Search Results (58)

Objectives: Recurrent vulvovaginal candidiasis (RVVC) is a difficult-to-treat infection, most likely due to the growth of Candida biofilms on the human vaginal epithelium. We assessed in vitro efficacy of conventional antifungals and complementary and alternative medicine (CAM) used in clinical settings, and sought for Candida biofilm-effective single or combination therapies. Methods: Standard broth microdilution assay and XTT (2,3-Bis-(2-Methoxy-4-Nitro-5-Sulfophenyl)-2H-Tetrazolium-5-Carboxanilide) assay were used for antifungal and anti-biofilm efficacies of three conventional antifungals, and selected CAM including boric acid, povidone-iodine, and allicin (garlic extract), against Candida clinical isolates grown at neutral and acidic pHs respectively. Fractional inhibitory concentration (FIC) indices were assessed to evaluate interactions between fluconazole and different CAM. Viable count-based cell enumeration and confocal laser scanning microscopy (CLSM) were performed to confirm the efficacy of single or combination therapies against Candida biofilms. Results: All selected conventional antifungals and CAM showed efficacies against planktonic Candida cells. Acidic vaginal microenvironments provided agent-specific protection to Candida cells against conventional antifungals and the CAM. Synergistic or additive interactions were observed between fluconazole at serum achievable concentrations and povidone-iodide at topically achievable concentrations against all tested Candida strains. Most antifungal agents except caspofungin had very limited activities against Candida biofilms. Combining fluconazole at 8 mg/L with povidone-iodine at 2048 mg/L effectively killed Candida biofilms in an acidic vaginal microenvironment to a level that is comparable to that of caspofungin. Conclusions: We provided robust in vitro evidence supporting the combinational use of oral fluconazole and topical CAM povidone-iodine against Candida biofilms in managing RVVC. Full article

Background: Recurrent vulvovaginal candidiasis (RVVC) is a chronic inflammatory disease primarily caused by Candida albicans (C. albicans). Its pathogenesis remains incompletely understood, and clinical management is challenged by recurrence and drug resistance. Ferroptosis, an iron-dependent form of programmed cell death driven by lipid peroxidation, has been implicated in various infectious and inflammatory diseases. However, its role in RVVC remains unclear, with a particular lack of evidence from clinical samples and animal experiments. Objective: This study aimed to investigate the association between RVVC and ferroptosis. First, we analyzed high-throughput sequencing data from human RVVC samples in the Gene Expression Omnibus (GEO) database to identify the expression profile of ferroptosis-related genes. Second, using an established murine model of chronic vulvovaginal candidiasis (CVVC), we validated changes in ferroptosis-related markers in vaginal tissues in vivo. Furthermore, an in vitro model of C. albicans-infected bone marrow-derived macrophages (BMDMs) was employed to explore the underlying mechanisms. This study provides experimental evidence for elucidating the pathogenesis of RVVC and exploring novel therapeutic strategies. Methods: The RVVC-related gene expression dataset GSE278036 was obtained from the GEO database. Differentially expressed genes (DEGs) were screened using the DESeq2 algorithm and intersected with ferroptosis-related genes from the FerrDb database to identify key targets. A protein–protein interaction (PPI) network was constructed using the STRING database and Cytoscape software, and hub genes were identified via the Betweenness centrality algorithm. Functional and pathway analyses, including gene set enrichment analysis (GSEA), Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and WikiPathways, were performed. Immune infiltration analysis characterized the immune microenvironment in RVVC patients. A CVVC mouse model was established in vivo, and a C. albicans-BMDMs infection model was established in vitro. The ferroptosis inhibitor ferrostatin-1 (Fer-1) was administered to investigate the pathological function and regulatory mechanisms of ferroptosis in RVVC at the molecular, cellular, and tissue levels. Results: Differential analysis identified 3132 DEGs in RVVC, which intersected with ferroptosis-related genes to yield 194 key targets. Among them, 20 hub genes were identified, including ferroptosis regulators and inflammatory factors. Functional enrichment analysis confirmed that these shared targets regulate RVVC pathology through a “ferroptosis-inflammation-immunity” multi-pathway network. Immune infiltration analysis revealed a specific immune disorder in RVVC patients characterized by “activation of the pro-inflammatory innate immune axis and suppression of the adaptive immune axis,” which was closely associated with ferroptosis-related genes. In vivo and in vitro experiments confirmed that C. albicans infection induced ferroptosis in vaginal tissues and macrophages, as manifested by lipid ROS accumulation, Fe²⁺ overload, GSH depletion, downregulation of GPX4 and SLC7A11, upregulation of ACSL4, 4-HNE, and MDA, and mitochondrial structural damage. Macrophages were identified as key target cells for ferroptosis, and their ferroptosis led to impaired antifungal function. Fer-1 treatment significantly inhibited ferroptosis, reduced vaginal histopathological damage and inflammatory cell infiltration, decreased fungal burden, downregulated abnormally elevated inflammatory factors, and restored Th1/Th2 immune balance. Furthermore, Fer-1 preserved macrophage viability and enhanced their antifungal killing capacity. Conclusions: This study provides the first evidence linking RVVC to ferroptosis through a combination of clinical data analysis and experiments, suggesting that ferroptosis is involved in its pathological process. These findings offer a new perspective for elucidating RVVC pathogenesis and developing targeted therapeutic strategies. Full article

