Imbalances in the vaginal flora during pregnancy elevate risks for adverse outcomes. While nifuratel targets pathogens, probiotics restore acidity, and robust cohort evidence supporting their combined safety and efficacy in restoring the microbiome remains limited. This retrospective cohort study assessed nifuratel-nystatin therapy plus Lactobacillus probiotics for restoring vaginal flora in pregnancy. Employing a retrospective cohort design, this study enrolled pregnant women diagnosed with vaginal microecological abnormalities between March 2023 and 2024. Participants were allocated via 1:1 propensity score matching (PSM) into two groups (n=58 each): a combination group receiving nifuratel-nystatin therapy combined with vaginal Lactobacillus probiotics and a monotherapy group receiving nifuratel-nystatin alone. All received a 7-day treatment. The primary endpoint was clinical effectiveness assessed one week post-treatment. Secondary outcomes included vaginal pH, Lactobacillus and bacterial diversity normalization rates, recurrence, adverse pregnancy outcomes and drug-related adverse events, monitored until delivery. Multivariate logistic regression identified independent predictors of treatment efficacy. After PSM, baseline characteristics were balanced (P>0.05). The comparative analysis indicated a superior overall efficacy for the combination therapy (93.1%) over monotherapy (77.6%), with statistical significance (P<0.01) Both groups exhibited reduced vaginal pH and increased rates of normalized Lactobacillus and bacterial diversity, with greater improvements observed in the combination group (P<0.01). The recurrence rate was significantly lower in the combination group (8.6% vs. 22.4%; P<0.05), as was the total incidence of adverse pregnancy outcomes (6.9% vs. 20.7%; P<0.01). No significant difference in adverse drug reactions was found. Multivariate analysis identified combination therapy as an independent protective factor for clinical efficacy (aOR=4.25, 95% CI: 1.42-12.71, P<0.05). In pregnant women with vaginal dysbiosis, combining nifuratel-nystatin with Lactobacillus probiotics safely enhances clinical efficacy, normalizes pH and flora and reduces adverse pregnancy outcomes.

CST-Based Molecular Profiling of Vaginal Microbiota Validated by qRT-PCR and 16S Amplicon Sequencing.

Artificial intelligence and machine learning in smart vaginal formulation development.

Consumer-directed female technology ("femtech") is rapidly expanding into pelvic floor and sexual health, yet clinicians have limited guidance on the safety, efficacy, and appropriate use of these products. The objective of this review is to summarize currently marketed female technology products relevant to pelvic floor disorders, sexual function, and vaginal health, and to highlight key evidence, limitations, and implications for clinical practice. This is a narrative review of commercially available technologies for urinary incontinence, sexual dysfunction, and vaginal health. Product categories include external and intravaginal pelvic floor muscle training devices, vibrators, and dilators for sexual function and pain, regenerative medicine interventions such as platelet-rich plasma injections, sexual wellness applications, and direct-to-consumer vaginal microbiome testing platforms. Pelvic floor devices and in-office energy therapies offer nonsurgical options for urinary incontinence, but high-quality, independent long-term data are scarce, and costs are often substantial. Sexual wellness devices, including vibrators and dilators, are increasingly recognized as therapeutic tools but are variably supported by clinical research. Early studies of platelet-rich plasma suggest potential benefit for sexual function, yet protocols and outcomes remain heterogeneous. Digital applications and microbiome tests expand access and patient engagement but frequently lack rigorous validation and may raise concerns regarding privacy, equity, and interpretation of results. Female technology offers novel opportunities to address pelvic floor and sexual health outside traditional care pathways. Urogynecologists should be familiar with these products, provide balanced counseling on benefits, risks, and cost, and advocate for independent trials to define their role within evidence-based care.

