Dietary amino acids, macronutrients, vaginal birth, and breastfeeding are associated with the vaginal microbiome in early pregnancy.

The vaginal microbiome is a key player in the etiology of spontaneous preterm birth. This study aimed to illustrate maternal environmental factors associated with vaginal microbiota composition and function in pregnancy. Women in healthy pregnancy had vaginal microbial sampling from the posterior vaginal fornix performed at 16 weeks gestation. After shotgun metagenomic sequencing, heatmaps of relative abundance data were generated. Community state type (CST) was assigned, and alpha diversity was calculated. Demography, obstetric history, well-being, exercise, and diet using food frequency questionnaires were collected and compared against microbial parameters. A total of 119 pregnant participants had vaginal metagenomic sequencing performed. Factors with strongest association with beta diversity were dietary lysine (adj-R2 0.113, P = 0.002), valine (adj-R2 0.096, P = 0.004), leucine (adj-R2 0.086, P = 0.003), and phenylalanine (adj-R2 0.085, P = 0.005, Fig. 2D). Previous vaginal delivery and breastfeeding were associated with vaginal beta diversity (adj-R2 0.048, P = 0.003; adj-R2 0.045, P = 0.004), accounting for 8.5% of taxonomy variation on redundancy analysis. Dietary fat, starch, and maltose were positively correlated with alpha diversity (fat +0.002 SD/g, P = 0.025; starch +0.002 SD/g, P = 0.043; maltose +0.440 SD/g, P = 0.013), particularly in secretor-positive women. Functional signature was associated with CST, maternal smoking, and dietary phenylalanine, accounting for 8.9%-11% of the variation in vaginal microbiome functional signature. Dietary amino acids, previous vaginal delivery, and breastfeeding history were associated with vaginal beta diversity. Functional signature of the vaginal microbiome differed with community state type, smoking, dietary phenylalanine, and vitamin K. Increased alpha diversity correlated with dietary fat and starch. These data provide a novel snapshot into the associations between maternal environment, nutrition, and the vaginal microbiome. IMPORTANCE: This secondary analysis of the MicrobeMom randomized controlled trial reveals that dietary amino acids, macronutrients, previous vaginal birth, and breastfeeding have the strongest associations with vaginal taxonomy in early pregnancy. Function of the vaginal niche is associated mainly by species composition, but smoking, vitamin K, and phenylalanine also play a role. These associations provide an intriguing and novel insight into the association between host factors and diet on the vaginal microbiome in pregnancy and highlight the need for further investigation into the complex interactions between the diet, human gut, and vaginal microbiome.

First Detection and Characterization of Smacovirus in the Human Vagina in Two Sequential Samples over a Twelve-Day Interval.

BACKGROUND: Smacovirus is a CRESS-DNA virus identified almost exclusively in transient fecal samples from various vertebrate species. OBJECTIVE: We evaluated human vaginal samples for the presence and maintenance of Smacovirus. METHODS: Viral metagenomics analysis was performed on vaginal samples collected from 28 apparently healthy women in New York City, USA. Twenty-one of the women provided duplicate samples over a 12-21-day interval. RESULTS: Phylogenetic analysis identified two samples from the same individual, collected over a twelve-day interval, that were positive for the complete Smacovirus genome. All detected sequence contigs belonged to a single variant of CRESS-DNA. CONCLUSIONS: The continuous presence of Smacovirus in the human vagina over a twelve-day period was identified.

Vaginal host response to polycarbonate urethane, an alternative material for the repair of pelvic organ prolapse.

