Vaginal Communities Research Articles

Vaginal microbiota in term pregnant women with differences in cervical ripeness revealed by 2bRAD-M

BackgroundCervical ripening is a multifactorial outcome, and the association between cervical ripening and vaginal microbiota remains unexplored in term primiparous women. A new sequencing technology, microbiome 2bRAD sequencing (2bRAD-M) that provides a higher level of species discrimination compared to amplicon sequencing. We applied 2bRAD-M to analyze the vaginal microbiota in a population with variations in cervical ripeness and to explore potential microbiota factors influencing cervical ripening.MethodsA total of 30 full-term primigravid women participated in this study, with 15 belonging to the low scoring group of cervical ripeness and 15 to the high scoring group. Clinical information was collected from the participants, and the vaginal microbiota and community structure of both groups were analyzed using 2bRAD-M sequencing. Microbiota diversity and differential analyses were conducted to explore potential factors influencing cervical ripening.ResultsA total of 605 species were detected. There was no difference in vaginal microbiota diversity between the two groups, and the vaginal microbial composition was structurally similar. In the two groups, Lactobacillus crispatus and Lactobacillus iners were identified as the two pivotal species through random forest analysis. Concurrent, extensive and close connections between species within the two groups were observed in the correlation analysis, influencing the aforementioned two species. Pairwise comparisons showed that Sphingomonas (P = 0.0017) and three others were abundant in high scoring group, while Alloprevotella (P = 0.0014), Tannerella (P = 0.0033), Bacteroides (P = 0.0132), Malassezia (P = 0.0296), Catonella (P = 0.0353) and Pseudomonas (P = 0.0353) and so on showed higher abundance in low scoring group. Linear discriminant analysis effect size identified 29 discriminative feature taxa.ConclusionFor the first time, vaginal microbiota was sequenced using 2bRAD-M. With a relatively simple structure, a more stable vaginal microbiota is associated with higher cervical ripeness, and certain microorganisms, such as Sphingomonas, may play a beneficial role in cervical ripening.

Use of an Ethinyl Estradiol/Etonogestrel Vaginal Ring Alters Vaginal Microbial Communities in Women with HIV.

HIV-1 antiretroviral therapy (ART) alters hormonal contraceptive levels delivered via intravaginal ring (IVR) in a regimen specific manner. We explored the role of the IVR on vaginal microbial communities, vaginal short chain fatty acids (SCFAs), vaginal HIV shedding, and the effect of vaginal microbes on hormone concentrations in cisgender women with HIV (WWH). Vaginal microbes were assessed by 16S RNA sequencing of weekly vaginal swabs, vaginal SCFA by mass spectrometry, HIV-1 shedding by nucleic acid amplification on vaginal aspirates, and bacterial vaginosis by Nugent scoring from 74 participants receiving an etonorgestrel/ethinyl estradiol (ENG/EE) intravaginal ring while on no ART (N=25), efavirenz-based ART (N=25), or atazanavir-based ART (N=24). At baseline, microbial communities of the 64 substudy eligible participants robustly classified as Lactobacillus crispatus--dominant (n=8), L. gasseri-dominant (n=2), L. iners-dominant (n=17), or mixed anaerobic communities (n=37). During IVR therapy, there was an increased probability of Lactobacillus-dominant community state types (CSTs) (odds-ratio=1.61, p=0.04). Vaginal CSTs were associated with Nugent scores. Bacterial vaginosis-associated bacteria were associated with significantly higher and L. iners with lower Nugent Scores (all p adj <0.1). Lactic acid levels were correlated with the relative abundance of Lactobacillus species (r2=0.574; p<0.001). Vaginal shedding of HIV-1 was less common in women with L. crispatus-dominant microbiomes (p=0.04). Mixed anaerobic vaginal communities modulated EE concentrations in a regimen-specific manner. Combined ENG/EE IVR therapy was associated with an increase in Lactobacillus-dominant vaginal microbial communities in WWH and may benefit those with bacterial vaginosis. EE levels were altered by the vaginal microbiota.

Edinburgh postpartum depression scores are associated with vaginal and gut microbiota in pregnancy

BackgroundPrenatal and postpartum depression may be influenced by the composition of host associated microbiomes. As such, the objective of this study was to elucidate the relationship between the human gut or vaginal microbiomes in pregnancy with prenatal or postpartum depression. Methods140 female participants were recruited at their first prenatal visit and completed the Edinburgh Postnatal Depression Scale (EPDS) to screen for depression and anxiety, in addition the EPDS was completed one month postpartum. Vaginal and stool biospecimens were collected in the third trimester, analyzed using 16S rRNA gene sequencing, and assessed for alpha and beta diversity. Individual taxa differences and clustering using the k-medoids algorithm enabled community state type classification. ResultsParticipants with higher postpartum EPDS scores had higher species richness and lower abundance of L. crispatus in the vaginal microbiota compared to those with lower EPDS scores. Participants with a higher prenatal EPDS score had lower species richness of the gut microbiome. Participants with a vaginal community state type dominated by L. iners had the highest mean prenatal EPDS scores, whereas postpartum EPDS scores were similar regardless of prenatal vaginal state type. LimitationsOur small sample size and participant's self-report bias limits generalizability of results. ConclusionsDepression in the prenatal and postpartum period is associated with the composition and diversity of the gut and vaginal microbiomes in the third trimester of pregnancy. These results provide a foundational understanding of the microbial relationships between maternal health and depression for identifying potential therapeutic treatments.

