Ethics Committee, Victoria; The University of Newcastle Human Research Ethics Committee, NSW; The University of NSW Human Research Ethics Committee, NSW; Family Planning NSW Research Ethics Committee, NSW [22], [26].
Results
Demographic and Behavioural Characteristics of the Study Population
Overall, 1116 women aged 16 to 25 years were recruited from 29 clinic-sites across Australia, with an overall response rate of 66% [22], [26]. A total of 1110 women (99.5%) agreed to participate in the BV sub-study. Of these 1093 (98.5%) provided a readable baseline slide and behavioural data, and were included in prevalence analyses. Nine hundred and sixty-four (88.2%) women with a baseline NS<7 continued in the cohort study, provided ≥1 slides and behavioural data, and contributed 871.20 person years of follow-up to incidence analyses.
Participants had a median age of 21 years, and were predominantly recruited from general practice (66%), Table 1. The majority (88%) were Australian-born, 24 (2%) were Aboriginal or Torres Strait Islander, 471 (44%) had completed tertiary studies, and 408 (38%) were employed. At baseline 638 (58%) women were using hormone-based contraception: 567 (89%) used the combined oral contraceptive pill (COCP), 5 (1%) an intra-vaginal ring, and 66 (10%) a progesterone implant or depot-injection. Seven women had an intra-uterine device; whether it was a levo-norgesterel device was unknown. In the prior year, one third of participants had ≥3 male sexual partners (MSP) and 103 (9%) ≥1 female partners (FSP).
Baseline Clinical and Laboratory Characteristics of the Study Population
At baseline, 129 women had a NS of 7–10 [11.8% (95%CI: 9.4–14.2)], Table 1. Fifty-nine women had intermediate flora (5.4%; 4.0–6.8), and 905 had normal flora (82.8%; 80.1–85.5). The prevalence of AF was 17.2% (15.1–19.5). At recruitment, 246 (23%) women reported an abnormal vaginal discharge and 184 (17%) an abnormal odour. Twelve women had concurrent BV and C.trachomatis[1.1%; 0.5–1.7)], and 4 concurrent BV and M.genitalium[0.4%; 0.0–0.7)], Figure 1.
Factors Associated with Prevalent BV and AF
By univariate analysis, women with prevalent BV were more likely to be recruited from sexual health/family planning services (OR = 1.7; 1.1–2.5), to have no tertiary education (OR = 2.2; 1.4–3.5), to be born overseas (OR = 1.6; 1.0–2.7), and in the prior year to report a FSP (OR = 2.7; 1.6–4.7) and ≥5 new MSPs (OR = 2.2; 1.2–3.9), Table 1. Current use of an oestrogen-containing contraception (OCC, defined as COCP or ring) was associated with a lower prevalence of BV (OR = 0.6; 0.4–0.8), while use of any hormone-based method was not (OR = 0.8; 0.5–1.2). Co-detection of C.trachomatis or M.genitalium had a borderline association with BV (OR = 2.0; 0.9–4.4). Women with prevalent BV were more likely to report an abnormal vaginal discharge (OR = 1.6; 1.1–2.4) or odour (OR = 3.0; 2.2–4.1).
By multivariate logistic regression analysis, women with prevalent BV were more likely to have had a FSP in the prior year (AOR = 2.1; 1.0–4.4), and no tertiary education (AOR = 1.9; 1.2–3.0). OCC-use was associated with decreased odds of BV (AOR = 0.6; 0.4–0.9), and there was a borderline association with co-detection of C.trachomatis or M.genitalium [AOR = 2.0; 0.9–4.6), Table 1. Clinical symptoms were not included in multivariate analyses.
The findings for prevalent AF did not greatly differ from BV. Women with prevalent AF were more likely in the last year to have had a FSP [AOR = 1.9; 1.0–3.6)], and ≥5 new MSPs (AOR = 1.8; 1.2–2.5), to be co-infected with C.trachomatis or M.genitalium (AOR = 2.1; 1.0–4.5) and to have no tertiary education (AOR = 1.6; 1.2–2.3); OCC-use decreased the odds of AF(AOR = 0.7; 0.5–1.0).
