Atopobium Vaginae Research Articles

Vaginal microbiome in early pregnancy and subsequent risk of spontaneous preterm birth: a case-control study.

To explore differences in the vaginal microbiome between preterm and term deliveries. Nested case-control study in 3D cohort (design, develop, discover). Quebec, Canada. Ninety-four women with spontaneous preterm birth as cases [17 early (<34 weeks) and 77 late (34-36 weeks) preterm birth] and 356 women as controls with term delivery (≥37 weeks). To assess the vaginal microbiome by sequencing the V4 region of the 16S ribosomal RNA (rRNA) gene in swabs self-collected during early pregnancy. Comparison of relative abundance of bacterial operational taxonomic units and oligotypes and identifying vaginal community state types (CSTs) in early or late spontaneous preterm and term deliveries. Lactobacillus gasseri/ Lactobacillus johnsonii (coefficient -5.36, 95% CI -8.07 to -2.65), Lactobacillus crispatus (99%)/ Lactobacillus acidophilus (99%) (-4.58, 95% CI -6.20 to -2.96), Lactobacillus iners (99%)/ Ralstonia solanacearum (99%) (-3.98, 95% CI -6.48 to -1.47) and Bifidobacterium longum/ Bifidobacterium breve (-8.84, 95% CI -12.96 to -4.73) were associated with decreased risk of early but not late preterm birth. Six vaginal CSTs were identified: four dominated by Lactobacillus; one with presence of bacterial vaginosis-associated bacteria (Gardnerella vaginalis, Atopobium vaginae and Veillonellaceae bacterium) (CST IV); and one with nondominance of Lactobacillus (CST VI). CST IV was associated with increased risk of early (4.22, 95% CI 1.24-24.85) but not late (1.63, 95% CI 0.68-5.04) preterm birth, compared with CST VI. Lactobacillus gasseri/L. johnsonii, L. crispatus/L. acidophilus, L. iners/R. solanacearum and B. longum/B. breve may be associated with decreased risk of early preterm birth. A bacterial vaginosis-related vaginal CST versus a CST nondominated by Lactobacillus may be associated with increased risk of early preterm birth. Largest study of its kind finds certain species of vaginal Lactobacillus + Bifidobacterium may relate to lower risk of preterm birth.

2408 Genital microbiomes of women with recurrent bacterial vaginosis and their regular male sexual partner

OBJECTIVES/SPECIFIC AIMS: Epidemiologic data suggest that BV is sexually transmitted with male partners colonized or infected with the responsible organism(s). The objective of this study was to compare the genital microbiota of women with recurrent BV and their regular male sexual partner using 16S rRNA gene sequencing and quantitative PCR targeting BV-candidate bacteria (Gardnerella vaginalis, Atopobium vaginae, BVAB1-3, Sneathia, Leptotrichia, and Megasphaera type I). METHODS/STUDY POPULATION: Women with recurrent BV (≥3 prior episodes, including a current episode) and their regular male partner participating in a BV treatment trial and providing genital specimens (women: vaginal; men: urethral, coronal sulcus, urine) at enrollment were included. Male specimens for each participant were pooled. 250 bp 16S rRNA V4 region PCR amplicons were sequenced and analyzed using the QIIME pipeline. Taxonomy was assigned using the RDP Classifier against a modified Greengenes database with additional vaginal taxonomies added. An average relative abundance cutoff of 0.5% was used for analysis. qPCR was also performed for specific BV-candidate bacteria. Spearman correlation coefficients were used to investigate associations between all genital bacteria in addition to BV-candidate bacteria between partnerships. To determine positive associations between partnerships, the Wilcoxon signed-rank test was used. RESULTS/ANTICIPATED RESULTS: In total, 45 partnerships were included. Mean partnership age was 31.3 (SD=7.9), 91.1% partnerships were African-American. The majority of partnerships (70.0%) reported condomless sex during the past 3 months. Regarding 16S data, 37 genital bacteria had an average relative abundance of ≥0.5%. The average Spearman correlation across all 45 partnerships was 0.28 (SD=0.27) (median=0.27, minimum=−0.21, maximum=0.84). Overall, a positive association of all genital bacteria existed across the partnerships (p&lt;0.0001). However, regarding specific BV-candidate bacteria, Spearman correlation tests for G. vaginalis, A. vaginae, Prevotella bivia, Megasphaera type I, BVAB1, and BVAB2 were nonsignificant. In contrast, Sneathia spp. were positively correlated between partnerships (r=0.37, p=0.01). With regards to qPCR results, RNA Cq analyses provided significant evidence for a linear association between male and females for only A. vaginae (r=0.52, p=0.006). DISCUSSION/SIGNIFICANCE OF IMPACT: In monogamous heterosexual couples in which the female has BV, the vaginal microbiota of women and the penile/urine microbiota of men were significantly correlated, particularly with regards to Sneathia spp. and A. vaginae, supporting the hypothesis that BV-associated bacteria are exchanged during sex.