Group B Streptococcus vaginal colonisation throughout pregnancy is associated with decreased Lactobacillus crispatus and increased Lactobacillus iners abundance in the vaginal microbial community.

Group B Streptococcus (GBS) asymptomatically colonises the vagina of up to 40% of pregnant women and can transmit to neonates during birth, causing neonatal pneumonia, sepsis, meningitis, and significant mortality. Vaginal GBS colonisation can be attributed to a range of host and bacterial factors, which may include the composition of the vaginal microbial community. There are few studies that have examined the vaginal community composition in relation to GBS colonisation throughout pregnancy. Here, we performed 16S rRNA sequencing (V3-V4) on vaginal swabs from women at 24- and 36-weeks' gestation, who were GBS culture-negative or GBS culture-positive at either 24 weeks or 36 weeks' gestation or at both timepoints. Vaginal swabs from 93 women were analysed; 46 women were culture-negative, 11 women GBS culture-positive at 24 weeks only, 21 women GBS culture-positive at 36 weeks only and 15 women GBS culture-positive at both timepoints on Brilliance GBS agar. V3-V4 16S rRNA gene amplicon sequencing demonstrated that in women that were GBS culture-positive at 36 weeks gestation only, G. vaginalis was significantly more abundant at 24-weeks' gestation despite a lack of significant changes in community richness between the 24- and 36-week samples. The vaginal microbial communities of women persistently colonised with GBS, had a significantly higher abundance of Lactobacillus iners, compared to other groups where L. crispatus, L. gasseri or L. jensenii were dominant. We have characterised the vaginal microbial community composition during pregnancy in relation to GBS colonisation status, in a longitudinal study for the first time. The most interesting finding was that in women that were persistently colonised with GBS throughout pregnancy, there was a significant increase in L. iners and significant reduction in L. crispatus abundance. Given the lack of detail of the role that the vaginal microbial community plays in GBS colonisation in the literature, it is imperative that the relationship between L. iners and GBS in this unique environmental niche is further investigated.

Abstract C057: Pilot study of vaginal microbiome profiles among Black and White endometrial cancer survivors

Abstract Background: Lower vaginal microbiome diversity, and specifically a Lactobacillus crispatis-dominant vagitype, is associated with more favorable health outcomes. Racial variation in the vaginal microbiome has been reported in some studies, including gynecologic cancers. Racial disparities in endometrial cancer 5-year survival are stark—62% for Black women vs. 83% for White women. Cancer treatments can acutely impact the microbiome of surrounding tissues with potential impacts on quality of life in survivorship, including sexual function. The objectives of this pilot research were to assess the feasibility of self-sampling and variation in vaginal microbiome communities by race, cancer treatment, and sexual function among endometrial cancer survivors. Methods: Endometrial cancer survivors (N=50, mean=11 months from diagnosis) enrolled in the Carolina Endometrial Cancer Study were mailed Genotek vaginal microbiome self-collection kits and the validated Sexual Function-Vaginal Changes Questionnaire (SVQ) before a nurse home visit. Microbiota profiles were characterized by bacteria 16S rRNA amplicon sequencing. Vagitypes were classified as Lactobacillus-dominant and other based on relative abundance (&amp;gt;50% vs. ≤50%) and Ward hierarchical clustering. Results: Overall, 48 vaginal samples were obtained. We excluded samples from women who reported antibiotic use within the last month (N=8) or were missing treatment data (N=6), leaving 34 samples. Of these, 47% were from Black survivors and 53% from White. Cancer treatments were categorized as surgery only (47%) vs. any chemotherapy or radiation (53%); and did not vary by race (p &amp;gt;.9). Black women did not have statistically significant different alpha (Shannon p=0.9) or beta (p=0.4) diversity, or a differential proportion of Lactobacillus dominant vagitype (59% vs. 41%, p=0.3) compared to White women. Alpha (Shannon p=0.04) and beta (p=0.004) diversity varied by cancer treatment with higher microbial diversity after chemotherapy or radiation compared to surgery alone. In the surgery only group, 63% of samples were Lactobacillus dominant, compared to 17% among the chemotherapy or radiation group (p=0.006). We examined cancer treatment associations in analyses restricted to the more favorable endometroid histology (N=25), as surgery alone is associated with endometrioid histology and better prognosis. In this subgroup, 57% (8/14) of women treated with surgery only had Lactobacillus-dominant vagitypes compared to 27% (3/11) in the chemotherapy/ radiation group (p=0.1). Lactobacillus-dominant vagitype was not associated with SVQ scales (global satisfaction; changes in intimacy or interest; dryness; pain). Conclusion: Self-collection of vaginal microbiome samples is feasible in cancer survivors. In our study, Lactobacillus-dominant vagitypes were less abundant among women who were treated with more intensive cancer therapies and did not vary by race or sexual function. Radiation and chemotherapy for endometrial cancer may affect vaginal microbiome composition after treatment ends. Citation Format: Mu Jin, Tope Keku, Nikki McCoy, Jordyn Brown, Jamie Hunter, Victoria Bae-Jump, M. Andrea Azcarate-Peril, Stephanie Engle, Andy Olshan, Hazel B Nichols. Pilot study of vaginal microbiome profiles among Black and White endometrial cancer survivors [abstract]. In: Proceedings of the 17th AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2024 Sep 21-24; Los Angeles, CA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2024;33(9 Suppl):Abstract nr C057.

