Vaginal Bacteria Research Articles

Exploring the antimicrobial properties of vaginal Lactobacillus crispatus against preterm birth-associated bacteria.

Lay summaryThe need to develop new treatments to prevent unprompted premature delivery before 37 weeks of pregnancy remains pressing and unmet. Bacteria (Lactobacillus species) that promote vaginal health produce biochemical compounds that prevent the growth of microbes such as Gardnerella vaginalis. Overgrowth of G. vaginalis can cause vaginal infection with smelly discharge and increase a woman’s risk of sexually transmitted infections and premature delivery. In this study, we examined how normal health-promoting (L. crispatus) and potentially harmful (G. vaginalis) vaginal bacteria interact in a laboratory setting. This was in order to observe natural and effective agent(s) from L. crispatus that can hinder the growth of G. vaginalis and accompanying immune response. We observed that L. crispatus clears G. vaginalis by itself and with several biochemical compounds that it produces. Such biochemical compounds can be developed into treatment for vaginal infections and premature delivery due to infection and inappropriate immune response.

Associations of First Void Urine Samples with Bacterial Vaginosis

Objective: The aim of this study was to investigate whether BV can be diagnosed from first-void urine (FVU). Study Design: Cross-sectional/case control study Place and Duration: Conducted at Hameed Latif Hospital Lahore, during from Sep, 2021 to Feb, 2022. Methods: There were 100 women had ages 20-65 years was presented in this study. Once informed written consent was obtained, participants' detailed demographic information was recorded. First void urine was collected from all women. According to Nugent's criteria, BV was indeed diagnosed. All of the qPCRs performed on the FVU and vaginal swabs for the selected vaginal bacteria (Atopobium vaginae, Prevotella spp., Gardnerella vaginalis, bacterial vaginosis-associated bacterium 2, Eggerthella-like bacterium, "Leptotrichia amnionii," Megasphaera type 1) had an area under the receiver operating characteristic (ROC) curve of >85%, All data was examined using SPSS 24.0. Results: In current study, mean age of the women was 28.11±15.68 years and had mean BMI 24.9±7.49 kg/m2. Majority of the females were nulliparous 67 (67%). Most of the females had no genital hair remove among 60 (60%). Among 100 females, 49 (49%) females had bacterial vaginosis, 40 (40%) had normal vaginal flora and 11 (11%) had intermediate flora. Nugent scoring was used to determine the sensitivity and specificity of seven assays for BV-associated bacteria in urine (A. vaginae, Prevotella spp., G. vaginalis, BVAB 2, Eggerthella-like bacterium, L. amnionii, and Megasphaera type 1). By qualitative detection, the assays were 80%-100% sensitive but only 8%-80% specific (presence or absence). The area under the curve (AUC) was > or = 91% for all seven bacterial species, indicating high diagnostic accuracy. Conclusion: Our research of ROC curves led us to the conclusion that FVU can be used to accurately diagnose BV. Combining qPCR findings for two bacteria, like Megasphaera type 1 and Prevotella spp., can improve sensitivity or specificity, respectively. Keywords: First Void Urine, BV, PCR, Nugnet scoring. 7-Assays

