Composition Of Vaginal Microbiome Research Articles

Effects of Dietary Quality on Vaginal Microbiome Composition Throughout Pregnancy in a Multi-Ethnic Cohort

Background: Vaginal Lactobacillus predominance is associated with improved vaginal health and reduced pregnancy complications. Little is known about how dietary quality may improve vaginal microbial composition or about dietary interventions that may promote Lactobacillus abundance. To understand the host factors affecting vaginal microbiota during pregnancy in a multi-ethnic cohort in Hawai`i. We hypothesize that better diet quality improves vaginal microbial composition, as represented by Lactobacillus abundance and depletion of anaerobic organisms. Methods: We compared comprehensive diet quality, as measured by the Healthy Eating Index-2015 (HEI-2015), to taxonomic classifications of bacteria present within the vagina. Participants of the four predominant ethnic groups in Hawai’i (Japanese, Filipino, Non-Hispanic White and Native Hawaiian) completed Quantitative Food Frequency Questionnaires (QFFQs) and collected vaginal swabs during each trimester. 16s rRNA amplicon sequencing (V2–V9 regions) was performed on vaginal samples. HEI-2015 scores and macro- and micronutrient intake were compared with the predominant species present using the Mann–Whitney-U test, PERMANOVA, and the Pearson correlation coefficient. A mixed-effects logistics regression model was used to predict the depletion of Lactobacillus species while accounting for confounding demographic factors. Results: Matched, longitudinal data for 40 participants demonstrated three predominant Lactobacillus species: L. crispatus, L. iners, and L. gasseri, with another subset of samples with anaerobic abundance. (Primarily, Atopobium vaginae, Prevotella, and Gardnerella vaginalis.) Non-Hispanic White participants had lower amounts of Lactobacillus iners compared to other racial and ethnic groups. HEI scores correlated with the chao index and observed species number primarily in the first trimester (r = 0.25, p < 0.05). Greater carbohydrate intake was associated with a higher abundance of L. crispatus, while lower carbohydrate intake trended towards more L. iners (0.056) and anaerobic species. Conclusions: Increased carbohydrate consumption and improved diet quality may be associated with beneficial vaginal microbial composition in pregnancy.

Abstract C017: Race-Specific Patterns of Vaginal Microbiome and HPV Infections Predict Risk of Precancerous Cervical Lesions

Abstract Background: Our previous research indicated that high-grade squamous intraepithelial lesions (HSIL), precursors to cervical cancer (CCa), are influenced by the vaginal microbiome (VMB) differentially by self-reported race. VMB enriched for certain anti-inflammatory Lactobacillus species confer protection against the risk of HSIL for white, but not Black women. Black women exhibit greater VMB diversity (associated with HPV tumorigenesis) and a lower prevalence of anti-inflammatory Lactobacillus species. But not all Black women with diverse VMB develop HSIL. Objective: Given HPV's essential role in cervical cancer, we aimed to assess the influence of sociodemographic, clinical, and microecological HPV attributes on HSIL risk within the context of the vaginal microbiome's variation by race. Methods: The VMB and HPV profiles of 1,168 women were analyzed alongside over 100 sociodemographic and clinical variables. Spearman’s correlation, Point-Biserial Correlation, chi-squared tests, and machine learning models, were employed to evaluate associations between sociodemographic, clinical, and microecological variables with HSIL, with a particular focus on the impact of race on these relationships. Results: The cohort was predominantly Black (72%), showing no significant race-based differences in HPV positivity (45%). However, HSIL incidence was nearly twice as high in Black (7%) compared to White women (4%). Women with HSIL had a higher prevalence of non-Lactobacillus-dominant VMB (68% vs. 51%) and were more likely to be HPV+ (73% vs. 44%) than those without HSIL. Co-infection with multiple HPV strains was associated with a higher risk of CIN3. Specifically, HPV16, HPV56, HPV58, HPV33, and HPV42 infections were significantly associated with higher CIN3 risks. The random forest model identified the relative abundance of Lactobacillus crispatus as the most essential factor in predicting CIN3 risk, followed by the number of HPV strains per sample. However, differential abundance analysis specifically indicated lower L. crispatus levels were associated with HSIL in white but not Black women. Conversely, HPV abundance was only associated with HSIL in Black women. Additionally, Black patients with 2 or more HPV co-infections had a higher likelihood of being diagnosed with CIN3+ compared to white counterparts. ML models highlighted different HPV-specific HSIL predictors by race: HPV number, and HPV 56, 16, and 33 in Blacks; sexual partners, HPV 90, 56, and 16 in whites. Conclusions: Our findings reveal race-specific interplays among individual risk factors, HPV infection, and VMB composition in HSIL development. The identification of key microbial and viral predictors highlights the importance of personalized approaches in preventing and managing HSIL, particularly among racially diverse populations. Citation Format: Katherine Y. Tossas, Bin Zhu, Katarzyna Tyc, Myrna Serrano, Jerome Strauss, Gregory Buck. Race-Specific Patterns of Vaginal Microbiome and HPV Infections Predict Risk of Precancerous Cervical Lesions [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 C017.

