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Background: An increasing number of studies indicate that the gut, cervical, and vaginal microbiota may play crucial roles in the development of cervical cancer (CC). However, the interactions between the microbiota and the host are yet unknown. To address this gap, a systematic review and meta-analysis were conducted to assess the microbiota alterations in a variety of body locations, including the gut and genital tract., Methods: Electronic searches of PubMed, Embase, Web of Science, and the Cochrane Library were conducted to retrieve eligible papers published from January 1, 2014, to January 1, 2024 (PROSPERO: CRD42024554433). This study was restricted to English-language studies reporting on alpha diversity, beta diversity, and relative abundance, as well as on patients with CC whose microbiota had been analyzed via next-generation sequencing technologies. To assess the risk of bias (RoB), we utilized the Newcastle‒Ottawa Quality Assessment Scale (NOS) and the ROBINS-I tool. For the meta-analysis, we employed Review Manager 5.4., Results: Thirty-six eligible studies were included in this review. The Chao1 index (SMD = 0.96, [95% CI: 0.71, 1.21], I 2  = 0%) and the Shannon index (SMD = 1.02, [95% CI: 0.53, 1.50], I 2  = 85%) values from vaginal samples were significantly greater in patients than in the controls. In the cervical samples, the Shannon index value (SMD = 1.29, [95% CI: 0.61, 1.97], I 2  = 93%) significantly increased, whereas the Chao1 index value did not significantly differ (SMD = 0.50, [95% CI: -0.46, 1.46], I 2  = 89%). The Shannon index value (SMD = 0.25, [95% CI: -0.22, 0.72], I 2  = 38%) did not significantly differ across the gut samples. The majority of studies (19/25) indicated that the patients and noncancer controls differed significantly in terms of beta diversity. Cancer-associated changes were observed, with a dramatic decrease in the Lactobacillus genus and significant increases in pathogenic bacteria, including the Anaerococcus, Peptostreptococcus, Porphyromonas, Prevotella, and Sneathia genera. Additionally, the impact of antineoplastic therapies on microbial diversity was inconsistently reported across several studies., Conclusion: This systematic review elucidates the microbiota alterations associated with the prevalence of CC and its response to anti-tumor therapies, aiming to provide insights for future research directions and precision medicine strategies to enhance women's quality of life., Prospero Registration: CRD42024554433., Competing Interests: Declarations. Ethics approval and consent to participate: Not applicable. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)

Background: Prenatal and postpartum depression may be influenced by the composition of host associated microbiomes. As such, the objective of this study was to elucidate the relationship between the human gut or vaginal microbiomes in pregnancy with prenatal or postpartum depression., Methods: 140 female participants were recruited at their first prenatal visit and completed the Edinburgh Postnatal Depression Scale (EPDS) to screen for depression and anxiety, in addition the EPDS was completed one month postpartum. Vaginal and stool biospecimens were collected in the third trimester, analyzed using 16S rRNA gene sequencing, and assessed for alpha and beta diversity. Individual taxa differences and clustering using the k-medoids algorithm enabled community state type classification., Results: Participants with higher postpartum EPDS scores had higher species richness and lower abundance of L. crispatus in the vaginal microbiota compared to those with lower EPDS scores. Participants with a higher prenatal EPDS score had lower species richness of the gut microbiome. Participants with a vaginal community state type dominated by L. iners had the highest mean prenatal EPDS scores, whereas postpartum EPDS scores were similar regardless of prenatal vaginal state type., Limitations: Our small sample size and participant's self-report bias limits generalizability of results., Conclusions: Depression in the prenatal and postpartum period is associated with the composition and diversity of the gut and vaginal microbiomes in the third trimester of pregnancy. These results provide a foundational understanding of the microbial relationships between maternal health and depression for identifying potential therapeutic treatments., Competing Interests: Declaration of competing interest None., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

The commensal microbiota influences the health, feeding efficiency, and reproductive performance of sows. The microbiota composition in the alimentary and genitourinary tracts and in colostrum/milk during pregnancy and lactation also impacts the microbiota and immune system, growth, and health of the piglets. Knowledge of the microbial compositions is important for evaluation of these effects and for discovering ways to improve the health and productivity of the sows. Oral, vaginal, and rectal mucosa and colostrum were sampled from 32 sows of variable parity in late pregnancy, and colostrum within 6 hours of delivery of the first piglet, on four commercial piglet-producing farms in Finland. Microbial compositions were analyzed by 16S rRNA gene amplicon sequencing. The most abundant genera of the oral microbiota were Rothia, Moraxella, and Streptococcus. The rectal microbiota was dominated by Clostridium sensu stricto 1. Streptococcus was the most abundant genus in the vagina and colostrum. Moderate differences in diversity and composition were observed between farms. The relative abundances of the genera Neisseria (MaAsLin 2 q = 0.002, ANCOMBC q = 0.005), Fusobacterium (MaAsLin 2 q = 0.008, ANCOMBC q = 0.04) and Bacteroides (MaAsLin 2 q < 0.005, ANCOMBC q = 0.06) were lower in oral samples and Romboutsia (MaAsLin 2 q = 0.07, ANCOMBC q = 0.05), Turicibacter (MaAsLin 2 q = 0.08, ANCOMBC q = 0.02) and Lachnospiraceae&#95;UCG&#95;007 (MaAsLin 2 q = 0.1, ANCOMBC q = 0.05) were higher in rectal samples of multiparous compared to primiparous sows. In vaginal samples there was a tendency of higher relative abundances of the genera Fusobacterium and Streptococcus in multiparous than primiparous sows. Among the differentially abundant taxa, F. necrophorum and F. nucleatum were identified in oral samples, F. gastrosuis and F. necrophorum in vaginal samples, and S. dysgalactiae in colostrum samples. This study provides a comprehensive overview of the mucosal and colostrum microbiota of periparturient sows during normal production conditions on Finnish commercial farms., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2025 Piirainen et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Ethnopharmacological Relevance: Jiawei Ermiao Granules (JWEMGs), a traditional Chinese herbal formulation, has been widely used in China for the treatment of human papillomavirus (HPV) infections. However, the underlying mechanisms through which it exerts its antiviral effects remain poorly understood., Aim of the Study: This study aimed to investigate the potential mechanisms by which JWEMGs modulate vaginal microecology and clear HPV infections, utilizing clinical trials, metagenomic sequencing, and in vitro models., Materials and Methods: Clinical indicators related to vaginal microecology, such as vaginal pH, cleanliness, Nugent score, Donders score, catalase, neuraminidase, and leukocyte esterase, were evaluated in 65 patients with high-risk HPV (HR-HPV) infection. The study examined the impact of two courses of oral JWEMGs on these clinical parameters. Additionally, metagenomic sequencing was performed on vaginal lavage samples from 33 patients to assess the alteration of the vaginal microbiome following JWEMGs treatment. Immunohistochemistry was used to detect ALPK1 expression in cervical exfoliated cells, and ELISA was employed to measure cytokine levels in vaginal lavage fluid. JWEMGs intervention was applied to HaCaT-HPV E6/E7 cells to evaluate its effects on restoring α-kinase 1 (ALPK1) expression and promoting the secretion of cytokines and chemokines., Results: Treatment with JWEMGs significantly improved several clinical indicators, including cleanliness, pH, Nugent score, Donders score, catalase, neuraminidase, and leukocyte esterase, in HR-HPV-infected patients. Furthermore, JWEMGs therapy led to an increased abundance of Lactobacillus species, especially Lactobacillus crispatus, and a marked reduction in Gardnerella species. JWEMGs treatment also significantly promoted ALPK1 expression in cervical exfoliated cells and augmented the secretion of key cytokines, including IL-6, IL-8, and TNF-α. In parallel, in vitro results showed that JWEMGs substantially enhanced IL-6, IL-8, TNF-α, CCL2, CCL5, and CCL7 secretion in HaCaT-HPV E6/E7 cells, which correlated with the activation of the ALPK1/NF-κB signaling pathway., Conclusion: In conclusion, JWEMGs treatment effectively remodels the vaginal microbiota and bolsters mucosal immunity in the lower genital tract, thereby improving the vaginal microecology in HR-HPV-infected individuals. In vitro findings further demonstrated that JWEMGs promote cytokine and chemokine expression, activating the ALPK1/NF-κB pathway., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Qingling Ren reports financial support was provided by the National Natural Science Foundation of China. Qingling Ren reports financial support was provided by the Chinese Clinical Medicine Innovation Center of Obstetrics, Gynecology, and Reproduction in Jiangsu Province. Xiu Li reports financial support was provided by the Postgraduate Research and Practice Innovation Program of Jiangsu Province. Chenxi Gu reports financial support was provided by the Postgraduate Research and Practice Innovation Program of Jiangsu Province. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2025 Elsevier B.V. All rights reserved.)

Clotrimazole (CLO) is a strong antifungal drug approved to treat vaginal candidiasis (VC). Nanosponges (NSs) were developed to maintain providing CLO in a steady pattern with amplified accumulation in the vaginal mucosa. The quasi-emulsion solvent diffusion method was utilized to prepare NSs. The optimized NSs selected by Design Expert® exhibited a production yield percent (PY%) of 60.10 ± 0.39 %, encapsulation efficiency percent (EE%) of 91.21 ± 0.59 %, particle size (PS) 275.50 ± 0.97 nm, polydispersity index (PDI) 0.425 ± 0.01, and zeta potential (ZP) of -27.40 ± 0.25 mV. The morphological results confirmed a spongy, porous structure. Fourier Transform Infrared Spectroscopy ensured the drug encapsulation. Differential scanning calorimetric studies showed no interaction between the excipients and CLO. The prepared NSs-loaded gel of optimized CLO-NSs was evaluated, the mucoadhesive strength (6065.85 ± 52.03 dyne/cm 2 ) with spermicidal activity of (0 % sperm motility/60 s). The ex-vivo deposition depicted significantly increased vaginal retention of CLO by 2.44-fold compared to Candistan® 2 % vaginal cream (the market product). Finally, the in-vivo study on rats demonstrated thesuperior efficacy of CLO-NSs gel relative to Candestan®, with significantly reduced inflammatory biomarkers and minimal histopathological alterations in the treatment of vaginal candidiasis with a high safety profile., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2025 Elsevier B.V. All rights reserved.)

Complement factor B (CFB) is a crucial component for the activation of the alternative pathway due to the formation of the C3 convertase with C3b, which further produces C3b to enhance the overall complement activity. Although Cfb is expressed not only in the immune tissues, but also in the reproductive tract, the physiological role of the alternative complement pathway in reproduction remains unclear. In this study, we addressed this issue by producing Cfb -knockout (KO) mice and analyzing their phenotypes. Sperm function, number of ovulated oocytes, and litter size were normal in KO mice. In contrast, the diversity of microbiomes in the gut and vaginal tract significantly increased in KO mice. Some serine protease activity in the serum from KO mice was lower than that of wild-type mice. Since the serum from KO mice showed significantly lower activity of the alternative complement pathway, CFB was found to be essential for this pathway. Our results indicate that although the alternative pathway is dispensable for normal fertility and development, it maintains the gut and vaginal microbiomes by suppressing their diversity and activating the alternative complement pathway.

