Lactobacilli In Women Research Articles

Microbial function and genital inflammation in young South African women at high risk of HIV infection

BackgroundFemale genital tract (FGT) inflammation is an important risk factor for HIV acquisition. The FGT microbiome is closely associated with inflammatory profile; however, the relative importance of microbial activities has not been established. Since proteins are key elements representing actual microbial functions, this study utilized metaproteomics to evaluate the relationship between FGT microbial function and inflammation in 113 young and adolescent South African women at high risk of HIV infection. Women were grouped as having low, medium, or high FGT inflammation by K-means clustering according to pro-inflammatory cytokine concentrations.ResultsA total of 3186 microbial and human proteins were identified in lateral vaginal wall swabs using liquid chromatography-tandem mass spectrometry, while 94 microbial taxa were included in the taxonomic analysis. Both metaproteomics and 16S rRNA gene sequencing analyses showed increased non-optimal bacteria and decreased lactobacilli in women with FGT inflammatory profiles. However, differences in the predicted relative abundance of most bacteria were observed between 16S rRNA gene sequencing and metaproteomics analyses. Bacterial protein functional annotations (gene ontology) predicted inflammatory cytokine profiles more accurately than bacterial relative abundance determined by 16S rRNA gene sequence analysis, as well as functional predictions based on 16S rRNA gene sequence data (p < 0.0001). The majority of microbial biological processes were underrepresented in women with high inflammation compared to those with low inflammation, including a Lactobacillus-associated signature of reduced cell wall organization and peptidoglycan biosynthesis. This signature remained associated with high FGT inflammation in a subset of 74 women 9 weeks later, was upheld after adjusting for Lactobacillus relative abundance, and was associated with in vitro inflammatory cytokine responses to Lactobacillus isolates from the same women. Reduced cell wall organization and peptidoglycan biosynthesis were also associated with high FGT inflammation in an independent sample of ten women.ConclusionsBoth the presence of specific microbial taxa in the FGT and their properties and activities are critical determinants of FGT inflammation. Our findings support those of previous studies suggesting that peptidoglycan is directly immunosuppressive, and identify a possible avenue for biotherapeutic development to reduce inflammation in the FGT. To facilitate further investigations of microbial activities, we have developed the FGT-DB application that is available at http://fgtdb.org/.DwvYvmP_Lqo1TfCUgVQTWuVideo

ОСОБЕННОСТИ МИКРОБИОЦЕНОЗА УРОГЕНИТАЛЬНОГО ТРАКТА ЖЕНЩИН С ВПЧ-АССОЦИИРОВАННЫМИ ПЛОСКОКЛЕТОЧНЫМИ ИНТРАЭПИТЕЛИАЛЬНЫМИ ПОРАЖЕНИЯМИ ШЕЙКИ МАТКИ

Objective: To reveal the features of the microbiocenosis of the cervical canal with the species identification of lactobacilli in women with HPV-associated squamous intraepithelial lesions of the cervix. Materials and methods: A survey of 80 patients of reproductive age with HPV-associated squamous intraepithelial lesions of the cervix (SIL), who had not received local and systemic antibacterial drugs at the time of the examination, was carried out. According to the results of morphohistological examination of cervical biopsies, this cohort of patients was divided into two groups. Group 1 included 40 patients with CIN II and CIN III, which corresponds to Нigh-grade squamous intraepithelial lesions (HSIL), group 2 — 40 patients with CIN I, which corresponds to Low-grade squamous intraepithelial lesions (LSIL). The comparison group (3rd) consisted of 30 clinically healthy women. All patients underwent a cytological study by the «liquid method» BD SurePath technology, genotyping of the human papillomavirus by the polymerase chain reaction method (6, 11, 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59), cultural studies of the microbiocenosis of the cervical canal (by the MALDI method — TOF mass spectrometry, in the VITEKMS analyzer), extended colposcopy. Conclusion. It was found that in all women with HPV-associated squamous intraepithelial lesions of the cervix, the microflora of the cervical canal is characterized by the presence of Gardnerella vaginalis, as well as a decrease in the detection rate and number of lactobacilli, with the presence of the facultative anaerobe Lactobacillus iners. In the structure of opportunistic microflora, the predominance of Enterococcus faecalis and Escherichia coli was demonstrated in HSIL, while for LSIL, representatives of the enterococcus group.