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.

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 (>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 >.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.

The In Vivo Effect of Water-Based Lubricants on the Vaginal Microbiome of Women from Varying Age Groups: Exploratory Analysis of a Randomized Controlled Trial.

Vaginal mucosa undergoes physiological changes across the lifespan, such as increased pH and reduced natural lubrication which are known to impact vaginal commensal microorganisms, hence increasing the chances of vaginal infections. An improved understanding of vaginal microbiome composition in different age groups and the effect of social behaviors, such as the use of personal lubricants, could facilitate the development of new strategies to maintain good vaginal health. The objective of this study was to assess the effect of water-based lubricants on the human vaginal microbiome. Fifty females from three age groups (18-29, 30-44, and 45-65 years) with mild-to-moderate vaginal dryness were randomized to one of five lubricants (four of which were formulated to meet expert guidance on osmolality and pH). Subjects entered the study at tolerance or treatment phase (vaginal intercourse minimum once a week using assigned lubricant). Four vaginal swabs per participant were sampled during pre-("baseline"), post-first ("2 h", "24 h") and post-last ("4 weeks") lubricant application to assess bacterial and fungal diversity via amplicon sequencing. Vaginal pH and relative humidity were measured at baseline, 2 h, and 24 h post-lubricant application. Relative bacteriome abundance was statistically compared between timepoints for each lubricant group. Vaginal moisture, age, BMI, and pH were correlated with bacteriome relative abundance. Lactobacilli and Gardnerella sp. Were predominant across participants. Repeated lubricant application did not significantly alter the vaginal bacteriome during 4 weeks of product use (p > 0.05) when measured by relative abundance and alpha-diversity index. Bacteriome diversity and abundance differed significantly between age groups at baseline whereas lactobacilli relative abundance was negatively associated with age and vaginal pH.

Effect of Waiting Period on Initial Adverse Vaginal Microbiome Composition in IVF-ICSI Patients.

Background/Objectives: In this observational prospective cohort study, conducted at the Fertility Centre of the University Hospital, Duesseldorf Germany, the spontaneous reversal capacity and the effect of waiting time on an adverse vaginal microbiome profile in subfertile patients were investigated. Methods: Vaginal swabs of 76 patients were obtained before starting a fertility treatment using a commercially available test to perform a microbiome analysis. Patients with a favorable microbiome profile ("medium" or "high profile") according to the manufacturer's algorithm proceeded with the fertility treatment. Patients with an unfavorable microbiome profile ("low profile") postponed their fertility treatment and were sampled up to four times in each successive cycle or until a shift to a more favorable profile was detected. Results: Initially, 54/76 subjects had a high or medium profile and 23/76 had a low profile. Within 3 months, 75% of patients with an initial low profile shifted to a more favorable profile (7/23 dropouts). The presence of Lactobacillus crispatus in the initial sample was associated with a higher likelihood of a spontaneous shift to a more favorable profile. Conclusions: The vaginal microbiome is subject to strong fluctuations. Even an unfavorable microbiome profile can develop into a favorable microbiome profile within a few months without treatment.

O-276 Implantation failure is not associated with seven different vaginal bacterial community-state types; however, a distinct signature, including Ureaplasma parvum, is strongly linked to treatment failure