Abstract C082: Descriptive study of the vaginal microbiome in Hispanic populations in Houston and Puerto Rico with high-risk normal and cervical dysplasia

Abstract Objective Cervical cancer is a leading cause of cancer-related deaths in women worldwide, leading to over 340,000 deaths in 2022. The majority of cervical cancers are caused by the persistent infection of high-risk human papillomavirus (HPV). In the United States, Hispanic women are at increased risk of cervical cancer development and are more likely to die of the disease in certain geographical locations. Furthermore, prior research implicated an association between the vaginal microbiota and the risk of HPV infection and cervical dysplasia. This descriptive study aims to explore the vaginal microbiome profiles of Hispanic or Latino women at risk for HPV-induced cervical dysplasia or with cervical dysplasia residing in Houston and in Puerto Rico. MethodsCervical swab samples were collected from Hispanic patients in Houston IRB 2019-1059 (N = 93) and Puerto Rico IRB 1050411 (N = 118) during routine clinical visits on a prospective collection protocol. Swabs were acquired at the patient’s initial visit and subjected to 16S V4 rRNA gene sequencing for microbiome profiling. Through HPV genotyping via PCR, patients were categorized into three groups: having HPV 16, other high-risk HPV types (18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, and 68), and “other”, comprised of low-risk HPV types, HPV negative, or unknown HPV status. Alpha and beta diversity were assessed using QIIME2. Microbiome composition was evaluated across different locations, HPV status, and disease status, followed by exploring alpha diversity between locations and disease status. The taxa species classification was performed using a custom classifier trained via a cervicovaginal-specific database. Samples were assigned to vaginal community state types (CSTs) using VALENCIA. Results Within the study cohort, 72% (N = 152) were classified as high-risk normal, and 28% (N = 59) had cervical dysplasia. HPV 16 was detected in 12% (N = 26), other high-risk HPV types in 33% (N = 70), while 55% (N = 115) of patients were classified as “other”. At the species level, the vaginal microbiota is dominated by Lactobacillus crispatus, Lactobacillus iners, and Trichomonas vaginalis as the top three abundant, regardless of the patient’s location, HPV status, and disease status. Alpha diversity between locations was 2.2 (1.4) for Houston and 1.6 (1.3) for Puerto Rico, as demonstrated by the Shannon Diversity Index. Alpha diversity was also observed between disease status, with a mean Shannon Diversity Index of 1.9 (1.4) for high-risk normal and 1.9 (1.1) for cervical dysplasia. The top three vaginal community state types were IV-B (32%), I (27%), and III (23%) for Houston, while the top three for Puerto Rico were III (34%), IV-B (31%), and I (21%). Conclusion The study examines the vaginal microbiome profiles of Hispanic women in Houston and Puerto Rico who are at risk for HPV-induced cervical dysplasia or already have cervical dysplasia. These findings provide insights into the vaginal microbiome composition, which plays a crucial role in cervical cancer development. Citation Format: D'Shaunique Walters, Molly El Alam, Timothy Harris, Tatiana Cisneros, Nadim Ajami, Jagannadha Sastry, Josefina Romaguera, Filipa Godoy-Vitorino, Stephanie Dorta-Estremera, Ann Klopp, Lauren Colbert. Descriptive study of the vaginal microbiome in Hispanic populations in Houston and Puerto Rico with high-risk normal and cervical dysplasia [abstract]. In: Proceedings of the 17th AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2024 Sep 21-24; Los Angeles, CA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2024;33(9 Suppl):Abstract nr C082.

A Metagenomics Pipeline to Characterize Self-Collected Vaginal Microbiome Samples.