Bacterial vaginosis, one of the most common pathologies in adult women, is defined as a chronic polymicrobial condition resulting from dysbiosis of the vaginal microbiota. Despite 70 % of patients being asymptomatic, diagnosis of this condition relies on clinico-microbiological tools. Despite being considered a gold standard technique, the Nugent score, established during microscopic examination of vaginal smears, has shown a lack of standardization and reproducibility. The discovery of new bacteria not included in the microscopic Nugent score, such as BVAB1,2,3, Atopobium sp., and Megaspheara sp., undetectable by conventional techniques, has highlighted a lack of specificity and sensitivity of the score currently provided to clinicians. This has spurred the development of new PCR kits to perform Nugent scores via molecular biology and address the shortcomings of microscopic techniques. This study has shed light on the contribution of molecular biology to the diagnosis of bacterial vaginosis, as well as the performance and limitations of the Allplex™ Bacterial Vaginosis kit marketed by Seegen® compared to microscopic examination.

Vaginitis is the most common problem afflicting women of childbearing age. However, the underlying etiological factors remain poorly understood, leading to recurrent vaginitis and constraining clinical management. Here, we explored whether the gut microbiota influences the risk of vaginitis by performing a two-sample Mendelian randomization analysis using the largest genome-wide association studies to date. Four gut taxa in genus levels were identified related to vaginitis: Candidatus Soleaferrea (inverse-variance weighted [IVW] odds ratio [OR] = 2.20, P = 0.026), Dialister (IVW OR = 2.62, P = 0.029), Lachnospiraceae UCG-008 (IVW OR = 0.41, P = 0.0067), and Ruminiclostridium 5 (IVW OR = 0.080, P = 1.42 × 10 -5 ). We further explored the mediation effect of the plasma metabolites by two-step Mendelian randomization (MR) and multivariable MR. The findings indicated that 3-phosphoglycerate and lysophosphatidylcholine antagonistically act against the two identified risk factors ( Candidatus Soleaferrea and Dialister , respectively) of vaginitis, thus appearing to confer protective effects against vaginitis. On the contrary, the elevation of arachidonate/pyruvate ratio and reduction in palmitate/myristate ratio mediated the protective effects of Lachnospiraceae UCG-008 against vaginitis. These findings support a potential causal role for gut microbiota in the development of vaginitis, thereby providing potential strategies for its prevention and intervention.IMPORTANCEVaginitis is the most common problem afflicting women of childbearing age. However, the underlying etiological factors remain poorly understood, leading to recurrent vaginitis and constraining clinical management. Besides, the human gut and vagina are important organs that are both colonized by thousands of microorganisms impacting human physiology and health. Whether there is an interplay between the microecosystems is intriguing and unclear. This study evaluated the potential causal relationship between the gut microbiota and vaginitis and suggested that specific types of gut microbiota may be the potential risk or protective factors of vaginitis mediated or suppressed by certain plasma metabolites. These findings provide treatment insights for vaginitis., Competing Interests: The authors declare no conflict of interest.

Background: The composition of the vaginal microbiota is closely linked to adverse sexual and reproductive health outcomes, due in part to effects on genital immunology. Compositional approaches such as metagenomic sequencing provide a snapshot of all bacteria in a sample and have become the standard for characterizing the vaginal microbiota, but only provide microbial relative abundances. We hypothesized that the addition of absolute abundance data would provide a more complete picture of host-microbe interactions in the female genital tract., Results: We analyzed cervicovaginal secretions from 196 female sex workers in Kenya and found that bacterial load was elevated among women with diverse, bacterial vaginosis (BV)-type microbiota and lower among women with Lactobacillus predominance. Bacterial load was also positively associated with proinflammatory cytokines, such as IL-1α, and negatively associated with chemokines, such as IP-10. The associations between bacterial load and immune factors differed across bacterial community states, but L. crispatus predominance was the only microbial community where higher bacterial load was not associated with higher proinflammatory cytokines. Total vaginal bacterial load was also a stronger predictor of the genital immune environment than BV by Nugent score, the current clinical standard, in the Kenya-based cohort and in a Uganda-based confirmatory cohort., Conclusions: Our results suggest that total vaginal bacterial load is at least as strong a predictor of the genital immune milieu as current BV clinical diagnostic tools, supporting exploration of the vaginal bacterial load as a predictor of adverse reproductive and sexual health outcomes. Video Abstract., Competing Interests: Declarations. Ethics approval and consent to participate: All participants provided written, informed consent and the study was approved by the Kenyatta National Hospital Ethics and Research Committee (KNH ERC P778/11/2018), the London School of Hygiene and Tropical Medicine (LSHTM) Ethics Committee (Approval number: 16229) and the University of Toronto ethics committee (Approval number: 37046). Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)

Background: Group B streptococcus (GBS) is a commensal bacterium of the digestive and genital tracts that can occasionally cause maternal and neonatal disease. GBS is particularly a burden in low-resource settings, where infections with HIV are also highly prevalent. This study investigates the impact of HIV status on GBS colonization and antibody levels., Methods: In Uganda, 90 nonpregnant women of childbearing age were followed for 3 months. Every 2 weeks, rectal and vaginal swabs were tested for GBS, and vaginal cups and blood were collected for measurement of GBS capsular polysaccharides (CPS) IgG using standardized assays., Results: Twenty-six of 90 women were living with HIV. Almost 51/90 women were GBS colonized at 1 or several visits. GBS colonization fluctuated in the rectal and vaginal sites. Most prevalent serotypes were Ia and III, with 33 individuals carrying different serotypes over time. Serum and vaginal CPS-IgG levels were stable over 12 weeks. In serum, for serotypes Ib-V, the geometric mean concentration (GMC) of CPS-IgG did not differ between HIV+ and HIV- participants. However, the GMC for serum CPS-Ia-IgG in the HIV+ group was 2.5 times lower than in the HIV- group (P = 0.038). Vaginal CPS-IgG was measurable in 5/26 (19%) HIV+ participants, and 32/64 (50%) HIV- participants., Conclusions: Despite fluctuating GBS colonization, antibody levels remained stable over 12 weeks. The level of CPS-Ia-specific IgG in serum was lower in women with HIV than in those without HIV. Vaginal CPS-specific IgG was not measurable in 81% of individuals with HIV., Competing Interests: The authors have no conflicts of interest to disclose., (Copyright © 2024 Wolters Kluwer Health, Inc. All rights reserved.)

The reproductive tract, as a lumen connected to the outside world, its microbial community is influenced by various factors. The changes in its microbiome are closely related to women's health. The destruction of the micro ecological environment will lead to various infections, such as Bacterial vaginosis, sexually transmitted infections, adverse pregnancy outcomes, infertility and tumors. In recent years, with the continuous development and progress of molecular biology, research on reproductive tract microbiota has become a clinical hotspot. The reproductive tract microbiota is closely related to the occurrence and development of female reproductive tract diseases such as vaginitis, pelvic inflammation, PCOS, cervical lesions, and malignant tumors. This article reviews the research on the relationship between vaginal microbiota and female reproductive tract diseases, in order to provide theoretical basis for the prevention and treatment of female reproductive tract diseases., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Objective: To identify a microbial signature for endometriosis for use as a diagnostic non-invasive biomarker., Design: Prospective cohort pilot study., Setting: Nepean Hospital and UNSW Microbiome Research Centre, Australia., Population: Sixty-four age- and sex-matched subjects (n = 19 healthy control (HC); n = 24 non-endometriosis (N-ENDO) and n = 21 confirmed endometriosis (ENDO)). All study participants, besides healthy controls, underwent laparoscopic surgical assessment for endometriosis, and histology was performed on excised lesions., Methods: Oral, stool and, vaginal samples were self-collected at a single time point for healthy controls, and preoperatively for patients undergoing laparoscopy. Samples underwent 16S rRNA amplicon sequencing, followed by bioinformatics analysis., Main Outcome Measures: Compositional differences between cohorts as identified by diversity analyses, and differentially abundant microbial taxa, as identified by LEfSE analysis., Results: The composition of the oral (adjusted p = 0.003), and stool (adjusted p = 0.042) microbiota is different between the three cohorts. Differentially abundant taxa are present within each cohort as identified by LEfSE analysis. Particularly, Fusobacterium was enriched in the oral samples of patients with moderate/severe endometriosis., Conclusions: Taxonomic and compositional differences were found between the microbiota in the mouth, gut and, vagina of patients with and without endometriosis and healthy controls. Fusobacterium was enriched in patients with moderate/severe endometriosis. Fusobacterium is noted as a key pathogen in periodontal disease, a common comorbidity in endometriosis. These findings suggest a role for the oral, stool and, vaginal microbiome in endometriosis, and present potential for microbial-based treatments and the design of a diagnostic swab., (© 2024 The Author(s). BJOG: An International Journal of Obstetrics and Gynaecology published by John Wiley & Sons Ltd.)

In vitro studies have reported the antimicrobial effect of an inoculum (L2) made from commercial probiotic strains of Lactobacillus spp. (Lactobacillus (L.) crispatus, L. brevis and L. gasseri) against some ruminants' pathogens. However, their possible beneficial effects on controlling the vaginosis caused by the use of intravaginal sponges for oestrus synchronisation have not been evaluated in in vivo models. Therefore, the purpose of this study was to perform an intravaginal double inoculation of this same non-lyophilised (non-freeze-dried) L2 inoculum, in 120 ewes from four dairy commercial ovine flocks using straws similar to those used in artificial insemination. Globally, in the entire study population, the fertility percentage (65%) of the inoculated groups increased by 10% compared to the control groups (55%). Specifically, increases in the percentages of pregnant sheep in flocks 1 and 4 (10.7 and 5.7% respectively; P > 0.05) and flock 3 (27.3%; P < 0.05) were observed in inoculated ewes. Overall, the administration of L2 on the day of intravaginal sponge insertion decreased the number of ewes with vaginitis by 11.13% at the time of sponge removal (T1). In the first flock, the administration of the probiotic positively modulated the microbiota with an increased richness and presence of Lactobacillales accompanied by a reduction in Mycoplasma spp. Genera such as Fusobacterium, Streptobacillus, Campylobacter or Trueperella were linked to the presence of vaginitis at T1. Genera belonging to Lactobacillales, including some genera previously reported to be beneficial for pregnancy, significantly increased at the time of pregnancy diagnosis but significantly decreased at T1. The results of this study suggest that the use of probiotics via intravaginal inoculation as a strategy to positively modulate the microbiota and vaginal inflammation, derived from the use of intravaginal sponges, is also associated with improved fertility. This novel approach could improve animal welfare in oestrus-synchronisation protocols and represents an alternative to the prophylactic use of antibiotics., (Copyright © 2025 The Author(s). Published by Elsevier B.V. All rights reserved.)