An exploratory pilot study evaluating the supplementation of standard antibiotic therapy with probiotic lactobacilli in south African women with bacterial vaginosis

BackgroundTo reduce acquisition and relapse of bacterial vaginosis (BV), lactobacilli must be maintained in the vaginal microbiome. Probiotic lactobacilli may aid this purpose. We investigated whether vaginal probiotics (containing Lactobacillus rhamnosus DSM 14870 and Lactobacillus gasseri DSM 14869) would result in vaginal colonisation with lactobacilli in women with and without BV.MethodsThis prospective, partially randomised, exploratory pilot study was conducted in Soweto, South Africa. Thirty-nine sexually-active, HIV negative women were enrolled from October 2014 to May 2016 into three arms. Women who did not have BV (Group 1, n = 13) self-administered probiotic capsules vaginally once daily for 30 days, then once a week until Day 190. Women diagnosed with BV were randomized into Group 2 (n = 12) or Group 3 (n = 14) and treated with the triple oral antibiotic combination for vaginal discharge syndrome per South African guidelines (cefixime 400 mg stat, doxycycline 100 mg BD for 7 days and metronidazole 2 g stat). Immediately after antibiotic treatment, women in Group 2 self-administered probiotic capsules vaginally once daily for 30 days then vaginally once a week until Day 190. Women in Group 3 were not given lactobacilli.ResultsDuring the study, L. rhamnosus DSM 14870 or L. gasseri DSM 14869, were isolated in 5/13 (38.5%) women in Group 1 compared to 10/12 (83.3%) women in Group 2 (p = 0.041). The 1-month and 6-month BV cure rates were similar (P > 0.05) between Group 2 (42 and 25%) compared to Group 3 (36 and 25%). In Group 2, no correlation was observed between the frequency of isolation of the two Lactobacillus strains and the 1-month or 6-month cure rate.ConclusionsSupplementation with vaginal probiotic capsules resulted in colonisation of the vagina by the Lactobacillus strains (L. rhamnosus DSM 14870 and L. gasseri DSM 14869) contained in the capsules. We observed low initial cure rates of BV after a stat dose of metronidazole and that the probiotic did not improve BV cure rates or alleviate recurrence which could be due to treatment failure or very limited power of the study.Trial registrationRegistered at the Pan African Clinical Trial Registry (www.pactr.org) on April 13, 2018 (retrospectively registered). Trial identification number: PACTR201804003327269.

542: Influence of vaginal microbiota on the pathophysiology of intra-amniotic infection and sterile-inflammation in preterm labor

Intra-amniotic infection and intra-amniotic inflammation (IAI) are leading responsible of spontaneous preterm delivery at early gestational ages being the ascending pathway the most common dissemination way. The influence of intra-amniotic infection and IAI on the vaginal diversity and Lactobacillus spp. abundance is poorly unknown. Our aim was to evaluate, in women with preterm labor (PTL), whether vaginal microbiota differs among different infectious/inflammatory phenotypes. Women admitted with PTL <34.0 weeks with an amniocentesis to rule out infection or IAI were included. Vaginal fluid was collected within 24 h after amniocentesis. Intra-amniotic infection was diagnosed based on amniotic fluid (AF) cultures or a positive AF PCR of 16S rDNA. IAI was defined based on AF IL-6 levels (≥13.4 ng/mL). Sequencing of 16S rRNA gene was performed using MiSeq platform. Data processing was performed using QIIME. Statistical analyses were performed using R and SPSS program (v24.0). Sixty-nine women were included being 26% women with intra-amniotic infection and 10% women with sterile IAI (negative culture but high levels of IL6). Only one woman with intra-amniotic infection had low levels of IL-6. Most common microorganisms isolated in the amniotic fluid were Ureaplasma spp. 37%. No differences were observed regarding progesterone administration or cerclage among groups. We found differences on ethnicity so sequencing data was adjusted for this variable. Higher diversity (Shannon index), (p = 0.02) and lower Lactobacillus spp. abundance (p = 0.01) were observed in women with intra-amniotic infection compared with the non-infection/non-IAI group. Using LEfSE test, we observed at genera level that Ureaplasma was enriched in women with infection, Haemophilus in women with sterile IAI and Lactobacillus in women with non-infection/non-IAI group. Vaginal diversity (OR = 2.8, CI 95% (1.1 – 7.1)) and Lactobacillus spp. abundance (OR = 0.98, CI 95% (0.97 – 0.99)) were found to be independent factors of preterm delivery < 34 weeks being Ureaplasma spp. an independent risk factor for preterm delivery < 32 weeks (OR = 1.7, CI 95% (1.0 – 2.7)). A higher vaginal diversity and lower Lactobacillus spp. abundance was observed in women with intra-amniotic infection being Lactobacillus spp. the microorganism dominated in women without infection or IAI.View Large Image Figure ViewerDownload Hi-res image Download (PPT)

