Quantitative bacteriology of the vaginal flora during the menstrual cycle

Wilks, Mark; Tabaqchali, Soad

doi:10.1099/00222615-24-3-241

Cited by 32 publications

(22 citation statements)

References 12 publications

(6 reference statements)

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“…Taken in sum, these data suggest the following: (i) there are distinct differences in the identities of the numerically dominant populations, thus suggesting that there is more than one kind of "normal" microbial community found in healthy women; (ii) the structures of the communities (at least as reflected by the most abundant populations) were comparatively invariant over time; and (iii) some communities (women 1 and 5) have phylotypes other than lactobacilli among the dominant populations. Conclusions i and ii are consistent with those of previous studies (3,19,30,53), whereas this represents the first instance where populations other than lactobacilli have been shown to be numerically dominant in the vaginal communities of healthy women. Additional studies are needed to verify this observation, since only a small number of women were included in this study.…”

Section: Optimization Of T-rflpsupporting

confidence: 81%

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Characterization of Microbial Communities Found in the Human Vagina by Analysis of Terminal Restriction Fragment Length Polymorphisms of 16S rRNA Genes

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Davis

et al. 2005

Appl Environ Microbiol

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To define and monitor the structure of microbial communities found in the human vagina, a cultivationindependent approach based on analyses of terminal restriction fragment length polymorphisms (T-RFLP) of 16S rRNA genes was developed and validated. Sixteen bacterial strains commonly found in the human vagina were used to construct model communities that were subsequently used to develop efficient means for the isolation of genomic DNA and an optimal strategy for T-RFLP analyses. The various genera in the model community could best be resolved by digesting amplicons made using bacterial primers 8f and 926r with HaeIII; fewer strains could be resolved using other primer-enzyme combinations, and no combination successfully distinguished certain species of the same genus. To demonstrate the utility of the approach, samples from five women that had been collected over a 2-month period were analyzed. Differences and similarities among the vaginal microbial communities of the women were readily apparent. The T-RFLP data suggest that the communities of three women were dominated by a single phylotype, most likely species of Lactobacillus. In contrast, the communities of two other women included numerically abundant populations that differed from Lactobacillus strains whose 16S rRNA genes had been previously determined. The T-RFLP profiles of samples from all the women were largely invariant over time, indicating that the kinds and abundances of the numerically dominant populations were relatively stable throughout two menstrual cycles. These findings show that T-RFLP of 16S rRNA genes can be used to compare vaginal microbial communities and gain information about the numerically dominant populations that are present.

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Section: Optimization Of T-rflpsupporting

confidence: 81%

“…The microflora of the lower genital tract in women has been studied in a variety of human populations by using different bacterial isolation and identification techniques (4,21,30,34,53). In healthy women, vaginal communities are typically dominated by various species of lactobacilli that number approximately 10 8 CFU per ml (or g) of vaginal secretion.…”

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Characterization of Microbial Communities Found in the Human Vagina by Analysis of Terminal Restriction Fragment Length Polymorphisms of 16S rRNA Genes

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Davis

et al. 2005

Appl Environ Microbiol

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“…Furthermore, vaginal flora are dynamic, and the level of lactobacilli measured in the present study may not reflect the level of lactobacilli present when the participant was infected with T. vaginalis; however, a study of vaginal flora has shown that the level of lactobacilli does not significantly change during the menstrual cycle [14].…”

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confidence: 76%

The Association betweenTrichomonas vaginalisInfection and Level of Vaginal Lactobacilli, in Nonpregnant Women

et al. 2007

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The effect that vaginal lactobacilli have on trichomoniasis is not known. At 3 clinics for treatment of sexually transmitted disease, we recruited 521 female patients with trichomoniasis and 176 control subjects. All participants underwent physical examinations and testing for sexually transmitted infection and completed questionnaires. little clinical and research attention, despite its public health importance.Lactobacillus species are the most prevalent organisms in normal human vaginal flora and may protect the vagina against infection [4]. The addition of lactobacilli to in vitro T. vaginalis culture slows growth of the protozoa, and T. vaginalis phagocytoses lactobacilli [5]. However, epidemiologic evidence is inconsistent [6][7][8][9], and the association between T. vaginalis and lactobacilli is not understood.We conducted a case-control study to assess the association between level of vaginal lactobacilli and T. vaginalis among nonpregnant women. Although this topic has been examined previously, this is the first study of nonpregnant women to have an adequate sample size, use the diagnostic reference standard, and comprehensively evaluate the effect that covariables have on the association between the level of vaginal lactobacilli and T. vaginalis. Clarifying this relationship could improve our understanding of T. vaginalis and may have implications for the treatment of trichomoniasis.Participants, materials, and methods. We conducted a prospective, case-control study in 3 health-department sexually transmitted disease clinics, located in Durham and Raleigh in North Carolina and Birmingham in Alabama, from November 2001 to July 2003. For every 3 women with positive wet mountmicroscopy results for T. vaginalis whom we enrolled, we recruited 1 woman with negative wet mount-microscopy results; this case:control ratio was selected as part of a larger study that focused on T. vaginalis-infection concordance between infected women and their male partners [10]. Female patients who were у18 years old, spoke English, and had not previously been prescreened or enrolled in the study were eligible for prescreening. Women were prescreened on the basis of wet mountmicroscopy results and T. vaginalis cultures. Prescreened patients with positive wet mount-microscopy results for T. vaginalis were recruited immediately and completed questionnaires at enrollment; prescreened patients with negative wet mount-microscopy results for T. vaginalis but with positive culture results for it were contacted and asked to return for treatment and/or counseling; during the follow-up visit, they were recruited for the study and completed questionnaires. The unmatched control group was selected from prescreened women whom wet-mount microscopy had found to be uninfected with T. vaginalis, but control subjects whom T. vaginalis cultures subsequently identified as having trichomoniasis were reclassified as case patients.Prescreened women who had a history of vaginal sex during Downloaded from https://academic.oup.