Background: Bacterial vaginosis (BV) is a significant global public health issue due to its high recurrence rate and association with various adverse health effects. Understanding the composition and dynamics of the vaginal microbiota (VMB) is essential for better understanding of vaginal health and for developing effective strategies to improve BV management. The study aimed to determine the composition and diversity of the VMB in Thai women with BV before and after metronidazole (MTZ) treatment, and in healthy women., Methods: Vaginal samples were collected from 20 women with BV (each sampled at three time points: pre-MTZ treatment, post-MTZ treatment and follow-up) and from 20 healthy women (each sampled once). The VMB was analyzed using 16S rRNA gene sequencing via next-generation sequencing on the Ion Torrent PGM platform., Results: The VMB in Thai women with BV was predominantly composed of Gardnerella, Prevotella and Fannyhessea (formerly Atopobium), while Lactobacillus dominated in healthy controls. Women with BV exhibited greater bacterial diversity and higher prevalence of anaerobic species compared to healthy women. There was higher diversity and abundance in the VMB from pre-MTZ samples, while post-MTZ and follow-up samples displayed lower diversity. In the follow-up stage, the VMB was divided into two subgroups: the larger cured subgroup, which shifted towards a Lactobacillus-dominated composition resembling healthy controls, and the small treatment-failure subgroup, which exhibited a Gardnerella-dominated profile similar to pre-MTZ., Conclusions: This study provides valuable insights into the structure and dynamics of the VMB in Thai women with BV before and after treatment, offering potential markers for predicting treatment outcomes., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2025 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

This study examined the relationship between the vaginal microbiome, HPV infection, and cervical intraepithelial neoplasia (CIN) in 173 women. Subjects were grouped by HPV status and cervical lesion severity, ranging from HPV-negative to CIN Grade 2 or higher. Using VALENCIA classification, the study identified different community state types (CSTs) of vaginal microbiota, with CST IV subtypes (Staphylococcus dominated) showing high diversity and increased pathogenic bacteria. Results showed a higher microbial diversity in women with CIN compared to HPV-negative women, with specific microbial profiles like Enterococcus faecalis and Staphylococcus aureus correlating with HPV infection severity. LEfSe and STAMP analyses revealed that particular bacterial species, such as Fannyhessea vaginae and Sneathia vaginalis, could serve as biomarkers for HPV and CIN status. Furthermore, Streptococcus and Staphylococcus were associated with HPV genotypes α7 and α9, respectively. These findings highlight how the composition of vaginal microbiota may influence HPV persistence and CIN progression, emphasizing the need for further research into microbial dysbiosis as a potential target for cervical cancer prevention and management., (© 2025 Wiley Periodicals LLC.)

Introduction: Despite national guidelines and use of intrapartum antibiotic prophylaxis (IAP), Streptococcus agalactiae (group B streptococci (GBS)) is still a leading cause of morbidity and mortality in newborns in Europe and the United States. The European DEVANI (Design of a Vaccine Against Neonatal Infections) program assessed the neonatal GBS infection burden in Europe, the clinical characteristics of colonized women and microbiological data of GBS strains in colonized women and their infants with early-onset disease (EOD)., Methods: Overall, 1083 pregnant women with a GBS-positive culture result from eight European countries were included in the study. Clinical obstetrical information was collected by a standardized questionnaire. GBS strains were characterized by serological and molecular methods., Results: Among GBS carriers included in this study after testing positive for GBS by vaginal or recto-vaginal sampling, 13.4% had at least one additional obstetrical risk factor for EOD. The five most common capsular types (i.e., Ia, Ib, II, III and V) comprised ~ 93% of GBS carried. Of the colonized women, 77.8% received any IAP, and in 49.5% the IAP was considered appropriate. In our cohort, nine neonates presented with GBS early-onset disease (EOD) with significant regional heterogeneity., Conclusions: Screening methods and IAP rates need to be harmonized across Europe in order to reduce the rates of EOD. The epidemiological data from eight different European countries provides important information for the development of a successful GBS vaccine., Competing Interests: Declarations. Conflict of interest: Financial interests: Margarit I, Maione D and Rinaudo D are employees of the GSK group of companies and hold shares in the GSK group of companies. Margarit I, Maione D and Rinaudo D are listed as inventor on patents owned by the GSK group of companies. Uffe B. Skov Sørensen and Mogens Kilian received personal consultancy fees from Suzhou VACMICRO Biotech Co., Ltd. Non-financial interests: Markus Hufnagel has been invited by Novartis to an advisory board meeting on juvenile idiopathic arthritis. Pierrette Melin has provided one scientific consultation for GSK vaccines. All other authors have no financial or non-financial interests to disclose. Ethical approval: The study was approved by local ethics committees of each participating institution, and informed consent for study participation has been obtained from the legal caregivers. The study was performed in accordance with the ethical standards as laid down in the 1964 Declaration of Helsinki and its later amendments or comparable ethical standards., (© 2024. The Author(s).)

Microbial infection is a significant health issue for humans. Despite the development of numerous antibiotics, the continuous rise of drug-resistant bacteria highlights the urgent need for new materials to combat these problems. In this study, four water-soluble quaternized cellulose (QC) derivatives with degrees of substitution (DS) ranging from 0.23 to 0.45 were synthesized homogeneously from cellulose carbamate (CC) in NaOH/ZnO aqueous solution. The QC derivatives exhibited broad-spectrum antibacterial activity against gram-negative/positive bacteria, fungi and drug-resistance bacteria. Models of bacterial vaginitis (BV) and vulvovaginal candidiasis (VVC) were used to evaluate the application of QC derivatives visually. Secretion smears and tissue section staining revealed that treatment with QC derivatives led to a reduction in mycelia and spores in the vagina and secretions in the VVC model, along with improved inflammation. In the BV model, vaginal secretions were reduced, clue cells in smears significantly decreased, and inflammation markedly improved. Additionally, cell experiments and staining of mouse organ tissue sections demonstrated that QC derivatives exhibited good biocompatibility. Therefore, using QC derivatives in flushing douches represents a novel approach for treating vaginitis and could serve as a benchmark for addressing other infectious diseases., Competing Interests: Declaration of competing interest All authors declare that they have no conflict of interest., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Introduction: Effective spontaneous preterm birth (sPTB) prevention is an urgent unmet clinical need. Vaginal depletion of Lactobacillus crispatus is linked to sPTB. This trial will investigate impact of an oral Lactobacillus spp. probiotic product containing an L. crispatus strain with other Lactobacilli spp., on the maternal vaginal and gut microbiome in pregnancies high-risk for sPTB., Methods: A double-blind, placebo-controlled, randomised trial will be performed at the National Maternity Hospital Dublin, Ireland. Inclusion criteria are women with history of sPTB or mid-trimester loss, cervical surgery (cone biopsy or two previous large-loop-excision-of-transformation-zone) or uterine anomaly. The intervention is oral supplementation for twelve weeks with probiotic or identical placebo. The probiotic will contains: ◦ 4 billion CFU Lactobacillus crispatus Lbv 88(2x10 9 CFU/Capsule) ◦ 4 billion CFU Lactobacillus rhamnosus Lbv 96(2x10 9 CFU/Capsule) ◦ 0.8 billion CFU Lactobacillus jensenii Lbv 116(0.4x10 9 CFU/Capsule) ◦ 1.2 billion CFU Lactobacillus gasseri Lbv 150(0.6x10 9 CFU/Capsule). Investigators and participants will be blinded to assignment., Results: The primary outcome is detectable L. crispatus in the vaginal microbiome after twelve weeks of treatment, measured using high-throughput DNA sequencing. A total of 126 women are required to detect a 25 % increase in detectable L. crispatus. Secondary outcomes include impact of intervention on the gut microbiome and metabolome, rate of sPTB and mid-trimester loss, neonatal outcomes and maternal morbidity., Conclusions: This randomised trial will investigate ability of an oral probiotic containing L. crispatus to increase its abundance in the vaginal microbiome, both directly by horizontal transfer and indirectly via microbiome and metabolome of the gut., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests:, (Copyright © 2024. Published by Elsevier Inc.)

The vaginal microbiota plays an important role in reproductive health, especially in the process of artificial insemination. The imbalance of microbiota may affect pregnancy outcomes. Therefore, this study aims to explore the composition of vaginal microbiota and its impact on artificial insemination pregnancy outcomes, combined with microscopic images and PCR fluorescence methods, in order to provide scientific basis for improving the pregnancy outcomes of husband sperm artificial insemination. This study included female patients who underwent artificial insemination with husband's sperm and collected reproductive tract samples. By observing the microbial community morphology in the sample under a microscope and analyzing microbial DNA using PCR fluorescence method, the microbial ecological status is evaluated. Among women with successful pregnancy, the proportion of beneficial bacteria (such as lactic acid bacteria) is higher, while the abundance of pathogenic bacteria is significantly reduced. Specific microbial markers detected by PCR fluorescence method are positively correlated with pregnancy rate. Therefore, the microecological state of the female reproductive tract has a significant impact on the pregnancy outcome of artificial insemination. Maintaining a good microecological balance, especially increasing the proportion of beneficial bacteria, can help improve the success rate of artificial insemination., Competing Interests: Declaration of competing interests The authors declare that there is no conflict of interests regarding the publication of this article., (Copyright © 2024. Published by Elsevier Inc.)

Purpose: Recurrent Vulvovaginal Candidiasis (RVVC) is a problematic clinical condition for which fluconazole treatment is commonly prescribed. This study investigated the interkingdom vaginal and gastrointestinal microbiomes of RVVC patients who use fluconazole intermittently or as longer-term maintenance therapy for symptom management and compared them to healthy controls., Methods: Vaginal swabs and fecal samples were collected. A novel interkingdom analysis was performed using 16 S rRNA and ITS1 gene sequencing to compare the diversity and taxonomic composition of vaginal microbiome (VMB) and gastrointestinal microbiome (GIMB)., Results: Twenty-seven women participated: 10 intermittent users and healthy controls and 7 maintenance therapy. The study revealed that microbiomes of fluconazole users do not differ in diversity metrics from healthy controls. RVVC patients using intermittent fluconazole displayed a higher abundance of vaginal C. albicans than healthy controls. Candida species pairings were not commonly observed between sites in individuals and, as such a fecal reservoir is unlikely to be implicated in recurrent symptomatology. In many of the RVVC non-Candida fungal spp. were identified in the vaginal microbiome. Users of fluconazole displayed elevations of the CST-I (Community State Type 1) associated bacterium L. crispatus. All participants displaying vaginal Candida spp. belonged to either bacterial CST-I or CST-III (Community State Type 3- L. iners associated)., Conclusion: To our knowledge, this is the first study to compare the interkingdom VMB-GIMB of women with RVVC using oral fluconazole. As fluconazole users in this study represent a typical RVVC population, trends observed in microbial abundance require further analysis to establish fluconazole's long-term microbiome safety. Examining the microbiome at both sites adds to the current understanding of microbial associated with the condition., Competing Interests: Declarations. Ethics approval and consent to participate: The study was performed in accordance with the Declaration of Helsinki (2000) for human studies. Ethics approval was obtained for the study from Griffith University Human Research Ethics Committee (GU Ref No: 2021/674). All participants provided written informed consent to participate. Clinical trial number: not applicable. Competing interests: The authors declare no competing interests., (© 2024. Crown.)