Vaginitis is a widespread issue for women worldwide, yet current diagnostic tools are lacking. Bacterial vaginosis (BV) is the most prevalent type of vaginitis, found in 10-50% of reproductive-aged women. Current diagnostic methods for BV rely on clinical criteria, microscopy, or the detection of a few microbes by qPCR. However, many vaginal infections lack a single etiological agent and are characterized by changes in the vaginal microbiome community structure (e.g., BV is defined as a loss of protective lactobacilli resulting in an overgrowth of anaerobic bacteria). Shotgun metagenomic sequencing provides a comprehensive view of all the organisms present in the vaginal microbiome (VMB), allowing for a better understanding of all potential etiologies. Here, we describe a robust VMB metagenomics sequencing test with a sensitivity of 93.1%, a specificity of 90%, a negative predictive value of 93.4%, and a positive predictive value of 89.6% certified by Clinical Laboratory Improvement Amendments (CLIA), the College of American Pathologist (CAP), and the Clinical Laboratory Evaluation Program (CLEP). We sequenced over 7000 human vaginal samples with this pipeline and described general findings and comparisons to US census data.

512 Divergence of the sow vaginal microbiome based on fertility status

Abstract The need for alternative biomarkers of reproductive potential in the pork industry is apparent as the current selection approach for sow fertility is not keeping up with the improvement of other production-related traits. To date, the vaginal microbiome has been overlooked as a source of potential biomarkers of sow fertility status. We aimed to elucidate a possible divergence of the vaginal microbiome between sows of differing fertility status and identify possible microbial biomarkers that classify the fertility potential of a sow. Microbial DNA was extracted from vaginal microbiome samples originating from highly reproductive sows (HRP, n = 41) with the number of piglets born alive ≥ 13 or infertile sows (INF, n = 22), that failed to become pregnant after two rounds of consecutive artificial insemination. Our 16S rRNA sequencing results revealed significant (P &amp;lt; 0.05) beta diversity divergence at the family and genus taxonomic levels between HRP and INF vaginal communities. INF vaginal communities displayed significantly greater (P &amp;lt; 0.05) alpha diversity at both the family and genus levels and had a significantly (P &amp;lt; 0.05) greater number of unique OTUs identified at the genus level. The composition of the vaginal microbiome diverges between HRP and INF sows, with INF sows experiencing a disruption of their communities via the colonization of opportunistic pathogens reducing the abundance of normal microbiota seen in HRP. The genera Streptococcus, Lachnospiraceae XPB1014 group, Staphylococcus and Treponema were selected to test as potential biomarkers due to their univariate significance, contribution to partial least squares discriminant analysis (PLS-DA) and selection in a recursive feature elimination model (RFE). The resultant support-vector machine model was diagnostic (achieving an ROC-AUC = 0.86), supporting the selected biomarkers as diagnostic markers for sow fertility status. Overall, there was an apparent shift from a normal vaginal microbiota in HRP sows to a dysbiotic vaginal microbiota in INF sows dominated by potentially pathogenic, more anaerobic and short-chain fatty acid (SCFA)-producing microbiota that may play damaging and pro-inflammatory roles in the vaginal canal. Lactobacillus, known to be the dominant genus in a majority of healthy human vaginal microbiomes, was not the dominating genus in either INF or HRP vaginal microbiomes, suggesting the uniqueness of pig vaginal microbiomes. This also emphasizes the need for pig-specific vaginal microbiome research. Investigations regarding the application of the potential vaginal microbial biomarkers identified in this study to a large, novel population of sow and gilt vaginal microbiome samples to test their on-farm accuracy are underway. In addition, the mechanistic and physiologic role of SCFAs and the bacteria that produce them in the sow vaginal microbiome should be further investigated to understand how they may influence or be a product of fertility status.

Vaginal microbiota, menopause, and the use of menopausal hormone therapy: a cross-sectional, pilot study in Chinese women.

To compare the vaginal microbiota of premenopausal and postmenopausal women and postmenopausal women undergoing menopausal hormone therapy (MHT) and examine the association between vaginal microbiota and genitourinary syndrome of menopause (GSM). This cross-sectional study classified 94 women aged 40 to 60 years into three groups: premenopausal (Pre, n = 32), postmenopausal (Post, n = 30), and postmenopausal women who received MHT orally (Post + MHT, n = 32). Neither the Pre nor the Post group received hormone therapy within the past 6 months. Postmenopausal women who received vaginal MHT were not included. Vaginal swabs were obtained, and microbial composition was characterized by 16S rRNA gene sequencing targeting the V3 to V4 region. Clinical data were collected and serum sex hormones were measured. The most bothersome symptom approach and vaginal health index were used to evaluate GSM. Mann-Whitney U or Kruskal-Wallis ANOVA followed by multiple comparison tests were performed for comparison between or across groups. The correlations between GSM symptom scores and vaginal microbiota were determined using Spearman's correlation analysis. The vaginal community of postmenopausal women was characterized by a decreased abundance of Lactobacillus (Post 18% vs Pre 69%); an increased abundance of several anaerobic bacteria, including Prevotella, Escherichia-Shigella, and Bifidobacterium; and a higher microbial diversity (P < 0.001 for Shannon and Simpson indexes) than those of premenopausal women. The vaginal community of postmenopausal women who received MHT had an increased abundance of Lactobacillus (54%) and lower microbial diversity (P < 0.001 for Shannon and Simpson indexes) than the postmenopausal women. The vaginal microbial community composition of the Pre group shared more similarity with that of the Post + MHT group (Adonis P = 0.051) than with that of the Post group (Adonis P < 0.001). A decreased abundance of Lactobacillus and high diversity in the vaginal community were found in women with moderate to severe GSM symptoms. Among Chinese postmenopausal women, those receiving MHT had higher Lactobacillus abundance but lower abundance of diverse anaerobes and diversity of the vaginal microbial community compared to non-MHT women. MHT in postmenopausal women may potentially contribute to reestablishing vaginal microbiota homeostasis. Findings in this pilot study, however, need to be examined in larger, prospective studies.

