Search Results (149)

Background/Objectives: This study investigated the effects of bovine lactoferrin (LF) on vaginal microbiota in healthy postmenopausal women. Methods: A randomized, double-blind, placebo-controlled trial was conducted. Healthy Japanese postmenopausal women were administered a placebo or bovine LF (300 mg/day) for 12 weeks. The primary endpoint was the Nugent score and the secondary endpoints were the Nugent sub-scores. The exploratory outcomes included vaginal immunoglobulin and urinary deoxypyridinoline levels. Results: Eighty participants were randomized (placebo, n = 40; LF, n = 40), of whom 8 were excluded. The remaining 72 participants (placebo group: n = 36; LF group: n = 36) were included in the per-protocol set. Although the Nugent score was lower in the LF group, the difference was not significant. The Gardnerella/Bacteroides Nugent sub-score was significantly lower in the LF group than in the placebo group. The vaginal immunoglobulin G1 levels were significantly higher in the LF group than in the placebo group. In addition, the urinary deoxypyridinoline levels were significantly lower in the LF group than in the placebo group. Adverse events were comparable between the groups and no adverse drug reactions were observed. Conclusions: Although oral LF supplementation did not significantly affect the primary endpoint (overall Nugent score), a lower Gardnerella/Bacteroides morphotype score and higher vaginal IgG1 levels were observed. As these findings were derived from secondary and exploratory analyses, respectively, they are considered hypothesis-generating and require confirmation in future studies. Full article

Human papillomavirus (HPV) drives cervical carcinogenesis, but viral infection alone is insufficient for malignant transformation. The vaginal microbiome may modulate HPV persistence and cervical intraepithelial neoplasia (CIN) progression, yet existing reviews remain inconclusive. This systematic review identifies specific vaginal microbiome risk factors associated with HPV and CIN and proposes plausible mechanisms linking dysbiosis to disease progression. Following (PRISMA) guidelines, databases were searched for studies published through October 2025. Thirty-three studies from 2013 to 2025 across 10 countries met the inclusion criteria, with 87.9% exhibiting low risk of bias. Most studies demonstrated marked Lactobacillus depletion alongside Prevotella and Gardnerella enrichment in HPV/CIN cases. Disease severity correlated with shifts toward high-diversity anaerobe communities, though alpha and beta diversity metrics varied across cohorts. Importantly, longitudinal data suggests a temporal sequence where baseline dysbiosis precedes clinical progression. Crucially, although the included cohorts directly observed and quantified these taxonomic shifts, the underlying mechanisms of disease progression, such as enzymatic degradation of mucosal barriers mediated by Gardnerella and immunosuppression driven by Fusobacterium, remain largely inferred from external in vitro literature. Lactobacillus iners emerged as a notable species-level risk factor in HPV persistence. Vaginal dysbiosis, characterised by Lactobacillus depletion and anaerobic expansion, is strongly correlated with HPV persistence and CIN progression, potentially facilitated through convergent barrier degradation and immune subversion. These findings highlight targets for microbiome-based risk stratification and therapeutic intervention. Full article

The cervicovaginal virome remains poorly characterized in relation to human papillomavirus (HPV) infection and bacterial community structure. We performed shotgun metagenomic sequencing of 311 cervicovaginal specimens from a Kazakhstani cohort spanning seven groups defined by HPV status and cervical cytology. Viral community composition differed according to HPV status, with increasing representation of the oncogenic Alphapapillomavirus 9 clade across more abnormal cytological categories. Predicted phage functional profiles also differed between NILM HPV-positive and NILM HPV-negative women. Integrase, excisionase, transcriptional repressor, anti-repressor Ant, and amidase annotations showed differential prevalence after false-discovery-rate correction. Auxiliary metabolic and host-interaction genes associated with nucleotide metabolism, DNA modification, anti-restriction functions, and toxin–antitoxin systems were also differentially represented. Stratification by bacterial community state type revealed contrasting predicted functional repertoires, with toxin–antitoxin-associated annotations enriched in Lactobacillus crispatus-dominated communities and anti-restriction-associated annotations enriched in Gardnerella vaginalis-dominated communities. These findings identify associations between HPV status, bacterial community structure, and cervicovaginal viral composition and predicted phage functions. Longitudinal and experimental studies are required to determine the directionality and biological activity of these associations. Full article