Ethnopharmacological Relevance: Vulvovaginal candidiasis (VVC) is a relatively common fungal infectious disease in the female reproductive tract. The pathogenesis of VVC not only involves Candida albicans (C. albicans) infection, but also the improper immune response of the vaginal mucosal immune system to the fungus. As a classical formula, Pulsatilla decoction has been proven to exert protective effect in both clinical and experimental research in VVC. However, the specific mechanism of Pulsatilla decoction in VVC remains elusive. This study investigated the mechanism of n-butanol extract of Pulsatilla decoction (BEPD) in treating VVC from the perspective of type I interferon signaling and related mitochondrial function., Materials and Methods: A VVC-rat model was developed using an estrogen-based method to evaluate the effectiveness of BEPD in treating VVC, and the therapeutic efficacy of BEPD against VVC was comprehensively evaluated by fungal morphology and burden, neutrophil numbers, histopathology, pro-inflammatory and anti-inflammatory cytokines production, and LDH level. Immunohistochemistry (IHC), immune fluorescence (IF), Western Blot (WB) and RT-qPCR assays were conducted to assess type I interferon signaling and mitochondria functions. Candidalysin-induced vaginal epithelial inflammation in vitro, as cellular models, was employed to detect the changes in type I interferon signaling and mitochondria function before and after BEPD-containing serum intervention., Results: BEPD could significantly improve the inflammation, reduce fungal loads and inhibit fungal growth, balance pro-inflammatory and anti-inflammatory cytokine levels in VVC model rats. The findings from IHC, IF, WB and RT-qPCR revealed that BEPD could promote type I interferon signaling and alleviate mitochondrial functional damage in VVC model rats, and BEPD-containing serum could play the same role in vitro., Conclusion: The study findings generally demonstrated that BEPD could improve the inflammation and correct the immune imbalance in VVC through regulation of type I interferon signaling and mitochondrial function., Competing Interests: Declaration of competing interest The author is an Editorial Board Member/Editor-in-Chief/Associate Editor/Guest Editor for this journal and was not involved in the editorial review or the decision to publish this article. The authors declare the following financial interests/personal relationships which may be considered as potential competing interests:The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Serotype M28 isolates of the bacterial pathogen the group A Streptococcus (GAS; Streptococcus pyogenes ), but not isolates of other serotypes, have a nonrandom association with cases of puerperal sepsis, a life-threatening infection that can occur in women following childbirth. In prior studies, we established that RD2, a pathogenicity island present in all M28 GAS isolates but mostly absent from other serotypes, is a factor in the M28-puerperal sepsis association. Here, we identified a significant reduction in the RD2 conjugation frequency in inter-serotype conjugation assays relative to intra-serotype assays. As isolates of most GAS serotypes produce an antiphagocytic hyaluronic acid capsule, while M28 isolates do not, we tested whether the capsule served as a barrier to RD2 acquisition or maintenance. The data showed that capsule production had no impact on the RD2 conjugation frequency or on the ability of RD2 to enhance vaginal colonization by GAS, but did inhibit the ability of RD2 to enhance GAS adherence to vaginal epithelial cell lines. Further molecular explanations for the inter-serotype barrier to RD2 conjugative transfer were investigated, and a conserved, chromosomally encoded Type I restriction-modification system was identified as being key. We also identified that RD2 modifies the GAS transcriptome, including mRNAs encoding virulence factors with adherence and dissemination roles, following exposure to human plasma. Our data provide insights into factors that contribute to the restriction of the RD2 pathogenicity island to discrete subsets of the GAS population., Competing Interests: The authors declare no conflict of interest.

Ethnopharmacological Relevance: Medicinal plants are frequently used in Caribbean traditional medicine as low-cost, culturally relevant treatments for women's health concerns, such as gynecological infections. These plants are typically applied topically, potentially affecting both pathogenic bacteria (e.g., Gardnerella vaginalis) and beneficial vaginal microbes (Lactobacillus spp.). However, few studies have examined the impact of these plants on both beneficial and pathogenic vaginal bacteria., Aim of the Study: Argemone mexicana, available in New York City and commonly used to treat gynecological infections by immigrants from the Dominican Republic, was investigated for its chemical variation and effects on the vaginal microbiota. We hypothesized that variations in the bioactivity of Argemone mexicana on Gardnerella vaginalis and Lactobacillus spp. are due to differences in antimicrobial compounds across different preparations., Materials and Methods: Untargeted and targeted metabolomic analysis using UPLC-qToF-MS and UPLC-TQD-MS were conducted on Argemone mexicana samples collected in New York City. Antimicrobial assays were used to assess the effects of Argemone mexicana samples on beneficial and pathogenic vaginal bacteria. ProGenesis QI and EZinfo were used for metabolomic analysis to link bioactivity with chemometric data., Results: UPLC-qToF-MS and statistical analyses showed that chemical variation correlated with plant tissue type and processing (dry or fresh samples). These differences were evident in antimicrobial screenings, where active plant samples were antimicrobial against pathogenic bacteria only, with no effect on beneficial Lactobacillus. Known antimicrobial benzoquinone alkaloids, such as berberine, were partly responsible for the observed microbiological activity. Berberine exhibited similar inhibition patterns, reduced biofilm formation, and trended towards higher concentration in active samples., Conclusions: Extracts of Argemone mexicana, a plant used in Caribbean women's health, did not have an effect on beneficial vaginal microbes, but did inhibit pathogenic Gardnerella vaginalis. This antimicrobial activity correlated with the chemical variation of berberine and other related alkaloids across traditional preparations of Argemone mexicana. These results may be relevant for treating gynecological infections, not only with this plant, but other berberine-containing taxa., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Background: The decision to use a particular test to diagnose patients presenting with symptoms of vaginitis and/or STI is based primarily on the prevailing standards of care in the clinic at which the patient evaluation takes place. As a result, laboratory testing of vaginal samples for these patients often involves either an STI or a vaginitis test, but rarely both options simultaneously, which complicates the diagnosis and management of concurrent infections., Methods: Using de-identified remnant vaginal specimens from symptomatic patients previously tested for STI ( Chlamydia trachomatis (CT), Neisseria gonorrhoeae (GC) and Trichomonas vaginalis (TV)) with the Becton Dickinson (BD) CTGCTV2 assay for BD MAX System, positivity for bacterial vaginosis (BV) and Candida spp (associated with vulvovaginal candidiasis (VVC)) were evaluated using the molecular-based BD MAX Vaginal Panel., Findings: The rate of STI/BV co-infection was 79.4% (227/286) in this symptomatic population, while that of STI/VVC was 27.0% (77/285). Women diagnosed with any one of the three STIs tested had an OR 2.86 (95% CI, 1.99, 4.11; p<0.0001) for a concurrent BV infection and OR 0.96 (95% CI, 0.67, 1.37; p=0.8085) for infection with Candida species., Conclusion: Our results suggest that women being tested for STI have a high prevalence of co-infection with BV and a lower, although appreciable, prevalence of co-infection with VVC. The detection of co-occurring vaginal infections can be facilitated by molecular testing using a single sample., Competing Interests: Competing interests: BVDP: Author’s institution received funding from BD Life Sciences for the conduct of this study and has also received study funding from Abbott, Cepheid, Cue, Hologic, MagIC, Rheonix and Roche. BVDP receives consulting fees and/honoraria from Abbott Rapid Diagnostics and Roche Molecular. SK, SP, VP and ET-C are employees of BD and own BD shares. PD and CA: no conflicts of interest., (© Author(s) (or their employer(s)) 2025. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ Group.)

Background: Genital inflammation increases HIV susceptibility and is associated with the density of pro-inflammatory anaerobes in the vagina and coronal sulcus. The penile urethra is a critical site of HIV acquisition, although correlates of urethral HIV acquisition are largely unknown. While Streptococcus mitis is a consistent component of the urethral flora, the presence of Gardnerella vaginalis has been linked with prior penile-vaginal sex and urethral inflammation. Here, we use a flow cytometry-based bacterial assay to quantify urethral IgA and IgG that bind G. vaginalis and S. mitis in a cross-sectional cohort of 45 uncircumcised Ugandan men and to evaluate their association with the urethral microbiome and local soluble immune factors., Results: Urethral antibodies binding both bacterial species were readily detectable, with G. vaginalis predominantly bound by IgA, and S. mitis equivalently by IgA and IgG. Gardnerella vaginalis-binding IgA was elevated in participants with detectable urethral Gardnerella, with the latter only present in participants who reported prior penile-vaginal sex. In contrast, detectable urethral S. mitis was not associated with sexual history or levels of S. mitis-binding IgA/IgG. The time from the last penile-vaginal sex was inversely correlated with the urethral concentrations of total IgA, G. vaginalis-binding IgA, and chemokines IL-8 and MIP-1β; these inflammatory chemokines were independently associated with higher total IgA concentration, but not with G. vaginalis-binding IgA., Conclusions: This first description of microbe-binding antibodies in the penile urethra suggests that urethral colonization by Gardnerella after penile-vaginal sex specifically induces a G. vaginalis-binding IgA response. Prospective studies of the host-microbe relationship in the urethra may have implications for the development of vaccines against sexually-transmitted bacteria. Video Abstract., Competing Interests: Declarations. Ethics approval and consent to participate: The Research and Ethics Committee (REC) of the Uganda Virus Research Institute in Entebbe, Uganda, and the Institutional Review Board at the University of Toronto in Canada provided review, ethical oversight, and approval of the study proposal. All study participants gave written informed consent in accordance with the Declaration of Helsinki. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)

Objectives: Azoles have been the mainstay of recurrent vulvovaginal candidiasis (RVVC) for many years. Because of a recent anecdotal increase in non- Candida albicans yeasts (NCAY) and azole-resistant C. albicans cases, their prevalence was calculated from cultures for yeasts in women with complicated/RVVC over 3 years., Methods: Retrospective data search of vaginal cultures from adult women in Leeds, UK between April 2018 and March 2021 was conducted. Samples with clinical details of complicated/RVVC had full yeast identification and antifungal susceptibility performed. Differences in prevalence between 12-month periods were determined using χ 2 tests., Results: Over the 3 years, cultures were performed on 5461 vaginal samples from women with clinical information indicating they had complicated/RVVC, RVVC, with 1828 (33.5%) growing yeasts.Over 85% of yeasts each year were C. albicans , however the proportion declined yearly with an increase in NCAY species. Nakaseomyces glabrata was the most frequent NCAY species isolated, increasing from 2.8% in 2018-19 to 6.8% in 2020-21. Total NCAY species increased from 6.0% in 2018-19 to 12.6% in 2020-21. Fluconazole-sensitive dose-dependant (SDD) and resistant isolates increased from 3.5% in 2018-19 to 7.7% in 2019-20 and 9.6% in 2020-21. Most resistance was in C. albicans and the majority of cases were seen in primary care. Most fluconazole non-sensitive isolates were either SDD or resistant to itraconazole (77% and 23%, respectively) and were intermediate or resistant to voriconazole (36.4% and 60%, respectively)., Conclusion: There was a significant increase in the prevalence of NCAY and fluconazole-resistant C. albicans in complicated/RVVC cultures over these 3 years. Successful treatment of such cases can be very challenging. The exact reasons for this increase remain unclear but it follows a policy change that encouraged a clinical diagnosis and empirical treatment of vulvovaginal candidiasis, rather than fungal culture, in primary care., Competing Interests: Competing interests: RCB has received fees from Pfizer Ltd for contributing to webinar presentions in 2023., (© Author(s) (or their employer(s)) 2025. No commercial re-use. See rights and permissions. Published by BMJ Group.)