Etiological and pathogenetic aspects of non-specific bacterial vaginosis

The aim of this review was the biological properties analysis of microorganisms known as the main contributors to the non-specific bacterial vaginosis development. The mechanisms of anaerobic vaginal microflora pathogenic action have been found out to be the main cause of non-specific inflammatory processes. Special attention was given to the genera of Mobilunkus spp., Atopobium spp., Prevotella sp. representatives , which were studied as vaginal flora due to the modern diagnostic methods and techniques development. The homeostasis of the vaginal ecosystem results from complex interactions and synergies among the host and different microorganisms that colonize the vaginal mucosa and healthy vaginal microbiota is generally dominated by Lactobacillus spp. Lactobacilli which have been known to be antagonistic toward a variety of microorganisms. In cases of bacterial vaginoses, the proportion of Lactobacilus spp sharply decreases, while the share of anaerobic microorganisms increases: Gardnerella vaginalis, Atopobium sp., Mobiluncus sp., Prevotella sp . The virulence factors of Gardnerella vaginalis pathogenicity are: its ability to form biofilms; the presence of genes responsible for cytotoxicity. The ability of Gardnerella vaginalis, Atopobium sp., Mobiluncus sp., Prevotella sp . to produce short chain volatile fatty acids isomers results in the vaginal mucosa irritation (itching or burning) and discharge with unpleasant odour in BV associated with these microorganisms. Atopobium vaginae , together with Mobiluncus spp . and Prevotella spp ., provoke the accumulation of pro-inflammatory cytokines. The whole mechanism leads to an inflammation worsening in BV and stimulates the chronic process development. Both microorganisms Mobiluncus spp . and A. vaginae are competitive lactobacilli in relation to the glycogen consumption. Conclusions. Analysis of literature shows that pathogens of non-specific bacterial vaginosis have wide range of pathogenicity factors. This provides pathogenesis and clinical signs of the disease.

Identification of Key Bacteria Involved in the Induction of Incident Bacterial Vaginosis: A Prospective Study.

The sequence of events preceding incident bacterial vaginosis (iBV) is unclear. African American women who have sex with women, who had no Amsel criteria and Nugent scores of 0-3, were followed for 90 days to detect iBV (defined as a Nugent score of 7-10 on at least 2-3 consecutive days), using self-collected vaginal swab specimens. For women with iBV (cases) and women maintaining normal vaginal flora (healthy women), 16S ribosomal RNA gene sequencing targeting V4 was performed. Longitudinal vaginal microbiome data were analyzed. Of 204 women screened, 42 enrolled; of these, 45% developed iBV. Sequencing was performed on 448 specimens from 14 cases and 8 healthy women. Among healthy women, Lactobacillus crispatus dominated the vaginal microbiota in 75%. In contrast, prior to iBV, the vaginal microbiota in 79% of cases was dominated by Lactobacillus iners and/or Lactobacillus jensenii/Lactobacillus gasseri. The mean relative abundance of Prevotella bivia, Gardnerella vaginalis, Atopobium vaginae, and Megasphaera type I became significantly higher in cases 4 days before (P. bivia), 3 days before (G. vaginalis), and on the day of (A. vaginae and Megasphaera type I) iBV onset. The mean relative abundance of Sneathia sanguinegens, Finegoldia magna, BV-associated bacteria 1-3, and L. iners was not significantly different between groups before onset of iBV. G. vaginalis, P. bivia, A. vaginae, and Megasphaera type I may play significant roles in iBV.

The direct and indirect association of cervical microbiota with the risk of cervical intraepithelial neoplasia.