Incidence and Risk Factors Associated with BV and AF
There were 82 cases of incident BV [9.4 (95% CI: 7.7–11.7)/100 person-years], and incidence was stable during follow-up at 9.3 (95% CI: 5.8–14.9)/100 person years over the first 6 months and 9.4 (95% CI:7.4–12.0)/100 person years over the last 6 months.
By univariate analysis, incident BV was associated with being employed (RR = 1.4; 1.0–2.0), and having a new MSP during follow-up (RR = 1.6; 1.2–2.3), Table 2. OCC-use in the 3-months prior to testing was associated with lower rates of incident BV (RR = 0.7; 0.5–1.0); use of any hormonal contraceptive was not (RR = 0.8; 0.5–1.2). Women with incident BV were more likely to report genital symptoms in the interval prior to testing including an abnormal vaginal discharge (RR = 1.9; 1.3–2.7), odour (RR = 2.9; 2.0–4.1), or abdominal pain (RR = 1.5; 1.1–2.1). By multivariate analysis, having a new MSP during follow-up was associated with incident BV (ARR = 1.5; 1.1–2.2); there was a borderline reduction in the risk of incident BV with OCC-use (ARR = 0.7; 0.5–1.1), Table 2.
There were 129 cases of incident AF during the study [14.8 (95% CI: 12.5–17.6)/100 person-years]. By multivariate analysis, having a new MSP during the study was associated with incident AF (ARR = 1.5; 1.1–2.0) and OCC-use was associated with a reduced risk of incident AF (ARR = 0.7; 0.5–1.0).
Discussion
This study presents findings on the prevalence and incidence of BV and AF among a large community-based cohort of Australian women. In this young cohort we found that exposure to a new male partner was associated with increased risk of incident BV and AF, while use of an OCC was associated with a reduced risk. The young age and low BV prevalence in participants, together with the stable BV incidence throughout the study, all facilitated the study of the association between recent sexual and contraceptive behaviours and the development of incident BV. These are the first data to give an estimation of the prevalence (12%) and incidence (9.4/100 person years) of BV in young Australian women in the community. Our findings indicate that sexual exposure to new partners is influential in the development of BV and AF, and that OCC appears to be associated with a reduced risk.
The prevalence and incidence of BV in this large cohort of young Australian women is significantly lower than that reported from the NHANES data [1], and the majority of prospective studies in heterosexual women [5], [6], [7], [8], [9], [10], [11], [12]. This is likely to be due to the young age of our participants and the fact that the majority were recruited from primary-care compared with those in published studies. Our prevalence data are more in keeping with a UK study of women attending general practices for cervical cytology (9%) [2], but notably higher than that found in 17–21 year old Australian university students (5%) [3]. Our data supports the previously reported association between BV and lower levels of education, which is likely to be more broadly reflective of socio-economic and health inequity, and greater levels of sexual risk behaviour [1], [28].
Incident BV and AF, in this young predominantly heterosexual cohort, were associated with report of a new male partner. A number of investigators have reported an association between the development of BV and exposure to a new male partner [5], [7], [9]. In a study of women recruited from an STI service, sex with a new partner was the only behaviour associated with incident BV [7]. In a cohort of female sex workers in West Africa, Nagot reported having ≥3 male sex partners in the prior week was associated with incident BV [9]. Using cohort and case cross-over analytical methods to reduce unmeasured confounding, Gallo found the only consistent association with incident BV in high-risk women was detection of spermatozoa on Gram stain [29]. In our cohort increased numbers of male partners in the year prior to enrolment was also associated with increased odds of prevalent AF but not BV. It is interesting to note the association reported in the meta-analysis between new or multiple male partnerships and prevalent and incident BV was of similar magnitude to that found in the current cohort (RR = 1.6; 95% CI: 1-5-1.8) [14].
It was not only exposure to male partners that conferred an increased risk of BV in this community cohort, but report of female partners in the prior year doubled the odds of prevalent BV and AF. The association between increased BV risk and female partnerships is well known but poorly understood. A higher prevalence of BV is consistently reported in lesbians compared to heterosexual women within the same communities, and high levels of concordance of vaginal flora has been reported between monogamous female partners [30], [31], [32]. Again the magnitude of the effect observed in our cohort was strikingly similar to that found by meta-analysis which showed report of any female partner(s) doubled the odds of BV (RR = 2.0; 95% CI 1.7–2.3) [14]. While these data are compelling, the fundamental question as to whether sexual activity with male or female partners contributes to the development of BV through transmission of BV associated-bacteria, or instead is impacting adversely on vaginal colonization with protective Lactobacillus species, remains unanswered. The association between BV and increased detection of STIs, which has been reported in a number of studies [5], [33], [34] provides further support for the strong relationship between BV and sexual activity. However, whether BV increases susceptibility to STIs, or BV and STIs are co-infections that result from shared risk factors is unknown.