Complications following surgical repair of pelvic organ prolapse (POP) with polypropylene mesh (PPM) are common. Recent data attributes complications, in part, to stiffness mismatches between the vagina and PPM. We developed a 3D printed elastomeric membrane (EM) from a softer polymer, polycarbonate urethane (PCU). EMs were manufactured with more material given the low inherent material strength of PCU. We hypothesized that the EMs would be associated with an improved host response as compared to PPM. A secondary goal was to optimize the material distribution (fiber width and device thickness) within EMs, in regards to the host response. EM constructs (2 × 1 cm2) with varied polymer stiffness, fiber width, and device thickness were implanted onto the vagina of New Zealand white rabbits for 12 weeks and compared to similarly sized PPMs. Sham implanted animals served as controls. Mixed effects generalized linear models were used to compare the effect of construct type accounting for differences in independent variables. EMs had an overall superior host response compared to PPM as evidenced by preservation of vaginal smooth muscle morphology (p-values<0.01), decreased total cellular response to construct fibers (p-values<0.001), and a reduced percent of macrophages (p-values<0.02) independent of how the material was distributed. Both PPM and EMs negatively impacted vaginal contractility and glycosaminoglycan (GAG) content relative to Sham (all p-values<0.001) with EMs having less of an impact on GAGs (p-values<0.003). The results suggest that softer PCU EMs made with more material are well tolerated by the vagina and comprises a future material for POP repair devices. STATEMENT OF SIGNIFICANCE: Prolapse is a debilitating condition in which loss of support to the vagina causes it and the organs supported by it to descend from their normal position in the pelvis. Surgical solutions to rebuild support involves the use of polypropylene mesh which is orders of magnitude stiffer than the vagina. This mismatch results in complications including exposure of the mesh into the vagina and pain. To provide an innovative solution for women, we have developed an elastomeric membrane from a soft polymer that matches the stiffness of the vagina. Here, we show in a rabbit animal model that this device incorporates better into the vagina and is associated with an overall improved host response as compared to polypropylene mesh.

Changes in vaginal cytokines concentrations during artificial insemination and natural service in beef heifers.

OBJECTIVE: Heifer development is crucial for the optimization of reproductive efficiency in beef production. Heifer development is largely influenced by nutrition. Nutritional status of these heifers can influence immunological responses that are crucial for reproduction. Commercial Angus heifers (n = 9) were utilized, in a pilot study, to elucidate the effects of sampling time (days 0, 7, 14, 21, 28, and 35), pregnancy status, and type of mating on individual cytokine concentrations and cytokine profiles in the vagina following breeding. RESULTS: Cytokine profiles were analyzed using MetaboAnalyst 5.0 and one-way ANOVAs were performed in R Studio to identify differences in individual cytokines based on sampling time, pregnancy status, and type of mating. Cytokine profiles were different (P = 0.05) 7 days after either mating type. Cytokines, IL-1ß, IL-17a, MCP-1, and TNFα were different based on the mating type and pregnancy status. Multiple cytokines, IL-1α, IL-1ß, IL-6, IL-8, IL-10, IL-17a, VEGFa, and MIP1α, were different between days regardless of pregnancy status. In conclusion, vaginal cytokines differ based on pregnancy status, type of mating, and time which may be indicative of vital pathways that need to be activated for pregnancy.

Reproductive Tract Microbial Transitions from Late Gestation to Early Postpartum Using 16S rRNA Metagenetic Profiling in First-Pregnancy Heifers.

Studies in recent years indicate that reproductive tract microbial communities are crucial for shaping mammals' health and reproductive outcomes. Following parturition, uterine bacterial contamination often occurs due to the open cervix, which may lead to postpartum uterine inflammatory diseases, especially in primiparous individuals. However, investigations into spatio-temporal microbial transitions in the reproductive tract of primigravid females remain limited. Our objective was to describe and compare the microbial community compositions in the vagina at late gestation and in the vagina and uterus at early postpartum in first-pregnancy heifers. Three swab samples were collected from 33 first-pregnancy Holstein Friesian heifers: one vaginal sample at gestation day 258 ± 4, and vaginal and uterine samples at postpartum day 7 ± 2. Each sample underwent 16S rRNA V4 region metagenetic analysis via Illumina MiSeq, with bioinformatics following Mothur MiSeq SOP. The reproductive tract bacterial communities were assigned to 1255 genus-level OTUs across 30 phyla. Dominant phyla, accounting for approximately 90% of the communities, included Proteobacteria, Firmicutes, Actinobacteria, Bacteroidetes, and Fusobacteria. However, the results revealed distinct shifts in microbial composition between the prepartum vagina (Vag-pre), postpartum vagina (Vag-post), and postpartum uterus (Utr-post). The Vag-pre and Utr-post microbial profiles were the most distinct. The Utr-post group had lower relative abundances of Proteobacteria but higher abundances of Bacteroidetes, Fusobacteria, and Tenericutes compared to Vag-pre, while Vag-post displayed intermediate values for these phyla, suggesting a transitional profile. Additionally, the Utr-post group exhibited lower bacterial richness and diversity compared to both Vag-pre and Vag-post. The unsupervised probabilistic Dirichlet Multinomial Mixtures model identified two distinct community types: most Vag-pre samples clustered into one type and Utr-post samples into another, while Vag-post samples were distributed evenly between the two. LEfSe analysis revealed distinct microbial profiles at the genus level. Overall, specific microbial markers were associated with anatomical and temporal transitions, revealing a dynamic microbial landscape during the first pregnancy and parturition. These differences highlight the complexity of these ecosystems and open new avenues for research in reproductive biology and microbial ecology.