Cervical microbiota composition is associated with cervical HPV infection and CIN severity. Previous studies only assessed the total association between cervical microbiota and HPV infections or CINs, and yet no study reported the direct and indirect associations between cervical microbiota and CINs mediated by HPV infection, respectively. The aim of this study was to investigate the direct and indirect associations between microbiotas and CIN severity. Cervical microbiota of 126 women with CIN 1− (normal cytology and CIN 1) and 40 with CIN 2+ (CIN 2 and CIN 3) were analyzed using Illumina sequencing based on the 16S rRNA gene. HPV was detected using a highly sensitive PCR primer set (SPF1/GP6+). Indirect effects of Pseudomonas stutzeri, Bacteroides fragilis, Lactobacillus delbrueckii, Atopobium vaginae, and Streptococcus agalactiae mediated by HPV infection on CIN status were observed. The directions of the direct and the indirect associations between CIN status and Ps. stutzeri were opposite. The directions of the direct and the indirect associations between CIN status and A. vaginae were the same. B. fragilis, L. delbrueckii, and S. agalactiae only had indirect association with CIN status. In summary, our study provided suggestive evidence that some microbial populations could have direct or indirect effects mediated by affecting HPV infection on CIN progression. Besides HPV infection, microbial community composition possibly plays a role in cervical carcinogenesis.

MODERN METHOD OF DETECTION OF TAXONOMIC COMPOSITION OF MICROBIALS IN URINARY WAYS OF WOMEN

In case of necessity to conduct a more qualitative examination of women with inflammatory processes, which are localized in the organs of the genitourinary system and in order to obtain an objective assessment of normal and pathogenic microflora in the urogenital pathways of a woman, a polymerase chain reaction method is proposed with the use of the diagnostic test of the system Femoflore-16. This system help identification of Lactobacillus spp., Enterobacterium spp., Streptococcus spp., Staphylococcus spp., Gardnerella vaginalis, Eubacterium spp., Fusobacterium spp., Veilonella spp., Clostridium spp., Mobiluncus spp., Peptostreptococcus spp., Atopobium vaginae, Mycoplasma hominis and genitalium, Ureaplasma (urealyticum and parvum), Candida spp. and perform complex analysis qualitative and quantitative composition of pathogenic and comditionaly pathogenis microorganismsare described. An original binary metal spoon probe is proposed for qualitative selection of the test material, tested in practice. This metal spoon probe is a cylindrical section of length 180 and a diameter of 2.0 mm made of stainless high quality chromium-nickel steel of the mark 12 KhNZA. The ends of the segment through the advancement of the formed permeable spoon-shaped terminals with a capacity of 0.025 and 0.05 ml. For the maximum reduction of the adhesive of the test material, the working surface of the probe of its surface has a roughness of 13-14 a-b classes according to the state standards 2.309.13. Sterile probe by way of (sterilization carried out by 96 ° ethanol or by heating) of local small pressure and scraping of the mucous membranes is easily carried out with a dosage capture of material in volume 0.025 or 0>0.5 ml. The volume of material taken with the probe is quite sufficient for the molecular-genetic method of investigation, in particular, the polymerase chain reaction. Clinical material is then placed in a prepared sterile test tube - eppendorf. After that, on the test tube, indicate the name of the patient, date and name of the analysis and send it to the laboratory.Described testimony to conduct a study and the requirements that a woman must observe before conducting a selection of material for PCR. The data on composition of microflora in different periods of a woman's life are given.

Bacterias relacionadas con vaginosis bacteriana y su asociación a la infección por virus del papiloma humano

Background and objectiveCervical cancer is an important health problem in our country. It is known that there are several risk factors for this neoplasm, and it has been suggested that cervical microbiome alterations could play a role in the development and progress of cancer. Bacterial vaginosis associated bacteria such as Atopobium vaginae and Gardnerella vaginalis has been suggested as potential risk factor for cervical lesions and cervical cancer. Material and methodsDNA from 177 cervical scraping samples was studied: 104 belonged to women without cytological or colposcopic alterations and 73 samples from precursor lesions with previous human papillomavirus (HPV) infection history. All samples were screened for Atopobium vaginae, Gardnerella vaginalis and HPV by PCR. ResultsHigh HPV prevalence was found in precursor samples, and 30% of samples without lesions were positive for HPV. Virtually all samples contained sequences of both bacteria, and interestingly, there was not HPV association observed; these results could suggest that these microorganisms could be part of the cervical microbiome in Mexican population. ConclusionsThe results obtained indicate that the bacteria analysed could be part of normal biome in Mexican women, suggesting a potential reconsideration of the pathogen role of these microorganisms.