Abstract Study question Is microbial diversity or vaginal bacterial composition associated with embryonic implantation in women undergoing frozen-thawed embryo transfer? Summary answer While global diversity measures of microbial composition do not correlate with positive implantation, specific taxa may serve as predictors of treatment outcomes. What is known already Clinical studies have indicated a reduced likelihood of embryo implantation in women diagnosed with microbial dysbiosis. The application of Next-Generation Sequencing for microbiota analysis has significantly advanced our understanding of vaginal microbiome composition. Recent studies have proposed distinct vaginal microbiome patterns as potential predictors of IVF treatment success or failure. However, the evidence regarding the biological basis of test algorithms (such as the independent association of distinct community state types with outcomes) and the reliability of diagnostic test performance (including replicability and external validity) in predicting implantation success or failure in women undergoing embryo transfer remains less robust. Study design, size, duration In a prospective, single-centre observational cohort study (05/2017-03/2019, NCT03507673, local ethical approval reference no.18-005), conducted at a university-affiliated centre for reproductive medicine in Germany, vaginal swab samples were collected from infertile, asymptomatic female patients (n = 260), aged 18-45, undergoing frozen-thawed embryo transfer. Patients, each contributing a single treatment cycle, received sequential oral estradiol and oral estradiol/dydrogesterone in an HRT regimen for FET. Vaginal swabs were taken from baseline visit before transfer day and stored at –80 °C. Participants/materials, setting, methods 16S rRNA gene amplicon sequencing was performed targeting V3/V4 hypervariable region using Illumina MiSeq platform. Raw data assembly, filtering and taxonomic assignment were performed using mothur against EzBioCloud database. Statistical analyses, and visualization were conducted using R. Community structure via hierarchical clustering, alpha and beta diversity, and indicator species via LefSe analysis were assessed. Main results and the role of chance Based on a hierarchical clustering using the 25 most abundant bacterial taxa, seven separate community state types (CSTs) being either dominated by species of the genus Lactobacillus (CST-A, CST-B, CST-C, CST-E, CST-G) or Gardnerella (CST-F) or presenting a variable community composition (CST-D) were identified. A community state type dominated by Lactobacillus fornicalis was found in a small fraction of women and enhanced resolution among the Gardnerella genus impacts CST definitions in this sample. Embryonic implantation had no significant association to either alpha diversity measures or CST distribution; likewise, the previously suggested ratio of relative abundances of Lactobacillus iners vs. Lactobacillus crispatus was not associated with implantation. Lactobacillus fornicalis is positively linked to implantation, while Lactobacillus iners and Ureaplasma parvum were linked to implantation failure: above 50% relative abundance of Lactobacillus fornicalis, was associated with 75% positive implantation. Conversely, just 0.5% relative abundance of Ureaplasma parvum was associated with a decrease in implantation rate to approximately 20%. LefSe and binary regression analysis identified a relatively rare (6% in our sample) microbiome signature, mostly from CST-C (Lactobacillus iners dominated) with a very low likelihood of implantation, with Ureaplasma parvum as the common denominator. Limitations, reasons for caution Studies on the vaginal microbiome in IVF face issues such as study design variability, small sample-sizes, limited control for temporal dynamics, and patient heterogeneity. Attempts at establishing causal relationships through interventional studies in IVF have been limited and inconclusive. Understanding the microbiome’s functional aspects is crucial for reproductive outcome elucidation. Wider implications of the findings Current study findings don’t confirm previously reported associations between common vaginal bacterial CSTs and outcome. Therefore, screening for vaginal microbiome composition in asymptomatic women with subfertility is not recommended outside research. Signatures/taxae with an independent and strong negative association with implantation are likely to be rare yet deserve further attention. Trial registration number NCT03507673

(129) WOMEN WHO REPORT PAIN WITH SEX HAVE VARYING MICROBIOME PROFILES BASED ON AGE

Abstract Introduction Even though the vaginal microbiome (VM) is a known contributor to vaginal epithelial health, the VM of women who report pain with sex has not been characterized. It is known that the VM shifts during menopause, therefore we segmented out analysis based on menopausal status. Objective An observational analysis comparing the vaginal microbiome composition and symptom profiles in individuals of both menopausal and reproductive age who experience moderate to severe pain with sex and those that do not report pain with sex. Methods This study involved the collection of self-reported health profiles and shotgun metagenomic analysis of vaginal samples from two distinct demographic groups: menopausal women and women of reproductive age (defined as non-pregnant, non-menopausal, and aged under 40 years). The focus was on identifying the microbial composition in individuals experiencing moderate to severe sexual intercourse pain and those without any reported pain, excluding samples from individuals with mild pain. An additional criterion for inclusion was a reported history of sexual activity within the preceding 30 days. The menopausal group comprised 157 individuals reporting pain and 367 individuals without pain. In the reproductive age cohort, there were 968 participants with pain and 968 age-matched controls without pain. Statistical analysis between the pain and control groups in both cohorts revealed no significant differences in body mass index (BMI, menopause p = 0.74, reproductive p = 0.19), reported hormone use (menopause p = 0.2, reproductive p = 0.24), or history of previous infections (menopause p = 0.29, reproductive p = 0.09). Results In both the menopausal and reproductive-aged cohorts, individuals experiencing pain during sexual intercourse exhibited a higher prevalence of symptomatic conditions, as illustrated in Figure 1 A & B. Reported symptoms included increased severity of burning, itchiness, vaginal and vulvar pain, erythema, dysuria, and vaginal dryness. A significant correlation was observed between these enhanced symptomatology and variations in vaginal microbiome compositions among the menopausal group, as depicted in Figure 2 A & B. Specifically, pain during sexual intercourse in menopausal women was linked to elevated vaginal microbiota (VM) diversity (p=0.047) and a reduction in lactobacilli levels (p=0.06). Conversely, in the reproductive-aged cohort, a higher VM diversity was noted in the control group (p=0.014), with no significant differences in Lactobacillus or Gardnerella concentrations between those experiencing pain and the control group. Conclusions Preliminary examination of the vaginal microbiome in women who self-report moderate to severe pain with sex reveals significant differences, particularly in menopausal women experiencing pain. These individuals exhibited a more diverse vaginal microbiome and lower levels of Lactobacillus. This study underscores the need for further research to explore the potential of targeted microbiome modifications as a therapeutic approach to alleviate pain symptoms in this demographic. Disclosure Yes, this is sponsored by industry/sponsor: Evvy. Clarification: Industry initiated, executed and funded study. Any of the authors act as a consultant, employee or shareholder of an industry for: Evvy.