Comparison of the Vaginal and Labial Dimensions of Ethnic Chinese and Western Nullipara.

INTRODUCTION AND HYPOTHESIS: Vaginal dimensions have clinical and surgical implications. We sought to quantify the differences between vaginal and labial dimensions in healthy ethnic Chinese and Western women with normal pelvic organ support. METHODS: This is a cross-sectional study of a convenience sample of ethnic Chinese nulliparas (n = 33) and Western nulliparas (n = 33) recruited for research purposes. For each subject, magnetic resonance imaging was used to quantify the vaginal and labial dimensions. Specifically, we identified the anterior and posterior vaginal wall, the outline of the cervix in the mid-sagittal and coronal planes, and the distance from the labia majora to the hymenal ring at the urethral meatus. RESULTS: There were significant differences in age and weight between groups. Substantial variation in vaginal and labial dimensions was found within each group. The vaginal and labial dimensions of ethnic Chinese women ranged from 9-21% smaller than those of Western women; In the ethnic Chinese group, increasing weight and BMI correlated with greater labial distance (r = 0.66 and r = 0.63 respectively); as did height and the distance from the vaginal opening to the cervical os (r = 0.5). In the Western group, only weight correlated with the labial distance (r = 0.51). CONCLUSIONS: Significant group differences in vaginal and labial dimensions were found, with the dimensions of Chinese nulliparas being up to 21% smaller than those of Western nulliparas.

Persistent Mullerian duct syndrome in a male child: A rare case report on the intersection of surgical importance and economic barriers in Syria.

INTRODUCTION: During embryology, the Mullerian duct develops later into the female reproductive tract, and it is usually obliterates in males by anti-Mullerian hormone. Mutations in the gene for anti-Mullerian hormone or anti-Mullerian hormone receptor will cause persistent Mullerian duct syndrome. To the best of our knowledge, this is the first case of this syndrome documented in the medical literature from Syria. PRESENTATION OF CASE: Herein, we report a case of a 3-year-old male with external male genitalia, who presented complaining mainly of urinary tract infection symptoms. The ultrasound showed the presence of both Mullerian and Wolffian duct derivatives. In fact, and unfortunately, the main obstacles to complete further investigations in this case were a lack of cooperation from parents and the costs of these investigations. We hope that this case report will make interesting additions to the medical literature. DISCUSSION: PMDS syndrome is a rare condition characterized by male pseudohermaphroditism, where individuals have male external features but also possess female reproductive organs. Diagnosis often involves imaging techniques like ultrasound and MRI to visualize Mullerian structures. Laboratory tests, including hormone level assessments, are crucial for confirming the diagnosis. Surgical intervention is the primary treatment approach to prevent complications. CONCLUSION: The most common presentations for PMDS include inguinal hernia, testicular tumors, cryptorchidism, and abdominal tumors. So, when treating an inguinal hernia, especially if there is cryptorchidism, the possibility of PMDD should be considered. Early diagnosis and investigations play a major role in effective management and prevention the complications of this syndrome.

Intrauterine Shaping of Fetal Microbiota.