RESULTS OF DETERMINING THE SPECIES COMPOSITION OF VAGINAL LACTOBACILLI USING REAL-TIME PCR TESTING AND BACTERIOLOGICAL METHOD

Aim. To compare the species composition of lactobacilli in women of reproductive age using real time PCR on urogenital samples before and after culturing microorganisms in Blood-Serum-Y^ast extract-Agar (BSYA). Materials and methods. Using real time PCR, we have examined the species composition of vaginal lactobacilli in 25 healthy women of reproductive age. Samples of urogenital swabs (endocervical and vaginal swabs) were studied. Species identification has been carried out twice: in the native clinical material and in the samples received after using microorganism culture technique in BSYA. Results. After culturing vaginal microorganisms in BSYA, L. iners predominance was found only in 2 cases (8%). At the same time, when using native clinical material this species predominance was discovered in 10 samples (40%). Conclusion. When culturing lactobacilli in BSYA, the growth of L. iners is stunted compared to other species of vaginal lactobacilli. The use of cultural testing turned out to be ineffective in case of predominance of L. iners, the species associated with an increased risk of vaginal dysbiosis.

Current views on vaginal lactobacilli in women of reproductive age

The paper presents the current data on the species diversity of lactobacilli in women of reproductive age and their role in maintaining the normal vaginal microbiocenosis. Attention was paid to genotypic features of some lactobacillus species and the mechanisms providing antimicrobial protection of the vaginal biotope. The controversial issue of the ability of Lactobacillus iners to ensure stability of the normal vaginal microbiota.

Diversity of Lactobacillus in vagina of vulvovaginal candidiasis

Objective To investigate the Lactobacillus species in the vaginas of vulvovaginal candidiasis and to assess the prevalence of each Lactobacillus species in vulvovaginal candidiasis. Methods 154 vaginal samples were analyzed, 92 of which were from fertile healthy women, and 62 of which were from women with vulvovaginal candidiasis; and species-specific PCR showed the prevalence of each Lactobacillus species Species-specific PCR was used to investigate the prevalence of each Lactobacillus species in healthy Chinese women and the women with vulvovaginal candidiasis. Results In women with vulvovaginal candidiasis: L. iners(6.5%), L. cripatus(79.0%), L. gasseri(37.1%), L. jensenii(74.2%), L. acidophilus(16.1%), L. brevis(19.4%), L. plantarum(1.6%), L. johnsonii(51.6%), L. fermentum(8.1%), L. salivarius(9.7%), L. reuter(1.6%), L. paracasei(8.1%), L. delbrueckii(3.2%); More than two different Lactobacillus species coexisted in 98% of women with vulvovaginal candidiasis, and no anyone species existed in 2% of them; In fertile women: L. iners(82.6%), L. cripatus(70.7%), L. gasseri(67.4%), L. jensenii(40.2%), L. acidophilus(39.1%), L. brevis(23.9%), L. plantarum(5.4%), L. rhamnosus(1.1%), L. paracasei(1.1%), L. reuter(1.1%)i, L. johnsonii(3.3%), L. fermentum(2.2%), L. salivarius(2.2%); More than two different Lactobacillus species coexisted in 97% of fertile women, and only one species existed in 3% of fertile women. Conclusions Species of lactobacillus in women with vulvovaginal candidiasis did not significantly reduced compared with healthy women. Lactobacillus inert may be a marker of the change of vaginal microenvironment; Lactobacillus crispatus is a dominant lactobacillus in the vaginal of fertile healthy women, pregnant women and women with vulvovaginal candidiasis. Key words: Vaginal Lactobacillus species; 16S rRNA gene

Is there a role for probiotics in the prevention of preterm birth?