Effects of dietary restriction and one-carbon metabolite supplementation during the first 63 days of gestation on the maternal gut, vaginal, and blood microbiota in cattle

BackgroundMaternal diet quality and quantity have significant impacts on both maternal and fetal health and development. The composition and function of the maternal gut microbiome is also significantly influenced by diet; however, little is known about the impact of gestational nutrient restriction on the bovine maternal microbiome during early gestation, which is a critical stage for maternal microbiome-mediated fetal programming to take place. The objective of the present study was to evaluate the impacts of diet restriction and one-carbon metabolite (OCM) supplementation during early gestation on maternal ruminal, vaginal, and blood microbiota in cattle. Thirty-three beef heifers (approx. 14 months old) were used in a 2 × 2 factorial experiment with main factors of target gain (control [CON]; targeted 0.45 kg/d gain vs restricted [RES]; targeted − 0.23 kg/d gain), and OCM supplementation (+ OCM vs − OCM; n = 8/treatment; except n = 9 for RES–OCM). Heifers were individually fed, starting treatment at breeding (d 0) and concluding at d 63 of gestation. Ruminal fluid and vaginal swabs were collected on d − 2, d 35, and d 63 (at necropsy) and whole blood was collected on d 63 (necropsy). Bacterial microbiota was assessed using 16S rRNA gene (V3–V4) sequencing.ResultsOverall ruminal microbiota structure was affected by gain, OCM, time, and their interactions. The RES heifers had greater microbial richness (observed ASVs) but neither Shannon nor Inverse Simpson diversity was significantly influenced by gain or OCM supplementation; however, on d 63, 34 bacterial genera showed differential abundance in the ruminal fluid, with 25 genera enriched in RES heifers as compared to CON heifers. In addition, the overall interaction network structure of the ruminal microbiota changed due to diet restriction. The vaginal microbiota community structure was influenced by gain and time. Overall microbial richness and diversity of the vaginal microbiota steadily increased as pregnancy progressed. The vaginal ecological network structure was distinctive between RES and CON heifers with genera-genera interactions being intensified in RES heifers. A relatively diverse bacterial community was detected in blood samples, and the composition of the blood microbiota differed from that of ruminal and vaginal microbiota.ConclusionRestricted dietary intake during early gestation induced significant alterations in the ruminal microbiota which also extended to the vaginal microbiota. The composition of these two microbial communities was largely unaffected by OCM supplementation. Blood associated microbiota was largely distinctive from the ruminal and vaginal microbiota.

Defining Vaginal Community Dynamics: daily microbiome transitions, the role of menstruation, bacteriophages, and bacterial genes

BackgroundThe composition of the vaginal microbiota during the menstrual cycle is dynamic, with some women remaining eu- or dysbiotic and others transitioning between these states. What defines these dynamics, and whether these differences are microbiome-intrinsic or mostly driven by the host is unknown. To address this, we characterized 49 healthy, young women by metagenomic sequencing of daily vaginal swabs during a menstrual cycle. We classified the dynamics of the vaginal microbiome and assessed the impact of host behavior as well as microbiome differences at the species, strain, gene, and phage levels.ResultsBased on the daily shifts in community state types (CSTs) during a menstrual cycle, the vaginal microbiome was classified into four Vaginal Community Dynamics (VCDs) and reported in a classification tool, named VALODY: constant eubiotic, constant dysbiotic, menses-related, and unstable dysbiotic. The abundance of bacteria, phages, and bacterial gene content was compared between the four VCDs. Women with different VCDs showed significant differences in relative phage abundance and bacterial composition even when assigned to the same CST. Women with unstable VCDs had higher phage counts and were more likely dominated by L. iners. Their Gardnerella spp. strains were also more likely to harbor bacteriocin-coding genes.ConclusionsThe VCDs present a novel time series classification that highlights the complexity of varying degrees of vaginal dysbiosis. Knowing the differences in phage gene abundances and the genomic strains present allows a deeper understanding of the initiation and maintenance of permanent dysbiosis. Applying the VCDs to further characterize the different types of microbiome dynamics qualifies the investigation of disease and enables comparisons at individual and population levels. Based on our data, to be able to classify a dysbiotic sample into the accurate VCD, clinicians would need two to three mid-cycle samples and two samples during menses. In the future, it will be important to address whether transient VCDs pose a similar risk profile to persistent dysbiosis with similar clinical outcomes. This framework may aid interdisciplinary translational teams in deciphering the role of the vaginal microbiome in women’s health and reproduction.9k3PVfrRnyX1V_8jfg2U5DVideo