Urogenital multiplex polymerase chain reaction (PCR) panels report Mycoplasma genitalium (MG) and Mycoplasma hominis (MH) similarly despite different clinical significance. We retrospectively analyzed 176,855 adult specimen-level laboratory records (2018–2022). MG and MH were compared using modified Poisson regression and a year-adjusted stacked interaction model. MG was detected in 4215 records (2.38%) and MH in 8024 (4.54%). MG prevalence fell from 4.48% (95% confidence interval (CI), 4.30–4.68) at 18–29 years to 0.21% (0.16–0.27) at ≥60 years, whereas MH fell from 5.70% to 2.39%. Female sex was associated with lower MG (adjusted prevalence ratio [aPR], 0.32) and higher MH (1.44). Age and sex gradients differed significantly (interaction PRs, 1.519 and 4.653; both p < 0.001), whereas no specimen-type interaction reached significance (all p ≥ 0.19). No other target was positive in 52.1% of MG-positive versus 54.5% of MG-negative records, but in 4.9% of MH-positive versus 55.7% of MH-negative records. MH co-detection was strongest with Gardnerella vaginalis (aPR, 9.94), Ureaplasma urealyticum (6.12), and Trichomonas vaginalis (5.48). These profiles support target-specific reporting. Cross-panel analytical differences cannot be excluded because MG and MH were measured on different panels. Full article

Multiplex PCR assays report Ureaplasma urealyticum (UU) and Ureaplasma parvum (UP) separately, but the results are often combined as “Ureaplasma positive.” We compared Gardnerella vaginalis (GV) co-detection in UU-only and UP-only adult specimen accession records and examined variation by sex and age. This retrospective study included 176,855 urogenital records from a Korean reference laboratory (2018–2022). The primary comparison (N = 46,319) used modified Poisson regression adjusted for sex, age group, specimen type, and collection year. GV co-detection ranged from 15.7% in UU−/UP− records to 86.0% in UU+/UP+ records. For UU−/UP+ versus UU+/UP− records, the adjusted prevalence ratio (aPR) was 1.06 (95% CI, 1.04–1.08). Sex-specific aPRs were 0.93 (0.91–0.95) in women and 1.13 (1.10–1.15) in men (p for interaction < 0.001); the sex-specific directions were reproduced in a urine-only analysis.. Age-related differences were nonmonotonic and exploratory. The overall association was small, and the use of different multiplex panels for UU and UP/GV means that cross-panel analytical differences could have contributed to, or potentially accounted for, the observed difference. These findings should therefore be interpreted as descriptive laboratory co-detection patterns rather than evidence of a biological interaction or clinically meaningful species-specific difference.. Full article