P-375 Optimising vaginal biospecimen collection

Abstract Study question Are menstrual tampons an acceptable and comparable method to lower vaginal swabs (LVS) for analysing the vaginal microbiome in non-pregnant women? Summary answer Tampons capture a higher bacterial load, volunteers are familiar with them, thus are well-accepted, and can be efficiently frozen at -20 °C prior to analysis. What is known already The vaginal microbiome plays a key role in women’s reproductive health. The application of investigating this niche allows a greater understanding of the impact of the vaginal microbiota in diverse reproductive disorders including infertility, recurrent miscarriage, spontaneous preterm birth and sexually transmitted infections. Fear and embarrassment related to physicians taking a traditional vaginal swab using a speculum results in many female genitourinary health appointments to be missed in the UK and recruitment challenges into vaginal microbiota research studies. Growing evidence suggests a menstrual tampon to be a superior alternative biospecimen collection device to a vaginal swab. Study design, size, duration This was a prospective, cross-sectional observational study. Twenty-one healthy volunteers (average age 30 years) were recruited over a period of 6 weeks at the Centre for Women's Health Research, University of Liverpool, an academic, tertiary referral centre. Participants/materials, setting, methods The volunteers consented to wearing a tampon for 20 minutes, taking 2 LVS and completing an acceptability questionnaire. Samples were halved for immediate processing and freezing at -20 °C for 2-4 weeks. Real-time PCR using Femoflor-16 was carried out to identify microorganisms present in vaginal microbiota. Microbiota was classified by total bacterial mass and relative abundance of lactobacillus species. Statistical analysis was carried out using GraphPad PRISM 5, Microsoft Excel 2019 and IBM SPSS Statistics 27. Main results and the role of chance Of the 84 samples, four had insufficient material for the purpose of analysis. Three of these were LVS samples and the fourth was a frozen tampon sample. Mean tampon total bacterial mass was higher for fresh and frozen samples versus the mean LVS total bacterial mass. The Femoflor algorithm gave identical vaginal microbiota characterisation between the fresh tampon and fresh LVS in 68.4% (13/19) of cases. 92% (12/13) samples with over 80% relative abundance of lactobacillus in the LVS also had over 80% relative abundance in the tampon. More discrepancy was seen between samples within the 20-80% and <20% relative abundance groups of lactobacillus in the LVS with 50% and 75% different classifications respectively. Importantly, only one case showed an LVS with lactobacillus >80% (normal) and a tampon with non-dominance of lactobacillus (<80%, dysbiosis). The questionnaires revealed that 90.5% of the volunteers highly rated using tampons (3 or more out of 5 on a Likert scale), with 52.4% rating tampon-use at 5/5. All participants (100%) stated that they would be happy to provide a tampon sample for clinical/research testing purposes in the future. Limitations, reasons for caution This study included 21 healthy volunteers; this is not necessarily representative of the entire UK population, but the tampon and LVS from the same volunteer were consistently comparable before and after freezing. We used a multiplex PCR assay to target common vaginal bacteria, inevitably some bacteria have not been characterised. Wider implications of the findings Self-collected tampons are suitable, well accepted biospecimen collection devices that may reduce barriers posed due to embarrassment, improve attendance in genitourinary medicine clinics and encourage participation in vaginal microbiota research. Freezing before processing reduces time and costs. A large study is necessary to ensure reproducibility and acceptability to patient population. Trial registration number Not Applicable

Models of Murine Vaginal Colonization by Anaerobically Grown Bacteria.

The mammalian vagina can be colonized by many bacterial taxa. The human vaginal microbiome is often dominated by Lactobacillus species, but one-in-four women experience bacterial vaginosis, in which a low level of lactobacilli is accompanied by an overgrowth of diverse anaerobic bacteria. This condition has been associated with many health complications, including risks to reproductive and sexual health. While there is growing evidence showing the complex nature of microbial interactions in human vaginal health, the individual roles of these different anaerobic bacteria are not fully understood. This is complicated by the lack of adequate models to study anaerobically grown vaginal bacteria. Mouse models allow us to investigate the biology and virulence of these organisms in vivo. Other mouse models of vaginal bacterial inoculation have previously been described. Here, we describe methods for the inoculation of anaerobically grown bacteria and their viable recovery in conventionally raised C57Bl/6 mice. A new, less stressful procedural method for vaginal inoculation and washing is also described. Inoculation and viable recovery of Gardnerella are outlined in detail, and strategies for additional anaerobes such as Prevotella bivia and Fusobacterium nucleatum are discussed.