The vaginal microbiome composition during pregnancy in a region compromising different ethnic origins.

The vaginal microbiota plays a significant role in pregnancy outcomes and newborn health. Indeed, the composition and diversity of the vaginal microbiota can vary among different ethnic groups. Our study aimed to investigate the composition of the vaginal microbiome throughout the three trimesters of pregnancy and to identify any potential variations or patterns in the Turkish population compromising mixed ethnicities. We conducted a longitudinal study to characterize the vaginal microbiota of pregnant women. The study included a total of 25 participants, and the samples were collected at each trimester: 11-13weeks, 20-24weeks and 28-34weeks gestation. Lactobacillus species were consistently found to be dominant in the vaginal microbiota throughout all trimesters of pregnancy. Among Lactobacillus species, L. crispatus had the highest abundance in all trimesters (40.6%, 40.8% and 44.4%, respectively). L. iners was the second most prevalent species (28.5%, 31% and 25.04, respectively). Our findings reveal that the dominant composition of the vaginal microbiota aligns with the CST-type I, commonly observed in the European population. This suggests that there are shared mechanisms influencing the microbial communities in the vagina, which are likely influenced by factors such as genetics, lifestyle, and cultural behaviors rather than ethnicity alone. The complex interplay of these factors contributes to the establishment and maintenance of the vaginal microbiota during pregnancy. Understanding the underlying mechanisms and their impact on vaginal health across diverse populations is essential for improving pregnancy outcomes. The study was approved by the Koc University Ethical Committee (no:2019.093.IRB2.030) and registered at the clinical trials.

Vaginal microbiota are associated with in vitro fertilization during female infertility.

The vaginal microbiome plays an essential role in the reproductive health of human females. As infertility increases worldwide, understanding the roles that the vaginal microbiome may have in infertility and in vitro fertilization (IVF) treatment outcomes is critical. To determine the vaginal microbiome composition of 1411 individuals (1255 undergoing embryo transplantation) and their associations with reproductive outcomes, clinical and biochemical features are measured, and vaginal samples are 16S rRNA sequenced. Our results suggest that both too high and too low abundance of Lactobacillus is not beneficial for pregnancy; a moderate abundance is more beneficial. A moderate abundance of Lactobacilluscrispatus and Lactobacillus iners (~80%) (with a pregnancy rate of I-B: 54.35% and III-B: 57.73%) is found beneficial for pregnancy outcomes compared with a higher abundance (>90%) of Lactobacillus (I-A: 44.81% and III-A: 51.06%, respectively). The community state type (CST) IV-B (contains a high to moderate relative abundance of Gardnerellavaginalis) shows a similar pregnant ratio (48.09%) with I-A and III-A,and the pregnant women in this CST have a higher abundance of Lactobacillus species. Metagenome analysis of 71 samples shows that nonpregnant women are detected with more antibiotic-resistance genes, and Proteobacteria and Firmicutes are the main hosts. The inherent differences within and between women in different infertility groups suggest that vaginal microbes might be used to detect infertility and potentially improve IVF outcomes.

Interactions between maternal parity and feed additives drive the composition of pig gut microbiomes in the post-weaning period

BackgroundNursery pigs undergo stressors in the post-weaning period that result in production and welfare challenges. These challenges disproportionately impact the offspring of primiparous sows compared to those of multiparous counterparts. Little is known regarding potential interactions between parity and feed additives in the post-weaning period and their effects on nursery pig microbiomes. Therefore, the objective of this study was to investigate the effects of maternal parity on sow and offspring microbiomes and the influence of sow parity on pig fecal microbiome and performance in response to a prebiotic post-weaning. At weaning, piglets were allotted into three treatment groups: a standard nursery diet including pharmacological doses of Zn and Cu (Con), a group fed a commercial prebiotic only (Preb) based on an Aspergillus oryzae fermentation extract, and a group fed the same prebiotic plus Zn and Cu (Preb + ZnCu).ResultsAlthough there were no differences in vaginal microbiome composition between primiparous and multiparous sows, fecal microbiome composition was different (R2 = 0.02, P = 0.03). The fecal microbiomes of primiparous offspring displayed significantly higher bacterial diversity compared to multiparous offspring at d 0 and d 21 postweaning (P < 0.01), with differences in community composition observed at d 21 (R2 = 0.03, P = 0.04). When analyzing the effects of maternal parity within each treatment, only the Preb diet triggered significant microbiome distinctions between primiparous and multiparous offspring (d 21: R2 = 0.13, P = 0.01; d 42: R2 = 0.19, P = 0.001). Compositional differences in pig fecal microbiomes between treatments were observed only at d 21 (R2 = 0.12, P = 0.001). Pigs in the Con group gained significantly more weight throughout the nursery period when compared to those in the Preb + ZnCu group.ConclusionsNursery pig gut microbiome composition was influenced by supplementation with an Aspergillus oryzae fermentation extract, with varying effects on performance when combined with pharmacological levels of Zn and Cu or for offspring of different maternal parity groups. These results indicate that the development of nursery pig gut microbiomes is shaped by maternal parity and potential interactions with the effects of dietary feed additives.