Background/Objectives: Certain strains of Lactobacillus acidophilus are widely used as probiotics. However, their functional potential for female reproductive tract health remains insufficiently characterized. While most studies have focused on individual strains, the distribution of putative probiotic-associated genes across the species remains unclear. This study aimed to evaluate the vaginal probiotic potential of Lb. acidophilus using population genomic analysis and comparative phenotypic characterization. Methods: Pan-genomic analysis was performed on 109 Lb. acidophilus genomes (107 public genomes and 2 vaginal isolates). Putative probiotic-associated gene clusters were identified by functional annotation, categorized into functional modules, and compared among ecological-origin groups. Two vaginal isolates (strains A2 and A3) were characterized in vitro for growth under different pH conditions, cell surface hydrophobicity, lactic acid and hydrogen peroxide production, antimicrobial activity, hemolysis, and antimicrobial susceptibility. Results: The Lb. acidophilus pan-genome was closed, with 1782 of 1902 gene clusters (93.69%) classified as core. Thirty-six putative probiotic-associated gene clusters were identified and grouped into four modules: environmental tolerance, adhesion/colonization, exopolysaccharide/biofilm synthesis, and nutrient metabolism/microbial competition. Thirty-four of the 36 gene clusters were present in all 109 genomes, and no general ecological origin-specific distribution pattern was observed. Three bacteriocin-related gene clusters were conserved across all genomes. A2 and A3 exhibited similar lactic acid production and growth patterns at pH 4–6. Both produced relatively low amounts of hydrogen peroxide compared with the reference strains. A3 showed higher cell surface hydrophobicity and moderate inhibition against Gardnerella vaginalis, while A2 showed no inhibition of this organism. Both isolates were non-hemolytic, and were susceptible to vancomycin and linezolid, resistant to clindamycin, and non-susceptible to daptomycin. No acquired antibiotic resistance genes were detected. Conclusions: Most putative probiotic-associated gene clusters were conserved across the Lb. acidophilus population, whereas A2 and A3 showed strain-dependent phenotypic differences. These findings support combining population genomic analysis with strain-level phenotypic testing when selecting Lb. acidophilus candidates for bacterial vaginal infections. Full article

Background: Genital tract infections and dysbiosis-related conditions contribute substantially to women’s sexual and reproductive morbidity. In South America, evidence on clinically relevant genital microorganisms remains fragmented across countries, populations, and diagnostic approaches. This systematic review and meta-analysis synthesized the prevalence of Chlamydia trachomatis, Neisseria gonorrhoeae, Trichomonas vaginalis, Gardnerella vaginalis, and Candida spp. among women in South America. Methods: We searched PubMed/MEDLINE, Web of Science, Cochrane Library, and SciELO for studies published from 1 January 2014 to 29 February 2024. Eligible studies reported prevalence data for at least one target microorganism in female populations from South American countries. Two reviewers independently screened studies, extracted data, and assessed methodological quality using Joanna Briggs Institute criteria for prevalence studies. Crude prevalence and random-effects pooled prevalence estimates with 95% confidence intervals were calculated. The review protocol was registered in PROSPERO (CRD420261324060). Results: Of 629 records identified, 51 studies from eight South American countries and French Guiana met the eligibility criteria for the systematic review, and 46 contributed to the main pooled prevalence meta-analysis. Substantial between-study heterogeneity was observed across all pathogen-specific analyses. Pooled prevalence was highest for G. vaginalis (28%), followed by Candida spp. (12%), C. trachomatis (7%), T. vaginalis (6%), and N. gonorrhoeae (3.2%). Conclusions: This review highlights the heterogeneous burden of clinically relevant genital microorganisms among women in South America and supports standardized diagnostic approaches, clearer phenotypic definitions, and stronger surveillance across underrepresented regional settings. Full article

Genitourinary syndrome of menopause (GSM) is a common chronic condition in postmenopausal women, yet biomarkers predicting response to local estrogen therapy remain insufficiently defined. In this prospective study, 216 postmenopausal women with GSM received local estradiol vaginal tablet therapy for 12 weeks. Baseline assessments included symptom evaluation, vaginal pH, vaginal maturation index (VMI), and quantitative real-time PCR measurement of six target microorganisms. Treatment response was defined as an improvement of at least 2 points in the most bothersome symptom (MBS) score. After 12 weeks, MBS score, vaginal pH, and VMI improved significantly, with increased abundance of L. crispatus, L. jensenii, and L. gasseri and decreased abundance of L. iners, Gardnerella, and Candida. Overall, 128 of 216 women responded (59.3%; 95% CI, 52.6–65.6%). Higher baseline L. iners abundance was independently associated with a lower probability of response (adjusted RR per log10-unit increase, 0.82; 95% CI, 0.70–0.95), with the highest tertile showing a lower response likelihood than the lowest (RR, 0.71; 95% CI, 0.54–0.93). These findings suggest that L. iners may be a promising candidate microbial marker for pretreatment risk stratification, although external validation is needed before clinical application. Full article