Models of Murine Vaginal Colonization by Anaerobically Grown Bacteria

The mammalian vagina can be colonized by many bacterial taxa. The human vaginal microbiome is often dominated by Lactobacillus species, but one-in-four women experience bacterial vaginosis, in which a low level of lactobacilli is accompanied by an overgrowth of diverse anaerobic bacteria. This condition has been associated with many health complications, including risks to reproductive and sexual health. While there is growing evidence showing the complex nature of microbial interactions in human vaginal health, the individual roles of these different anaerobic bacteria are not fully understood. This is complicated by the lack of adequate models to study anaerobically grown vaginal bacteria. Mouse models allow us to investigate the biology and virulence of these organisms in vivo. Other mouse models of vaginal bacterial inoculation have previously been described. Here, we describe methods for the inoculation of anaerobically grown bacteria and their viable recovery in conventionally raised C57Bl/6 mice. A new, less stressful procedural method for vaginal inoculation and washing is also described. Inoculation and viable recovery of Gardnerella are outlined in detail, and strategies for additional anaerobes such as Prevotella bivia and Fusobacterium nucleatum are discussed.

Vaginal microbiota as an unusual cause of spondylodiscitis.

The incidence of extra pelvic infections due to vaginal microflora bacteria has increased as growth media and methods of isolation have improved. However, bone infections seem to be still relatively rare, and little is known about their risk factors, clinical presentation, treatment and final outcome. We describe here a spondylodiscitis due to Gardnerella vaginalis, Atopobium vaginae, Peptostreptococcus indolicus and Prevotella amnii, anaerobic bacteria from vaginal microbiota. Our patient had no obvious predisposing factor and recovered after antibiotic treatment. To our knowledge, this case is the first reported spondylodiscitis caused by polymicrobial vaginal flora in a healthy, immunocompetent woman.

How were Lactobacillus species selected as single dominant species in the human vaginal microbiota? Coevolution of humans and Lactobacillus

The optimal microbiota in the vagina for women’s health is dominated by single species of Lactobacillus. However, the mechanism by which these species become dominant remains unknown. Therefore, understanding this mechanism is important to establish new treatments, such as probiotics, for vaginal dysbiosis-related conditions. To date, at least 5 hypotheses have been proposed to explain the vaginal dominance of Lactobacillus. However, these hypotheses do not address why a “single” species of Lactobacillus becomes dominant in the vaginal microbiota. First, we evaluated the effect of Lactobacillus culture supernatant on the biomass concentration of each different strain and species of Lactobacillus. The results indicated that the biomass concentrations were not significantly affected by the culture supernatant from other vaginal Lactobacillus species. Then, we discussed host factors that select the vaginal Lactobacillus species and the antagonism of vaginal bacteria. We also assess the mechanism of coevolution of humans and Lactobacillus. Here, we present the hypothesis that host factors such as antimicrobial peptides and proteins (AMPs) and immunoglobulins could play important roles in the selection of specific species of Lactobacillus in the vagina.

Distinct gut and vaginal microbiota profile in women with recurrent implantation failure and unexplained infertility

BackgroundFemale reproductive tract dysbiosis impacts implantation. However, whether gut dysbiosis influences implantation failure and whether it accompanies reproductive tract dysbiosis remains scantly explored. Herein, we examined the gut-vaginal microbiota axis in infertile women.MethodsWe recruited 11 fertile women as the controls, and a cohort of 20 infertile women, 10 of whom had recurrent implantation failure (RIF), and another 10 had unexplained infertility (UE). Using amplicon sequencing, which employs PCR to create sequences of DNA called amplicon, we compared the diversity, structure, and composition of faecal and vaginal bacteria of the controls with that of the infertile cohort. Of note, we could only sequence 8 vaginal samples in each group (n = 24/31).ResultCompared with the controls, α-diversity and β-diversity of the gut bacteria among the infertile groups differed significantly (p < 0.05). Taxa analysis revealed enrichment of Gram-positive bacteria in the RIF group, whereas Gram-negative bacteria were relatively abundant in the UE group. Strikingly, mucus-producing genera declined in the infertile cohort (p < 0.05). Hungatella, associated with trimethylamine N-oxide (TMAO) production, were enriched in the infertile cohort (p < 0.05). Vaginal microbiota was dominated by the genus Lactobacillus, with Lactobacillus iners AB-1 being the most abundant species across the groups. Compared with the infertile cohort, overgrowth of anaerobic bacteria, associated with vaginal dysbiosis, such as Leptotrichia and Snethia, occurred in the controls.ConclusionThe gut microbiota had little influence on the vaginal microbiota. Gut dysbiosis and vaginal eubiosis occurred in the infertile women, whereas the opposite trend occurred in the controls.