VagiBIOM Lactobacillus suppository improves vaginal health index in perimenopausal women with bacterial vaginosis: a randomized control trial

Bacterial vaginosis (BV) can cause vaginal dysbiosis that may influence general vaginal health and pregnancy complications. Balancing vaginal microbiome using Lactobacillus spp. may be a new way to prevent and treat mild BV. We conducted a randomized, double-blind, placebo-controlled pilot study aimed at evaluating the effect of the product VagiBIOM, a multi-Lactobacillus vaginal suppository, on peri- and premenopausal women with BV in restoring vaginal pH and overall vaginal health by resetting the vaginal microbiome composition. Sixty-six peri- and premenopausal women with BV symptoms were randomized with a 2:1 ratio to be treated with VagiBIOM or placebo suppositories. Vaginal pH, VAS itching score, total Nugent score, and vaginal health index (VHI) were measured. Vaginal microbiome changes before and after the treatment were analyzed by 16S rRNA sequencing and bioinformatics analysis. After 4 weeks of intervention with VagiBIOM or a placebo, the mean score for vaginal pH, VAS itching, and total Nugent score was significantly decreased from the baseline. Compared to the baseline scores, the VHI scores improved significantly following 28-day intervention (p < 0.001). Our results revealed two Lactobacillus species, L. hamsteri, and L. helveticus, as indicator species occurring differentially in the VagiBIOM-treated group. Furthermore, the regression and species network analyses revealed significant bacterial associations after VagiBIOM treatment. Lactobacillus hamsteri was positively associated with the Nugent score and negatively associated with vaginal pH. L. iners and L. salivarius were positively and inversely associated with VHI. As is typical, Bacteroides fragilis was positively associated with vaginal pH and negatively associated with the Nugent score. Interestingly, the Lactobacillus spp. diversity improved after VagiBIOM treatment. The VagiBIOM suppository treatment for peri- and premenopausal women with BV significantly relieved vaginal itching by decreasing vaginal pH and Nugent scores and improving the overall VHI after 4 weeks’ intervention. This effect was primarily the result of VagiBIOM improving vaginal Lactobacillus diversity.Trial Registration ClinicalTrials.gov registration: NCT05060029, first registration 09/28/2021: Title: A Pilot Study to Evaluate the Efficacy and Safety of Lactobacillus Species Suppositories on Vaginal Health and pH.

Abstract B127: The impact of sociodemographic and lifestyle factors on the vaginal microbiome and cervical cancer disparities in Native American women