Impact of contraceptive initiation on vaginal microbiota

BackgroundData evaluating the impact of contraceptives on the vaginal microbiome are limited and inconsistent.ObjectiveWe hypothesized that women initiating copper intrauterine device use would have increased bacterial vaginosis and bacterial vaginosis-associated microbes with use compared to women initiating and using hormonal contraceptive methods.Study DesignVaginal swabs (N = 1047 from 266 participants seeking contraception) for Nugent score determination of bacterial vaginosis and quantitative polymerase chain reaction analyses for assessment of specific microbiota were collected from asymptomatic, healthy women aged 18-35 years in Harare, Zimbabwe, who were confirmed to be free of nonstudy hormones by mass spectrometry at each visit. Contraception was initiated with an injectable (depot medroxyprogesterone acetate [n = 41], norethisterone enanthate [n = 44], or medroxyprogesterone acetate and ethinyl estradiol [n = 40]), implant (levonorgestrel [n = 45] or etonogestrel [n = 48]), or copper intrauterine device (n = 48) and repeat vaginal swabs were collected after 30, 90, and 180 days of continuous use. Self-reported condom use was similar across all arms at baseline. Quantitative polymerase chain reaction was used to detect Lactobacillus crispatus, L jensenii, L gasseri/johnsonii group, L vaginalis, L iners, Gardnerella vaginalis, Atopobium vaginae, and Megasphaera-like bacterium phylotype I from swabs. Modified Poisson regression and mixed effects linear models were used to compare marginal prevalence and mean difference in quantity (expressed as gene copies/swab) prior to and during contraceptive use.ResultsBacterial vaginosis prevalence increased in women initiating copper intrauterine devices from 27% at baseline, 35% at 30 days, 40% at 90 days, and 49% at 180 days (P = .005 compared to marginal prevalence at enrollment). Women initiating hormonal methods had no change in bacterial vaginosis prevalence over 180 days. The mean increase in Nugent score was 1.2 (95% confidence interval, 0.5–2.0; P = .001) in women using copper intrauterine devices. Although the frequency and density of beneficial lactobacilli did not change among intrauterine device users over 6 months, there was an increase in the log concentration of G vaginalis (4.7, 5.2, 5.8, 5.9; P = .046) and A vaginae (3.0, 3.8, 4.6, 5.1; P = .002) between baseline and 30, 90, and 180 days after initiation. Among other contraceptive groups, women using depot medroxyprogesterone acetate had decreased L iners (mean decrease log concentration = 0.8; 95% confidence interval, 0.3–1.5; P = .004) and there were no significant changes in beneficial Lactobacillus species over 180 days regardless of contraceptive method used.ConclusionCopper intrauterine device use may increase colonization by bacterial vaginosis–associated microbiota, resulting in increased prevalence of bacterial vaginosis. Use of most hormonal contraception does not alter vaginal microbiota.

Bacterial vaginosis: epidemiologic, clinical and diagnostic updates

Background and aims. Bacterial vaginosis (BV) is one the more frequently identified genital syndrome among childbearing aged women. The basic condition that generates this condition is a modification in the vaginal microbiota. The aim of this paper is to briefly review the current status of the art of BV and to report the results of a pilot study performed with an innovative PCR based technique. Materials and Methods. 36 samples of vaginal fluid routinely submitted for the diagnosis of BV to the Unit of Microbiology – GRHL were comparatively evaluated by standard techniques and with the HP-Vaginiti e Vaginosi NLM kit that simultaneously detects in a quantitative way specific DNA from Candida (albicans, glabrata; krusei, tropicalis), Gardnerella vaginalis, Lactobacillus spp. and Atopobium vaginae. Results and conclusions. Candida spp. has been identified in 8 samples with culture and in 15 with the molecular test. 29 G. vaginalis were found by PCR whereas only in 7 samples a specific prescription for this microbe was present (of which 4 positive). A. vaginae has been identified in 20 samples by the molecular approach and Lactobacillus spp. was identified in 19 samples (by culture) and in 32 by PCR. The overall diagnosis of BV was made in 9 patients by standard techniques and in 7 by applying the molecular approach. (Cohen’s kappa test: 0,84). The findings of this study clearly demonstrate that the joint use of the routine culture- based techniques with the multiplex PCR methods amplifies by far the sensitivity of the overall diagnostic workflow of BV.