Healthy Vaginal Microbiota and Influence of Probiotics Across the Female Life Span.

Vaginal microbiota plays a central role in women’s health and reproduction. Vaginal microbiota is dynamic and shaped by hormonal shifts in each stage of a woman’s life from pre-puberty to postmenopause. Current research has mainly focused on vaginal bacterial and fungal members of the community and emphasized their role in disease. However, the impact of balanced vaginal microbiota on health and its interaction with the host is yet poorly understood. High abundance of vaginal lactobacilli is most strongly associated with health, but the concept of health may vary as vaginal dysbiosis may be asymptomatic. Furthermore, there is a lot of variation between ethnic groups in terms of dominating vaginal bacteria. Probiotic lactobacilli could be a safe and natural means to balance and maintain healthy vaginal microbiota. Research evidence is accumulating on their role in supporting women’s health throughout life. This review describes the current literature on vaginal microbiota, the major factors affecting its composition, and how the communities change in different life stages. Furthermore, we focused on reviewing available literature on probiotics and their impact on vaginal microbiota and health.

MinION, a portable long-read sequencer, enables rapid vaginal microbiota analysis in a clinical setting

BackgroundIt has been suggested that the local microbiota in the reproductive organs is relevant to women's health and may also affect pregnancy outcomes. Analysis of partial 16S ribosomal RNA (rRNA) gene sequences generated by short-read sequencers has been used to identify vaginal and endometrial microbiota, but it requires a long time to obtain the results, making it unsuitable for rapid bacterial identification from a small specimen amount in a clinical context.MethodsWe developed a simple workflow using the nanopore sequencer MinION that allows high-resolution and rapid differentiation of vaginal microbiota. Vaginal samples collected from 18 participants were subjected to DNA extraction and full-length 16S rRNA gene sequencing with MinION.ResultsThe principal coordinate analysis showed no differences in the bacterial compositions regardless of the sample collection method. The analysis of vaginal microbiota could be completed with a total analysis time of approximately four hours, allowing same-day results. Taxonomic profiling by MinION sequencing revealed relatively low diversity of the vaginal bacterial community, identifying the prevailing Lactobacillus species and several causative agents of bacterial vaginosis.ConclusionsFull-length 16S rRNA gene sequencing analysis with MinION provides a rapid means for identifying vaginal bacteria with higher resolution. Species-level profiling of human vaginal microbiota by MinION sequencing can allow the analysis of associations with conditions such as genital infections, endometritis, and threatened miscarriage.

Associations Between Vaginal Bacteria and Bacterial Vaginosis Signs and Symptoms: A Comparative Study of Kenyan and American Women.

BackgroundBacterial colonization and associations with bacterial vaginosis (BV) signs and symptoms (Amsel criteria) may vary between populations. We assessed relationships between vaginal bacteria and Amsel criteria among two populations.MethodsKenyan participants from the placebo arm of the Preventing Vaginal Infections (PVI) trial and participants from a Seattle-based cross-sectional BV study were included. Amsel criteria were recorded at study visits, and the vaginal microbiota was characterized using 16S rRNA gene sequencing. Logistic regression models, accounting for repeat visits as appropriate, were fit to evaluate associations between bacterial relative abundance and each Amsel criterion.ResultsAmong 84 PVI participants (496 observations) and 220 Seattle participants, the prevalence of amine odor was 25% and 40%, clue cells 16% and 37%, vaginal discharge 10% and 52%, elevated vaginal pH 69% and 67%, and BV 13% and 44%, respectively. BV-associated bacterium 1 (BVAB1) was positively associated with all Amsel criteria in both populations. Eggerthella type 1, Fannyhessea (Atopobium) vaginae, Gardnerella spp., Sneathia amnii, and Sneathia sanguinegens were positively associated with all Amsel criteria in the Seattle study, and all but discharge in the PVI trial.ConclusionsCore vaginal bacteria are consistently associated with BV signs and symptoms across two distinct populations of women.