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

Candida albicans is the primary agent of acute vulvovaginal candidiasis (VVC) and its recurrent form (RVVC). Local innate immunity contributes to both defense and pathogenesis during vaginal Candida infection, where epithelial β-defensins (BD) constitute key components of the mucosal barrier. We previously reported that epithelial BD-1 expression is dynamically modulated during murine and human vaginitis, revealing strain-dependent and stimulus-specific regulation but leaving the host pathways involved unresolved. This study functionally defines the contribution of key immune pathways to epithelial antimicrobial peptide regulation. Using a murine model of VVC and the virulent C. albicans strain SC5314, we aimed to evaluate the immune signaling pathways governing the temporal regulation of epithelial BD-1 and BD-3 expression during vaginal infection. In wild-type mice, both defensins displayed a biphasic pattern: early induction followed by attenuation as infection progressed. Genetic loss-of-function approaches revealed that NLRP3/IL-1β signaling is required for early BD-1 induction, whereas IL-17RA signaling preferentially supports sustained BD-3 expression. Together, these findings establish a causal and temporal link between host immune signaling and epithelial defensin regulation and reveal a transient subversion of mucosal defenses by C. albicans. This work advances understanding of epithelial innate immunity, defining distinct temporal programs for BD-1 and BD-3 and identifying NLRP3/IL-1β and IL-17RA signaling as key pathways shaping mucosal defensin expression. Full article

Vulvovaginal candidiasis (VVC) represents a widespread gynaecological challenge, affecting approximately 75% of women at some point during their reproductive years, with a significant subset progressing to recurrent forms (RVVC). Classical azoles and polyenes remain the cornerstone of therapy. However, their clinical utility is undermined by the rise of azole-resistant non-Candida albicans species, the capacity of Candida to form biofilms, and a complex variety of host-related factors that complicate disease expression and therapeutic response. This narrative review provides a critical up-to-date examination of the therapeutic landscape, integrating current diagnostic algorithms with pharmacological strategies for both acute, recalcitrant and recurrent VVCs. We assess the efficacy and safety of established antifungal agents alongside the breakthrough introduction of novel drug classes, with a particular interest in the oral triterpenoid ibrexafungerp and the tetrazole oteseconazole, which offer new mechanisms of action for cases that fail to respond to standard regimens. Furthermore, we address the management of a special clinical scenarios, including pregnancy and lactation, and explore promising emerging innovative approaches such as mucoadhesive formulations, immunomodulatory approaches, and alternative non-antifungal therapies. Ultimately, this review aims to support clinical decision-making by balancing the accessibility and user-friendliness of conventional treatments with the targeted precision offered by modern antifungal agents. Full article