The vaginal microbiome is often interpreted through static taxonomic patterns. Yet microbial composition alone does not explain why some communities resist perturbation, recover after disruption, or transition toward dysbiosis. This narrative review synthesizes evidence that vaginal microbiome stability is shaped by endocrine phase, epithelial substrate availability, microbial functional capacity, mucosal tone and candidate host modifiers. High-estrogen states, particularly pregnancy, are associated with epithelial maturation, glycogen accumulation, low vaginal pH, and Lactobacillus-dominant communities, whereas postpartum, lactational, menopausal, and other hypoestrogenic states are associated with reduced epithelial support and increased vulnerability to diverse anaerobe-rich configurations. We review the linking of the estrogen–glycogen–Lactobacillus axis, focusing on microbial functions involved in glycogen degradation, lactate production and biofilm persistence, and host pathways that may modify mucosal responsiveness. Direct human genotype-to-vaginal-microbiome stability evidence remains limited; therefore, host genetic features are treated as candidate modifiers rather than validated clinical predictors. We propose a conceptual multi-omic hierarchy for organizing endocrine, epithelial, microbial, immune, temporal, and candidate host-modifier domains relevant to vaginal microbiome resilience. This framework is hypothesis-generating and requires longitudinal, phase-resolved human validation before quantitative prediction or clinical application. Full article

Vaginal dysbiosis, particularly bacterial vaginosis (BV), is associated with altered immune responses that may influence the formation of extracellular traps (ETs). This study aimed to characterize neutrophil and macrophage extracellular traps (NETs and METs) in vaginal discharge samples obtained from women with normal microbiota (NM) and BV, with particular emphasis on Gardnerella vaginalis (GV) detection. An ex vivo analysis was performed using vaginal smears from 14 patients previously classified according to the Nugent criteria. Immunofluorescence assays targeting neutrophil elastase (NE), citrullinated histone H3 (Citr-H3), CD15, and CD68 were conducted, and quantitative image analysis was performed using the TissueFAXS and StrataQuest platforms. NETs were classified into three morphotypes: spread (spr), diffuse (diff), and aggregated (agg). BV samples exhibited a substantially higher mean NET count than NM samples (842.43 vs. 91.86). The number of diffNETs was significantly higher in BV samples than in NM samples (p = 0.004), whereas GV-positive samples showed increased sprNET abundance compared with that in negative samples (248 vs. 8; p < 0.05). CD68⁺ cell counts were significantly higher in BV samples (p = 0.026), whereas no significant differences in NE or Citr-H3 fluorescence intensity were observed between groups. MET structures were also identified, suggesting macrophage involvement in the local immune response. Collectively, these findings indicate that vaginal dysbiosis and GV presence are associated with enhanced NET formation and distinct morphotype distributions, supporting a role for ETs in the immunopathology of BV. Full article