Bacterial vaginosis (BV), characterized by an imbalance in the vaginal microbiota, is a prevalent condition among women of reproductive age and a risk factor for human immunodeficiency virus, sexually transmitted infections, and preterm birth. BV is generally considered to induce mucosal inflammation, but the specific pathways and cell types involved are not well characterized. This prospective study aimed to assess associations between microbial changes and mucosal immune responses in BV patients. Therefore, samples from 20 premenopausal women with BV and treated with metronidazole were analyzed. Vaginal swabs, menstrual cup, and endocervical cytobrush samples were collected before treatment, weekly for four weeks, and at 2, 4, and 6 months for Nugent scoring, immune cell populations and cytokine analysis. Of 105 study intervals, 27 (25.7%) showed improvement in Nugent category, 61 (58.1%) remained unchanged, and 17 (16.2%) worsened. Improvement correlated with decreased monocytes (p = 0.005), while worsening was linked to increased monocytes (p < 0.001) and dendritic cells (p = 0.02). B cells (p = 0.02) and IFN-γ-induced chemokines - IP-10 (p = 0.007), MIG (p = 0.049), and ITAC (p = 0.005) - were associated with improvement. In conclusion, although the T-cell-associated chemokines IP-10, ITAC, and MIG were strongly associated with improvements in Nugent category, our findings indicate that antigen-presenting cells, particularly monocytes, show the most dynamic response to shifts in the vaginal microbiota in patients with BV., Competing Interests: Declarations. Competing interests: Dr. Mitchell serves on the scientific advisory board of Concerto Bio, and Ancilia Biosciences, and receives royalties from Up to Date. The remaining authors declare no conflict of interest., (© 2025. The Author(s).)

Background: Tuberculosis remains an infectious disease of global concern, with potential impacts on respiratory and intestinal microbiota owing to prolonged broad-spectrum antibiotic therapy. Despite its potential to cause infertility, the vaginal microbiota of women with genital tuberculosis remains poorly understood. We comprehensively analyzed the vaginal microbiota in Chinese women with genital tuberculosis., Results: We recruited women with pelvic (n = 28), endometrial (n = 16), and pulmonary (n = 12) tuberculosis as the research group, and healthy women (n = 11) as the control group. Vaginal discharges were collected for metagenomic analysis of its microbiota. The alpha diversity of the vaginal microbiota in women with genital tuberculosis was slightly higher than that in healthy women, though the difference was not statistically significant (P = 0.23). Similarly, no significant differences in alpha diversity were observed between women with genital and pulmonary tuberculosis (P = 0.82) or between those with pelvic and endometrial tuberculosis (P = 0.82). Notably, the lowest alpha diversity was recorded six months to one year after initiating anti-tuberculosis treatment, with this decline being statistically significant (P = 0.023). The dominance of Lactobacillus iners in the vaginal microbiota was more common in women with genital tuberculosis than that of Lactobacillus crispatus. Furthermore, the abundance of short-chain fatty acid -producing anaerobes, such as Actinomycetes, Streptococcus, and Finegoldia, were significantly increased. Short-chain fatty acid precursor pathways, including the ko03010 ribosome pathway, ko00970 aminoacyl-tRNA synthesis, ko00230 purine metabolism, ko00240 pyrimidine metabolism, and ko00010 glycolysis gluconeogenesis pathway, were significantly upregulated in women with endometrial tuberculosis., Conclusions: Extrapulmonary tuberculosis, particularly genital tuberculosis and its associated vaginal dysbiosis impacts female fecundity. Vaginal dysbiosis is more pronounced when M. tuberculosis invades the endometrium. Given the effect of antibiotics on vaginal flora, probiotic combined interventions could be used as a future research direction., Clinical Trial Number: Not applicable., Competing Interests: Declarations. Ethics approval and consent to participate: All participants provided their informed consent before participating in the study. The study was conducted in accordance with the Declaration of Helsinki, and the protocol was approved by the Ethics Committee of Beijing Obstetrics and Gynecology hospital, Capital Medical University (2022-KY-064-01). Consent for publication: Not applicable. Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)

Objective : During pregnancy, vulvovaginal infections (VVIs), including abnormal vaginal flora (AVF), bacterial vaginosis (BV), and vulvovaginal candidiasis (VVC), are associated with serious complications and discomfort. We aimed to elucidate the effectiveness of oral probiotics in secondary prevention of VVIs in pregnant women. Study design : A multicenter prospective randomized, double-blind, placebo-controlled trial was conducted at three medical centers between 2016 and 2021. Women who complained of vaginal symptoms with positive smear for AVF/BV and/or candida were treated with antibiotics or an antimycotic agent, respectively. After confirmation of VVI eradiation by repeated vaginal smear, the women were divided into a research group, receiving two capsules/day of oral probiotic formula containing Bifidobacterium bifidum , Bifidobacterium lactis , Lactobacillus ( L. ) acidophilus , L. paracasei , L. rhamnosus and Streptococcus thermophilus (>6 × 10 9 CFU/capsule), and a control group, receiving a placebo (two capsules/day) until delivery. At least once a month or following complaints, a vaginal smear was taken to assess vaginal microbiota. If VVIs were found, they were treated with antibiotics/antimycotics, and eradication was assessed by a repeated vaginal smear. Lactobacilli vaginal colonization, including the specific strains from the probiotic capsules, were detected using the matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI TOF-MS). The primary outcome was the rate of women who developed VVI during the study period until delivery. Results : Twenty-three and twenty-four women were analyzed in the probiotic and placebo cohorts, respectively. There was no difference in the rate of any VVI between the probiotic and placebo cohorts (16 (67%) versus 11 (48%), respectively; p = 0.19), time until first infection or pregnancy outcomes. The lactobacilli strains that colonized the vagina were similar at baseline and following probiotic or placebo administration. No woman was detected with vaginal colonization of the strains from the capsule, although the probiotics were taken for about 4 months. Conclusions : The oral probiotic product tested in this study did not reduce the recurrence rate of VVIs in pregnant women following eradication.

Background: A dysbiotic vaginal microbiome (VMB) is associated with clinical conditions such as bacterial vaginosis (BV) and an increased risk of human immunodeficiency virus (HIV-1) infection. Considering the high prevalence of BV among African, Caribbean and Black (ACB) women, we conducted a prospective, randomized, open-label phase 1 clinical trial to determine the feasibility, safety and tolerability of administering low-dose estrogen, probiotics or both in combination to improve vaginal health and decrease HIV-1 susceptibility., Methods: ACB women aged 18-49 from the Greater Toronto Area (GTA) were randomized to one of four study arms: intravaginal estradiol (Estring©; 7.5mg/day); a vaginal probiotic (RepHresh™ Pro-B™) administered twice daily; a combination of Estring© and vaginal RepHresh™ Pro-B™ (twice daily); or the Estring© and oral RepHresh™ Pro-B™ (twice daily), for a duration of 30 days. Feasibility was evaluated through enrolment, retention, and adherence rates, while safety and tolerability were determined by a pre- and post-treatment blood panel and reported adverse events (AEs)., Results: Overall, 63 ACB women were screened, 50 were enrolled and received the intervention while 41 completed the study, resulting in 80% enrollment and 82% retention rates. Overall adherence to the study protocol was high at 93%, with an adherence of 92% for RepHresh™ Pro-B™ and 97% for Estring©. A total of 88 AEs were reported by 29 participants which were mild (66/88; 75%) and largely resolved (82/88;93%) by the end of the study, with no serious AEs (SAEs) noted. In addition, a panel of safety blood markers measured pre- and post-intervention confirmed no clinically significant changes in blood chemistry or blood cell count., Conclusion: Overall, the administration of intravaginal estrogen and/or probiotics in pre-menopausal ACB women is feasible, safe, and well tolerated., Trial Registration: The trial was registered with Clinicaltrials.gov (NCT03837015) and CIHR HIV Clinical Trials (CTN308)., Competing Interests: I have read the journal’s policy and the authors of this manuscript have the following competing interests: GR developed the probiotic strains GR-1 and RC-14 but has had no financial interest in them for 15 years. GR consults for Seed, a company producing probiotic strains not used in this study. He was not paid salary for any roles with the company. All the other authors have declared no conflict of interest. This does not alter our adherence to PLOS One policies on sharing materials and data.”, (Copyright: © 2025 Gill et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Bacterial vaginosis (BV) is an abnormal gynecological condition caused by the overgrowth of specific bacteria in the vagina. This study aims to develop a novel method for BV detection by integrating surface-enhanced Raman scattering (SERS) with machine learning (ML) algorithms. Vaginal fluid samples were classified as BV positive or BV negative using the BVBlue Test and clinical microscopy, followed by SERS spectral acquisition to construct the data set. Preliminary SERS spectral analysis revealed notable disparities in characteristic peak features. Multiple ML models were constructed and optimized, with the convolutional neural network (CNN) model achieving the highest prediction accuracy at 99%. Gradient-weighted class activation mapping (Grad-CAM) was used to highlight important regions in the images for prediction. Moreover, the CNN model was blindly tested on SERS spectra of vaginal fluid samples collected from 40 participants with unknown BV infection status, achieving a prediction accuracy of 90.75% compared with the results of the BVBlue Test combined with clinical microscopy. This novel technique is simple, cheap, and rapid in accurately diagnosing bacterial vaginosis, potentially complementing current diagnostic methods in clinical laboratories., Importance: The accurate and rapid diagnosis of bacterial vaginosis (BV) is crucial due to its high prevalence and association with serious health complications, including increased risk of sexually transmitted infections and adverse pregnancy outcomes. Although widely used, traditional diagnostic methods have significant limitations in subjectivity, complexity, and cost. The development of a novel diagnostic approach that integrates SERS with ML offers a promising solution. The CNN model's high prediction accuracy, cost-effectiveness, and extraordinary rapidity underscore its significant potential to enhance the diagnosis of BV in clinical settings. This method not only addresses the limitations of current diagnostic tools but also provides a more accessible and reliable option for healthcare providers, ultimately enhancing patient care and health outcomes., Competing Interests: The authors declare no conflict of interest.

Studies in humans indicate that certain Chlamydia trachomatis serovars are more pathogenic than others. Specifically, several studies concluded that serovars from the C-complex are more pathogenic than those from the B-complex, although there are reports that do not support this finding. To investigate these results in an animal model, the eight genitourinary C. trachomatis serovars were tested in two strains of mice: C3H/HeN and BALB/c. These two strains of mice were investigated because C3H/HeN is more susceptible to Chlamydia muridarum infections than BALB/c, indicative of differences in their immunogenetic background. Mice were infected transcervically with 10 5 inclusion forming units of each of the C. trachomatis serovars, and vaginal cultures were collected. To determine the pathogenicity and its impact on fertility, at week seven post-infection, female mice were caged with male mice. In the C3H/HeN mice, significant differences in vaginal shedding and fertility were observed between serovars from the B-complex (D and E) and those from the C-complex (H, I, J) and B- and C-related complexes (G, F, and K). The animals infected with serovars F, G, H, I, J, and K shed less but had significantly more infertility than the mice infected with serovars D or E. The experiments in the BALB/c mice, however, did not show major differences in pathogenicity between the eight C. trachomatis serovars. These results support the findings in humans and emphasize the critical importance of the immunogenetic background of the host on the outcome of C. trachomatis infections. The data imply that management of C. trachomatis -infected patients may require a more personalized approach.