Development of a Multiplex Real-Time Quantitative PCR Assay for Detecting Vaginal Microbiota in Chinese Women - China, 2021-2022.

The Nugent score, limited by subjectivity and personnel requirements, lacks accuracy. Establishing a precise and simple molecular test is therefore essential for detecting vaginal microbiota compositions and evaluating vaginal health. We evaluated the vaginal health of Chinese women using quantitative polymerase chain reaction (qPCR) to target Lactobacillus crispatus (L. crispatus), L. iners, Gardnerella vaginalis (G. vaginalis), Atopobium vaginae (A. vaginae), and Megasphaera phylotype1. bacterial vaginosis (BV)-related bacteria shared a fluorescent channel. Using 16S rDNA sequencing as a reference standard, we evaluated and validated the diagnostic accuracy of the qPCR assay. Both qPCR and 16S rDNA sequencing demonstrated 90.5% concordance in segregating vaginal community state type (CST), as visualized through heatmaps and PCoA. Spearman's correlation analysis revealed strong correlations between the two methods in calculating the RA of L. crispatus (CST I), L. iners (CST III), and BV-related bacteria (CST IV), with coefficients of 0.865, 0.837, and 0.827, respectively. Receiver operating characteristic analysis showed that qPCR had significant diagnostic accuracy for CST I, CST III, and CST IV (molecular BV), with area under the curve values of 0.967, 0.815, and 0.950, respectively, indicating strong predictive power. Vaginal health can be evaluated using a single qPCR amplification experiment, making the multiplex qPCR assay a highly accurate tool for this purpose.

Evaluation of fecal microbiota of late gestation sows in relation to pelvic organ prolapse risk.

Sow mortality in the U.S. swine industry has increased in recent years, for which pelvic organ prolapse (POP) is a major contributor, accounting for 21% of all sow mortality. Dysbiosis of microbial communities has been associated with disease and reproductive dysfunction in several species, and previous studies have shown changes in vaginal microbiota in sows with increased risk for POP during late gestation. However, there is insufficient knowledge surrounding the potential relationship between fecal microbiota and POP in sows. Therefore, the study objective was to identify differences in sow fecal microbiota and determine if fecal and vaginal microbial communities are correlated in relation to POP risk. Sows were evaluated for POP risk using an established perineal scoring system, with a perineal score (PS) of 1 (PS1) presuming little to no risk of POP to a PS of 3 (PS3) presuming high risk of POP. In the current study, 2,864 sows were scored during gestation week 15, and 1.0%, 2.7%, and 23.4% of PS1, PS2, and PS3 sows, respectively, subsequently experienced POP. Fecal swabs (n = 215) were collected between gestation days 108-115, DNA was extracted, and 16S rRNA gene amplicon sequencing libraries were analyzed using mothur, phyloseq and SAS in reference to PS and POP outcome. Additionally, co-occurrence networks were constructed using CoNet to compare fecal and vaginal microbiota from the same cohort of sows and identify correlations between different taxa. Differences in fecal community composition (PERMANOVA; P < 0.05), structure (alpha diversity measurements; P < 0.05), and 13 individual operational taxonomic units (OTUs) were revealed between PS1 and PS3 assigned sows. No differences in fecal microbiota were detected as a result of POP outcome. However, the abundances of several taxa were correlated across sample collection sites, suggesting the fecal and vaginal microbial communities may be related to one another. Collectively, fewer differences in the fecal microbiota exist in sows with differing risk for POP compared to the vaginal microbiota, suggesting the vaginal microbiome may be more relevant in relation to POP outcome, although correlations between fecal and vaginal communities may provide insight for strategies to combat POP.

Evidence of Lactobacillus strains shared between the female urinary and vaginal microbiota.

Lactobacillus species are common inhabitants of the 'healthy' female urinary and vaginal communities, often associated with a lack of symptoms in both anatomical sites. Given identification by prior studies of similar bacterial species in both communities, it has been hypothesized that the two microbiotas are in fact connected. Here, we carried out whole-genome sequencing of 49 Lactobacillus strains, including 16 paired urogenital samples from the same participant. These strains represent five different Lactobacillus species: L. crispatus, L. gasseri, L. iners, L. jensenii, and L. paragasseri. Average nucleotide identity (ANI), alignment, single-nucleotide polymorphism (SNP), and CRISPR comparisons between strains from the same participant were performed. We conducted simulations of genome assemblies and ANI comparisons and present a statistical method to distinguish between unrelated, related, and identical strains. We found that 50 % of the paired samples have identical strains, evidence that the urinary and vaginal communities are connected. Additionally, we found evidence of strains sharing a common ancestor. These results establish that microbial sharing between the urinary tract and vagina is not limited to uropathogens. Knowledge that these two anatomical sites can share lactobacilli in females can inform future clinical approaches.