Background/Objectives: This study aimed to investigate the genital pathogen profile and co-infection dynamics using multiplex RT-qPCR, specifically evaluating the independent associations with age, sex, and HIV status. Methods: Data from 1217 patients who underwent a sexually transmitted infection (STI) panel study at the Microbiology Laboratory of Marmara University Pendik Training and Research Hospital between January 2024 and December 2025 were retrospectively reviewed. Pathogen detection was performed using a commercial kit (Bioeksen, Istanbul, Türkiye) with the multiplex RT-qPCR method. The independent effects of HIV positivity, age, and sex on pathogen frequency and co-infection were analyzed using logistic regression models; results were evaluated using adjusted odds ratio (aOR) and 95% confidence interval (CI). Results: Any-pathogen positivity was detected in 57.8% of patients, with Ureaplasma spp. (36.9%) and Gardnerella vaginalis (32.5%) being the most prevalent. While overall pathogen positivity did not differ significantly by HIV status (p = 0.158), HIV-positive patients exhibited distinct microbiological architectures. Higher positivity rates were observed in women in both groups, and a strong correlation was found between sex and the presence of infection (p < 0.001). Multivariate analysis revealed that HIV positivity was independently associated with an over fivefold increase in HSV-2 detection (aOR = 5.09, 95% CI: 1.47–17.65; p = 0.010). Furthermore, HIV-positive individuals were significantly enriched for complex polymicrobial patterns involving three or more pathogens (p = 0.008). Conversely, male sex was independently associated with a substantially lower risk of co-infection (aOR = 0.17, 95% CI: 0.12–0.22; p < 0.001), and increasing age showed an inverse relationship with co-infection frequency (aOR = 0.98 per year, 95% CI: 0.97–0.99; p = 0.001). Conclusions: This study adds to current epidemiological evidence by showing that genital pathogen distribution is shaped by age- and sex-related heterogeneity, with flora-associated co-infections predominating in women and more classical STI-related agents occurring more often in men. Our findings suggest that multiplex RT-qPCR provides value not only for broad pathogen detection but also for identifying demographic- and HIV-associated co-infection patterns that may support stratified screening and targeted clinical management. Full article

Cervicovaginal dysbiosis is an established co-factor of high-risk human papillomavirus (HPV) persistence and cervical neoplastic development, yet most studies address the bacterial compartment in isolation, leaving fungal communities and bacterial–fungal cross-kingdom interactions underexplored, particularly in Central Asian populations. We performed shotgun metagenomic sequencing (mNGS) of cervicovaginal samples from 311 Kazakhstani women undergoing routine cervical screening. HPV status was determined using combined PCR and mNGS methods, and cervical screening was completed using liquid-based cytology (NILM, ASC-US, LSIL, ASC-H). Bacterial, viral, and fungal taxa were profiled from a single shotgun dataset with Kraken2 pipeline. Bacterial community state types (CSTs) were determined based on dominant bacterial species, functional gene content was annotated against KEGG using eggNOG, and covariate-adjusted associations were estimated using MaAsLin3. Mycobiome β-diversity differed significantly by HPV status (p = 0.003). In particular, Candida positivity was significantly associated with HPV presence and with high-risk α-9 HPV in cytologically normal (NILM) samples (OR = 3.6, [1.6–9.6], p ≤ 0.001). Covariate-adjusted analysis was consistent with this positive association (q < 0.05). Concurrently, among CSTs, Lactobacillus iners-dominated CST III and dysbiotic Gardnerella vaginalis-dominated CST IV showed a 3-fold higher Candida albicans prevalence (p < 0.01). Further analysis demonstrated that, functionally, both of these CSTs had depleted capacity for lactate metabolism (ko00620, p < 0.0001) and, in particular, for the genetic capacity for pyruvate-dependent H₂O₂ generation (half that of the L. crispatus-dominated CST I). These findings support L. iners as a metabolically permissive rather than protective Lactobacillus and suggest cross-kingdom functional signatures as candidate biomarkers for HPV acquisition and persistence in Central Asia, a region previously absent from the cervicovaginal microbiome literature. Full article

Background: Chronic endometritis (CE) is a subtle, often asymptomatic endometrial inflammation marked by CD138⁺ plasma cell infiltration and linked to recurrent implantation failure (RIF), recurrent pregnancy loss (RPL), and unexplained infertility. Emerging evidence implicates endometrial microbiome dysbiosis in CE. Objective: To systematically review and conduct meta-analysis on the association between CE and endometrial microbiome alterations and their reproductive implications. Methods: We searched MEDLINE, Embase, Web of Science, Scopus, Cochrane CENTRAL, and Google Scholar for studies diagnosing CE via CD138 immunostaining, assessing microbiota with molecular techniques. Data extraction, quality assessment, and meta-analysis were performed. Results: Twenty-two studies including 4022 women were analyzed. CE was associated with reduced prevalence of Lactobacillus-dominated microbiota and increased detection of non-Lactobacillus species, particularly Streptococcus spp., Enterococcus spp., Escherichia coli, Staphylococcus spp., Ureaplasma spp., and Gardnerella vaginalis. In the meta-analysis (2947 women), Enterococcus spp. and Ureaplasma spp. were significantly more prevalent in women with CE, whereas Streptococcus spp., E. coli, Staphylococcus spp. and G. vaginalis showed non-significant trends. Only E. coli and Streptococcus spp. showed significant heterogeneity between-studies. Conclusions: CE is linked to microbial dysbiosis with reduced Lactobacillus dominance and enrichment of potentially pathogenic taxa, notably Enterococcus and Ureaplasma spp. These findings suggest that the endometrial microbiome contributes to chronic inflammation and adverse reproductive outcomes, yet heterogeneity and limited evidence call for standardized diagnostics and robust trials before clinical implementation. Full article