Background: Gardnerella is a key pathogen in bacterial vaginosis (BV), but the role of the different Gardnerella species remains unclear. We investigated the role of four Gardnerella species, as well as Fannyhessea vaginae , Lactobacillus crispatus and L. iners in BV., Methods: From 331 pregnant women from the Democratic Republic of the Congo, BV was diagnosed using Nugent scoring and a cervicovaginal lavage was used to quantify G. leopoldii , G. piotii , G. swidsinskii , G. vaginalis, F. vaginae, L. crispatus and L. iners by qPCR. Univariate associations between these species and clinical outcomes were assessed. A logistic regression model and ROC curves were calculated to determine the best diagnostic marker for BV., Results: Here, L. iners (75.8%) was the most prevalent species and G. vaginalis (36.0%) the most common Gardnerella species. All investigated Gardnerella spp. were prevalent (50.9-57.9%) in women with (asymptomatic) BV. Univariate analysis revealed no significant associations with clinical symptoms of BV, while F. vaginae (positive Whiff test, high pH), G. vaginalis (high pH) and L. crispatus (low pH) were associated with signs of BV. G. piotii was associated with markers of urinary tract infection. Women with L. iners had higher odds of delivering preterm. ROC analyses showed that F. vaginae was the best marker for BV (AUC 0.81), and the combined model further increased the diagnostic performance (AUC 0.90)., Conclusion: All Gardnerella species were involved in BV, although none were associated with the most important clinical symptoms of BV and none emerged as a superior molecular marker for BV., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2025 Himschoot, Mulinganya, Rogier, Bisimwa, Kampara, Kujirakwinja, Mongane, Mubalama, Callens, Vaneechoutte and Cools.)

Introduction: The vaginal microbiota is a complex and dynamic micro-ecosystem that plays a pivotal role in protecting the host from various pathogens. Previous studies have investigated the diversity of the vaginal microbiome and its association with health outcomes, particularly the development of HPV-related disorders. This study aimed to investigate the correlation between the vaginal microbiota, HPV infection, cervical intraepithelial neoplasias (CINs), and cervical cancers in 69 women., Methods: DNA was extracted from vaginal samples, followed by HPV genotyping through PCR and sequenced of the16S rRNA gene., Results: Our results revealed that Lactobacillus was the predominant bacterium across all groups, with prevalence rates of 60.2% in women with HPV+, 63.9% in CINI, 97.7% in CINII, 52.0% in CINIII, 36.9% in cervical cancer, and 70.9% in NILM (normal cytology). Additionally, an elevated proportion of Gardnerella was identified as a high-risk bacterium associated with HPV infection, potentially contributing to the progression of cervical lesions. High-risk HPV genotypes, particularly HPV16, 52, and 33, were found to be more prevalent among women with HPV+, CIN, and cervical cancer. We also observed significantly higher alpha diversity in the vaginal microbiome of women with HPV+ and CIN, as indicated by increased Sobs, Shannon, Ace, and Chao indices, compared to the NILM group., Conclusion: These findings suggest that HPV infection and its associated pathological conditions are closely linked to alterations in the vaginal microbiome. This underscores the need for further research to unravel the intricate relationship between HPV genotype infections and vaginal microbiota, which could pave the way for new diagnostic and therapeutic approaches., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2025 Li and Wu.)

Background: Aerobic vaginitis (AV) is a state of abnormal vaginal microbiota, which is associated with increased numbers of aerobic, enteric bacteria and inflammation of the vaginal epithelium. Anti-microbial treatment combined with anti-inflammatory therapy could be useful in the treatment of this condition. It is known that calcitriol, the active form of vitamin D, plays an important role in modulating the immune response in several inflammatory diseases. The aim of this preliminary study was to evaluate in vitro the influence of calcitriol on the immune response of human vaginal epithelial cells to bacterial infection. Moreover, we assessed the anti-bacterial properties of calcitriol, as well as its synergistic activity with antibiotics that are used for the treatment of AV., Results: Human vaginal A431 epithelial cells were treated with calcitriol (100 nM) and then stimulated with thermally inactivated strains of AV-associated bacteria (Escherichia coli, Enterococcus faecalis, Streptococcus agalactiae), or first stimulated with heat-treated bacteria and then incubated with calcitriol (30 nM). After 24 h, 48 h, and 72 h post-infection, culture supernatants were collected, and levels of pro-inflammatory cytokines IL-6 and IL-1β were analyzed using enzyme-linked immunosorbent assays (ELISA). As it turned out, calcitriol treatment pre- or post-bacterial stimulation of vaginal epithelial cells significantly decreased the levels of IL-6 and IL-1β compared to cells stimulated only with bacteria. Downregulation of pro-inflammatory cytokines was more frequently significant when cells were pre-treated with calcitriol. Additionally, this study evaluated the anti-bacterial properties and synergistic activity of calcitriol with antibiotics by determining the Minimal Inhibitory Concentration (MIC) using colorimetric, Resazurin-based microdilution method detecting the active metabolism of bacteria. As a result, calcitriol by itself did not exhibit clinically relevant anti-bacterial activity; however, when combined with antibiotics, calcitriol significantly reduced the MIC values., Conclusions: The results of this preliminary study confirm that calcitriol treatment influences cytokine-mediated immune response during infection and indicate that it can be effective in enhancing antibiotic activity. Such properties could be very useful in the treatment of inflammatory diseases. However, conclusions regarding the potential use of calcitriol in the treatment of AV require further confirmation in in vivo studies as well as in well-designed clinical trials., Competing Interests: Declarations. Ethics approval and consent to participate: Clinical trial number: not applicable. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)

Introduction: For years, the placenta was believed to be sterile, but recent studies reveal it hosts a unique microbiome. Despite these findings, significant questions remain about the origins of the placental microbiome and its effects on pregnancy and fetal health. Some studies suggest it may originate from the vaginal tract, while others indicate that oral bacteria can enter the maternal bloodstream and seed the placenta. However, research analyzing the vaginal, oral, and placental microbiomes within the same cohort is lacking. Additionally, it's unclear whether the placental microbiome differs between healthy pregnancies and those with complications like preterm birth (PTB), which remains a leading cause of neonatal morbidity and mortality worldwide., Methods: In this study, we performed 16S rRNA gene sequencing to investigate the composition of the oral and placental microbiome in samples collected from 18 women who experienced PTB and 36 matched controls who delivered at term (TB), all of whom were part of the Molecular Signature in Pregnancy (MSP) study. We leveraged on the multisite microbiome sampling from the MSP participants and on our previously published vaginal microbiome data to investigate the potential origins of the placental microbiome and assess whether its composition varies between healthy and complicated pregnancies., Results and Discussion: Our analysis revealed distinct profiles in the oral microbiome of PTB subjects compared to those who delivered at term. Specifically, we observed an increased abundance of Treponema maltophilum, Bacteroides sp, Mollicutes, Prevotella buccae, Leptotrichia, Prevotella&#95;sp&#95;Alloprevotella, in the PTB group. Importantly, Treponema maltophilum species showed higher abundance in the PTB group during the second trimester, suggesting its potential use as biomarkers. When we assessed the placenta microbiome composition, we found that Firmicutes, Bacteroidetes, Actinobacteria, and Proteobacteria were the most dominant phyla. Interestingly, microorganisms such as Ureaplasma urealyticum were more abundant in PTB placenta samples. Our findings suggest that the placenta microbiome could originate from the oral or vaginal cavities, with a notable increase in the crosstalk between the vaginal and placental sites in cases of PTB. Specifically, our data revealed that in PTB cases, the placental microbiome exhibited a closer resemblance to the vaginal microbiome, whereas in term pregnancies, the placental microbiome was similar to the oral microbiome., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2025 Saadaoui, Djekidel, Murugesan, Kumar, Elhag, Singh, Kabeer, Marr, Kino, Brummaier, McGready, Nosten, Chaussabel, Terranegra and Al Khodor.)

Aims: Vulvovaginal candidiasis (VVC) is a common women's health issue, with rising antifungal resistance. This study was aimed to prepare and evaluate the efficacy of a lipid nanoparticle-based vaccine in a murine model of VVC., Materials and Methods: Dried and reconstituted vesicles containing C. albicans antigens (DRNPs-Ca-Ags) vaccine, formulated with phosphatidylcholine and cholesterol-based lipid nanoparticles via film hydration and freeze-drying. The safety evaluation of DRNPs-CaAgs was conducted by determining hepatic (AST, ALT) or renal (BUN, creatinine) biomarkers. Female mice were immunized with DRNPs-CaAgs or Alum-CaAgs, and immune responses were evaluated via antibody titers, IgG isotypes, and splenocyte proliferation. Protective efficacy of vaccine formulations was assessed through fungal burden, biofilm formation, cytokine levels, and histopathological analysis of vaginal tissues., Key Findings: Mice vaccinated with DRNPs-CaAgs showed significantly enhanced immune responses, with higher antibody titers and IgG2a levels as compared to the Alum-CaAgs group. Vaginal fungal burden was dramatically reduced (665 ± 78 CFUs in DRNPs-CaAgs immunized group vs. 12,944 ± 3540 CFUs in Alum-CaAgs group, p < 0.01). Biofilm formation decreased by 45 % (p < 0.05), and inflammatory cytokines were significantly lowered. Histopathological analysis revealed minimal tissue damage in DRNPs-CaAgs vaccinated mice., Significance: The findings suggest DRNPs-CaAgs as a promising vaccine for VVC, eliciting strong immunity, reducing fungal load, and minimizing inflammation. While the reliance on a murine model is a limitation, future clinical trials are essential to evaluate its efficacy and safety in humans, offering a potential strategy to combat drug-resistant infections and improve women's reproductive health., Competing Interests: Declaration of competing interest There is no conflict of interest to declare., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Background: Human papillomavirus (HPV) is a viral infection, and its acquisition and persistence are significantly influenced by the vaginal microbiota. Understanding and comparing the vaginal microbiome of HPV infected women in Andaman and Nicobar Islands is crucial., Methods: The study involved collecting vaginal swabs and extracting DNA using the QIAamp DNA Minikit. The DNA was then subjected to PCR amplification to confirm HPV infection. illumina NovaSeq 6000 platform was utilized to perform sequencing utilizing 2 x 250 paired end chemistry. Taxonomic analysis was performed and Bacterial abundance plots were generated and samples were grouped based on demographic parameters, pap test diagnosis, and genotypes. To assess diversity, samples were rarefied to 49,000 sequence reads per sample, and alpha and beta diversity metrics were calculated., Results: The study analyzed the presence of 21 assigned phyla, with Firmicutes, Actinobacteria, Bacteriodetes, and Proteobacteria emerging as the predominant taxa. At the genus level, Lactobacillus and Gardnerella dominated across all samples. Gardnerella was significantly more abundant in HPV-positive (22.40%) compared to HPV-negative samples (10.04%). Symptomatic group of HPV-positive samples had Gardnerella , and unclassified Coriobacteriaceae being dominant. In terms of bacterial diversity, the study found statistically significant association when comprising individuals aged 21 to 30 years to those aged 31 to 40 years., Conclusion: Most research concluded that exposure to HPV can boost bacterial diversity in vagina compared to healthy women, increasing the risk of cervical cancer development. Current study highlights the importance of vaginal microbiome associated with high and low risk HPV, various age group as well as the symptomatic and asymptomatic cases of HPV infected women in South Andaman., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2025 Parvez, Vijayakumar, Vins, Ramaiah, Anbarasu, Biswas, Beniwal, Kaur and Muruganandam.)