Mechanisms resulting from the physiological immaturity of the digestive system in children delivered before 32 weeks of gestation and, in particular, different interactions between the microbiome and the body have not been fully elucidated yet. Next-generation sequencing methods demonstrated the presence of bacterial DNA in the placenta and amniotic fluid, which may reflect bacterial populations that initiate intestinal colonization in utero. Numerous studies confirmed the hypothesis stating that intestinal bacteria played an important role in the pathogenesis of necrotizing enterocolitis (NEC) early- and late-onset neonatal sepsis (EONS and LONS). The model and scale of disorders within the intestinal microbiome are the subject of active research in premature infants. Neonatal meconium was primarily used as an indicator defining the environment in utero, as it is formed before birth. Metagenomic results and previous data from microbiological bacterial cultures showed a correlation between the time from birth to sample collection and the detection of bacteria in the neonatal meconium. Therefore, it may be determined that the colonization of the newborn's intestines is influenced by numerous factors, which may be divided into prenatal, perinatal, and postnatal, with particular emphasis put on the mode of delivery and contact with the parent immediately after birth. Background: The aim of this review was to collect available data on the intrauterine shaping of the fetal microbiota. Methods: On 13 March 2024, the available literature in the PubMed National Library of Medicine search engine was reviewed using the following selected keywords: "placental microbiome", "intestinal bacteria in newborns and premature infants", and "intrauterine microbiota". Results: After reviewing the available articles and abstracts and an in-depth analysis of their content, over 100 articles were selected for detailed elaboration. We focused on the origin of microorganisms shaping the microbiota of newborns. We also described the types of bacteria that made up the intrauterine microbiota and the intestinal microbiota of newborns. Conclusions: The data presented in the review on the microbiome of both term newborns and those with a body weight below 1200 g indicate a possible intrauterine colonization of the fetus depending on the duration of pregnancy. The colonization occurs both via the vaginal and intestinal route (hematogenous route). However, there are differences in the demonstrated representatives of various types of bacteria, phyla Firmicutes and Actinobacteria in particular, taking account of the distribution in their abundance in the individual groups of pregnancy duration. Simultaneously, the distribution of the phyla Actinobacteria and Proteobacteria is consistent. Considering the duration of pregnancy, it may also be concluded that the bacterial flora of vaginal origin dominates in preterm newborns, while the flora of intestinal origin dominates in term newborns. This might explain the role of bacterial and infectious factors in inducing premature birth with the rupture of fetal membranes.

Creation of a Neovagina with Single Staged Uterine Anastomosis.

Introduction: While vaginal agenesis most often occurs with an absent or rudimentary, nonfunctioning uterus, it may also occur with a fully developed uterine body. In these scenarios, anastomosis of the functional uterus to a neovagina allows for both egress of menstrual blood as well as potential preservation of fertility: case reports exist of spontaneous conception following creation of a neovagina. However, prior attempts at anastomosis have all included delayed surgery with anastomosis to the uterus several months following the creation of the neovagina. Case Description: The patient was a 17-year-old female who presented with amenorrhea several years after thelarche. After noting a blind ending vagina and a 46XX karyotype, ultrasound and MRI revealed an apparently normal uterus with questionable presence of a cervix, and polycystic appearing ovaries, which may have accounted for the patients minimal hematometra. A multidisciplinary team including gynecology, urology, plastic surgery, and colorectal surgery was organized for creation of a neovagina and attempted anastomosis to the normal appearing uterus. In a single staged robotic procedure, a peritoneal neovagina created in a modified Davydov technique was successfully connected to the uterus. A foley catheter was placed in the uterine cavity to allow for canalization. Diagnostic hysteroscopy six weeks after surgery confirmed a canal into the uterus, and the patient reported ongoing cyclical bleeding with the use of oral contraceptives several months after surgery. Conclusions: In vaginal agenesis with a functional upper reproductive tract, peritoneal neovaginas may be successfully anastomosed to the uterus in a single stage robotic approach.

Clinical Applicability of Microbiota Sampling in a Subfertile Population: Urine versus Vagina.

The urogenital microbiota is increasingly gaining recognition as a significant contributor to reproductive health. Recent studies suggest that microbiota can serve as predictors for fertility treatment outcomes. Our objective was to investigate the degree of similarity in microbial composition between patient-collected urine and vaginal samples in a subfertile population. We enrolled women of reproductive age (20-44 years) diagnosed with subfertility and requiring in vitro fertilization (IVF) or IVF with intracytoplasmic sperm injection (IVF-ICSI) treatment. They self-collected both mid-stream urine samples and vaginal swabs before commencing the IVF or IVF-ICSI procedure. All samples were analysed using the intergenic spacer profiling (IS-pro) technique, a rapid clinical microbiota analysis tool. The main outcome measures were the degree of similarity of microbial composition between the two different, but simultaneously collected, samples. Our findings revealed a high correlation (R squared of 0.78) in microbiota profiles between paired urine and vaginal samples from individual patients. Nevertheless, the urinary microbiota profiles contained fewer species compared to the vaginal microbiota, resulting in minor but distinguishable differences. Furthermore, different subfertility diagnoses appeared to be associated with differences in microbial profiles. A noteworthy observation was the exclusive presence of Escherichia coli (E. coli) in both samples of women diagnosed with male factor subfertility. In conclusion, since urinary microbiota profiles seem to represent a diluted version of the vaginal microbiota, vaginal microbiome sampling to predict fertility treatment outcome seems preferable. To enhance the success of fertility treatments, further research is needed to gain deeper insights into a putative causal role of microbiota in the mechanisms of subfertility.