Insight into the ecology of vaginal bacteria through integrative analyses of metagenomic and metatranscriptomic data

BackgroundVaginal bacterial communities dominated by Lactobacillus species are associated with a reduced risk of various adverse health outcomes. However, somewhat unexpectedly, many healthy women have microbiota that are not dominated by lactobacilli. To determine the factors that drive vaginal community composition we characterized the genetic composition and transcriptional activities of vaginal microbiota in healthy women.ResultsWe demonstrate that the abundance of a species is not always indicative of its transcriptional activity and that impending changes in community composition can be predicted from metatranscriptomic data. Functional comparisons highlight differences in the metabolic activities of these communities, notably in their degradation of host produced mucin but not glycogen. Degradation of mucin by communities not dominated by Lactobacillus may play a role in their association with adverse health outcomes. Finally, we show that the transcriptional activities of L. crispatus, L. iners, and Gardnerella vaginalis vary with the taxonomic composition of the communities in which they reside. Notably, L. iners and G. vaginalis both demonstrate lower expression of their cholesterol-dependent cytolysins when co-resident with Lactobacillus spp. and higher expression when co-resident with other facultative and obligate anaerobes. The pathogenic potential of these species may depend on the communities in which they reside and thus could be modulated by interventional strategies.ConclusionsOur results provide insight to the functional ecology of the vaginal microbiota, demonstrate the diagnostic potential of metatranscriptomic data, and reveal strategies for the management of these ecosystems.

Dynamics of vaginal microbiota in women of reproductive age during the menstrual cycle

Objective: To investigate the dynamic changes of vaginal microbiota in different phases of menstrual cycle in healthy Chinese women of childbearing age. Methods: A total of 11 healthy women of childbearing age with regular menstruation, who had physical examination in the Gynecology Clinic of Beijing Obstetrics and Gynecology Hospital from September to December 2020 were randomly selected as research subjects. Vaginal secretions were collected during menstrual phase (2nd-3rd day), mid-follicular phase (7th-8th day), and mid-luteal phase (21st-22nd day) for microbiota analysis through metagenomic sequencing. Results: (1) Vaginal microbiota species were the most diverse in menstrual phase and the least in follicular phase, observing dominant vaginal bacteria gradually changing to Lactobacillus from menstrual phase to follicular phase and then to luteal phase. (2) The dynamic evolution of vaginal microbiota from menstrual phase to follicular phase and then to luteal phase was divided into: no change in dominant bacteria, replacement of dominant bacteria, changes in the proportion of dominant bacteria, and recurrence of dominant bacteria (non-Lactobacillus-dominance appeared again in luteal phase after returning to normal Lactobacillus-dominance in follicular phase). (3) Prevotella, especially Prevotella_bivia, was significantly higher during menstrual phase. Conclusions: Healthy vaginal microbiota should be relatively stable, but also have the ability of dynamic change and self-recovery. Prevotella plays a central role among opportunistic pathogens in the vagina, whose function remains to be investigated.

Molecular study to some Virulance gene of Staphylococcus aureus and Pseudomonas aeruginosa bacteria isolated from women suffered from bacterial vaginitis