Use of an OCC in this cohort was associated with a reduced risk of prevalent BV and AF, and incident AF, after adjusting for confounding factors including numbers of sexual partners. This predominantly reflected COCP use in this cohort, and when use of any hormonal contraceptive, which included progesterone-only methods, was examined there was no significant association. This association has been evident in a number of published studies. Hormonal contraceptives, mainly combined methods, have been reported to be protective against prevalent [18], [19], [20], incident [6], [35] and recurrent [17], [19], [21] BV. While there are a number of confounding factors that may explain this association it is consistently evident in the literature, and many analyses including our own, have adjusted for known confounders such as numbers of sexual partners and educational level. There are also biologically plausible explanations for this apparent association. It has been hypothesized that oestrogen increases the glycogen-content of epithelial cells, a substrate for Lactobacillus species to generate lactic acid, which appears to be a potent inhibitor of BV-associated bacteria [36]. It is possible that contraceptive use influences the vaginal immune response, with Cherpes reporting an association between hormonal contraceptive use and altered vaginal immunity in BV [37]. Clearly further studies are need to determine if and how OCC may provide protection against BV or whether the observed association is due to unmeasured confounding.
There were a number of limitations to our study. Participants were more likely to be Australian-born, better educated, and more sexually-active than the background Australian population of this age [22], [38]. As a consequence, our study may not be representative of all Australian 16–25 year old women. Two thirds of recruits were derived from general practice and the remainder from sexual health/family planning services. BV prevalence was slightly higher in recruits from family planning/sexual health services compared to general practices, however, the BV incidence did not differ between these sites, and care was taken to adjust all analyses for clinic type. The inclusion of sexual health/family planning services in this cohort may have some impact on the generalizability of prevalence and incidence estimates, however, it is less likely to effect the measures of association (relative risks). Overall, 80–90% of young Australian women visit a general practice clinic each year [39], so this study population should provide a reasonable sample of young women in the community in this age group. Another important limitation is that the precision of our estimates of BV incidence reflect the availability of event data, which was constrained by six-monthly sample collection. Studies employing Gram stain analysis and more sensitive molecular methods, such as quantitative PCR and pyrosequencing, have shown that the vaginal microbiota can be highly dynamic. Dramatic changes in lactobacillus and BV-associated bacterial species, and rapid fluctuation in Nugent scores can occur in some individuals over a menstrual cycle, although longitudinal studies have also shown that vaginal bacterial communities can be more stable in some women than others [40], [41], [42], [43], [44], [45]. These studies indicate that factors such as menstruation and sexual behaviours may influence the stability of the vaginal microbiota in women, however the clinical significance of these fluctuations is not currently known. Importantly, the broad sampling interval in this study means cases of BV could have been missed, which may have lead to an under-estimate of the incidence rate. Of note, there was a significant association between self-report of abnormal vaginal discharge and/or odour and diagnosis of incident BV by the Nugent method, indicating that cases of incident infection that were detected were more likely to represent clinically significant BV.
Overall the strengths of this study include the diverse range of geographical location and socio-economic status from which participants were recruited, the high recruitment and retention rates, and inclusion of a high proportion of under 21 year old women (42%), a group that is difficult to engage in research. A further strength is that the young age, the low BV prevalence, and the stable incidence of BV throughout the study all facilitated the study of the relationship between recent behaviours and incident BV.