Background: Group B streptococcus (GBS) colonization in pregnant women is associated with adverse perinatal outcomes, including stillbirth. This meta-analysis investigated the relationship between maternal rectovaginal GBS colonization and the risk of stillbirth., Methods: We conducted a comprehensive literature search across several databases, including PubMed, Embase, Web of Science, Wanfang, and China National Knowledge Infrastructure, covering studies published from the inception of the database until September 9, 2024. The search focused on observational studies comparing the risk of stillbirth in pregnant women with and without rectovaginal GBS colonization. Results were summarized using odds ratios (ORs) and 95% confidence intervals (CIs), and a random-effects model was used to account for potential heterogeneity., Results: A total of ten studies comprising 121,195 pregnant women were included in the analysis. The pooled results indicated no significant overall association between maternal rectovaginal GBS colonization and the risk of stillbirth (OR: 1.66, 95% CI [0.95-2.91], p = 0.08; I 2 = 84%). However, sensitivity analyses revealed a significant association in studies that included intrapartum antibiotic prophylaxis (IAP) (OR: 1.36, 95% CI [1.02-1.80], p = 0.03). Subgroup analyses demonstrated a significant association between maternal rectovaginal GBS colonization and stillbirth risk in retrospective studies (OR: 2.62, p = 0.04) and in studies employing multivariate analysis (OR: 2.11, p = 0.04)., Conclusions: While the meta-analysis did not find a significant overall association between maternal rectovaginal GBS colonization and stillbirth, significant associations were noted under specific conditions, such as studies using IAP, retrospective designs, and multivariate analyses. Further research is needed to clarify these associations., Competing Interests: The authors declare that they have no competing interests., (© 2025 Wang et al.)

Lactic acid bacteria (LAB), traditionally consumed as fermented foods, are now being applied to the medical field beyond health-functional food as probiotics. Therefore, it is necessary to continuously discover and evaluate new strains with suitable probiotic characteristics, mainly focusing on safety. In this study, we isolated eight new strains from postmenopausal vaginal fluid using culturomics approaches, an emerging area of interest. Data showed that most strains possessed significant cell surface hydrophobicity (≥ 76%), auto-aggregation capacity (17 to 61%), strong adhesion activity (8 to 34%), and excellent resistance to gastric acid, bile salt, and digestive enzyme, enhancing their survival in the gastrointestinal tract. Moreover, the strains exhibited functional characteristics, including substantial antibacterial activity with a minimal inhibitory concentration (MIC) ranging from 12.5 to 50%. They also harbored bacteriocins genes, produced short-chain fatty acids (acetate and propionate), exhibited significant phagocytic activity, possessed high antioxidative properties, rapidly depleted sodium nitrite, and exhibited proteolysis and β-glucosidase activity. In addition, heat-killed LAB strains significantly reduced the gene expressions of proinflammatory cytokines such as IL-β, IL-6, and iNOS in macrophages. Safety assessment revealed no cytotoxicity in macrophage cell lines. All strains tested negative for biogenic amine or H 2 O 2 production, displayed no gelatinase or hemolytic activity, lacked virulence genes or detrimental enzymes, and displayed antibiotic susceptibility. In summary, these newly isolated strains demonstrate excellent probiotic functionality with a strong focus on safety, making them promising candidates for future drug development in the relevant fields., Competing Interests: Declarations. Conflict of interest: There are no financial conflicts of interest for the authors to disclose. Ethical Approval: The Soonchunhyang University Cheonan Hospital Ethics Committee IRB-approved samples (IRB number 2019-10-017-005) were used in the study., (© 2025. The Author(s).)

To investigate the efficacy and mechanisms of fractional CO2 laser treatment for Vaginal Relaxation Syndrome (VRS) combined with recurrent bacterial vaginitis. Patients with VRS and recurrent bacterial vaginitis were randomly assigned to an intervention group (n = 60) receiving fractional CO2 laser therapy in addition to metronidazole, or a control group (n = 60) receiving metronidazole alone. Post-treatment assessments included vaginal relaxation, vaginal health index (VHI) scores, lactobacilli distribution, vaginal pH, recurrence rates, and the correlation between lactobacilli distribution and VHI. (1) Vaginal relaxation improved significantly in the intervention group compared to both pre-treatment and the control group (P = 0.000). (2) VHI scores and lactobacilli distribution improved in both groups post-treatment, with significantly higher scores in the intervention group (P = 0.000). (3) The intervention group exhibited a significantly lower vaginal pH compared to pre-treatment and the control group (P = 0.001). (4) Recurrence rates were significantly lower in the intervention group (8.33%) compared to the control group (36.67%) (P = 0.000). A strong positive correlation was found between lactobacilli distribution and VHI in the intervention group (r = 0.79, P < 0.001). Fractional CO2 laser combined with conventional therapy improves VRS and recurrent bacterial vaginitis outcomes, reduces recurrence, and enhances vaginal microecology.Trial registration: Chinese Clinical Trial Registry, Registration number ChiCTR2400094125., Competing Interests: Declarations. Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)

Background: Vaginal colonization by Candida can lead to vulvovaginal candidiasis, which is the second most prevalent vaginal condition globally. It is frequently associated with sepsis and adverse neonatal outcomes in pregnant women. This issue is worsening in Sub-Saharan Africa, including Ethiopia. However, evidence of the existing problem is very scarce yet crucial. Thus, this study aimed to determine the vaginal colonization and vertical transmission of Candida species and their associated factors among pregnant women and their neonates in public health facilities of northeast Ethiopia., Methods: A facility-based cross-sectional study was conducted at selected public health facilities in Dessie town from April 1 to June 30, 2023, among 348 pregnant women and their newborns, using convenience sampling techniques. Socio-demographic, and clinical-related data were collected using a pre-tested, semi-structured questionnaire. Vaginal swab samples from pregnant women and pooled swabs from the external ear, nasal area, and umbilical areas of the newborns were collected and transported using Amies transport media. The samples were inoculated into Sabouraud Dextrose Agar for isolation, followed by inoculation onto a standard CHROM agar Candida plate for species identification, and a germ-tube test confirmed pseudophyphae of C.albicans. Data was entered into Epi Data version 4.6.0 software and exported and analyzed by SPSS version 25.0. A stepwise logistic regression model was used to identify the associated factors. Variables with p < 0.05 and their 95% confidence interval were considered statistically significant., Result: A total of 348 pregnant women attending vaginal delivery were included in the study. The maternal and neonatal colonization rates of Candida species were 14.1% (49/348) and 6.3% (22/348), respectively. The overall proportion of vertical transmission of Candida species was 44.9% (22/49, 95% CI: 41.2, 49.7). Among Candida isolates, 63.3% (31/49) were C. albicans and 36.7% (18/49) were C. krusei. Gestational diabetes mellitus (AOR: 4.2, 95% CI: 1.23-38.6, P = 0.047) and HIV (AOR: 1.58, 95% CI: 1.11-6.12, P = 0.049) were independently associated with maternal colonization of Candida species. Moreover, rural residence (AOR = 3.6, 95% CI: 1.37-9.5, P = 0.010) and maternal age above 28 years (AOR = 2.39, 95% CI: 1.97-5.89, P = 0.048) were independently associated with vertical transmission of Candida species., Conclusion: The findings of this study highlight the need for effective screening and treatment of Candida colonization during antenatal care., Competing Interests: Declarations. Ethical approval and consent to participate: Ethical clearance was obtained from the ethical review Board of the College of Medicine and Health Sciences, Bahir Dar University (Protocol number 749/2023). Permission for the study was granted in a letter from DCSH and DHC, where the study was conducted. After briefly describing the significance of the study, written informed consent from the pregnant mother and assent for the newborns were obtained. Confidentiality of the data was maintained. Finally, based on a microbiologically confirmed positive result, the clinician responsible for the participant was informed and the participant was treated with an appropriate treatment protocol. This study was conducted in accordance with the Declaration of Helsinki. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests. Clinical trial number: Not applicable., (© 2024. The Author(s).)

The colonization of Enterococcus in the female vagina leads to neonatal and pediatric enterococcal septicemia. Linezolid (LZD) is a kind of mainstream drug for treating multidrug-resistant Gram-positive infections. OptrA is the main LZD-resistance gene at Enterococci in human isolates. It is essential to explore the prevalence of optrA -carrying Enterococcus in vaginal secretions of late pregnant women and the drug resistance of optrA . From May to June 2023, this study recruited 340 volunteers in late pregnancy (35-40 weeks of pregnancy) to provide non-repetitive vaginal discharge samples. Luria-Bertani broth and florfenicol (10 µg/mL) were used to enrich bacteria. Enterococci was identified through time-of-flight mass spectrometry. Additionally, antimicrobial susceptibility, polymerase chain reaction, and next-generation sequencing assays were applied to this study. Fifty-four optrA -carrying Enterococcus strains were obtained, the proportion of the whole vagina of late pregnant women was 15.88% (54 out of 340), and Enterococcus faecalis account the highest proportion. All optrA -carrying Enterococcus were resistant to at least three drugs. Tetracycline, chloramphenicol, erythromycin, and LZD have higher bacterial resistance rates. Genetic environment analysis revealed that IS 1216E , fexA , and erm (A) may synergistically exert multidrug resistance with optrA . It is necessary to strengthen the surveillance of optrA -carrying Enterococcus in pregnant women. This study provides scientific support for controlling hospital infections and managing antibiotic-resistant bacteria, and provides a scientific basis for rational clinical medication.IMPORTANCEThe disruption of cervicovaginal microbiota homeostasis is considered a key factor in causing imbalance in the microenvironment, leading to inflammation, transmission of infections, and illness. Enterococcus is considered a major cause of healthcare-related infections globally. It has resistance to multiple antimicrobial drugs, which pose significant challenges for clinical treatment. Therefore, it is crucial to assess the prevalence of optrA -carrying Enterococcus in vaginal secretions of late pregnant women and the drug resistance of optrA . This study detected 15.88% of optrA -carried Enterococci in 340 pregnant women. Furthermore, we found that optrA -carrying Enterococcus strains are highly resistant to tetracycline, chloramphenicol, erythromycin, and Linezolid. Additionally, genetic environment analysis revealed that IS1216E , fexA , and erm (A) may synergistically exert multidrug resistance with optrA . This study provides scientific support for controlling hospital infections and managing antibiotic-resistant bacteria and provides a scientific basis for rational clinical medication., Competing Interests: The authors declare no conflict of interest.

Introduction: Although evidence exists on the impact of microbiota on pregnancy outcomes in many high-resource settings, there is a lack of research in many low-resource settings like Ethiopia. This study aims to fill this gap by studying the gut and vaginal microbiota changes throughout pregnancy and assess how these changes relate to pregnancy outcomes among a cohort of pregnant women in eastern Ethiopia., Methods and Analysis: Vaginal and stool samples will be collected using DNA/RNA Shield Collection kits three times starting at 12-22 weeks, 28-36 weeks and at birth (within 7 days). Postnatally, newborns' skin swabs (at birth) and rectal swabs will be obtained until 2 years of age. Moreover, breast milk samples at birth and 6 months and environmental samples (water, indoor air and soil) will be collected at enrolment, birth, 6, 12 and 24 months post partum. DNA will be extracted using Roche kits. Metagenomic sequencing will be performed to identify metataxonomic profiling and assess variations in microbial profiles, and α and β diversity of the microbiota. Information on socioeconomic, behavioural, household and biological factors will be collected at enrolment. The collected data will be coded, entered into EpiData 3.1 and analysed using Stata 17., Ethics and Dissemination: The Institutional Health Research Ethics Review Committee (Ref No. IHRERC/033/2022) of Haramaya University, Ethiopia has approved this study ethically. Written informed consent regarding the study and sample storage for biobanking will be obtained from all participants. Results will be published in international peer-reviewed journals, and summaries will be provided to the study funders. Clinical study data will be submitted to Data Compass (https://datacompass.lshtm.ac.uk/), and molecular profiles of the microbiome and whole-genome sequences will be submitted to the European Nucleotide Archive (https://www. ebi.ac.uk/ena). Requests for data should be directed to daberaf@gmail.com. The decision to share data will be made by the study steering committee under the College of Health and Medical Sciences, Haramaya University, Ethiopia., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2025. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ Group.)