Preterm birth (PTB) continues to be a global health challenge. An over-production of inflammatory cytokines and chemokines, as well as an altered maternal vaginal microbiome has been implicated in the pathogenesis of inflammation/infection-associated PTB. Lactobacillus represents the dominant species in the vagina of most healthy pregnant women. The depletion of Lactobacillus in women with bacterial vaginosis (BV) has been associated with an increased risk of PTB. It remains unknown at what point an aberrant vaginal microbiome composition specifically induces the cascade leading to PTB. The ability of oral or vaginal lactobacilli probiotics to reduce BV occurrence and/or dampen inflammation is being considered as a means to prevent PTB. Certain anti-inflammatory properties of lactobacilli suggest potential mechanisms. To date, clinical studies have not been powered with sufficiently high rates of PTB, but overall, there is merit in examining this promising area of clinical science.

Characterization of vaginal lactobacilli in women after kidney transplantation

Limited number of publications described vaginal microflora after kidney transplantation. Our PubMed search revealed only 18 publications including words “vaginal bacteria & kidney transplant” in the period of 1978–2011. The aim of this study was to characterize lactobacilli isolated from vaginal swabs of women after kidney transplantation, compared with healthy women. Eighteen renal transplant recipients (mean age 36.1) and 20 healthy women (mean age 36.0) were evaluated. Lactobacilli were cultured on MRS and Columbia blood agars. Biochemical identification with API 50 CHL (bioMerieux, Marcy L’Etoile, France) and multiplex PCR according to Song et al. was performed. Lactobacilli were tested for production of H2O2. Minimal inhibitory concentrations (MICs) of selected antimicrobial agents were determined with E-tests (bioMerieux, Marcy L’Etoile, France) and interpreted with CLSI and EUCAST criteria. No bacterial vaginosis was found among studied women. Two strains of group I were identified as Lactobacillus delbrueckii; 18 strains as Lactobacillus gasseri and 15 strains as Lactobacillus crispatus. Only 3 strains from group II were not identified by species-specific mPCR. Group IV was represented with 2 unidentified strains. Vaginal lactobacilli isolated from healthy women represented more homogenous group compared with heterogenous renal transplant recipients. Biochemical identification of lactobacilli by API 50 CHL kits was concordant with mPCR results only in 7 cases (17.5%), all 7 strains were identified as L. crispatus. Majority (93%) of lactobacilli were H2O2 producers. All isolated lactobacilli (100%) demonstrated high resistance to metronidazole (MIC > 256 μg/ml). Only 2 strains resistant to vancomycin (MICs: 32 and 256 μg/ml respectively), in the study and control group, and one to moxifloxacin (MIC = 32 μg/ml), were found. Resistance to metronidazole and vancomycin was concordant in CLSI and EUCAST (2010) criteria. Although significant differences between lactobacilli isolated from vaginas of kidney transplant and healthy women were not demonstrated, we demonstrated strains resistant to metronidazole, vancomycin and moxifloxacin in groups of examined women. Our study was performed on a small group of kidney transplant recipients and further more detailed molecular studies on a larger group of patients are required to confirm our results.