To systematically review the current literature on gynecologic function and dysfunction in transmasculine and gender-diverse individuals in the context of testosterone therapy, with a focus on menstrual suppression, contraceptive needs, pelvic pain, vulvovaginal changes, and sexual health. PubMed, Embase, and Web of Science databases were searched through September 2025 using search terms related to transmasculine individuals, assigned female at birth, reproductive organs, and testosterone and gender-affirming hormone therapy. No restrictions on publication year were applied. Studies were included if they reported on the impact of exogenous testosterone on gynecologic or sexual (dys)function and contraceptive use and choices, in transmasculine and gender-diverse individuals. Only original human research studies were eligible. Studies focusing on fertility, reproductive outcomes, malignancy, histology, animal models, or individuals with differences of sex development were excluded. All included studies were critically appraised using Joanna Briggs Institute quality assessment tools, and findings were synthesized narratively. Fifty-seven studies were included. Testosterone was generally effective in achieving menstrual suppression, although breakthrough bleeding and ovulatory activity occurred in a substantial proportion of individuals. Contraceptive needs were frequently unmet, partly due to misinformation and provider-related barriers. Pelvic pain was commonly reported, with varied etiologies. While testosterone often increased sexual desire, dyspareunia and genital discomfort were frequently described. Vaginal microbiome alterations and epithelial changes were observed, although their clinical implications remain unclear. Testosterone-based gender-affirming hormone therapy has diverse effects on gynecologic function in transmasculine and gender-diverse individuals, including both physiologically expected as well as underexplored effects. Clinicians should adopt an individualized and affirming approach to care, while further research is needed to understand long-term outcomes, improve assessment tools, and close gaps in inclusive gynecologic healthcare.

Vaginal microbiota dysbiosis has been associated with female reproductive health and infertility. This study aimed to compare vaginal microbiota profiles according to fertility and ovarian induction status among Thai women. In this cross-sectional study, vaginal samples were collected from Thai reproductive-aged women during their periovulatory period. The 16S rRNA gene was amplified and sequenced to ascertain the composition of the vaginal microbiota among 92 women (30 fertile and 62 infertile women). Vaginal microbiota profiles at the phylum and genus levels were compared among fertile women, infertile women before ovulation induction, and infertile women after ovulation induction. The dominant vaginal microbial phyla across all groups were Firmicutes, Actinobacteria, Bacteroidetes, and Fusobacteria. Lactobacillus was the predominant genus across all groups. However, exploratory differential abundance analyses identified several differentially enriched taxa across fertile, infertile (pre-ovulation induction), and infertile (post-ovulation induction) groups. No significant differences in alpha or beta diversity were observed between groups. Our findings did not demonstrate significant differences in overall vaginal microbial diversity between fertile and infertile Thai women. However, exploratory differential abundance analyses identified several taxa that may warrant further investigation. Larger prospective studies are needed to better clarify the potential role of vaginal microbiota in reproductive health and infertility.

Bacterial vaginosis (BV) is a common vaginal dysbiosis caused by Gardnerella vaginalis, a facultative anaerobic bacillus. The failure of conventional antibiotics and recurrence of bacterial vaginosis call for alternative novel therapeutic strategies. Antimicrobial peptides (AMPs) provide a targeted, resistance-sparing alternative with their broad-spectrum activity and distinct mode of action. Two AMPs, i.e., TCCP-1 (cyclic) and ZMLP-2 (linear), were designed in silico from proteome sequences of Thymbra capitata and Zataria multiflora already available in NCBI. The designed peptides were chemically synthesized, evaluated for their antibacterial activity, cytotoxicity, hemolytic effects and mechanism of action against G.vaginalis. TCCP-1, a cyclic peptide with an MIC of 1.95µg/mL against G. vaginalis showed minimal cytotoxicity even at 100µg/mL, which is much higher than its MIC value (1.95µg/mL). TCCP-1 maintained high cell viability at lower concentrations, while a concentration-dependent reduction in viability was observed at higher concentrations. In contrast, ZMLP-2, a linear AMP, showed weak antimicrobial activity with an MIC of 100µg/mL, exhibited a moderate reduction in cell viability (~ 70-75%) when tested at 100µg/mL or a concentration below its MIC. Both peptides showed the disruption of bacterial membranes and, therefore, support the re-establishment of healthy vaginal flora. More significantly, TCCP-1 demonstrated efficient antimicrobial activity against G.vaginalis along with decreased cytotoxicity, making it an excellent candidate for future in vivo studies and possible clinical uses. Thus, plant-derived AMPs could prove to be useful, targeted, and sustainable alternatives to BV prevention while treating both resistance and recurrence.

Oral metronidazole versus clindamycin to treat bacterial vaginosis in pregnancies at risk for preterm labor.