Conclusion
These are the first data to give an estimation of the prevalence and incidence of BV in young Australian women in the community. The consistent manner in which sexual risk factors are identified in association with BV, in populations that differ widely in their geographical and socioeconomic circumstances, lends considerable weight to the concept that sexual activity with both men and women is highly influential if not integral to the development of BV. However, it appears likely that sexual and contraceptive practices influence not just the development of BV and AF but also recurrence. A recent treatment trial showed no benefit from combining oral metronidazole with vaginal clindamycin over oral metronidazole alone. BV recurrence in this cohort was instead strongly associated with remaining with the same pre-treatment sexual partner and inconsistent condom use, while use of oestrogen-containing contraceptives was associated with a reduced risk of recurrence [17], [19], [21]. Debate will continue as to whether the consistent association between BV and sexual activity is a direct result of transmission of putative agent(s) that have an aetiological role in the development and recurrence of BV, or whether sexual activity itself impacts adversely on vaginal colonization with protective Lactobacillus species thereby facilitating BV development and recurrence. We are only just beginning to understand the diversity of the vaginal microbiota in healthy women [41], and have yet to fully understand the effects of sexual behaviours on it, and if and how use of hormone-based contraceptives may influence a woman’s vaginal microbiota and susceptibility to BV. Clearly, identifying potentially modifiable behaviours that are associated with the development and recurrence of BV within large cohort studies is an integral step to the development and evaluation of targeted interventions to improve prevention and treatment approaches for BV.
Acknowledgments
The authors would like to acknowledge the staff at all the clinics and all the women who participated in the study. All BV microscopy was done by Technical Officer Amanda Liebelt, Medical Scientist Ena Ribic & Emeritus Microbiologist Helen McDonald at the Microbiology and Infectious Diseases Department, SA Pathology at Women’s and Children’s Hospital, Adelaide.
Author Contributions
Conceived and designed the experiments: CB JW JH CF M. Chen BD KM ST SG MP LG FB JK HB. Performed the experiments: CB JW JH CF M. Chen BD KM ST SG MP LG FB JK HB SW EU M. Currie. Analyzed the data: CB JH JW CF. Contributed reagents/materials/analysis tools: CB JW JH CF M. Chen BD KM ST SG MP LG FB JK HB SW EU M. Currie. Wrote the paper: CB JW JH CF M. Chen BD KM ST SG MP LG FB JK HB SW EU M. Currie.
References
- 1. Allsworth JE, Peipert JF (2007) Prevalence of bacterial vaginosis: 2001–2004 national health and nutrition examination survey data. Obstet Gynecol 109: 114–120.
- 2. Lamont RF, Morgan DJ, Wilden SD, Taylor-Robinson D (2000) Prevalence of bacterial vaginosis in women attending one of three general practices for routine cervical cytology. Int J STD AIDS 11: 495–498.
- 3. Fethers KA, Fairley CK, Morton A, Hocking JS, Hopkins C, et al. (2009) Early sexual experiences and risk factors for bacterial vaginosis. J Infect Dis 200: 1662–1670.
- 4. Chico RM, Mayaud P, Ariti C, Mabey D, Ronsmans C, et al. (2012) Prevalence of malaria and sexually transmitted and reproductive tract infections in pregnancy in sub-Saharan Africa: a systematic review. Jama 307: 2079–2086.
- 5. Gallo MF, Macaluso M, Warner L, Fleenor ME, Hook EW 3rd, et al (2012) Bacterial vaginosis, gonorrhea, and chlamydial infection among women attending a sexually transmitted disease clinic: a longitudinal analysis of possible causal links. Ann Epidemiol 22: 213–220.
- 6. Avonts D, Sercu M, Heyerick P, Vandermeeren I, Meheus A, et al. (1990) Incidence of uncomplicated genital infections in women using oral contraception or an intrauterine device: a prospective study. Sex Transm Dis 17: 23–29.
- 7. Schwebke J, Desmond RA (2005) Risk Factors for bacterial vaginosis in women at high risk for sexually transmitted diseases. Sex Transm Dis 32: 654–658.
- 8. McClelland RS, Lavreys L, Katingima C, Overbaugh J, Chohan V, et al. (2005) Contribution of HIV-1 infection to acquisition of sexually transmitted disease: a 10-year prospective study. J Infect Dis 191: 333–338.
- 9. Nagot N, Ouedraogo A, Defer MC, Vallo R, Mayaud P, et al. (2007) Association between bacterial vaginosis and Herpes simplex virus type-2 infection: implications for HIV acquisition studies. Sex Transm Infect 83: 365–368.