Recurrent vulvovaginal candidiasis (RVVC) affects 5–8% of women of reproductive age. Host genetic factors, particularly single nucleotide polymorphisms (SNPs) in Toll-like receptors (TLRs), may influence RVVC susceptibility by impairing vaginal mucosal antifungal immunity. The aim of this study was to assess the effect of SNPs in genes encoding TLRs on RVVC susceptibility. Τhe distribution of TLR2 Arg753Gln and TLR4 Asp299Gly/Thr399Ile polymorphisms in Greek women, including RVVC (n = 63), first-episode VVC (n = 37), Gardnerella vaginalis vaginitis (GV, n = 36) patients, and healthy controls (n = 61), was investigated using TaqMan SNP genotyping. Genotype and allele frequencies were analyzed under allelic and dominant models, with odds ratios (ORs), 95% confidence intervals (CIs), and linkage disequilibrium assessed. TLR4 Asp299Gly and Thr399Ile heterozygotes were significantly more frequent in RVVC patients compared with controls and affected RVVC susceptibility (OR: 5.57, 95% CI: 1.17–26.56, p: 0.0172; OR: 4.92, 95% CI: 1.02–23.78, p: 0.0306, respectively). No associations were observed for TLR2 Arg753Gln or for any SNP with GV or first-episode VVC. TLR4 variants co-segregated, indicating a haplotype effect. TLR4 haplotypes, rather than TLR2 polymorphism, confer increased RVVC susceptibility, supporting a genetically distinct, mucosal immunity-driven pathogenesis. Larger, ethnically diverse studies with functional assays are warranted to validate these findings and guide personalized prevention and treatment strategies. Full article

Background: Antimicrobial resistance (AMR) is a global healthcare threat. Traditional largely non-selective antibiotics produce side effects due to the natural host microbiome being modified creating a loss in homeostasis. In women, AMR is a cause of acute generational impact. For example, bacterial vaginosis (BV), the most common gynecological infection in reproductive-age women, is a serious public health concern due to its high rates of recurrence, secondary infections, and reproductive issues; and two currently prescribed antibiotics for BV do not fully resolve the symptoms. Objective: The strong need for innovative, potent, safe, and selective therapeutics has prompted a search for such bioactive molecules in milk. Resulting from 200 million years of evolutionary pressure, mammalian lactation not only nourishes infants, but it has also been under relentless Darwinian selective pressure to provide protection from a variety of infections. Methods: Computationally designed human-milk-inspired peptides (AMPs) were tested in standard microbicidal assays for activity against BV pathogens, and evaluated for stability and safety. Results: Several AMPs are bactericidal towards Gardnerella vaginalis, a major BV-associated pathogen, and other BV-associated pathogens. Some novel AMPs do not impact the viability of key lactobacilli linked to a healthy vaginal microbiome. These stable, membrane-acting cationic AMPs reduce inflammation during an infection assay and are safe in EpiVag organoid tissues. Conclusions: AMPs can address concerns like non-selectivity and antibiotic resistance—thereby addressing AMR. Lead AMPs from this study offer a promising solution for the development of novel therapeutics for the treatment of BV, which may reduce the burden of AMR. Full article