Symptomatic vulvovaginal candidiasis (VVC) affects around three-quarters of women at least once in their lifetime. Around 10% of these women will experience prolonged or recurrent vulvovaginal candidiasis (RVVC), which fails to respond, despite following recommended therapy. Most commonly prescribed therapy involves suppression therapy—usually for two weeks—which aims at eliminating symptoms by frequent administration of antifungals, followed by maintenance (weekly/monthly) therapy for up to six months. However, following cessation of maintenance therapy, around 50% of these women experience relapse. The vaginal ecology of RVVC can be characterized, and it is thought that biofilms and/or the development of antifungal resistance prevent adequate resolution. However, hypersensitivity may also confound management. This narrative review was performed to identify key studies that examine the management of VVC and the challenges of current prolonged antifungal therapy. It identifies gaps that show it remains important to investigate microbiological findings in RVVC and how these may inform rational choices in therapy in an era of rising antimicrobial resistance. Hope exists, as studies of the vaginal microbiome highlight that the type of microbiota may influence the level of inflammation and reduce symptomatology. Future research will continue to explore whether a personalized medicine approach can promote healthy vaginal ecology and prevent the debilitating long-term effects of RVVC. Full article

Vulvovaginal candidiasis (VVC) is a prevalent vaginal infection predominantly attributed to Candida albicans. A considerable proportion of women experience more than three episodes of VVC annually, a condition referred to as recurrent vulvovaginal candidiasis (RVVC). It is estimated that RVVC affects more than 130 million women globally each year and has a substantial negative impact on their quality of life, resulting in physical discomfort, psychological distress, and social stigma. Nevertheless, not all individuals who develop VVC progress to RVVC, suggesting that genetic variation may play a critical role in host susceptibility. The present review aims to evaluate the associations between genetic predispositions—specifically polymorphisms in Toll-like receptors 2 and 4 (TLR2, TLR4)—and RVVC. TLRs are essential for detecting pathogen-associated molecular patterns (PAMPs) and initiating immune responses. During RVVC episodes, Candida undergoes a reversible transition from the yeast form to the hyphal form, resulting in alterations in surface PAMPs, which are subsequently recognized by innate immune receptors expressed on vaginal epithelial cells. Polymorphisms in these receptors may modulate individual susceptibility to RVVC. This review examines the literature on the impact of specific polymorphisms in TLR2 and TLR4 on fungal recognition and infection. Furthermore, the interactions between TLRs and other elements of the innate immune system have also been explored. A deeper understanding of how genetic variability in immune receptors influences infection susceptibility could pave the way for personalized therapeutic strategies for RVVC, potentially involving immunomodulatory agents or antifungal treatments tailored to an individual’s genetic profile. Full article

Background: Gynecological infections, including bacterial vaginosis, vulvovaginal candidiasis, and recurrent urinary tract infections, represent a major clinical burden and are often complicated by biofilm formation and antimicrobial resistance. Novel non-antibiotic strategies are urgently needed. We previously demonstrated the antimicrobial activity of silver-bound titanium dioxide (TiAB) against multidrug-resistant bacteria isolated from dermatological infections. Objectives: We evaluated whether TiAB, at concentrations used in marketed medical devices, exerts antibacterial and antifungal effects against clinically relevant vaginal isolates by determining Minimum Inhibitory Concentration/ Minimum Bactericidal and Fungicidal Concentration (MIC, MBC/MFC), and time–kill kinetics. Methods: A total of 73 clinical isolates were collected from vaginal swabs, including Staphylococcus aureus (MSSA, MRSA), Escherichia coli (ESBL+ and non-ESBL), Klebsiella pneumoniae, Enterococcus spp., Streptococcus agalactiae, and Candida albicans. Minimum inhibitory concentrations (MICs) and minimum bactericidal/fungicidal concentrations (MBCs/MFCs) were determined by broth microdilution, and bactericidal activity was confirmed by time-kill assays. Results: TiAB exhibited potent activity against Gram-negative bacteria, with median MIC values of 1–2% (w/v) for E. coli and K. pneumoniae. Gram-positive isolates, including S. agalactiae and Enterococcus spp., showed higher MIC values (2–4%). Candida albicans displayed fungistatic inhibition at 4%. Time-kill assays confirmed rapid bactericidal effects for Gram-negative isolates within 8 h at 2× MIC, while Gram-positive bacteria required prolonged exposure. Conclusions: These findings extend previous evidence of TiAB’s antimicrobial properties to gynecological pathogens, supporting its potential as a topical, non-antibiotic option for managing vaginal infections in an era of rising antimicrobial resistance. Further in vivo validation is warranted. Full article