Effects of dietary restriction and one-carbon metabolite supplementation during the first 63 days of gestation on the maternal gut, vaginal, and blood microbiota in cattle.

BACKGROUND: Maternal diet quality and quantity have significant impacts on both maternal and fetal health and development. The composition and function of the maternal gut microbiome is also significantly influenced by diet; however, little is known about the impact of gestational nutrient restriction on the bovine maternal microbiome during early gestation, which is a critical stage for maternal microbiome-mediated fetal programming to take place. The objective of the present study was to evaluate the impacts of diet restriction and one-carbon metabolite (OCM) supplementation during early gestation on maternal ruminal, vaginal, and blood microbiota in cattle. Thirty-three beef heifers (approx. 14 months old) were used in a 2 × 2 factorial experiment with main factors of target gain (control [CON]; targeted 0.45 kg/d gain vs restricted [RES]; targeted - 0.23 kg/d gain), and OCM supplementation (+ OCM vs - OCM; n = 8/treatment; except n = 9 for RES-OCM). Heifers were individually fed, starting treatment at breeding (d 0) and concluding at d 63 of gestation. Ruminal fluid and vaginal swabs were collected on d - 2, d 35, and d 63 (at necropsy) and whole blood was collected on d 63 (necropsy). Bacterial microbiota was assessed using 16S rRNA gene (V3-V4) sequencing. RESULTS: Overall ruminal microbiota structure was affected by gain, OCM, time, and their interactions. The RES heifers had greater microbial richness (observed ASVs) but neither Shannon nor Inverse Simpson diversity was significantly influenced by gain or OCM supplementation; however, on d 63, 34 bacterial genera showed differential abundance in the ruminal fluid, with 25 genera enriched in RES heifers as compared to CON heifers. In addition, the overall interaction network structure of the ruminal microbiota changed due to diet restriction. The vaginal microbiota community structure was influenced by gain and time. Overall microbial richness and diversity of the vaginal microbiota steadily increased as pregnancy progressed. The vaginal ecological network structure was distinctive between RES and CON heifers with genera-genera interactions being intensified in RES heifers. A relatively diverse bacterial community was detected in blood samples, and the composition of the blood microbiota differed from that of ruminal and vaginal microbiota. CONCLUSION: Restricted dietary intake during early gestation induced significant alterations in the ruminal microbiota which also extended to the vaginal microbiota. The composition of these two microbial communities was largely unaffected by OCM supplementation. Blood associated microbiota was largely distinctive from the ruminal and vaginal microbiota.

Exploring the Pathological Journey of Superficial Spreading Squamous Cell Carcinoma of the Cervix Into the Endometrium.

The superficial extension of cervical squamous cell carcinoma (SCC) into the endometrium by replacing the endometrial glands is rare, as it normally spreads by invading the stroma or by lymphatic invasion. We present a case of a postmenopausal female complaining of vaginal discharge followed by vaginal bleeding. Microscopy showed a superficial spreading SCC of the cervix extending superficially into the endometrium with focal myometrial invasion. Carcinoma in situ changes were observed in the vagina. Based on the Fluhmann criteria and ancillary immunohistochemical testing, it was concluded to be an extension of cervical SCC and not primary endometrial carcinoma. The importance of this entity has not been given because of its low incidence.

Reconstruction of Cloacal Defect: Switch Operation - A New Approach.

Introduction: Reconstruction of a cloacal defect in a girl is often difficult and complicated.[1] This is most often done either by the sacro-perineal route or through the posterior sagittal route.[2] The procedures may involve total mobilization of the cloaca with or without the creation of a lower vagina with the help of a loop of vascularized bowel.[3] In our approach, such defects can be corrected using the lower anorectum for the creation of a lower vagina and abdominoperineal pull-through of the proximal divided bowel. This method can be used both in short and long common channel cloaca. Materials and Methods: We have used this procedure in one patient of posterior cloaca and four patients of anterior cloaca. These operations were done in patients of 1 year and 6 months to 4 years. Only one patient is waiting for the closure of the colostomy. Results: In all the cases, the urethra, the vagina, and the anus were successfully separated. Conclusion: This operation restores normal pelvic anatomy and physiology with minimal mutilation of the pelvic floor.

Vaginal artery pseudoaneurysm after a hysterectomy: A case report and review of the literature.

