An analysis of the vaginal microflora in women positive for Group B Streptococcus during the third trimester of pregnancy

An analysis of the vaginal microflora in women positive for Group B Streptococcus during the third trimester of pregnancy | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article An analysis of the vaginal microflora in women positive for Group B Streptococcus during the third trimester of pregnancy Tong Zhang, Chenglong Tong, Jialin Wang, Shuang Gao, Kangyi Li, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6487350/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 26 Jul, 2025 Read the published version in BMC Microbiology → Version 1 posted 10 You are reading this latest preprint version Abstract Background Presently, 20–40% of pregnant women are colonized with Streptococcus agalactiae , which is commonly referred to as Group B Streptococcus (GBS). Numerous studies have demonstrated the association of GBS infection with adverse pregnancy outcomes and neonatal infectious diseases. However, few studies have explored the complex interactions between GBS and other reproductive tract microbes. Method This study employed a retrospective case‒control design. The research subjects included 62 pregnant women at 35–37 weeks of gestation who received treatment at Shenyang Women and Infants Hospital between November 1, 2022, and July 1, 2024. Chi-square tests and multiple logistic regression analyses were performed to identify factors associated with genital tract colonization in GBS patients. Additionally, reproductive tract swabs from 53 pregnant women were subjected to 16S rRNA microbiome analysis using the Illumina NovaSeq platform. Results Our analysis revealed that factors such as premature rupture of membranes, preterm delivery, diabetes mellitus, vaginal cleanliness, elevated leukocyte count in the vaginal discharge, and fungal infection were associated with an increased risk of GBS colonization. Significant differences in the composition of the reproductive microflora were observed across different GBS infection statuses, with notably greater species diversity in the GBS culture-positive group. At the genus level, Lactobacillus was the predominant genus in the GBS culture-positive group, followed by Gardnerella, Streptococcus, and Bifidobacterium. In contrast, in the GBS culture-negative group, Lactobacillus and Gardnerella remained the most abundant genera but were followed by Prevotella, Bacteroides, and others. Furthermore, GBS reproductive tract colonization was positively correlated with the presence of other microorganisms within the microenvironment. Conclusion GBS colonization is positively correlated with the presence of other microorganisms in the reproductive tract microenvironment during late pregnancy. This association may contribute significantly to severe complications, including reproductive tract inflammation, preterm delivery, and premature rupture of membranes in the later stages of pregnancy. group B Streptococcus pregnant vaginal microflora vagina Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Background Group B Streptococcus (GBS), or Streptococcus agalactiae , is an opportunistic pathogen that can affect pregnant women and colonize the gastrointestinal and reproductive tracts [ 1 – 3 ]. Studies have demonstrated that 20–40% of pregnant women globally carry GBS, with variations observed across different racial backgrounds and regional populations. Generally, the prevalence of GBS infection is more pronounced in low- and middle-income countries. For example, while a whole-genome sequencing analysis conducted in Nagasaki Prefecture, Japan, in 2024 revealed a GBS carriage rate of 19% among pregnant women [ 4 ], and a contemporaneous study in the UK indicated that 20–25% of pregnant women in the region were carriers of GBS [ 5 ], a prospective cross-sectional study in Malaysia reported GBS colonization in 41.0% (235 out of 573) of participants [ 6 ], and a study in Qatar reported GBS infection rates as high as 39.2% among bacteraemia-positive women [ 7 ]. Additionally, a study in Ethiopia reported a GBS colonization rate of 19.08% (162 out of 849) [ 8 ]. However, there is limited research on GBS colonization in the reproductive tract during late pregnancy in China. In general, GBS colonization may present as mild and asymptomatic [ 3 ]. However, emerging evidence suggests that even asymptomatic GBS colonization involves bacterial factors that can facilitate transmission and increase virulence [ 9 ]. GBS colonization is closely associated with infectious disease in both mothers and newborns [ 10 , 11 ] and increases the risk of preterm birth [ 12 ], premature rupture of membranes [ 13 , 14 ], neonatal infectious diseases [ 15 , 16 ], neonatal sepsis [ 17 , 18 ], neonatal pneumonia [ 8 , 10 , 11 , 19 ], neonatal meningitis [ 20 , 21 ], and neonatal neurodevelopmental disorders [ 22 ]. Nevertheless, the pathogenesis of GBS colonization remains incompletely understood, particularly in terms of the mechanisms by which GBS transitions from a harmless commensal to a pathogenic organism. The genital tract microbiome plays a crucial role in modulating local immune responses, defending against pathogens, and maintaining immune tolerance to physiological changes [ 23 ]. Moreover, the microbiome across different body sites defines host metabolic characteristics and significantly contributes to host immune homeostasis and resistance to invading pathogens [ 24 ]. Research has demonstrated that probiotic supplementation during the third trimester of pregnancy can reduce rectal GBS colonization in the reproductive tract and increase perinatal safety [ 13 , 25 ]. Further investigating changes in the reproductive tract microbiome can provide deeper insights into the complex interactions between GBS and other microorganisms, thereby facilitating a more comprehensive understanding of how GBS contributes to disease onset and progression. In summary, GBS infection may be associated with alterations in the microflora of the reproductive tract. However, the potential interaction between GBS colonization and the composition of the reproductive tract microbiome remains poorly understood [ 24 ], and the current research exhibits several limitations, including unclear confounding factors, low sampling rates for GBS, limited data on GBS-infected populations in China, and the inability of classical culture-based methods to detect low-abundance or unculturable microorganisms. Consequently, there is no consensus regarding the relationship between reproductive tract microecological imbalance and GBS infection, necessitating further investigation. High-throughput sequencing technology offers a novel approach to explore the distribution and function of reproductive tract microflora. Therefore, in this study, we have carried out a retrospective case‒control analysis using 16S rRNA next-generation sequencing to examine the reproductive tract microbiome of third-trimester pregnant women. Specifically, we compared the microbiomes of 22 GBS culture-positive and 31 GBS culture-negative women with third-trimester pregnancies, analysed the microbial diversity of the microbiomes and investigated the functional pathways associated with GBS carriage to elucidate its clinical significance and potential impact. Methods Sample source and grouping This retrospective case‒control study examined pregnant women at 35–37 weeks of gestation who were treated at Shenyang Women and Infants Hospital between November 1, 2022, and July 1, 2024. The study included 31 patients in the GBS culture-positive group (hereafter referred to as the SY group) and 31 patients in the GBS culture-negative group (hereafter referred to as the DZ group). All participants provided informed consent, and this study was approved by the institutional ethics committee. Instruments and reagents The NovaSeq 6000 sequencing platform (Illumina, USA), Agilent 2100 Bioanalyzer (Agilent, USA), Qubit 2.0 nucleic acid and protein quantifier (Thermo Field, USA), and Qsep400 high-throughput nucleic acid protein analysis system (Bioptic, Taiwan) were used. A TGuide S96 genomic DNA extraction kit (Tiangen Biochemical Technology, Beijing) and an MGISP-960 high-throughput automated sample preparation system (Huada Zhizao Technology, Shenzhen) were used. Data analysis was conducted using IBM SPSS Statistics 27 (International Business Machines Corporation, USA). Inclusion criteria and exclusion criteria GBS culture was performed, and 31 patients who tested positive for GBS infection were categorized into the GBS-positive group. An additional 31 patients tested during the same period who did not have GBS infection were classified into