Background: Recurrent vaginitis in conjunction with urinary tract infection (RV/UTI) in perimenopausal women is a common clinical condition that impacts both doctors and patients. Its pathogenesis is not completely known, but the urogenital microbiota is thought to be involved. We compared the urogenital and gut microbiotas of perimenopausal women experiencing RV/UTI with those of age-matched controls to provide a new microbiological perspective and scheme for solving clinical problems., Results: Fifty women of perimenopausal age who were diagnosed with RV/UTI and 50 age-matched healthy controls were enrolled. The urogenital and intestinal microbiota were analyzed via 16S ribosomal RNA gene sequencing by collecting samples from the mouth, anus, urine, cervix, and upper and lower vaginal ends. Among the microbiota of healthy perimenopausal women, the mouth had the highest richness, whereas the anus and mouth had the highest levels of diversity. Compared with those in healthy controls, in the microbiota of patients with RV/UTI, the evenness of the upper vaginal end, anus and cervix significantly increased, whereas the richness and diversity of the cervix significantly decreased. Lactobacillus accounted for 40.65% of the bacteria in the upper vaginal end and 39.85% of the bacteria in the lower vaginal end of healthy women of perimenopausal age, and there were no significant differences in Lactobacillus abundance among the patients with RV/UTI. The relative abundances of 54 genera and 97 species were significantly different between patients and healthy individuals, particularly in the cervix and urine. A total of 147 predicted pathways were significantly different between patients and healthy controls, with the microbiota of the anus exhibiting the greatest number of functional changes, followed by the urine microbiota. A random forest model composed of 16 genera in the lower vaginal end had the highest discriminatory power (AUC 81.48%) to predict RV/UTI., Conclusions: Our study provides insight into the nature of the urogenital and intestinal microbiota in perimenopausal women, and reveals significant changes in the microbiota in patients with RV/UTI. This information will help characterize the relationship between the urogenital microbiota and RV/UTI, potentially aiding in the development of diagnostic and therapeutic strategies., Competing Interests: Declarations. Ethics approval and consent to participate: The study was conducted in accordance with the Declaration of Helsinki and approved by the Medical Ethics Committee of Shanghai Fourth People’s Hospital (SYLL2023124). All specimens were collected after obtaining written informed consent from participants. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

The presence of high-risk human papillomavirus (HR-HPV) contributes to the development of cervical lesions and cervical cancer. Recent studies suggest that an imbalance in the cervicovaginal microbiota might be a factor in the persistence of HR-HPV infections. In this study, we collected 156 cervicovaginal fluid (CVF) of women with HR-HPV infection, which were divided into three groups (negative for intraepithelial lesions = 78, low/high-grade squamous intraepithelial lesions = 52/26). We performed metabolomics and 16 S rRNA sequencing to identify changes in metabolites and cervicovaginal microbiota among patients with HR-HPV infection and varying grades of cervical lesions. We detected 164 metabolites and 389 flora types in the three groups. Ten CVF metabolites-N-methylalanine, phenylacetaldehyde, succinic acid, 2-3-dihydroxypyridine, DL-p-hydroxylphenyllactic acid, gluconic acid lactone, guanine, glucose-6-phosphate, erythrose, and sucrose showed significant associations with disease severity and distinct separation patterns in HR-HPV-infected patients with LSIL and HSIL, with an area under the curve of 0.928. The most abundant microbial communities in the CVF were Gardnerella. Gardnerella was found to be associated with increased levels of succinic acid, thereby highlighting distinct metabolic profiles. These findings suggest that the development of cervical lesions resulting from persistent HR-HPV infection is associated with significant alterations in systemic metabolism and shifts in the cervicovaginal microbiota, providing valuable insights into the metabolic and microbiota changes linked to disease severity., Competing Interests: Declarations. Competing interests: The authors declare no competing interests. Ethical statement: This study was conducted in accordance with the Declaration of Helsinki. The experimental procedures complied with relevant privacy protection laws and guidelines. Ethical approval was obtained from the Ethics Committee of the Affiliated Hospital of Nanjing University of Chinese Medicine (approval number 2021NL-025-03). All experiments were carried out in accordance with the relevant guidelines and regulations, and informed consent was obtained from each participant., (© 2025. The Author(s).)

Background: Candidiasis can be present as a cutaneous, mucosal, or deep-seated organ infection, which is caused by more than 20 types of Candida spp., with C. albicans being the most common. Hence, this work aimed to estimate some virulence factors, including phospholipase and biofilm formation, in some Candida spp., Methods: A total of eighty-six specimens were collected from patients with oral and vaginal candidiasis and subjected into different examinations, including cultural characteristic (on Sabouraud Dextrose Agar SDA and chromogenic Candida agar) and microscopic examination and germ tube formation (GT)to isolate Candida spp. In addition, the egg-yolk agar plate method was used to determine the extracellular phospholipase production, and the microtiter plate method was used to determine biofilm formation of Candida spp. Vitek Compact equipment was used to identify the highest phospholipase and biofilm-producers of Candida spp., Results: As a result of all examinations, 58.1% (n = 50/86) of isolates of Candida spp. were obtained, including 26.7% (n = 23/86) isolates of Candida spp. from oral cavity and 31.3% (n = 27/86) isolates of Candida spp. from vaginal cavity. These isolates included 58% (n = 29/50) C. albicans, 10% (n = 5/50) C. glabrata, 6% (n = 3/50) C. parasilosis, 6% (n = 3/50) C. krusei, 6% (n = 3/50) C. lusitaniae, 6% (n = 3/50) C. kefyr, 6% (n = 3/50) C. tropicalis, and 2% (n = 1/50) C. ciferrii. To quantify extracellular phospholipase production, the egg-yolk agar plate method was utilized. The results indicated that the majority of isolates (n = 33; 66%) were phospholipase-strong producers, 18% (n = 9) of isolates were phospholipase-moderate producers, 5% (n = 10) were phospholipase-weak producers, and 6% (n = 3) were non-phospholipase producers. Microtiter plate method was utilized to estimate formation of biofilm by Candida spp. obtained from vaginal and oral cavities. The majority of Candida spp. isolates (n = 32; 64%) were biofilm-strong producers, followed by 30% (n=15) moderate-biofilm producers and 6% (n = 3) weak-biofilm producers. The results of VITEK 2 system indicated that the probability of C. albicans, C. krusei, C. kefyr, C. tropicalis, C. lusitaniae, C. glabrata, and C. ciferrii was 98, 95, 94, 91, 85, 93, and 85 %, respectively., Conclusions: Candida albicans was the most frequent isolate among all isolates.

Patients with obstructed hemivagina and ipsilateral renal anomaly (OHVIRA) syndrome sometimes present with distinct symptoms related to coexisting urogenital abnormalities. We present a case of refractory abscess formation in a blind-ended remnant ureter associated with ipsilateral renal agenesis. A 15-year-old patient with OHVIRA syndrome, who had a blind-ended remnant ureter, underwent obstructed hemivagina opening 18 months after menarche due to heavy genital bleeding and abdominal pain after the end of menstruation. Four years later, the patient presented with recurring fever and abdominal pain due to a refractory abscess in the ureter. Although the continuity between the ureter and the previously opened hemivagina was not identified, the same bacteria were detected in the abscess and vaginal discharge, indicating that an ascending bacterial infection of the vagina may cause refractory abscess formation in a blind-ended remnant ureter. Open ureterectomy with partial bladder resection was needed to resolve the symptoms., (© 2024 Japan Society of Obstetrics and Gynecology.)

Background: Composition of the vaginal microbiome in pregnancy is associated with adverse maternal, obstetric, and child health outcomes. Therefore, identifying sources of individual differences in the vaginal microbiome is of considerable clinical and public health interest. The current study tested the hypothesis that vaginal microbiome composition during pregnancy is associated with an individual's experience of affective symptoms and stress exposure., Methods: Data were based on a prospective longitudinal study of a medically healthy community sample of 275 mother-infant pairs. Affective symptoms and stress exposure and select measures of associated biomarkers (diurnal salivary cortisol, serum measures of sex hormones) were collected at each trimester; self-report, clinical, and medical records were used to collect detailed data on socio-demographic factors and health behavior, including diet and sleep. Vaginal microbiome samples were collected in the third trimester (34-40 weeks) and characterized by 16S rRNA sequencing. Identified taxa were clustered into three community clusters (CC1-3) based on dissimilarity of vaginal microbiota composition., Results: Results indicate that depressive symptoms during pregnancy were reliably associated with individual taxa and CC3 in the third trimester. Prediction of functional potential from 16S taxonomy revealed a differential abundance of metabolic pathways in CC1-3 and individual taxa, including biosynthetic pathways for serotonin and dopamine. We did not find robust evidence linking symptom- and stress-related biomarkers and CCs., Conclusions: Our results provide further evidence of how prenatal psychological distress during pregnancy alters the maternal-fetal microbiome ecosystem that may be important for understanding maternal and child health outcomes., Competing Interests: Declaration of competing interest The authors declare that they have no competing interests., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Chlamydia trachomatis (Ct) is the most common bacterial sexually transmitted infection globally, and a vaccine is urgently needed to stop transmission and disease. Chlamydial Protease Activity Factor (CPAF) is an immunoprevalent and immunodominant antigen for CD4 T cells and B cells, which makes it a strong vaccine candidate. Due to the tolerogenic nature of the female genital tract (FGT) and its lack of secondary lymphoid tissue, effective induction of protective cell-mediated immunity will likely require potent and safe mucosal adjuvants. To address this need, we produced CPAF in a cell-free protein synthesis platform and adjuvanted it with the TLR9-agonist CpG1826, a synthetic cyclic-di-AMP (CDA) STING (stimulator of interferon genes) agonist ADU-S100, and/or the squalene oil-in-water nanoemulsion, AddaS03. We determined that intranasal immunization with CPAF plus ADU-S100 was well tolerated in female mice, induced CD4 T cells characterized by TNFα production alone or in combination with IL-17 A or IFNγ, significantly reduced bacterial shedding, and shortened the duration of infection in mice intravaginally challenged with Chlamydia muridarum. These data demonstrate the potential for CDA as a mucosal adjuvant for vaccines against Chlamydia genital tract infection., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Daisy Arroyo reports financial support was provided by Vaxcyte, Inc. Tony Bruno reports financial support was provided by Vaxcyte, Inc. Manhong Wu reports financial support was provided by Vaxcyte, Inc. James Rozzelle reports financial support was provided by Vaxcyte, Inc. Jeff Fairman reports financial support was provided by Vaxcyte, Inc. Taylor B. Poston has patent Chlamydial Protease-Like Activity Factor and Adjuvant Compositions And Uses Thereof pending to Assignee. Toni Darville has patent Chlamydial Protease-Like Activity Factor and Adjuvant Compositions And Uses Thereof pending to Assignee. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)