The epidemiology of vulvovaginal candidiasis (VVC) in Ecuador remains poorly reported and outdated. We therefore conducted a 12-month prospective survey to assess the aetiology and antifungal resistance patterns among symptomatic Ecuadorian patients. VVC diagnosis was confirmed by microscopic examination and culture. Isolates were identified by biochemical and molecular methods. In vitro antifungal susceptibilities to amphotericin B, clotrimazole, fluconazole, itraconazole, miconazole, and nystatin were determined by CLSI methods. Among 195 women, 71 VVC episodes were recorded (36.4%), whereof 56 (28.7%) had acute VVC (AVVC) and 15 (7.7%) had recurrent VVC (RVVC). The predominant species was Candida albicans, isolated in pure culture from 45 AVVC (80.3%) and 9 RVVC patients (60%), and in mixed culture from 7 AVVC (12.5%) and 3 RVVC patients (20%). Candida glabrata and Saccharomyces cerevisiae were also isolated in AVVC and RVVC patients, but Candida parapsilosis and Candida famata were only isolated from AVVC. Fluconazole- and miconazole-resistant C. albicans isolates were recovered from 5 (8.9%) and 24 (42.9%) of 56 AVVC patients, respectively, and from 1 (8.3%) and 5 (41.7%) of 12 RVVC patients, respectively. Fluconazole and miconazole resistance is relevant in Ecuador, emphasising the need for targeted antifungal strategies. Full article

Background/Objectives: Fungal colonization and biofilm formation on intrauterine device (IUD) strings are known to contribute to recurrent infections and decreased contraceptive efficacy. This study aims to develop a novel approach to prevent Candida reservoir and biofilm formation on IUD strings, thereby lowering the risk of IUD-associated vulvovaginal candidiasis (VVC). Methods: Cervicovaginal samples were collected from human cervix using a sterile cytobrush, avoiding microbial contamination. Cytological examination using the Papanicolaou method was performed to detect the presence of Candida. The antifungal effect of the essential oils (EOs) was determined by broth dilution and disk diffusion methods. Antifungal and biofilm inhibitory effects of Thymus zygis (Tz) EO-coated IUD strings were determined by agar diffusion and crystal violet binding assays, while fungal growth on the coated strings was assessed using Scanning Electron Microscopy (SEM) and Energy-Dispersive X-ray (EDX) analysis. Results: Tz EO exhibited significantly lower minimum inhibitory concentration (MIC ≤ 0.06 µL/mL) and minimum fungicidal concentration (MFC = 0.24 µL/mL) values compared to Melaleuca alternifolia (Ma) EO (MIC > 0.24 µL/mL, MFC = 1.95 µL/mL), along with larger zones of inhibition (ZOI) against both Candida albicans (110.0 ± 6.0 mm vs. 91.3 ± 7.0 mm) and Candida glabrata (84.0 ± 13.1 mm vs. 50.0 ± 9.2 mm), indicating a stronger antifungal potential. On IUD strings coated with 4% (40 μL/g) Tz EO in hypromellose ointment, the biofilm formation of both C. albicans and C. glabrata strains was inhibited by 58.9% and 66.7%, respectively, as confirmed by SEM and EDX. Conclusions: Tz EO-coated IUD strings effectively inhibit Candida growth, suggesting a promising natural strategy to reduce recurrent IUD-associated fungal infections. However, before these results can be translated to clinical practice, additional research is needed. Future investigations may encompass an extended number of Candida isolates, stability and release studies of the EO in relation to the formulation, toxicity to vaginal mucosa, epithelial cells and sperm motility, and the effect on vaginal microbiotia. Full article