Особенности структуры патологии шейки матки у беременных женщин

The aim of study was to determine the frequency and structure of cervical pathology and its relationship with the vaginal microbiota in 113 pregnant women in period from 22 to 36 weeks. Microscopy of vaginal smears, cytological examination of the cervical epithelium, determination of the microbial mass of vaginal secretion using test system Femoflor-17 and detection of human papillomavirus DNA (HPV) in real time was conducted. Statistical processing was performed with calculation of median, mean, standard deviation, confidence interval and Pearson correlation. HPV type 16 was found in 23.8%+4.2% (95% CI: 16.8-30.8%), other high-oncogenic types of HPV in 39.8%+5% (95% CI: 30.8-48.8%) of patients, while HPV type 18 was absent in all examined women. In 48 (55%) women the titer of microbial flora exceeded 104. Atopobium vaginae was detected in 43.7%+3.2% (95% CI: 39.7-47.7%) of pregnant women. According to the results of сytology, the inflammatory was found 27: 56,2%%+2,3% (95% DI: 51.9-60.4%) of pregnant women. Inclusion cytological smears type ASCUS was in 11: 9,7%+2,9% (95% CI: 4.7-14.7%) of women, which correlated with the detection of a combination of anaerobic and aerobic flora (r=0.76).

Modern approach to treatment of a recurrent bacterial vaginosis at women of the reproductive period

Bacterial vaginosis is closely associated with increase in frequency of inflammatory diseases of woman's pelvis organs, infertility, spontaneous abortions, preterm labor, contaminating fetus, postnatal inflammatory complications and cervical neoplasia. Goal of the study was to improve treatment of relapsed bacterial vaginosis in women of reproductive age and asses the efficacy of the pathogenetic combination therapy. Materials and methods. In our research we observed 40 female patients, at the age of 18-35 years who were revealed relapsed bacterial vaginosis according to clinical and laboratory examination. We studied their anamnesis, clinical manifistation, microscopic examination of cervical discharge to detect bacterial vaginosis. Also we carried out polymerase chain reaction to identify conditionally pathogenic microbes (Femoflor-16) and pH-metry of vagina contents with colpotest. Results. Our research revealed increased quantity or lack of a normal microflora (Lactobacillus spp.) in all patients. Streptococcus spp. 45.00%; Staphylococcus spp. 40.00%; Atopobium vaginae 80.00%; Gardnerella vaginalis + Prevotella bivia Porphyromonas (titer106) 70.00% were identified. Combination therapy allows to restore a vaginal ecosystem and prevents bacterial vaginosis relapse over period of 12 months in our patients. Conclusion. Polymerase chain reaction in real time (Femoflor-16) with vaginal pH-metry are the basic laboratory examination methods to confirm bacterial vaginosis. Use of a combination therapy allows to restore a vaginal ecosystem and prevents bacterial vaginosis relapse over period of 12 months in our patients.

Tranzitorna bakterijemija prouzrokovana Atopobium-om vaginae nakon carskog reza - prikaz slučaja

Introduction: Genital microbiome of woman is still a puzzle to this day. It consists of a large number of different bacterial species and phenotypes that make up the normal genital flora, while in the cases of its change there is a predominance of certain, especially anaerobic species, which is the case with bacterial vaginosis. These types are mostly rare causing invasive infections, but under certain conditions can lead to severe infections, such as multiple parts of the genital tract, and to an unwanted outcome in the pregnancy. Case report: This paper presents an extremely rare case of transient bacteremia caused by Atopobium vaginae type in a patient following a cesarean section where empirical therapy gave a positive effect and the patient recovered and released home with healthy newborn. Conclusions: During the clinical work it is important to be aware of the different anaerobic species present in the genital tract of women microbiome like Atopobium vaginae and should adjust the empirical therapy, which was the case here.

Toll-like receptor variants and cervical Atopobium vaginae infection in women with pelvic inflammatory disease.