Molecular methods to describe the spectrum and dynamics of the vaginal microbiota

The human vagina hosts a collection of microbes that is distinct from other human surfaces and mucosal sites, with reduced microbial diversity that is likely driven by the acidic environment. The microbial ecosystem of the vagina is dominated by lactobacilli in women without bacterial vaginosis (BV), and is characterize by increased species richness, diversity, and evenness in women with BV. The use of molecular, cultivation-independent methods to describe the bacterial biota of the human vagina has revealed many novel putative anaerobes in women with BV, and has demonstrated the almost ubiquitous nature of Lactobacillus iners which is found in most women regardless of BV status. A variety of molecular tools are being employed to study the vaginal microbiota, and each approach has distinct advantages and disadvantages that are reviewed. Longitudinal studies have demonstrated that the vaginal microbiota can be highly dynamic, with dramatic shifts in bacterial composition and concentrations in response to numerous endogenous and exogenous factors.

Low levels of faecal lactobacilli in women with iron-deficiency anaemia in south India

Fe deficiency in women contributes significantly to maternal and child morbidity in India. The intestinal bacterial flora may facilitate absorption of Fe from the caecum and proximal colon. The present study investigated the possibility that intestinal microbiota of anaemic young women may differ from that of women with normal Hb levels. The microbiota was quantified by real-time PCR in faeces of eight anaemic (Hb ≤ 100 g/l) and twenty-six normohaemic (Hb ≥ 120 g/l) women aged 18-25 years. Sequences of 16S ribosomal DNA (rDNA) specific to Bifidobacterium genus, Lactobacillus acidophilus group, Bacteroides-Prevotella-Porphyromonas group, Clostridium leptum group and Eubacterium rectale were amplified and expressed (as relative difference) relative to the universally conserved bacterial 16S rDNA sequences. Dietary intakes of energy, carbohydrate, fibre and Fe were ascertained by maintenance of a diet diary for a week. Faecal lactobacilli were significantly lower in anaemic women (median 6.6 × 10(-8), relative difference compared with total bacteria) than in the reference group (2.9 × 10(-6); P = 0.001, unpaired t test with logarithmic transformation). There was no difference between the two groups with respect to any of the other bacteria that were examined. Intakes of energy, carbohydrate, fibre, Fe and milk were similar in both the groups. Fe deficiency in young women in south India was associated with low levels of lactobacilli in the faeces. The relationship between lactobacilli and Fe deficiency needs to be explored further.

Metabolic properties of lactobacilli in women experiencing recurring episodes of bacterial vaginosis with vaginal pH ≥ 5

While 60% of women experiencing recurring episodes of bacterial vaginosis (BV) with vaginal pH >or= 5 are depleted of resident probiotic lactobacilli, the remainder carry one or more strains of lactobacilli. Their ability to make D-lactic acid is, however, low (3.94 +/- 0.72 mM/L) compared to the D-lactic acid produced by strains from healthy vagina with vaginal pH approximately 4 (8.04 +/- 1.07 mM/L) culture supernatant of 0.5 McFarland concentration (P < 0.001).

Hydrogen peroxide producing lactobacilli in women with vaginal infections

Objective Hydrogen peroxide producing lactobacilli are isolated from the vaginas of a majority of healthy reproductive age women. Their toxic and inhibitory effect against the overgrowth of pathogens in the vagina is documented by in vitro studies. Clinical studies concerning the role of hydrogen peroxide producing lactobacilli in vaginal infections are controversial. Study design The aim of this study was to isolate lactobacilli from women with major vaginal infections: bacterial vaginosis (BV), vulvovaginal candidiasis (VVC) and trichomoniasis (TV), and from women with normal flora (NF), to test their ability to produce hydrogen peroxide, to quantitate lactobacilli and to evaluate factors related to the colonization of vagina with hydrogen peroxide producing lactobacilli. The study group comprised 108 women: 27 with VVC, 26 with BV, 20 with TV and 35 with NF. Data was analysed with Fisher exact test, χ 2-test, and ANOVA. Results Lactobacilli were isolated from 81.5% of women with VVC, 58% of women with BV ( p = 0019), 55% of women with TV ( p = 0.015) and from 86% of women with NF. The majority of isolates of lactobacilli from women with VVC, TV and NF produced hydrogen peroxide (77%, 63.5% and 80%, respectively), compared with women with BV where only 26.5% of isolates produced hydrogen peroxide ( p = 0.01). Quantitative analysis showed that in an overwhelming majority of women colonized with lactobacilli, their count was >10 7 CFU/ml. Upon evaluation of behavioural and demographic characteristics, we found that smokers had a lower incidence of isolation of hydrogen peroxide producing lactobacilli ( p = 0.018). Our results showed that women with BV lack lactobacilli, especially hydrogen peroxide producing ones. Women with TV had a lower rate of lactobacilli. In women with VVC, there was neither significant difference in the isolation of lactobacilli, nor in their ability to produce hydrogen peroxide, compared to women with NF. Conclusion Our results point out that hydrogen peroxide producing lactobacilli could protect against the development of BV, but not against VVC and TV.