Vulvovaginal candidiasis (VVC) is a prevalent mucosal infection primarily caused by Candida albicans, affecting up to 75% of women of reproductive age. VVC therapeutic management is increasingly challenged by antifungal resistance, host toxicity, and high recurrence rates. Lactobacillus species, the dominant members of healthy vaginal microbiota, are known to antagonize pathogens and maintain mucosal homeostasis by producing antimicrobial metabolites and modulating epithelial responses. However, few studies have explored the antagonistic and anti-virulence properties of Lactobacillus strains isolated from asymptomatic women, particularly their activity against clinical C. albicans isolates. This study aimed to characterize vaginal Lactobacillus strains and their cell-free supernatants (CFS) isolated from asymptomatic women and evaluate their antifungal and anti-virulence activities against clinical isolates of C. albicans. Forty-two clinical Lactobacillus isolates were identified, of which eight inhibited the growth of the tested pathogens. However, among the clinical isolates, only L. fermentum JAC 231 and Lactiplantibacillus plantarum MDBL 269 inhibited all the fungal isolates tested. Based on these results, L. fermentum ATCC 23271 (positive control) and the clinical strains L. fermentum JAC 231 and L. plantarum MDBL 269 were selected for detailed analyses of their ability to interfere with key virulence factors of C. albicans, including hyphal morphogenesis, biofilm formation and disruption, adhesion, and the secretion of extracellular enzymes. The selected strains and their CFS displayed antagonistic activity: they reduced pathogen adhesion, co-aggregated with yeast cells, impaired biofilm formation, and destabilized preformed biofilms. In addition, whole-genome sequencing combined with antimicrobial peptide gene-cluster prediction using BAGEL4 and antiSMASH revealed two biosynthetic gene clusters in L. fermentum JAC 231 and seven in L. plantarum MDBL 269, providing a plausible genetic basis for the antimicrobial effects observed.

Gut, vaginal, and urinary microbiome alterations in women with genitourinary syndrome of menopause: A systematic review.

Objective: to study the features of the vaginal microbiome in women with postmenopausal OVA and to conduct a comparative analysis of the microbiota in groups with varying degrees of severity of symptoms. Materials and methods: a prospective study was conducted involving 212 naturally postmenopausal patients (age 48–59 years) with moderate IVA. The degree of symptoms was assessed using the VSQ questionnaire and the vaginal Health index. The state of microbiocenosis was analyzed by PCR “Femoflor 16”. The patients were divided into three groups according to the severity of symptoms: mild (≤5 symptoms, n=85), moderate (6-10, n=94) and severe (≥11, n=33). Results: a clear correlation was found between the severity of the symptoms of IVA and the nature of dysbiotic changes. In the group with severe symptoms, 90.9% of patients were diagnosed with moderate or severe aerobic/mixed dysbiosis with a predominance of Gardnerella vaginalis, Prevotella bivia, Porphyromonas spp. and Streptococcus spp., as well as a significant decrease in lactobacillus levels (median — 10^3.3 CFU/ ml). In the group with mild symptoms, 65.9% had normocenosis or conditional normocenosis, but the total bacterial mass was reduced (median — 10^4–5 CFU/ml). In 15.4% of patients, “silent atrophy” (objective signs of IVA in the absence of complaints) with a variable state of the microbiota was detected. Conclusion: the severity of clinical manifestations of BBA in postmenopausal women is associated with specific changes in the vaginal microbiota: a pronounced symptom complex correlates with dysbiosis and inhibition of lactobacillar flora. The data obtained substantiate the need for a comprehensive diagnosis,