The study included 250 vaginal swabs were collected from women patients clinical symptoms of bacterial vaginitis and they have come to the women's and children's hospital in Ramadi and ages (17- 45 years) nearly, to Investigation of the bacterial species positive and negative gram stain were Infected of the vagina, and also To determination the appearance and molecularly of virulence factors. The vaginal swabs were cultured, 86 samples were found to give positive growth of 34.4%, but (164 sample) 65.6% were negative. The growing isolates were subjected to bacteriological and biochemical tests. The isolates of positive gram stain 48 about 55.81% were having character of Staphylococcus epidermidis 18 about 20.93% and Staphylococcus aureus (9 (10.46%), Streptococcus pneumoniae 7 nearly 8.13%, Staphylococcus saprophyticus 6 about 7.97%, Streptococcus pyogenes 4, (4.65%) and Bacillus subtilis only 3, (3.48%) and Streptococcus viridans was only 1 isolate about 1.16% . The isolates of negative gram stain were recorded 38 percentage 44.18% . The results of the tests a bacteriological and biochemical were recorded Escherichia coli 15 isolates percent 17.44% and 14 of Klebsiella pneumoniae 16.27% , Pseudomonas aeruginosa was 6 percent 7.97%, Gardnerella vaginalis 2 about 2.32% , and Proteus mirabilis was one isolate only percent 1.16% . The polymerase chain reaction (PCR) technique were proved, the exoA and exoS genes were encodes toxin production. algD was encoded the production of the capsule which were found in Pseudomonas aeruginosa percent 100% for the three genes. In Staphylococcus aureus found that (hla) gen that encodes production of alpha-hemolycin 66.66% and finbA which encodes the production of (adhesion protein) 100%, as well as investigate some of the virulent factors of the vaginal bacteria, which included their ability to produce the enzyme hemolysis, urease and protease.

Enhanced IgA coating of bacteria in women with Lactobacillus crispatus-dominated vaginal microbiota

BackgroundImmunoglobulin A (IgA) plays an important role in maintaining a healthy intestinal microbiome, but little is known about the interaction between local immunoglobulins and the vaginal microbiome. We assessed immunoglobulins (unbound and bound to bacteria), their association with vaginal microbiota composition and the changes over time in 25 healthy women of reproductive age.ResultsIn both Lactobacillus crispatus-dominated and non-L. crispatus-dominated microbiota, IgA and IgG (unbound and bound to bacteria) were higher during menses (T = 1) compared to day 7‑11 (T = 2) and day 17‑25 (T = 3) after menses onset. The majority of vaginal bacteria are coated with IgA and/or IgG. Women with L. crispatus-dominated microbiota have increased IgA coating of vaginal bacteria compared to women with other microbiota compositions, but contained less IgA per bacterium. Presence of a dominantly IgA-coated population at T = 2 and/or T = 3 was also strongly associated with L. crispatus-dominated microbiota. In women with non-L. crispatus-dominated microbiota, more bacteria were uncoated. Unbound IgA, unbound IgG, and bound IgG levels were not associated with microbiota composition.ConclusionsIn conclusion, L. crispatus-dominated vaginal microbiota have higher levels of bacterial IgA coating compared to non-L. crispatus-dominated vaginal microbiota. Similar to its regulating function in the intestinal tract, we hypothesize that IgA is involved in maintaining L. crispatus-dominated microbiota in the female genital tract. This may play a role in L. crispatus-associated health benefits.3F84r7gHjdTjEchicFxFSRVideo abstract

A Capillary Electrophoresis Method for Analyzing the Diversity of Vaginal Microbiome