Abstract Cervical cancer morbidity and mortality are continually experienced by Native American women compared to white women. Human papillomavirus (HPV) is the most common sexually transmitted infection and can cause cervical cancer in women when HPV is not cleared by an individual. Depletion of Lactobacillus species in the vaginal microbiome (VMB) is linked to HPV acquisition and persistence of HPV infection and development/progression of cervical neoplasm. Most Lactobacillus spp. protect the host against invading pathogens of the genital mucosa, including the cervix. Changes in the VMB composition to dysbiotic anaerobes decrease an individual’s ability to clear HPV, increasing susceptibility to malignancy. Depletion of Lactobacilli is associated with race and ethnicity, despite research showing genetics to be statistically similar across ethnicities. However, there may be other factors that drive changes in the microbiome, including demographic, socio-economic, lifestyle and sexual &amp; reproductive health factors. Here, we aim to determine which demographic, socioeconomic, lifestyle and sexual &amp; reproductive health factors impact the microbial community within the VMB, regardless of race and ethnicity. These factors have not been studied in the context and inclusion of NA women. In this study, 31 women at the Native American for Community Action Clinic in Flagstaff, Arizona, U.S. were enrolled, and participants completed self-reported surveys. The survey assessed the individual’s socioeconomic, demographics, lifestyle, reproductive health, as well as their knowledge on HPV and VMB. Vaginal swabs were also collected for microbiome analysis via 16S rRNA sequencing. Surveys were collected and stored in REDCap. Surveys were statistically analyzed using t-test, Fisher’s exact, or Chi-square tests. Most women identified as NA (n=16), white (n=13), and Hispanic (n=5). We identified bacterial profiles of 28 of the 31 participants to be Lactobacillus-dominant (n=16) and non-Lactobacillus-dominant (n=12) and stratified them into two groups. Significant differences were observed, suggesting association of multiple people in a household (p=0.0228), lower level of education (p=0.0093), and high parity (p=0.0283) to undesirable non-Lactobacillus-dominant VMB. It is noteworthy that there were no differences between groups regarding race/ethnicity, age, body mass index, marital status, employment status, household income, hrHPV positivity, vaccination status, use of contraceptives, sexual activity, number of partners and knowledge of HPV and VMB. Yet, the non-Lactobacillus group tended to have more women with high-risk HPV infection. Ongoing analysis includes global health and perceived stress data in relation to the VMB and HPV status. Upon full analysis, we will report on trends between the VMB composition and sociodemographic, lifestyle factors and quality of life. The findings will improve ways for people to find solutions to decrease HPV infections and cervical cancer in Native American populations. Citation Format: Tawnjerae Joe, Paweł Łaniewski, Tristen Eddie, Skyler Bordeaux, Verity Quiroz, Haiyan Cui, Denise Roe, Donna Peace, J. Gregory Caporaso, Naomi Lee, Melissa M. Herbst-Kralovetz. The impact of sociodemographic and lifestyle factors on the vaginal microbiome and cervical cancer disparities in Native American women [abstract]. In: Proceedings of the 16th AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2023 Sep 29-Oct 2;Orlando, FL. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2023;32(12 Suppl):Abstract nr B127.

The vulvar microbiome in lichen sclerosus and high-grade intraepithelial lesions.

The role of the vulvar microbiome in the development of (pre)malignant vulvar disease is scarcely investigated. The aim of this exploratory study was to analyze vulvar microbiome composition in lichen sclerosus (LS) and vulvar high-grade squamous intraepithelial lesions (HSIL) compared to healthy controls. Women with vulvar lichen sclerosus (n = 10), HSIL (n = 5) and healthy controls (n = 10) were included. Swabs were collected from the vulva, vagina and anal region for microbiome characterization by metagenomic shotgun sequencing. Both lesional and non-lesional sites were examined. Biophysical assessments included trans-epidermal water loss for evaluation of the vulvar skin barrier function and vulvar and vaginal pH measurements. Healthy vulvar skin resembled vaginal, anal and skin-like microbiome composition, including the genera Prevotella, Lactobacillus, Gardnerella, Staphylococcus, Cutibacterium, and Corynebacterium. Significant differences were observed in diversity between vulvar skin of healthy controls and LS patients. Compared to the healthy vulvar skin, vulvar microbiome composition of both LS and vulvar HSIL patients was characterized by significantly higher proportions of, respectively, Papillomaviridae (p = 0.045) and Alphapapillomavirus (p = 0.002). In contrast, the Prevotella genus (p = 0.031) and Bacteroidales orders (p = 0.038) were significantly less abundant in LS, as was the Actinobacteria class (p = 0.040) in vulvar HSIL. While bacteria and viruses were most abundant, fungal and archaeal taxa were scarcely observed. Trans-epidermal water loss was higher in vulvar HSIL compared to healthy vulvar skin (p = 0.043). This study is the first to examine the vulvar microbiome through metagenomic shotgun sequencing in LS and HSIL patients. Diseased vulvar skin presents a distinct signature compared to healthy vulvar skin with respect to bacterial and viral fractions of the microbiome. Key findings include the presence of papillomaviruses in LS as well as in vulvar HSIL, although LS is generally considered an HPV-independent risk factor for vulvar dysplasia. This exploratory study provides clues to the etiology of vulvar premalignancies and may act as a steppingstone for expanding the knowledge on potential drivers of disease progression.