Hydrogen Peroxide Producing Lactobacilli in Women with Cervical Neoplasia

It is well known that human papillomavirus (HPV) is the main cause of cervical neoplasia, and hydrogen peroxide-producing lactobacilli are the most important microorganisms for maintaining the balance of the vaginal ecosystem. The purpose of our study was to investigate the relationship of hydrogen peroxide-producing lactobacilli, cervical neoplasia and high-risk HPV. We enrolled 1138 women with abnormal cervical smears or cervicograms who were referred to the department of Obstetrics and Gynecology at Chonnam National University Medical School. In all of them, 1,138 vaginal swabs were collected for the qualitative assay of hydrogen peroxide producing lactobacilli and 150 cervical swabs were used for the HPV hybrid capture II test without regard to the subjects' pregnancy status. In the non-pregnant women, 880 cervical biopsies and/or loop electrosurgical excision procedures were performed for making the histological diagnosis. There was no significant difference not only between the distribution of H(2)O(2) producing lactobacilli and the cervical histology, but also between the distribution of H(2)O(2) producing lactobacilli and the positivity for high-risk HPV. Both cervical neoplasia and high-risk HPV may not be influenced by the existence of hydrogen peroxide producing lactobacilli in the vagina.

Prevalence of bacterial vaginosis and vaginal flora changes in peri- and postmenopausal women.

Our aim was to evaluate the prevalence of bacterial vaginosis and decrease in lactobacillus colonization in women 40 years old or older in relation to menopausal status by evaluation of Gram-stained smears. A total of 1,486 smears from Italian Caucasian women aged 40 to 79 years were examined. Women were classified as follows: fertile (regular cycles) (n = 328), perimenopausal (irregular cycles) (n = 237), and postmenopausal (n = 921), including 331 women on estroprogestinic hormone replacement therapy (HRT). The prevalences of bacterial vaginosis (assessed as a Nugent score of >or=7) in fertile (9.8%) and perimenopausal (11.0%) women were not statistically different, whereas the prevalence was significantly lower overall in postmenopausal women (6.0%) (P = 0.02). Specifically, 6.3% of postmenopausal women without HRT and 5.4% of postmenopausal women with HRT were positive for bacterial vaginosis. The Nugent score system was not adequate for evaluating the normal and intermediate vaginal flora in women over the age of 40 years. High numbers of peri- and postmenopausal women had no lactobacilli and no bacterial-vaginosis-associated microorganisms. This nonpathological absence of lactobacilli in women with a Nugent score of 4 was scored as 4*, and this group was considered separately from the intermediate flora group. A score of 4* was obtained for 2.1% of fertile women, 11.4% of perimenopausal women, 44.1% of postmenopausal women without HRT, and 6.9% of postmenopausal women with HRT. The physiological reduction in lactobacillus colonization of the vagina in postmenopausal women does not cause an increase in bacterial-vaginosis prevalence. Reversion of lactobacillus flora to premenopausal levels due to HRT does not increase the prevalence of bacterial vaginosis in postmenopausal women.

Hydrogen peroxide-producing lactobacilli in women with and without vaginal discharge: evidence for a role in vaginitis prevalence

Hydrogen peroxide-producing lactobacilli in women with and without vaginal discharge: evidence for a role in vaginitis prevalence

Postoperative infection after legal abortion in relation to the different lactobacilli in women without bacterial vaginosis

Postoperative infection after legal abortion in relation to the different lactobacilli in women without bacterial vaginosis