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“…LB may also inhibit opportunist bacteria through the production of bacteriocins, immune stimulation and via nutrient and stearic competition. There is also substantial interest in the role of hydrogen peroxide (H The normal vagina has a pH of 3.5 -6.0 [4] and is anaerobic, with pO 2 and pCO 2 lower and higher, respectively, than atmospheric levels; menstruation increases pH [9,12] and oxygen tension [8]. Studies of LB characteristics that may protect the vagina should address these conditions.…”

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confidence: 99%

Growth and Acidification by Vaginal Lactobacilli in Anaerobic Liquid Medium Over the pH Range 5.5 – 8.0

Wade¹,

Graver²

2010

J Bacteriol Parasitol

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The flora of the healthy vagina is simple and usually dominated by one or a few species of lactobacilli (LB) [7,10]. LB maintains the acidity of the vagina and are critical for vaginal health. Bacterial vaginosis (BV) is the most prevalent vaginal syndrome with major public health implications worldwide [6] and is characterized by a depletion of vaginal LB. Recurrence of BV is seen in approximately 30% of affected females and LB-containing probiotic preparations have been evaluated for treatment and prevention. A means of quantifying the growth and acidification potential of LB over a range of pH values would be useful for intra-and inter-species comparisons of LB considered as potential probiotic candidates. To address this, we studied three vaginal LB under anaerobic conditions in liquid medium over a range of pHs.A defined liquid medium rendered anaerobic with 'Oxyrase for Broth' (Oxyrase Inc, Ohio, USA) was prepared and used as described previously [11] at initial pHs 5.5, 6.0, 6.5, 7.0, 7.5 and 8.0. In brief, media with and without buffer -and for each of these, with and without 1.25g/L human hemoglobin (Hb) -were adjusted to the required initial pH using 1M HCl before filter sterilizing through a 0.22μm filter. The buffers used were 30mM MES for pH 5.5 -6.5 and 30mM MOPS for pH 7.0 -8.0. Media were pre-reduced in 7mL screwtop plastic bijoux (Bibby Sterilin) by incubation at 37 o C for at least 2h before inoculation [11]. Suspensions of Lactobacillus crispatus NCTC 4505 (LC), Lactobacillus gasseri ATCC 9857 (LG), Lactobacillus jensenii ATCC 25258 (LJ) and Fusobacterium nucleatum ATCC 25586 (FN) in phosphate-buffered saline (PBS) were prepared such that 100 -200L added to 7mL of medium gave 10 4 cfu/mL. Controls for each medium at each initial pH included: uninoculated media to measure the pH effect of Oxyrase; rezasurin sodium at 0.002g/L as a reduction indicator; and FN to demonstrate ability to support anaerobic growth. Also, each strain was inoculated in to MES-and MOPS-containing media adjusted to pH 6.6 -which is at the overlap of the buffers' ranges -to identify any potential inhibition by the buffers. To provide growth curves for each strain in unbuffered medium, each was inoculated in to medium without buffer adjusted to pH 6.6. Bijoux were incubated statically at 37 o C and counts done after vortexing for 5s on an undiluted aliquot and serial 10 -1 dilutions in PBS on blood agar (Oxoid, UK) using a spiral plater (Don Whitley, Shipley, UK) at inoculation and at 25h, at which time pH was measured. For growth curves, additional samples were taken at 15h and 20h. Plates were incubated in 5% CO 2 for 24 -48h for LB and in anaerobic jars for 48h for FN. All chemicals were from Sigma, Poole, UK and all experiments done at least in triplicate. The log 10 cfu/mL change in colony counts between time 0 and 25h were calculated for all medium-bacterium combinations and the pH change over 25h calculated using the uninoculated Oxyrase-containing control as baseline. For each strain the Wilcoxon Rank-sum test wa...

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Quantitative bacteriology of the vaginal flora during the menstrual cycle

Cited by 32 publications

References 12 publications

Characterization of Microbial Communities Found in the Human Vagina by Analysis of Terminal Restriction Fragment Length Polymorphisms of 16S rRNA Genes

Characterization of Microbial Communities Found in the Human Vagina by Analysis of Terminal Restriction Fragment Length Polymorphisms of 16S rRNA Genes

The Association betweenTrichomonas vaginalisInfection and Level of Vaginal Lactobacilli, in Nonpregnant Women

Growth and Acidification by Vaginal Lactobacilli in Anaerobic Liquid Medium Over the pH Range 5.5 – 8.0

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