Candida albicans is the primary etiological agent of vulvovaginal candidiasis (VVC), a widespread fungal infection affecting millions of women worldwide. Although often self-limiting, VVC can become recurrent or severe, significantly impacting quality of life. The pathogenesis of C. albicans is driven by key virulence factors, including hyphal transformation, biofilm formation, and immune evasion, which all facilitate persistence and resistance to host defenses. Epidemiological data indicate that up to 75% of women experience at least one episode of VVC, with 5–10% developing recurrent vulvovaginal candidiasis. The condition typically presents with vaginal itching, burning, erythema, edema, and an abnormal discharge. Diagnosis relies on both clinical presentation and microbiological confirmation; however, misdiagnosis remains common due to symptom overlap with other vaginal infections and conditions in general. Azole antifungals remain the cornerstone of treatment; however, increasing resistance (particularly in non-albicans Candida species) poses substantial therapeutic challenges. Consequently, the emergence of antifungal-resistant strains underscores the need for novel treatment strategies, including probiotics and natural antifungal agents. Preventive measures—including maintaining vaginal microbiota balance, avoiding unnecessary antibiotic usage, and improving hygiene practices—play a pivotal role in reducing disease burden due to C. albicans. Given the rising incidence of VVC and the burden of recurrent cases, further research is essential to develop targeted therapeutic interventions. This comprehensive review highlights the evolving epidemiology, pathogenesis, and clinical challenges of C. albicans-associated VVC, emphasizing the need for improved diagnostic strategies, alternative therapeutic approaches, and targeted preventive measures to reduce disease burden and enhance patient outcomes. Full article

Background and purpose: Vulvovaginal candidiasis (VVC), caused by Candida albicans (C. albicans), is exacerbated by oxidative stress and uncontrolled inflammation. Pathogens like C. albicans generate reactive oxygen species (ROS) to enhance virulence, while host immune responses further amplify oxidative damage. This study investigates the antioxidant and antifungal properties of Hyssopus cuspidatus Boriss volatile extract (SXC), a traditional Uyghur medicinal herb, against fluconazole-resistant VVC. We hypothesize that SXC’s bioactive volatiles counteract pathogen-induced oxidative stress while inhibiting fungal growth and inflammation. Methods: GC-MS identified SXC’s major bioactive components, while broth microdilution assays determined minimum inhibitory concentrations (MICs) against bacterial/fungal pathogens, and synergistic interactions with amphotericin B (AmB) or fluconazole (FLC) were assessed via time–kill kinetics. Anti-biofilm activity was quantified using crystal violet/XTT assays, and in vitro studies evaluated SXC’s effects on C. albicans-induced cytotoxicity (LDH release in A431 cells) and inflammatory responses (cytokine production in LPS-stimulated RAW264.7 macrophages). A murine VVC model, employing estrogen-mediated pathogenesis and intravaginal C. albicans challenge, confirmed SXC’s in vivo effects. Immune modulation was assessed using ELISA and RT-qPCR targeting inflammatory and antioxidative stress mediators, while UPLC-MS was employed to profile metabolic perturbations in C. albicans. Results: Gas chromatography-mass spectrometry identified 10 key volatile components contributing to SXC’s activity. SXC exhibited broad-spectrum antimicrobial activity with MIC values ranging from 0.125–16 μL/mL against bacterial and fungal pathogens, including fluconazole-resistant Candida strains. Time–kill assays revealed that combinations of AmB-SXC and FLC-SXC achieved sustained synergistic bactericidal activity across all tested strains. Mechanistic studies revealed SXC’s dual antifungal actions: inhibition of C. albicans hyphal development and biofilm formation through downregulation of the Ras1-cAMP-Efg1 signaling pathway, and attenuation of riboflavin-mediated energy metabolism crucial for fungal proliferation. In the VVC model, SXC reduced vaginal fungal burden, alleviated clinical symptoms, and preserved vaginal epithelial integrity. Mechanistically, SXC modulated host immune responses by suppressing oxidative stress and pyroptosis through TLR4/NF-κB/NLRP3 pathway inhibition, evidenced by reduced caspase-1 activation and decreased pro-inflammatory cytokines (IL-1β, IL-6, TNF-α). Conclusions: SXC shows promise as a broad-spectrum natural antimicrobial against fungal pathogens. It inhibited C. albicans hyphal growth, adhesion, biofilm formation, and invasion in vitro, while reducing oxidative and preserving vaginal mucosal integrity in vivo. By disrupting fungal metabolic pathways and modulating host immune responses, SXC offers a novel approach to treating recurrent, drug-resistant VVC. Full article