Toll-like (TLR) receptor genetic variants have been implicated in bacterial vaginosis (BV). We determined whether TLR variants are associated with fastidious BV-associated microbes that are linked with infertility following pelvic inflammatory disease (PID). Sneathia spp., Atopobium vaginae, BVAB1, and Ureaplasma urealyticum were measured in 250 women from the PID Evaluation and Clinical Health (PEACH) study. Relative risk (RR) and 95% confidence intervals (CI) were calculated adjusting for chlamydia and gonorrhea. Principal component analysis was used to adjust for population stratification. A false discovery rate q-value of 0.05 was significant. TLR2-1733C>A (P=.003) and TLR2-616A>G (P=.004) were associated with cervical A.vaginae. TLR2-1733C>A and TLR6-438C>T were associated with A.vaginae detection in the endometrium, but this was not significant after adjustment for multiple comparisons (FDR q-value=0.06). Host gene variants in TLR2 signaling pathways were modestly associated with cervical A.vaginae in women with clinical PID.

Bacterial communities forming the vaginal micro-ecosystem in norm and in bacterial vaginosis

Background. Bacterial vaginosis (BV) is disturbance of the vaginal microbiota, characterized by displacement of lactobacilli with anaerobic bacteria and capable of adversely affecting women’s reproductive health. In the development of BV, a wide spectrum of bacteria substantially differing in their properties is involved. Grouping vaginal bacterial communities into clusters, or types of microbiocenosis, might contribute to understanding of pathogenic mechanisms and elaboration of effective tools for diagnostics and therapy of the disease.&#x0D; Aim. Determination and comparative analysis of clusters of vaginal bacterial communities in norm and in BV.&#x0D; Materials and methods. Women of reproductive age were enrolled in the study. For the diagnosis of BV, the Nugent score was used. Vaginal swab samples from all women were analyzed with the test Femoflor-16, intended for evaluation of the vaginal microbiocenosis using multiplex quantitative real-time PCR. Two-step cluster analysis was applied for grouping bacterial communities. Differences between the clusters were evaluated using pairwise comparisons.&#x0D; Results. Of 280 women enrolled in the study, 172 had normal microflora, 27 – intermediate microflora, 81 – BV. In cluster analysis, 270 samples valid in PCR testing were included. All the vaginal bacterial communities were grouped into 4 clusters. Cluster 1 (n = 171) included cases when the vaginal microflora consisted mostly of lactobacilli. Cluster 2 (n = 11) encompassed cases of domination of aerobic microflora: Enterobacteriaceae, Streptococcus and Staphylococcus. Clusters 3 (n = 57) and 4 (n = 31) were connected with BV and included cases of prevailing of facultative anaerobes (Gardnerella vaginalis, Atopobium vaginae) and obligate anaerobes (Sneathia/Leptotrichia/Fusobacterium, Megasphaera/Veillonella/Dialister, Lachnobacterium/Clostridium), respectively. Nearly all cases of cluster 1 belonged to the category of normal microflora of the Nugent score. The majority of bacterial communities of cluster 2 matched intermediate microflora, cluster 3 – BV category with a score of 7 or 8, cluster 4 – BV category with a score of 9 or 10. The clusters differed significantly in vaginal рН, with the highest values observed for cluster 4.&#x0D; Conclusions. Vaginal bacterial communities are grouped into 4 main clusters, characterized by domination of lactobacilli, aerobes, facultative anaerobes or obligate anaerobes. The clusters belong to different categories of the Nugent score and differ significantly in vaginal pH.