186 Examining the Vaginal Microbiome During Gilt Reproductive Development

Abstract Gilt reproductive failure is a common challenge in the US swine industry. Indeed, as many as three million breeding females are removed from swine herds each year, resulting in an annual replacement rate of 40%. Furthermore, about 20% of gilts selected for replacement never successfully enter the reproductive herd. The vaginal microbiome may play a role in gilt reproductive success; this study compared the vaginal bacterial composition from eight gilts at two pre-pubertal time points (d70 and d130 of age). Vaginal swabs were collected and analyzed using the 16S rRNA gene through PCR-amplification of the V1-V3 regions from vaginal microbial genomic DNA, followed by Illumina MiSeq 2X300 sequencing. Seven of the gilts achieved first estrus between d 164 and d 190 days of age based on physical signs of vulva swelling and locked stance in the presence of a boar. One female had no visible signs of estrus before 210 d of age. Beta diversity analysis by PCoA revealed that samples clustered into two separate groups according to time points. Alpha diversity indices (Sobs, Ace and Chao) were numerically decreased between d 70 to d 130. A comparative analysis of the most highly represented Operational Taxonomic Units (OTUs) using the non-parametric Kruskal-Wallis sum-rank test and Wilcoxon pairwise test identified 12 OTUs that varied in abundance (P &amp;lt; 0.05) between the two time points. Notably, OTU SD_Ssd-00001, predicted to be a strain of Lactobacillus amylovorus (99.82% sequence identity), was greatest at d 70 (21.33%), then significantly decreased at d 130 (7.67%). In contrast, OTU SD_Ssd-01393, corresponding to an uncultured species of Prevotellaceae (closest relative: Prevotella disiens, 92.48%), was in less abundance at d 70 (4.87%), but became the most abundant OTU at d 130 (18.19%). The vaginal OTUs identified in this study included not only known bacterial species, such as members of the Lactobacillus genus which have been previously found to be normal and beneficial residents of the human vagina, but also uncharacterized bacterial species. These results are indicative of distinct vaginal microbiome composition between d 70 and d 130 of age and suggest that certain bacterial species could potentially be used as indicators of reproductive potential in gilts.

A citizen-science-enabled catalogue of the vaginal microbiome and associated factors

Understanding the composition and function of the vaginal microbiome is crucial for reproductive and overall health. Here we established the Isala citizen-science project to analyse the vaginal microbiomes of 3,345 women in Belgium (18–98 years) through self-sampling, 16S amplicon sequencing and extensive questionnaires. The overall vaginal microbiome composition was strongly tied to age, childbirth and menstrual cycle phase. Lactobacillus species dominated 78% of the vaginal samples. Specific bacterial taxa also showed to co-occur in modules based on network correlation analysis. Notably, the module containing Lactobacillus crispatus, Lactobacillus jensenii and Limosilactobacillus taxa was positively linked to oestrogen levels and contraceptive use and negatively linked to childbirth and breastfeeding. Other modules, named after abundant taxa (Gardnerella, Prevotella and Bacteroides), correlated with multiple partners, menopause, menstrual hygiene and contraceptive use. With this resource-rich vaginal microbiome map and associated health, life-course, lifestyle and dietary factors, we provide unique data and insights for follow-up clinical and mechanistic research.

Cervical and Vaginal Microbiomes in Early Miscarriages and Ongoing Pregnancy with and without Dydrogesterone Usage.

Emerging evidence suggests that the reproductive tract microbiota is a key modulator of local inflammatory and immune pathways throughout pregnancy and may subsequently impact pregnancy outcomes. In this study, our objective was to analyze the cervical and vaginal microbiomes during early pregnancy among three groups: women with healthy ongoing pregnancies, women undergoing dydrogesterone treatment, and those who experienced miscarriages. The experiment involved 51 women at 8-11 weeks of gestation. The microbiome was examined using 16S rRNA sequencing on the Ion Torrent PGM platform. Across all groups, Lactobacillus iners was predominant, suggesting that the vaginal community type CST III is common among the majority of participants. Notably, our data highlighted the significant roles of Gardnerella vaginalis and Mycoplasma girerdii in the pathogenesis of early miscarriage. Conversely, L. iners and Bifidobacterium longum have a protective effect in early pregnancy. Moreover, dydrogesterone intake appeared to influence notable differences between the cervical and vaginal microbiomes. Overall, our study enhanced our understanding of the cervical and vaginal microbiome composition in the eastern European population during early pregnancy.

Relationship between maternal Group B Streptococcal colonization and gestational vaginal microbiome composition: A pilot study

PurposeGroup B Streptococcus (GBS) colonization and vaginal microbiome (VMB) dysbiosis are associated with adverse perinatal outcomes. However, the role of GBS colonization in maternal VMB remains unclear. Herein, we aimed to investigate this relationship and identify additional pathogens associated with GBS colonization for potential implications in understanding their clinical significance. Materials and methodsVaginal swab samples were obtained before delivery from nine women with normal pregnancies for GBS detection and 16S rRNA gene sequencing. The diversity analysis and community state types clustering were used to compare the GBS-positive vs. GBS-negative groups. ANCOM-BC was implemented to identify differentially abundant microbes (DAMs) associated with GBS colonization. The correlation and receiver operating characteristic analysis were used to evaluate the relationship between DMAs and clinical parameters. ResultsThere were 6/9 (66,7%) GBS-negative pregnant women. The α-diversity index (p ​= ​0.71 for observed operational taxonomic units and p ​= ​0.90 for Shannon diversity), β-diversity index (p ​= ​0.583), and community state types clustering (p ​= ​0.23) were not significantly different between the GBS-positive and -negative groups. Four DAMs, namely, Actinomyces, Shigella, Fenollaria, and Gemella, were significantly associated with GBS colonization, reflecting the dynamicity of the gestational VMB. Their abundances were negatively correlated with birth weight and had acceptable discriminating ability in premature membrane rupture (area under the curve, 0.9). ConclusionsDespite the absence of significant effects on overall VMB composition, our preliminary results investigated that maternal GBS colonization related to high abundance of four pathogens with potential clinical utility as microbial signatures.