Recurrent vulvovaginal candidiasis (RVVC), predominantly caused by Candida albicans, represents a global health issue, particularly in developing regions. This study explores the antifungal potential of aqueous leaf extract of Schinus weinmanniifolia Mart. ex Engl., a native Latin American plant. The extract was evaluated for phytochemical composition, antifungal efficacy, and safety profile. Phytochemical analyses identified six major compounds, including shikimic acid, gallic acid, and methyl gallate, with antioxidant and antimicrobial properties. The extract showed potent antioxidant activity, with IC₅₀ values between 1.52–5.51 µg/mL. It strongly inhibited C. albicans, with a minimum inhibitory concentration (MIC) of 1.95 µg/mL, and was active against other yeasts (MIC 0.48–62.5 µg/mL). The growth kinetics assay revealed reduced C. albicans viability after 12 h at 2 × MIC versus the positive control. Scanning electron microscopy confirmed reduced fungal counts without morphological damage. The extract impaired C. albicans virulence, reducing germ tube formation by 75.49% and hyphal transition by 84.34%, outperforming fluconazole. Biocompatibility assays showed it is non-hemolytic (IC₅₀ > 1000 µg/mL), non-mutagenic, and highly selective for fungal cells (SI = 512.82), suggesting minimal human cell toxicity. In conclusion, the extract combines strong antifungal activity and favorable safety, with cost-effective preparation suitable for traditional medicine in resource-limited regions. Full article

Recurrent vulvovaginal candidiasis (RVVC) is a common condition that affects women of reproductive age. The etiology of RVVC remains largely unknown, but it is believed to be associated with changes in vaginal microbiota composition. This study investigates the vaginal microbiota in 57 women with RVVC and 38 healthy controls. Bacterial DNA was analyzed using high-throughput 16S rRNA gene sequencing, and Candida and Saccharomyces species were determined by PCR. RVVC cases had a higher prevalence of Nakaseomyses glabratus (former Candida glabrata) compared to controls. Alpha diversity metrics were similar between groups, but beta diversity analysis revealed significant differences in vaginal microbiota composition. The Firmicutes abundance was altered in RVVC cases, with genus Bifidobacterium and phylum Actinobacteriota being more abundant than in the controls. At the genus level, Lactobacillus dominated controls using antibiotics, while Bifidobacterium was higher in cases with no antibiotic intake. Our study provides evidence that Nakaseomyses glabratus (former Candida glabrata) is a significant pathogen in RVVC, while Candida albicans was more prevalent in healthy women. The vaginal microbiota composition differs significantly between the two groups, with distinct patterns of bacterial abundance and changes in Firmicutes abundance. Full article

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 × 10⁹ 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. Full article