THE CLINICAL AND MICROBIOLOGICAL FEATURES OF BACTERIAL VAGINOSIS

Aim : to determine the clinical and microbiological features of bacterial vaginosis on the basis of the microbiological examination, assessment of the cytokine status in female patients. Material and methods . The article presents the results of the complex clinical and microbiological examination of 86 female patients of the fertile age. 30 (34.88 ± 5.14 %) patients were diagnosed bacterial vaginosis on the basis of revelation and identification of DNA of Gardnerella vaginalis, Atopobium vaginae, Lactobacillus spp. and the total number of bacteria. The control group consisted of 56 (65.12 ± 5.14 %) patients without bacterial vaginosis. The work also presents the results of the microbiological analysis of the material obtained from the cervical canal and endometrium. All the patients underwent blood tests for detection of the inflammatory reaction - interleukins IL-1, IL-2, tumor necrosis factor (TNF-α), interferon (γ-IFN). Results . The prevalence of bacterial vaginosis among the patients of the reproductive age was 34.8 %. The pathology of the reproductive function was found with equal frequency in the patients of both the groups. Disorders of the immune status in favor of pro-inflammatory cytokines were not diagnosed. The clinical and laboratory criteria made it possible to diagnose bacterial vaginosis in 3.49 % of the patients, the use of PCR diagnosis - in 34.88 %. The concentration of lactobacillus spp. is reliably lower in the patients with bacterial vaginosis, than in the control group (p = 0.0085). As for the concentrations of Gardnerella vaginalis and Atopobium vaginae the groups do not significantly differ. Only 4 (13.33 ± 6.21 %) patients (χ = 5.51, p = 0.02) in the main group detected sexually transmitted infections, which should be taken into account while performing the diagnostic activities. Conclusion. The identification of DNA of certain kinds of microorganisms give an opportunity to assess the state of vaginal microcenosis and the degree of its malfunction even in the absence of clinical and other laboratory signs of bacterial vaginosis. The malfunction of the biocenosis of the genital tract is not associated with disorders of the reproductive function and does not lead to a change in the cytokine status. Patients with bacterial vaginosis are in the risk group for development of sexually transmitted infections, which should be taken into account while performing the diagnostic activities.

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

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

Effect of the combined preparation Terzhinan® on microorganisms isolated from the urogenital tract of women. In vitro experiment

Introduction. The study is in vitro experiment, aimed to determine antibiotic susceptibility of microorganisms isolated from the urogenital tract of women to the combination of substances included in the preparation Terzhinan.&#x0D; Methods. In total, 516 microorganism strains isolated from the vagina of reproductive age women have been tested: Candida spp. (n = 83), bacteria of the family Enterobacteriaceae (n = 138), Streptococcus agalactiae (n = 72), Enterococcus spp. (n = 147), Staphylococcus spp. (n = 37), Actinomyces urogenitalis (n = 7), Gardnerella vaginalis (n = 27), Atopobium vaginae (n = 5). Suspensions of each strain in a volume of 0.5 ml (0.5 according to McFarland) were applied onto culture medium (Muller-Hinton Agar or 5% Blood Muller-Hinton Agar). Furthermore, serial dilutions of Terzhinan were applied with a calibrated loop onto three sectors. Petri dishes were then incubated at 37 °С for 24 h. The results were evaluated visually, by measuring growth inhibition zones.&#x0D; Results. The majority of Candida albicans isolates were susceptible (S) to the preparation, using both undiluted and diluted (10 and 100 times) preparation. Terzhinan without dilution and in the dilution 1:10 was 100% effective against all tested bacteria of the family Enterobacteriaceae. Susceptibility of gram positive bacteria to the antibiotics included in Terzhinan, was 74.1%. All isolates of staphylococci, including Staphylococcus aureus, were susceptible. The frequency of susceptible strains of Streptococcus agalactiae and Enterococcus spp. was 70%. All clinical isolates of Gardnerella vaginalis and Atopobium vaginae were susceptible to the combinations of antibiotics included in the preparation Terzhinan, which is in agreement with data on its high efficiency in the treatment of bacterial vaginosis.&#x0D; Conclusions. In in vitro experiment, the combined preparation Terzhinan showed high activity against major groups of microorganisms isolated from the vagina including pathological conditions, such as bacterial vaginosis and vulvovaginitis of different etiology.

The urinary microbiota of men and women and its changes in women during bacterial vaginosis and antibiotic treatment

BackgroundThe urinary microbiota is similarly complex as the vaginal and penile microbiota, yet its role as a reservoir for pathogens and for recurrent polymicrobial biofilm diseases like bacterial vaginosis (BV) is not clear.ResultsHere, we analysed the urinary microbiota of healthy men and women and compared it with that of women during BV and after antibiotic treatment using next-generation sequencing of the 16S rRNA gene V1-V2 regions. Eight different community types, so called urotypes (UT), were identified in healthy humans, all of which were shared between men and women, except UT 7, dominated in relative abundance by Lactobacillus crispatus, which was found in healthy women only. Orally applied metronidazole significantly reduced Shannon diversity and the mean relative abundance of Gardnerella vaginalis, Atopobium vaginae, and Sneathia amnii, while L. iners increased to levels twofold higher than those found in healthy women. Although individual urine microbial profiles strongly responded to the antibiotic, the healthy community could not be restored. The correlation between urinary and vaginal fluid microbiota was generally weak and depending on UT and BV status. It was highest in UT 1 in acute BV (59% of samples), but after metronidazole treatment, only 3 out of 35 women showed a significant correlation between their urinary and vaginal microbiota composition.ConclusionsUrethra and bladder thus harbor microbial communities distinct from the vagina. The high abundance of BV related species in the urine of both men and women suggests that urine may act as a reservoir of pathogens and contribute to recurrence.Trial registrationClinicalTrials.gov, NCT02687789