16S ribosomal RNA (rRNA) gene sequence analysis is the common method to identify the bacteria in human vaginal flora. While specific DNA primers were designed to target 16S rRNA gene sequences, DNA fragment analysis using capillary electrophoresis can obtain more accurate qualitative and quantitative information on the microbiome. This study aimed to assess the ability of capillary electrophoresis method to analyze the diversity of vaginal microbiome and provide a theoretical basis for the accurate gene detection of vaginal flora. We collected 75 vaginal secretion samples from female outpatients aged 25 - 50, who had undergone rou-tine gynecologic examinations in Fujian provincial hospital from March 2021 to April 2021. Clinical diagnosis was based on the results of microscopic examination of Gram-stained specimens and biochemical tests of bacteria (pH value, catalase, leukocyte esterase, sialidases, β-glucuronidase, and acetylglucosaminidase). Vaginal secretion samples were collected and then total bacterial DNA was extracted. We used six pairs of fluorescent dye tagged specific primers that were designed based on the 16S rRNA genes of four Lactobacillus species (L. iners, L. crispatus, L. jensenii, L. gasseri), Gardnerella vaginalis, and Atopobium vaginae. PCR products of six species of bacteria were detected and analyzed by a 3130 Genetic Analyzer. Seventy-five samples were divided into two groups according to the vaginal microbiome evaluation, including 50 cases which had vaginal bacteria balance and 25 cases which had vaginal bacteria disorder. PCR amplification of 16S ribosomal RNA genes of L. iners, L. crispatus, L. jensenii, L. gasseri, Gardnerella vaginalis, and Atopobium vaginae was successfully performed on the DNA extracted from vaginal secretion samples. Four Lac-tobacillus species were detected in 4 - 33 cases of "Balance" group, and Gardnerella vaginalis was detected in 23 cases of "Disorder" group and, simultaneously, Atopobium vaginae was detected in 20 cases. Based on the technique of DNA fragment analysis using capillary electrophoresis method, the most common vaginal bacteria in Chinese healthy women are L. iners and L. crispatus. Gardnerella vaginalis and Atopobium vaginae are the most common pathogenic bacteria detected in the patients who had vaginal bacteria disorder. Using capillary electrophoresis method to detect the vaginal bacteria will be useful for accurate identifica-tion of vaginal microbiome. There will be an application value to find out the composition of the vaginal microbiome rapidly and detect specific gene markers to identify potential pathogenic bacteria when women are at risk of serious illness before they develop obvious symptoms.

Lactobacilluscrispatus dominance in the vaginalmicrobiome reduces the occurrence of spontaneouspreterm birth in women with a short cervical length

Introduction: The majority of pregnant women with a short cervix will deliver at term and, thus, may unnecessarily receive advanced monitoring and treatment. It is still necessary to define more accurately which sub-population of women with a short cervix is at elevated risk for early delivery. Objective: To determine if vaginal microbiome composition influenced the rate of spontaneous preterm birth in women with a short cervical length. Methods: In an exploratory, observational prospective study, vaginal secretions were obtained from 591 women at 21–24 week gestation. Vaginal microbiome composition was determined by analyzing the V1–V3 region of the bacterial 16S ribosomal RNA gene. Results: Lactobacillus crispatus was numerically dominant in the vagina in 41.7% of subjects, followed by L. iners in 32% and Gardnerella vaginalis in 12%. In women whose cervix was ≤25mm, the sensitivity to predict an spontaneous preterm birth was 11.8%. However, when L. crispatus was not the dominant vaginal bacterium, this sensitivity increased to 81.8%. Similarly, in women with a cervical length ≤30mm, the sensitivity to predict an spontaneous preterm birth increased from 21.7 to 78.3% when L. crispatus was not the dominant vaginal bacterium.In women with a prior spontaneous preterm birth and a cervix ≤25 or ≤30mm, L. crispatus dominance was also associated with a reduced rate of spontaneous preterm birth in the current pregnancy (p&lt;0.001). Conclusion: In pregnant women with a cervix ≤25mm or ≤30mm, the risk for an spontaneous preterm birth is increased if L. crispatus is not dominant in the vagina.

English

The aim of this study is to determine the distribution of the genotypic pathogenicity traits of vaginal high-risk infectious bacteria (HRIB) collected in the CHU-MEL of Cotonou (Benin). To achieve this, a recto-vaginal swab of 42 pregnant women in the third trimester of pregnancy was collected. Species identification was carried out by specific biochemical tests. Antimicrobial susceptibility was tested according to the microbiology standard recommendation. Macrolide resistance genes in Gram-positive bacteria and virulence genes in Escherichia coli were investigated by polymerase chain reaction (PCR). E. coli is the most isolated species (14.7%) followed by Klebsiella pneumoniae (11.8%). Mono-microbial carriage was 55.9%. Gram-negative antibiotic susceptibility shows strong resistance to beta lactam. While Gram-positive bacteria showed strong resistance to beta-lactamine, tetracycline and macrolides with cMLSB (70.4%), iMLSB (3.7%) and M (25.9%) phenotypes. ErmB and ermTR were not detected in Gram-positive bacteria but mef(A/E) was detected at a high. Virulence genes in E. coli were detected and fimA was the most common (52.2%) followed by sfa/foc (30.4%) and cnf1 (13.0%). NeuC and ibeA have not been detected. The hvgA virulence gene was detected in S. agalactiae at a rate of 61.54%. These results demonstrate the importance of introducing antenatal screening for HRIB to improve obstetric and neonatal management in Benin. &nbsp; Key words: Pregnant woman, vaginal swab, neonatal infection, virulence factor.