Vaginal anaerobes are associated with cervicitis: A case-control study

Cervicitis is associated with important reproductive sequelae. Primary causes include chlamydia and gonorrhoea, but a known sexually transmitted infection (STI) is not identified in >50% of cases (i.e. STI-negative cervicitis). Bacterial vaginosis (BV) and specific BV-associated bacteria have also been associated with cervicitis, but data are limited. We investigated the association between STI-negative cervicitis and vaginal microbiota composition. This was a case-control sub-study of the OhMG study conducted at the Melbourne Sexual Health Centre. Cases were women with cervicitis who tested negative for STIs (STI-negative cervicitis, n=64). Controls were STI-negative asymptomatic women attending for STI-screening (n=128). The vaginal microbiota was characterised using 16S rRNA gene sequencing. Vaginal community state types were compared between cases and controls using logistic regression. Differential abundance analysis was performed to identify taxa associated with STI-negative cervicitis. STI-negative cervicitis cases were more likely than controls to have a Lactobacillus-deficient non-optimal microbiota (adjusted-odds-ratio 2.55, 95% CI 1.18-5.50). Compared to controls, cases had increased abundance of four BV-associated bacteria (Gardnerella, Fannyhessea vaginae, Prevotella bivia, Dialister micraerophilus) and decreased abundance of optimal lactobacilli. We report a positive association between non-optimal vaginal microbiota composition and STI-negative cervicitis. Specific anaerobic BV-associated bacteria may represent infectious causes of cervicitis.

Longitudinal profiles of the vaginal microbiota of pre-, peri-, and postmenopausal women: preliminary insights from a secondary data analysis.

Menopause is often accompanied by lowered Lactobacillus spp. relative abundance and increased abundance of diverse anaerobic/aerobic bacteria in the vaginal microbiota due in part to declines in estrogen. These microbiota are associated with urogenital symptoms and infections. In premenopause, vaginal microbiota can fluctuate rapidly, particularly with menstrual cycles and sexual activity; however, the longitudinal dynamics of vaginal microbiota are understudied in peri- and postmenopause. We described vaginal community stability across reproductive stages. Pre- (n = 83), peri- (n = 8), and postmenopausal (n = 11) participants provided twice-weekly mid-vaginal samples (total, 1,556; average, 15 per participant) over 8 weeks in an observational study. Composition of the vaginal microbiota was characterized by 16S rRNA gene amplicon sequencing, and a community state type (CST) was assigned to each sample. Clustering of longitudinal CST profiles, CST transition rates, duration of low-Lactobacillus/high bacterial diversity CSTs, and other metrics of bacterial community dynamics were assessed across reproductive stages. The proportion of participants with longitudinal CST profiles characterized by low-Lactobacillus CSTs was similar among pre- (38.6%), peri- (37.5%), and postmenopausal (36.4%) participants (P = 0.69). CST transition rates between consecutive samples were 21.1%, 16.7%, and 14.6% for pre-, peri-, and postmenopausal participants, respectively (P = 0.49). Low-Lactobacillus CST tended to persist for at least 4 weeks, irrespective of reproductive stage. Findings from this small yet frequently sampled cohort revealed vaginal bacterial fluctuations over 8 weeks that were similar across reproductive stages. Larger and longer-term studies based on these preliminary data could provide insights into the influence of microbiota dynamics on urogenital outcomes during menopause.

The interrelation between microbial immunoglobulin coating, vaginal microbiota, ethnicity, and preterm birth

BackgroundVaginal microbiota composition is associated with spontaneous preterm birth (sPTB), depending on ethnicity. Host-microbiota interactions are thought to play an important underlying role in this association between ethnicity, vaginal microbiota and sPTB.MethodsIn a prospective cohort of nulliparous pregnant women, we assessed vaginal microbiota composition, vaginal immunoglobulins (Igs), and local inflammatory markers. We performed a nested case–control study with 19 sPTB cases, matched based on ethnicity and midwifery practice to 19 term controls.ResultsOf the 294 included participants, 23 pregnancies ended in sPTB. We demonstrated that Lactobacillus iners-dominated microbiota, diverse microbiota, and ethnicity were all independently associated with sPTB. Microbial Ig coating was associated with both microbiota composition and ethnicity, but a direct association with sPTB was lacking. Microbial IgA and IgG coating were lowest in diverse microbiota, especially in women of any ethnic minority. When correcting for microbiota composition, increased microbial Ig coating correlated with increased inflammation.ConclusionIn these nulliparous pregnant women, vaginal microbiota composition is strongly associated with sPTB. Our results support that vaginal mucosal Igs might play a pivotal role in microbiota composition, microbiota-related inflammation, and vaginal community disparity within and between ethnicities. This study provides insight in host-microbe interaction, suggesting that vaginal mucosal Igs play an immunomodulatory role similar to that in the intestinal tract.-XF_JK898P7idx1NXSVZ7eVideo