the GBS-negative group. Using the Illumina NovaSeq sequencing platform, paired-end libraries were constructed and sequenced. Nine patients in the GBS culture-positive group whose sequencing data did not include streptococcus were excluded from the analysis. Statistical methods Chi-square tests and multiple logistic regression analyses were employed to identify factors associated with GBS infection, with statistical significance set at P < 0.05. Sociodemographic and clinical characteristics of the study participants were collected through a thorough review of medical records. Bioinformatics analysis Trimmomatic (version 0.33) was utilized for quality filtering of the raw data, whereas Cutadapt (version 1.9.1) was employed for the identification and removal of primer sequences. The paired-end reads were subsequently assembled using USEARCH (version 10), followed by chimaera detection and removal using UCHIME (version 8.1). This process yielded high-quality sequences suitable for further analysis. Sequence characteristic analysis was conducted using USEARCH (version 10.0) and QIIME2 (version 2020.6). For all samples, operational taxonomic units (OTUs) were defined by clustering at a similarity threshold of 97%. Results Demographic data and 16S sequence characteristics The microorganisms present on reproductive tract swabs collected from 53 pregnant women were sequenced using the Illumina NovaSeq platform for 16S rRNA microbiome analysis. The cohort comprised 31 GBS culture-negative participants and 22 GBS culture-positive participants. The subjects ranged in age from 22 to 46 years, with a mean age of 29.96 ± 4.84 years. The mean age of the GBS-positive participants was 29.45 ± 2.61 years, and that of the GBS-negative participants was 30.32 ± 5.96 years. There was no statistically significant difference in age between the two groups (P > 0.05). The average parity for the tested patients was 1.15 ± 0.41, 1.23 ± 0.43, and 1.10 ± 0.40, respectively, with no statistically significant difference between the groups (P > 0.05). The mean gravidity for the subjects was 2.74 ± 1.36, 2.97 ± 1.66, and 2.41 ± 0.67, respectively, with no statistically significant difference between the groups (P > 0.05). Additionally, the chi-square test revealed significant differences in vaginal cleanliness (P = 0.003) and fungal infection (P = 0.042) between the groups. As shown in Table 1, adverse pregnancy outcomes, such as premature rupture of membranes and preterm delivery, gestational diabetes, abnormal vaginal cleanliness (grade III or IV) in the third trimester, elevated leucorrhoeal white blood cell count, and concurrent fungal infection, were associated with an increased risk of GBS infection (P < 0.05). This study also revealed that, compared with lower uterine segment caesarean section, vaginal delivery was associated with a lower risk of GBS infection. Factors such as anaemia and twin pregnancy were observed to potentially increase the risk of GBS infection, although these differences did not reach statistical significance. Table 1. Risk factors for GBS colonization Group Number of cases Percentage P value OR (95% CI) Number of foetuses Single birth 48 90.6 0.162 Twins 5 9.4 2.84 (0.34–23.7) Delivery mode Lower uterine caesarean section 30 56.6 0.096 Vaginal delivery 14 26.4 1.8 (0.49–6.65) Others 9 17 3.5 (0.62–19.68) Premature rupture of membranes No 44 83 < 0.001 Yes 9 17 1.99 (0.47–8.45) Premature delivery * No 44 83 < 0.001 Yes 9 17 1.16 (0.27–4.9) Anaemia No 29 54.7 0.25 Mild anaemia 16 30.2 1.42 (0.29–6.81) Moderate anaemia 8 15.1 1.67 (0.3–9.27) Diabetes * No 39 73.6 < 0.001 Yes 14 26.4 2.38 (0.69–8.26) Cleanliness of leucorrhoea * Ⅰ/Ⅱ 36 67.9 5/HPF 12 22.6 6.46 (1.5–27.9) Fungal infection * No 43 81.1 0.034 Yes 10 18.9 4.36 (0.98–19.34) Abbreviations: HPF: high-power field of view. Notes: * P value < 0.05. The species composition of the samples was elucidated through a series of bioinformatics processes, including filtering, clustering or denoising, species annotation, and abundance analysis of the reads. A total of 3,877,318 paired-end reads were obtained from 53 samples. After quality control and splicing, a total of 3,502,966 clean reads were generated, with each sample yielding at least 43,459 clean reads, with an average of 66,094 clean reads per sample. Distribution of the microecological flora A Venn