Key clinical message: This was the first report of a pseudoaneurysm in a vaginal artery after hysterectomy, unlike other published studies that were of pseudoaneurysms in uterine or vaginal arteries after delivery. Abstract: A 51-year-old woman presented with massive vaginal bleeding 7 days after a hysterectomy, which caused hemoglobin to drop. The patient was suspicious of having a vaginal artery pseudoaneurysm according to the sonography. Her bleeding was stopped after the ligation of her left internal iliac artery.

Shaping Microbiota During the First 1000 Days of Life.

Given that the host-microbe interaction is shaped by the immune system response, it is important to understand the key immune system-microbiota relationship during the period from conception to the first years of life. The present work summarizes the available evidence concerning human reproductive microbiota, and also, the microbial colonization during early life, focusing on the potential impact on infant development and health outcomes. Furthermore, we conclude that some dietary strategies including specific probiotics and other-biotics could become potentially valuable tools to modulate the maternal-neonatal microbiota during this early critical window of opportunity for targeted health outcomes throughout the entire lifespan.

Vaginal Sensors.

The development and market emergence of vaginal sensors have begun to demonstrate their impact on women's healthcare. Until recently, in limited cases, these sensors have exhibited their capabilities in diagnosing and monitoring disorders of the vaginal tract during different stages of women's lives. This Perspective is a compilation of what has been accomplished so far in the landscape of vaginal sensors. The text explores the diverse types of vaginal sensor technologies, their applications, and their potential impact on women's healthcare. The review introduces the anatomy of the vagina and cervix and categorizes vaginal sensors that have been developed, highlighting the technologies and potential applications. The paper covers biomarkers of the vaginal tract and discusses their importance in maintaining the overall characteristics of the vaginal system. The text also explores the clinical implications of vaginal sensors in pregnancy monitoring, disease detection, and sexual health management. In the final step, the manuscript provides future perspectives and possibilities that can be incorporated in the emerging field of vaginal sensors.

Advancements in three-dimensional bioprinting for reproductive medicine: a systematic review.

Reproductive failure due to age, genetics and disease necessitates innovative solutions. While reproductive tissue transplantation has advanced, ongoing research seeks superior approaches. Biomaterials, bioengineering and additive manufacturing, such as three-dimensional (3D) bioprinting, are harnessed to restore reproductive function. 3D bioprinting uses materials, cells and growth factors to mimic natural tissues, proving popular for tissue engineering, notably in complex scaffold creation with cell distribution. The versatility which is brought to reproductive medicine by 3D bioprinting allows more accurate and on-site applicability to various problems that are encountered in the field. However, in the literature, there is a lack of studies encompassing the valuable applications of 3D bioprinting in reproductive medicine. This systematic review aims to improve understanding, and focuses on applications in several branches of reproductive medicine. Advancements span the restoration of ovarian function, endometrial regeneration, vaginal reconstruction, and male germ cell bioengineering. 3D bioprinting holds untapped potential in reproductive medicine.

Lactobacilli Have an Opposite Effect on the Resistance to Oxidative Damage of HPV-Infected Compared with Uninfected Vaginal Epithelial Cells.

To study how indigenous or probiotic-introduced lactobacilli affect the sensitivity (estimated as the proportion of surviving, apoptotic, and nonapoptotic deaths) of vaginal epithelial cells obtained from HPV-negative and HPV-positive patients to oxidative damage. The tendency to resist oxidative damage in vaginal epithelial cells of 147 HPV-positive and 59 HPV-negative patients with physiological or suboptimal levels of Lactobacillus was evaluated. Adaptation of cell to curb the oxidative damage in 146 HPV positive and 41 HPV negative with probiotic (Lacticaseibacillus rhamnosus Lcr35) supplementation and without was studied. Resistance of epithelial cells to damage was measured by the ratio of surviving, apoptotic, and dead nonapoptotic cells after three times of hydrogen peroxide treatment using a kit containing annexin V-fluorescein in combination with propidium iodide. If uninfected epithelial cells were in an environment with a physiological level of lactobacilli for significant duration, then these cells were more resilient to damage, and if they lost their viability, it was mainly due to apoptosis. Probiotic therapy also increased the resistance of uninfected epithelial cells to damage. HPV-infected epithelial cells were less resistant to damage at normal levels of lactobacilli compared with Lactobacillus deficiency. In HPV-positive patients with Lactobacillus deficiency, probiotic therapy decreased the resistance of infected epithelial cells to damage; the increase in cell death was mainly due to apoptosis.