Presence and Persistence of Putative Lytic and Temperate Bacteriophages in Vaginal Metagenomes from South African Adolescents.

The interaction between gut bacterial and viral microbiota is thought to be important in human health. While fluctuations in female genital tract (FGT) bacterial microbiota similarly determine sexual health, little is known about the presence, persistence, and function of vaginal bacteriophages. We conducted shotgun metagenome sequencing of cervicovaginal samples from South African adolescents collected longitudinally, who received no antibiotics. We annotated viral reads and circular bacteriophages, identified CRISPR loci and putative prophages, and assessed their diversity, persistence, and associations with bacterial microbiota composition. Siphoviridae was the most prevalent bacteriophage family, followed by Myoviridae, Podoviridae, Herelleviridae, and Inoviridae. Full-length siphoviruses targeting bacterial vaginosis (BV)-associated bacteria were identified, suggesting their presence in vivo. CRISPR loci and prophage-like elements were common, and genomic analysis suggested higher diversity among Gardnerella than Lactobacillus prophages. We found that some prophages were highly persistent within participants, and identical prophages were present in cervicovaginal secretions of multiple participants, suggesting that prophages, and thus bacterial strains, are shared between adolescents. The number of CRISPR loci and prophages were associated with vaginal microbiota stability and absence of BV. Our analysis suggests that (pro)phages are common in the FGT and vaginal bacteria and (pro)phages may interact.

Bacterial Profile and Antimicrobial Susceptibility Pattern of Aerobic Vaginal Pathogens in Gynae Patients Visiting to Khyber Teaching Hospital Peshawar

OBJECTIVES: &#x0D; The aim of this study was to determine the prevalent aerobic vaginal bacteria and their antibiogram to commonly prescribed antibiotics for the treatment of aerobic vaginitis (AV).&#x0D; METHODOLOGY:&#x0D; A total of 200 high vaginal swabs (HVS) samples were collected from different AV suspected patients visiting Khyber Teaching Hospital (KTH) and processed for identification of bacterial isolates followed by antibiotic susceptibility patterns as per CLSI protocols.&#x0D; RESULTS:&#x0D; Out of 200 clinical samples, 70 (35%) HVS isolates yielded bacterial growth. Of the isolates, E.coli was the common pathogen 36 (51.4%) followed by S.aureus 20 (28.5%), Enterobacter spp 08 (11.4%), Pseudomonas spp 04 (5.7%) and Citrobacter spp 02 (2.8%). The highest prevalence was observed in the age group of 21-35 years (31.4%) followed by age groups 16-20 years (25.7%) and 26-30 years. S.aureus isolates (n=20) were resistant to ciprofloxacin (90%), cephradine (70%), erythromycin (70%), gentamicin (50%) and cefotaxime (40%) while 1 (5%) of each isolate was resistant to methicillin and vancomycin. Majority of the gram-negative isolates (n=50) were resistant to cotrimoxazole, cephalosporins, quinolones, aminoglycosides and susceptible to carbapenems, tigecycline, sulbactam and tazobactam.&#x0D; CONCLUSION:&#x0D; Aerobic vaginitis should be treated very selectively in order not to kill the beneficial bacteria. Before treating AV, the causative agents should be accurately identified and tested for drug susceptibility patterns and empirical antibiotic therapy should be avoided.