diagram (Figure 1) was constructed to illustrate the presence of both shared and unique operational taxonomic units (OTUs) between the GBS culture-negative group and the GBS culture-positive group. A total of 16,495 OTUs were identified in the GBS culture-negative group, whereas 15,740 OTUs were found in the GBS culture-positive group. Specifically, 2,023 OTUs were common to both groups, leaving 14,472 unique OTUs in the GBS culture-negative group and 13,717 unique OTUs in the GBS culture-positive group. The specie diversity was lower in the GBS culture-positive group than in the GBS culture-negative group. However, in addition to these differences, the overall composition of the vaginal microflora in pregnant women from both groups also exhibited many similarities. As illustrated in the species distribution histogram (Figure 2), Firmicutes was the predominant phylum in the core reproductive tract microbiome, followed by Actinobacteria , Bacteroidetes , Proteobacteria , and Acidobacteria , in both groups. At the genus level, Lactobacillus was the most abundant, followed by Gardnerella . However, significant variations were observed in the proportions of microecological flora components across the different groups. Firmicutes constituted 72% and Actinomyces constituted 8% of the total species in the GBS-positive group, whereas Firmicutes accounted for 46% and Actinomyces 31% in the GBS-negative group (P<0.01). Additionally, Lactobacillus and Gardnerella represented 65% and 4% of the bacteria in the GBS-negative group, respectively, whereas they represented 31% and 22% of the bacteria in the GBS-positive group, respectively (P<0.01). Furthermore, our analysis revealed greater species diversity in the GBS culture-positive group than in the GBS culture-negative group. In addition to the aforementioned species, the GBS culture-positive group presented greater abundances of Verrucomicrobia , Chloroflexi , Myxococcus , and Blastomonas . Conversely, the GBS culture-negative group presented greater abundances of Chloroflexi , Verrucomicrobia , Blastomonas , and Myxococcus . At the genus level, the GBS culture-positive group presented greater abundances of Streptococcus , Bifidobacterium , Atopobium , Prevotella , Lactobacillus mucosus reuteri , Corynebacterium , and Bacteroides . In contrast, the GBS culture-negative group presented greater abundances of Prevotella , Bacteroides , Atopobium , Corynebacterium , Lactobacillus reuteri , Streptococcus , and Bifidobacterium . Alpha and beta diversity The microbial community in the reproductive tract in the GBS-positive group presented significantly greater diversity and unevenness than that in the GBS-negative group, with a notable reduction in the abundance of Lactobacillus. Therefore, we conducted further analysis of the differences in reproductive tract microbial composition across the different groups. Alpha diversity analysis further revealed that the microbiota of the GBS culture-positive group presented greater species richness (Figure 3); however, the difference between the groups was not statistically significant. For example, the Shannon index was not significantly different between the DZ group and the SY group (3.6 vs. 4.2, P = 0.47). Similarly, the differences in the Simpson index (0.6 vs. 0.7, P = 0.11), PD whole-tree index (20.0 vs. 19.4, P = 0.85), ACE index (631.7 vs. 812.1, P = 0.45), and Chao 1 index (635.1 vs. 815.0, P = 0.45) values also did not reach statistical significance. Principal component analysis (Figure 4) demonstrated that the first principal component accounted for a substantial proportion of the total variation (54.66%). We subsequently calculated various distance metrics, including binary Jaccard, Bray‒Curtis, and unweighted UniFrac, to quantify the dissimilarity between samples and derive the beta diversity values. Our analysis revealed significant differences in species composition. Specifically, PERMANOVA utilizing the Bray‒Curtis and weighted UniFrac algorithms indicated that the differences between the two groups were statistically significant (P < 0.01). This analysis revealed significant differences in the composition of the reproductive tract microbiomes between the third-trimester GBS culture-positive and GBS culture-negative groups (Figure 5). Additionally, intergroup differences were more pronounced than intragroup