Recent research has shown that the COVID-19 virus changes the gastrointestinal microbiome as it is a bacteriophage [1], leading to immune dysregulation and bowel dysbiosis [2]. This creates an environment where opportunistic pathogens thrive while beneficial bowel flora diminish [3]. There is an association among the bowel, vaginal and bladder microbiomes where the bacteria from the bowel are found in the bladder [4]. Urinary tract infections (UTIs) are among the most common bacterial infections, typically caused by Escherichia coli (E. coli), Enterococcus faecalis (E. faecalis), Klebsiella spp, and Proteus mirabilis (P. mirabilis) in women [5]. Changes in the bowel microbiome lead to changes in the bladder microbiome [6]. We aimed to evaluate whether the composition of observed uropathogens differed across the different phases of the COVID-19 pandemic. This retrospective study analysed positive urine cultures from women before, during and after the COVID-19 pandemic. The leading three causative uropathogens with a count over 103cfu/mL were reported. There were 5543 positive cultures with pre-COVID-19 from 1 January 2019 to 30 January 2020, n = 204, during from 1 February 2020 to 4 May 2023, n = 2518, and after from 5 May 2023 to 1 February 2025, n = 2821. In total, 10,456 bacteria were identified. After May 2023, E. coli prevalence dropped from 27 to 21%, and Pseudomonas spp decreased from 6 to 2% (p value < 0.001). In contrast, Klebsiella spp incidence rose from 8 to 12%. Corynebacterium spp (p < 0.05) and Acinetobacter spp (p < 0.001) were increasingly isolated in symptomatic UTIs after May 2023. The bacterial organisms found in urinary tract infections detected before and subsequent to the onset of COVID-19 have changed. This may reflect an interaction between changes in the bowel microbiome and pathological urinary tract infections.

Broadly neutralizing antibodies (bNAbs) are a promising HIV prevention strategy due to their potent antiviral activity and potential for long-acting protection. While the vaginal microbiome can influence mucosal immunity and the efficacy of topical interventions, its effect on the pharmacokinetics of systemically administered bNAbs remains unclear. Forty-two women were included in a retrospective analysis of the CAPRISA 012B clinical trial evaluating passive immunization for HIV prevention. Vaginal microbiota were profiled using 16S rRNA gene sequencing and classified into three community state types (CSTs): CST I (Lactobacillus crispatus-dominated), CST III (Lactobacillus iners-dominated), and CST IV (diverse, non-Lactobacillus-dominated). Mucosal concentrations of CAP256V2LS were measured from Soft-cup® cervicovaginal fluid using the Meso Scale Discovery (MSD) platform with electrochemiluminescence (ECL) detection. Longitudinal analyses assessed CST stability, transitions, and associations with mucosal antibody pharmacokinetics. Lactobacillus-dominated CSTs were most frequent (CST I, 5.3%; CST III, 57.9%), whereas BV-associated CST IV subtypes were less common (CST IV-A, 2.6%; CST IV-B, 34.2%). Lactobacillus-dominated communities were generally stable, while high-diversity CST IV communities were more dynamic, with transitions toward Lactobacillus-dominated states observed over time. Despite these microbial shifts, mucosal bNAb kinetic patterns appeared broadly similar across CSTs. Within the limits of this exploratory analysis, we did not observe clear evidence of an association between CST composition and the timing or magnitude of mucosal bNAb accumulation. These observations were descriptive and not derived from inferential statistical or pharmacokinetic modelling analyses. In this exploratory sub-analysis, cervicovaginal microbiome composition was not clearly associated with differences in mucosal concentrations of systemically administered bNAbs. These findings suggest that systemic bNAb delivery may achieve measurable genital tract exposure across diverse vaginal microbial communities; however, larger studies incorporating inferential pharmacokinetic and immunological analyses are needed to confirm these observations and exclude subtle microbiome-associated effects.

Currently, live biotherapeutic products (LBP) have been widely used to improve gut microbiota, enhance immunity, and promote digestion. Engineering bacteria, as one form of LBP, have made breakthrough progress in the treatment of diseases, but their effectiveness and specific mechanisms in treating vaginal infections are not clear. Here, we innovatively constructed Escherichia coli Nissle 1917-attB-IFN-α2b (EcN-IFN-α2b) expressing antiviral interferon alpha-2b (IFN-α2b) using genome integration technology, and evaluated its ability to reduce human papillomavirus (HPV) viral load and the underlying mechanisms in an NSG mouse surrogate model of HPV. Our results showed that EcN-IFN-α2b had good growth performance, acid resistance, antioxidant ability, and antimicrobial activity in vitro. EcN-IFN-α2b also significantly reduced the HPV viral load (M vs. EcN-IFN-α2b = 1.003 vs. 0.643, p < 0.01), and effectively alleviated the levels of interleukin 1β (IL-1β) (M vs. EcN-IFN-α2b, p < 0.01), interleukin 6 (IL-6) (M vs. EcN-IFN-α2b, p < 0.01), and tumor necrosis factor α (TNF-α) (M vs. EcN-IFN-α2b, p < 0.05) in the vagina. Further mechanistic studies suggested that EcN-IFN-α2b may reduce HPV viral load by activating the JAK/STAT/cGAS/STING signaling pathway. Additionally, we found that EcN-IFN-α2b altered the composition of vaginal microbiota, especially the relative abundance of vaginal pathogens such as Vibrionaceae_Vibrio, Streptococcus, and Staphylococcaceae_Staphylococcus, and the relative abundance of beneficial Lactobacillus. This study systematically evaluated the effects of the novel engineered strain EcN-IFN-α2b in reducing HPV viral load, alleviating associated inflammation, and restoring vaginal microbiota in an NSG mouse surrogate model, providing a new strategy for the future clinical treatment of HPV infection.