O-264 Vaginal Microbiota Transplantation (VMT) for treatment of vaginal dysbiosis without the use of antibiotics – A Randomized Controlled Trial in healthy women with vaginal dysbiosis

Abstract Study question Can vaginal microbiota transplantation with shotgun-verified eubiotic vaginal microbiome over three menstrual cycles, convert vaginal dysbiosis to eubiosis in healthy women? Summary answer The RCT is ongoing until it is by March 2023. Results will be ready for presentation at EHSRE conference. What is known already Vaginal dysbiosis covers imbalances in the vaginal flora, caused by the composition of bacteria, viruses, and vaginal fungi. A large proportion of women who have vaginal dysbiosis do not experience any symptoms. Dysbiosis occurs in about 16-20% of all women. Vaginal dysbiosis is associated with infertility, euploid pregnancy loss, preterm labour or bacterial vaginosis. Treatment of vaginal dysbiosis consists of antibiotic treatment, and/or probiotics. Vaginal transplantation with eubiotic vaginal bacterial flora in combination with antibiotics has successfully been performed in four out of five recipients in an earlier study, but no study has been performed without use of antibiotic pretreatment. Study design, size, duration Randomized, controlled, double-blinded trial with a randomization ratio 3:1 to receive either a eubiotic microbiome transplant or placebo. 320 healthy women between 18 and 40 years, was enrolled for screening of vaginal dysbiosis. 30 donors with a eubiotic vaginal microbiome and 48 recipients with a dysbiotic microbiome were identified. The trial began in June 2021 and will end in March 2023 Participants/materials, setting, methods Vaginal microbiome composition was assed by next-generation ShotGun sequencing. To qualify as donor bacterial DNA from a vaginal swab had to show a bacterial composition of at least 80% lactobacilli and less than 5% pathogenic bacteria. To qualify as recipient vaginal microbiome composition should be with at least 20% pathogenic bacteria and no more than 10% lactobacilli. The recipient could have up to three attempts of VMT treatments with a follow-up period of 6 months. Main results and the role of chance We expect the RCT to end in March 2023, and the results of this trial will be presented at the ESHRE annual meeting . If we can show engraftment of a eubiotic microbiome transplant without the use of pre-treatment with antibiotics it could be a potential treatment of vaginal dysbiosis. Limitations, reasons for caution As this is the first VMT without antibiotic pretreatment we did not have reliable data for our power calculation and a negative result may be due to lack of statistical power. Wider implications of the findings Effective treatments for vaginal dysbiosis are urgently needed and VMT may be one such treatment strategy. Trial registration number NCT04855006

Vaginal Microbiome Dysbiosis is Associated with the Different Cervical Disease Status

Vaginal microbiome composition was demonstrated to be associated with cervical disease. The colonization characteristics of vaginal microbes and their association with the different cervical disease status, especially cervical cancer (CC), are rarely investigated. In this cross-sectional study, we characterized the vaginal microbiome of women with different status of cervical diseases, including 22 NV + (normal tissue with HPV infection), low-grade squamous intraepithelial lesion (LSIL, n = 45), high-grade squamous intraepithelial lesion (HSIL, n = 36) and CC (n = 27) using bacterial 16S DNA sequencing. Thirty HPV-negative women with normal tissue were used as the control group. We found that higher diversity of microbiome with gradual depletion of Lactobacillus, especially L. crispatus, was associated with the severity of cervical disease. High-risk HPV16 infection was associated with higher microbiome diversity and depletion of Lactobacillus in high-grade cervical diseases (i.e. HSIL and CC). The CC group was characterized by higher levels of Fannyhessea vaginae, Prevotella, Bacteroides, Finegoldia, Vibrio, Veillonella, Peptostreptococcus, and Dialister. Co-occurrence network analyses showed that negative correlations were exclusively observed between Lactobacillus and other bacteria, and almost all non-Lactobacillus bacteria were positively correlated with each other. In particular, the most diverse and complex co-occurrence network of vaginal bacteria, as well as a complete loss of L. crispatus, was observed in women with CC. Logistic regression model identified HPV16 and Lactobacillus as significant risk and protective factors for CC, respectively. These results suggest that specific Lactobacillus species (e.g. L. crispatus and L. iners) can be used as important markers to target prevention measures prioritizing HPV16-infected women and other hrHPV-infected women for test, vaccination and treat initiatives.