- 10. Cherpes TL, Hillier SL, Meyn LA, Busch JL, Krohn MA (2008) A delicate balance: risk factors for acquisition of bacterial vaginosis include sexual activity, absence of hydrogen peroxide-producing lactobacilli, black race, and positive herpes simplex virus type 2 serology. Sex Transm Dis 35: 78–83.
- 11.
Ness RB, Kip KE, Soper DE, Stamm CA, Rice P, et al.. (2006) Variability of Bacterial Vaginosis Over 6- to 12-Month Intervals. Sex Transm Dis Publish Ahead of Print.
- 12. Hawes SE, Hillier SL, Benedetti J, Stevens CE, Koutsky LA, et al. (1996) Hydrogen peroxide-producing lactobacilli and acquisition of vaginal infections. J Infect Dis 174: 1058–1063.
- 13.
Fethers KA, Fairley CK, Morton A, Hocking JS, Fehler G, et al.. (2011) Low Incidence of Bacterial Vaginosis in Cohort of Young Australian Women. Sex Transm Dis.
- 14. Fethers KA, Fairley CK, Hocking JS, Gurrin LC, Bradshaw CS (2008) Sexual risk factors and bacterial vaginosis: a systematic review and meta-analysis. Clin Infect Dis 47: 1426–1435.
- 15. Hellberg D, Nilsson S, Mardh PA (2000) Bacterial vaginosis and smoking. Int J STD AIDS 11: 603–606.
- 16. Klebanoff MA, Nansel TR, Brotman RM, Zhang J, Yu KF, et al. (2010) Personal hygienic behaviors and bacterial vaginosis. Sex Transm Dis 37: 94–99.
- 17. Bradshaw CS, Morton AN, Hocking J, Garland SM, Morris MB, et al. (2006) High recurrence rates of bacterial vaginosis over the course of 12 months after oral metronidazole therapy and factors associated with recurrence. J Infect Dis 193: 1478–1489.
- 18. Calzolari E, Masciangelo R, Milite V, Verteramo R (2000) Bacterial vaginosis and contraceptive methods. Int J Gynaecol Obstet 70: 341–346.
- 19. Riggs M, Klebanoff M, Nansel T, Zhang J, Schwebke J, et al. (2007) Longitudinal association between hormonal contraceptives and bacterial vaginosis in women of reproductive age. Sex Transm Dis 34: 954–959.
- 20. Shoubnikova M, Hellberg D, Nilsson S, Mardh PA (1997) Contraceptive use in women with bacterial vaginosis. Contraception 55: 355–358.
- 21.
Bradshaw CS, Vodstrcil LA, Hocking JS, Law M, Pirotta M, et al.. (2013) Recurrence of bacterial vaginosis is significantly associated with post-treatment sexual activities and hormonal contraceptive use. Clin Infect Dis Jan 11. In press.
- 22. Walker J, Fairley CK, Urban E, Chen MY, Bradshaw C, et al. (2011) Maximising retention in a longitudinal study of genital Chlamydia trachomatis among young women in Australia. BMC Public Health 11: 156.
- 23. Nugent RP, Krohn MA, Hillier SL (1991) Reliability of diagnosing bacterial vaginosis is improved by a standardized method of gram stain interpretation. J Clin Microbiol 29: 297–301.
- 24. Nelson DB, Bellamy S, Gray TS, Nachamkin I (2003) Self-collected versus provider-collected vaginal swabs for the diagnosis of bacterial vaginosis: an assessment of validity and reliability. J Clin Epidemiol 56: 862–866.
- 25. Yoshida T, Deguchi T, Ito M, Maeda S, Tamaki M, et al. (2002) Quantitative detection of Mycoplasma genitalium from first-pass urine of men with urethritis and asymptomatic men by real-time PCR. J Clin Microbiol 40: 1451–1455.
- 26. Walker J, Tabrizi SN, Fairley CK, Chen MY, Bradshaw CS, et al. (2012) Chlamydia trachomatis incidence and re-infection among young women–behavioural and microbiological characteristics. PLoS One 7: e37778.
- 27. Carlin JB, Wolfe R, Coffey C, Patton GC (1999) Analysis of binary outcomes in longitudinal studies using weighted estimating equations and discrete-time survival methods: prevalence and incidence of smoking in an adolescent cohort. Stat Med 18: 2655–2679.