Vaginal polycyclic aromatic hydrocarbons (PAHs), HPV infection, and vaginal microbiome in firefighters: A cross-sectional study.

BackgroundTestosterone-associated vulvovaginitis is an under-recognised cause of genital symptoms in transmasculine patients receiving testosterone as gender-affirming hormone therapy, with 5-years prevalence 64-78%. BASHH's Gender and Sexual Minorities Special Interest Group convened sexual health, gender, gynaecological and primary care experts and patient representatives to formulate a first-in-world best practice statement.MethodWe searched Medline, Embase, CINAHL, UpToDate, Trip Medical Database and Cochrane, to April 2025, regarding testosterone's impact on transmasculine people's vaginal histology and microbiomes, symptoms, and management. Papers on cis- or transgender women, editorials, and study protocols were excluded. Included studies were assessed using the Critical Appraisal Skills Programme tool. Recommendations for best practice were graded using GRADE methodology where possible, and supplemented by expert group consensus where evidence was limited.Results14 papers were included, of which 11 were descriptive/non-management focused. Testosterone is associated with vaginal Lactobacilli depletion, increased speciation, pH changes, atrophy, inflammation, and epithelial cell differentiation shift. The diagnosis of testosterone-associated vulvovaginitis is clinical; symptom severity does not reliably correlate with any of the above, nor other markers such as hormone levels. Two small case series and one cohort study (total n = 12) discussed treatment; topical estrogen reduced vaginal symptoms and increased Lactobacilli, with some refractory inflammatory cases requiring topical clindamycin +/- topical steroids. Presumed efficacy and safety are extrapolated from topical estrogen use in cisgender menopausal women.ConclusionSpecific evidence for testosterone-associated vulvovaginitis management is lacking; we suggest a culturally competent approach, considering it as a differential in any transmasculine people on testosterone with genitourinary symptoms. Offer examination (if possible) to exclude infection or dermatoses; offer first-line treatment with topical vaginal estrogens. Adjunctive/alternative measures include emollients and lubricants; topical clindamycin +/- topical steroids for refractory cases or aerobic/desquamative vaginitis. Long-term maintenance therapy and review is advised. Robust trans-specific clinical trials are needed to strengthen the evidence and optimise long-term outcomes.

Recent studies from Western countries suggest an association between cervicovaginal microbiome (CVM) and HPV persistence. This study aims to identify CVM diversity within HPV-negative/positive women and to establish an association between HPV infection and CVM in Indian women. This was a pilot, hospital-based, prospective study conducted at a tertiary care center in India over 6 months. Seventy-two participants underwent HPV genotyping, yielding 20 HPV-positive samples. Of these, 16 samples with good-quality DNA, along with 16 matched HPV-negative samples, were identified for CVM analysis. Sequencing of bacterial 16S ribosomal RNA region and bioinformatic analysis were performed in 32 samples. Baseline characteristics were similar for HPV-positive and HPV-negative women. Out of 16 HPV-positive women, 9 had HPV-16 genotype, one had HPV-16&18, and 6 had other genotypes. Seven patients had cervical cancer on histopathology, 7 had NILM, 1-ASCUS, and 1-ASC-H on cytology. HPV-negative women's CVM had a larger abundance of lactobacillus species than HPV-positive women (43.24% vs 2.44%). Whereas CVM among HPV-positive women was characterized by a greater abundance of anaerobes, such as Prevotella (12.46% vs. 4.59%), Atopobium (6.66% vs. 1.98%), Anaerococcus (5.27% vs. 0.57%), and Porphyromonas (2.98% vs. 0.67%), compared with HPV-negative women. The current study from the Sub-Himalayan region of India characterizes the diversity of CVM in HPV-positive and HPV-negative women. This makes a significant geographical contribution to vaginal microbiome research worldwide.