Treatment failure of bacterial vaginosis is not associated with higher loads of Atopobium vaginae and Gardnerella vaginalis

Cervicovaginal Atopobium vaginae and Gardnerella vaginalis are strongly associated with bacterial vaginosis (BV) and are the main components of vaginal biofilms. The low efficacy of BV treatment with metronidazole may be due to the presence of such biofilms. Thus, the aim of this study was to compare the pretreatment cervicovaginal loads of A. vaginae and G. vaginalis for women who restored normal flora and those who persisted with BV after a full course of oral metronidazole. In this cross-sectional study, 309 reproductive-aged women were recruited in a primary health care service in Botucatu, Brazil. Cervicovaginal samples were tested for genital tract infections, microscopic classification of local microbiota and molecular quantification of A. vaginae and G. vaginalis. All the participants with concurrent cervicovaginal infections (n=64) were excluded. A total of 84 out of 245 (34.3 %) women had BV at enrolment and 43 (51.2 %) of them completed the treatment and returned for follow-up. Evaluation of the vaginal microbiota at follow-up showed that 29 (67.4 %) women restored normal vaginal flora, while 14 (32.6 %) still had BV. The pretreatment loads of G. vaginalis were lower in women with treatment failure (P=0.001) compared to those who successfully restored normal flora. The loads of A. vaginae did not differ between the groups. Although G. vaginalis produces several virulence factors and its loads correlate positively with those of A. vaginae, higher cervicovaginal quantities of these bacteria are not associated with treatment failure of BV after oral metronidazole.

Criteria for diagnosis of bacterial vaginosis using the test Femoflor-16

Background. Bacterial vaginosis is disturbance of the balance of the vaginal microflora, associated with a number of infectious diseases of the urogenital tract and adverse pregnancy outcomes. In this country, for the detection of vaginal dysbiotic conditions, the test Femoflor-16 (DNA-Technology, Moscow) is widely used, however interpretation algorithms of this test do not include the category of BV.&#x0D; Aim. The study aimed to elaborate diagnostic criteria for the detection of BV using Femoflor-16 test.&#x0D; Materials and methods. Women of reproductive age addressing a gynecologist with vaginal discharge were enrolled in the study. For clinical diagnosis of BV, the Amsel criteria were used, laboratory analysis for BV was performed via microscopic investigation of vaginal discharge using the Nugent score. Samples of vaginal discharge from all women were analyzed with the test Femoflor-16, intended for characterizing vaginal microbiocenosis using multiplex quantitative real-time PCR.&#x0D; Results. A total of 280 women were included in the study. BV was diagnosed in 86 women (31%) using the Amsel criteria, and in 81 women (29%) using the Nugent score. All groups of anaerobic bacteria included in Femoflor-16 test were shown to be associated with BV, with the exception of bacteria of the genus Mobiluncus, which are detected together with phylogenetically related but not BV-associated bacteria of the genus Corynebacterium. A low amount of lactobacilli (&lt; 10% of total bacterial load) coupled with an elevated amount of Gardnerella vaginalis/Prevotella bivia/Porphyromonas (&gt; 1%) and/or Eubacterium (&gt; 2%) and/or Sneathia/Leptotrichia/Fusobacterium (&gt; 0.1%) and/or Megasphaera/Veillonella/Dialister (&gt; 0.1%) and/or Lachnobacterium/Clostridium (&gt; 0.1%) and/or Peptostreptococcus (&gt; 0.1%) and/or Atopobium vaginae (&gt; 0.2%) detected BV with a sensitivity of 99% and specificity of 93%.&#x0D; Conclusions. Criteria for BV diagnosis using the test Femoflor-16 have been elaborated, which enable to detect BV or exclude it with a sensitivity of 99% and specificity of 93%. These criteria for BV and criteria of the test manufacturers for severe anaerobic dysbiosis determine to a large extent the same category of the vaginal microbiocenosis.