- 28. Allsworth JE (2010) Bacterial vaginosis–race and sexual transmission: issues of causation. Sex Transm Dis 37: 137–139.
- 29. Gallo MF, Warner L, King CC, Sobel JD, Klein RS, et al. (2011) Association between semen exposure and incident bacterial vaginosis. Infect Dis Obstet Gynecol 2011: 842652.
- 30. Marrazzo JM, Koutsky LA, Eschenbach DA, Agnew K, Stine K, et al. (2002) Characterization of vaginal flora and bacterial vaginosis in women who have sex with women. J Infect Dis 185: 1307–1313.
- 31. Bailey JV, Farquhar C, Owen C (2004) Bacterial vaginosis in lesbians and bisexual women. Sex Transm Dis 31: 691–694.
- 32. Berger BJ, Kolton S, Zenilman JM, Cummings MC, Feldman J, et al. (1995) Bacterial vaginosis in lesbians: a sexually transmitted disease. Clin Infect Dis 21: 1402–1405.
- 33. Wiesenfeld HC, Hillier SL, Krohn MA, Landers DV, Sweet RL (2003) Bacterial vaginosis is a strong predictor of Neisseria gonorrhoeae and Chlamydia trachomatis infection. Clin Infect Dis 36: 663–668.
- 34. Brotman RM, Klebanoff MA, Nansel TR, Yu KF, Andrews WW, et al. (2010) Bacterial vaginosis assessed by gram stain and diminished colonization resistance to incident gonococcal, chlamydial, and trichomonal genital infection. J Infect Dis 202: 1907–1915.
- 35. Baeten JM, Nyange PM, Richardson BA, Lavreys L, Chohan B, et al. (2001) Hormonal contraception and risk of sexually transmitted disease acquisition: results from a prospective study. Am J Obstet Gynecol 185: 380–385.
- 36. O’Hanlon DE, Moench TR, Cone RA (2011) In vaginal fluid, bacteria associated with bacterial vaginosis can be suppressed with lactic acid but not hydrogen peroxide. BMC Infect Dis 11: 200.
- 37. Cherpes TL, Marrazzo JM, Cosentino LA, Meyn LA, Murray PJ, et al. (2008) Hormonal contraceptive use modulates the local inflammatory response to bacterial vaginosis. Sex Transm Infect 84: 57–61.
- 38. Smith AM, Rissel CE, Richters J, Grulich AE, de Visser RO (2003) Sex in Australia: the rationale and methods of the Australian Study of Health and Relationships. Aust N Z J Public Health 27: 106–117.
- 39. Hocking JS, Walker J, Regan D, Chen MY, Fairley CK (2008) Chlamydia screening–Australia should strive to achieve what others have not. Med J Aust 188: 106–108.
- 40. Brotman RM, Ravel J, Cone RA, Zenilman JM (2010) Rapid fluctuation of the vaginal microbiota measured by Gram stain analysis. Sex Transm Infect 86: 297–302.
- 41. Gajer P, Brotman RM, Bai G, Sakamoto J, Schutte UM, et al. (2012) Temporal dynamics of the human vaginal microbiota. Sci Transl Med 4: 132ra152.
- 42. Schwebke JR, Morgan SC, Weiss HL (1997) The use of sequential self-obtained vaginal smears for detecting changes in the vaginal flora. Sex Transm Dis 24: 236–239.
- 43. Srinivasan S, Liu C, Mitchell CM, Fiedler TL, Thomas KK, et al. (2010) Temporal variability of human vaginal bacteria and relationship with bacterial vaginosis. PLoS One 5: e10197.
- 44. Thoma ME, Gray RH, Kiwanuka N, Aluma S, Wang MC, et al. (2011) The short-term variability of bacterial vaginosis diagnosed by Nugent Gram stain criteria among sexually active women in Rakai, Uganda. Sex Transm Dis 38: 111–116.
- 45. Thoma ME, Gray RH, Kiwanuka N, Wang MC, Sewankambo N, et al. (2011) The natural history of bacterial vaginosis diagnosed by gram stain among women in Rakai, Uganda. Sex Transm Dis 38: 1040–1045.