Probiotic supplements are marketed for diverse health benefits, yet species inclusion often lacks functional rationale. Our survey of 352 over-the-counter probiotic products available in the USA revealed 36 unique microbial species. However, there is no clear link between species inclusion and the intended health benefit. Here, to address this gap, we developed HaPaPro, a collection of 1,012 genome-scale metabolic models spanning pathogenic, probiotic and host-associated bacteria, constructed from publicly available genome sequences. Flux balance analysis revealed that probiotic species fail to capture the metabolic diversity of host-associated microbes. Focusing on vaginal health, we computationally identified vaginal microbes with metabolic profiles overlapping Gardnerella vaginalis. In vitro spent media assays using 11 vaginal isolates showed variable inhibition of G. vaginalis, primarily driven by D-lactic acid production, which was also produced by non-Lactobacillus species. These findings highlight the need for function-based probiotic design and demonstrate a scalable framework integrating metabolic modelling with experimental validation.

The principal therapeutic challenge in vulvovaginal candidiasis (VVC) is that the non-selective nature of conventional antifungal drugs, which frequently perturb vaginal microecological homeostasis and disrupt Lactobacillus barrier, lead to the emergence of recurrent infection and drug resistance. This study aims to develop novel antifungal agents capable of efficiently and selectively eradicating pathogenic fungi while protecting and promoting the growth of Lactobacilli, with real-time monitoring capabilities and significant potential for clinical application. Harnessing the principle of charge-competition, we engineered a cationic amphiphilic aggregation-induced emission luminogens (AIEgens, named as TPE-ET), where hydrophobic chain length served as a key determinant governing membrane affinity, aggregation propensity, and antimicrobial selectivity. This design empowered potent eradication of Candida albicans (C. albicans) while concomitantly favoring the proliferation of beneficial Lactobacilli. Moreover, TPE-ET disrupted C. albicans biofilms and suppressed virulence genes related to adhesion, invasion, and drug resistance. In a murine VVC model, TPE-ET reduced fungal burden by over 90%, facilitating the repair of damaged vaginal epithelium and the reconstitution of a Lactobacillus-dominat vaginal microbiome. Remarkably, TPE-ET outperformed clotrimazole in restoring healthy microecological balance, as manifested by diminished Proteobacteria abundance alongside increased Firmicutes (notably Lactobacillus) and Bacteroidetes. Mechanistic studies revealed that TPE-ET exerted its remarkable antifungal activity by targeting the mitochondrial inner membrane, disrupting the metabolism-inflammation axis and eliciting mitochondrial dysfunction. Collectively, this dual merits of membrane charge-selective targeting and AIEgens-mediated visualization established an innovative therapeutic strategy for VVC, featuring superior efficacy, exquisite selectivity, and real-time monitoring capability with significant clinical potential.

The vaginal microbiome is essential for women's health, yet its genomic diversity and interaction with the host remain incompletely characterized. Here we present the Global Vaginal Metagenome-assembled Genomes catalog, an extensive repository of vaginal microbial genomes generated by integrating 10,665 in-house Chinese metagenomes, with 2,967 publicly available metagenomes and 1,433 bacterial isolates. The catalog comprises 65,055 genomes from 890 prokaryotes, 11 eukaryotes and 6,590 viral taxonomic units, many not represented in public reference databases. We investigate virus-bacteria interactions, revealing conserved phages-host associations. We then identify substantial intraspecies genomic and functional variations displaying population-specific patterns. A metagenome-genome-wide association study identifies seven host genetic loci associated with vaginal species at study-wide significance and replicated in at least one independent cohort, notably connecting the gene OPRK1 with the potential pathogen Ureaplasma urealyticum. In summary, our research provides a comprehensive reference for future studies on genotype-phenotype interplay within the human vaginal microbiome.