Host factors are associated with vaginal microbiome structure in pregnancy in the ECHO Cohort Consortium

Using pooled vaginal microbiota data from pregnancy cohorts (N = 683 participants) in the Environmental influences on Child Health Outcomes (ECHO) Program, we analyzed 16S rRNA gene amplicon sequences to identify clinical and demographic host factors that associate with vaginal microbiota structure in pregnancy both within and across diverse cohorts. Using PERMANOVA models, we assessed factors associated with vaginal community structure in pregnancy, examined whether host factors were conserved across populations, and tested the independent and combined effects of host factors on vaginal community state types (CSTs) using multinomial logistic regression models. Demographic and social factors explained a larger amount of variation in the vaginal microbiome in pregnancy than clinical factors. After adjustment, lower education, rather than self-identified race, remained a robust predictor of L. iners dominant (CST III) and diverse (CST IV) (OR = 8.44, 95% CI = 4.06–17.6 and OR = 4.18, 95% CI = 1.88–9.26, respectively). In random forest models, we identified specific taxonomic features of host factors, particularly urogenital pathogens associated with pregnancy complications (Aerococcus christensenii and Gardnerella spp.) among other facultative anaerobes and key markers of community instability (L. iners). Sociodemographic factors were robustly associated with vaginal microbiota structure in pregnancy and should be considered as sources of variation in human microbiome studies.

Relationship Between Vaginal and Gut Microbiome on Stress and Depression in Pregnancy [ID 2683607

INTRODUCTION: Pregnancy is a period of significant stress and weight gain; however, the effect of the vaginal and gut microbiome on pregnancy, depression, stress, and the neonate is not completely understood. METHODS: An IRB-approved prospective cohort study was performed for pregnant individuals and their neonates. The Perceived Stress Scale (PSS) and Center for Epidemiologic Studies Depression Scale (CES-D) surveys were administered in the antepartum and postpartum patient, vaginal and fecal microbiome samples were collected, and neonate umbilical cord blood and fecal specimens were also collected. Using full-length 16S rRNA sequencing, fecal and vaginal community composition was characterized. Additionally, possible covariates such as body mass index, cohabitation status, educational attainment, dietary intake, public/private insurance status, chronic health conditions, and pregnancy complications were captured. RESULTS: The study sample consisted of 35 pregnant individuals, average age 30 years, and half primiparous. Increased early pregnancy stress (PSS greater than or equal to 14) was associated with increased abundance of fecal taxa, and increased stress from late pregnancy through postpartum was associated with increased abundance of fecal microbial pathogens. Depressed participants (CES-D greater than or equal to 16) experienced steeper decreases in prenatal vaginal microbiome diversity. In late pregnancy, maternal stress and depression scores were associated with elevated maternal inflammatory markers (CCL2). At delivery, neonatal umbilical CCL2 concentration was negatively correlated with relative abundance of maternal fecal lactobacilli. CONCLUSION: These findings highlight the relationship between stress and depression in pregnancy, the microbiome of the pregnant patient, and inflammatory markers in the pregnant patient and the neonate.

Menstrual Blood as a Diagnostic Specimen for Human Papillomavirus Genotyping and Genital Tract Infection Using Next-Generation Sequencing as a Novel Diagnostic Tool.

Menstrual blood (MB) is a convenient specimen type that can be self-collected easily and non-invasively by women. This study assessed the potential application of MB as a diagnostic specimen to detect genital tract infections (GTIs) and human papillomavirus (HPV) infections in women. Genomic DNA was extracted from MB samples. Pacific Bioscience (Pacbio) 16S ribosomal DNA (rDNA) high-fidelity (HiFi) long-read sequencing and HPV PCR were performed. MB samples were collected from women with a pathological diagnosis of CIN1, CIN2, CIN3 or HPV infection. The sensitivity and positive predictive value (PPV) of high-risk HPV detection using MB were found to be 66.7%. A shift in vaginal flora and a significant depletion in Lactobacillus spp. in the vaginal microbiota communities were observed in the MB samples using 16S rDNA sequencing. In this study, we demonstrated that MB is a proper diagnostic specimen of consideration for non-invasive detection of HPV DNA and genotyping using PCR and the diagnosis of GTIs using metagenomic next-generation sequencing (mNGS). MB testing is suitable for all women who menstruate and this study has opened up the possibility of the use of MB as a diagnostic specimen to maintain women's health.