variations (Table 2). Together, these data clearly indicate a positive correlation between GBS reproductive tract colonization and the presence of other microorganisms within this microenvironment. Table 2. Beta diversity analysis Index ANOSIM PERMANOVA R P SumsOfSqs MeanSqs F.Model R 2 P value binary_jaccard 0.05 0.09 0.49 0.49 1.01 0.02 0.27 bray_curtis * 0.23 < 0.01 1.33 1.33 3.29 0.06 < 0.01 unweighted_unifrac 0.05 0.10 0.34 0.34 1.55 0.03 0.13 weighted_unifrac * 0.18 < 0.01 0.83 0.83 4.61 0.08 < 0.01 Abbreviations: F.Model: F-test value; MeanSqs: mean square error; R: degree of difference; R 2 : The degree of explanation for differences between sample groups; SumsOfSqs: total variance. Notes: * P 0: difference between groups. R > 0.75: large difference. R > 0.5: moderate difference. R > 0.25: small difference. If R = 0 or is close to 0, there is no difference between groups. If R < 0, the within-group difference is significantly greater than the between-group difference. A larger R 2 indicates a greater degree of explanation for between-group differences. Next, we aimed to investigate which bacterial taxa contribute to the differences in reproductive tract microecological populations under various GBS infection states. Linear discriminant analysis (LDA) conducted on samples from different groups (Figure 6) revealed that Actinomycetaceae , Bifidobacteriaceae , and Streptococcaceae , particularly Streptococcus agalactiae , were significantly associated with GBS-positive status (LDA > 4). Conversely, Lactobacillus inertus and Firmicutes (specifically Bacilli and Lactobacillaceae ) were closely correlated with GBS-negative status (LDA > 4). Analysis of variance (ANOVA) revealed significant differences in the populations of Actinomycetaceae , Firmicutes , Bacilli , Lactobacillaceae , Bifidobacteriaceae , and Gardnerella under different GBS infection states (P < 0.05). Additionally, G-TEST analysis corroborated these findings, revealing significant differences in the abundance of Lactobacillus , Gardnerella , Microbacterium , and Corynebacterium among the different groups (P < 0.05). Functional changes in the reproductive tract microbiota To investigate the functional capabilities of the reproductive tract microbiota in women during the third trimester of pregnancy, we conducted a comprehensive analysis of the dataset involving various gene function predictions. These predictions included KEGG pathway analysis, COG function classification, BugBase phenotype prediction, and FAPROTAX ecological function prediction. Our findings revealed significant differences in the relative abundance percentages of metabolic pathways between the GBS culture-positive group and the GBS culture-negative group (p=0.0056). Notably, the RNA processing and modification metabolic pathway differed significantly between the two groups (p=0.0019). Additionally, there were statistically significant differences (p<0.01) in the chemical heterotrophy, animal parasitism/symbiosis, fermentation, mammalian intestinal microbial functions, and human intestinal microbial functions pathways. Furthermore, the GBS culture-positive group presented a marked increase in aerobic capacity and biofilm formation ability, whereas the ability to harbour mobile elements decreased. Discussion In this study, no significant differences were observed between the GBS-positive and GBS-negative groups in terms of age, maternal history, or other fundamental sociodemographic characteristics. In addition to known correlates such as premature rupture of membranes, preterm delivery, and gestational diabetes, this study identified additional factors associated with an increased risk of genital GBS colonization in pregnant women during the third trimester. These factors include the cleanliness of vaginal discharge, the leukocyte count in vaginal secretions, and the presence of concurrent fungal infections. Through an in-depth analysis of the distribution of reproductive tract microbial populations in pregnant women with different GBS infection statuses, this study identified both common and distinct characteristics between the GBS culture-negative and GBS culture-positive groups. The GBS culture-positive group presented greater species diversity, reduced homogeneity, and significantly decreased Lactobacillus abundance. The reproductive tract microbiome composition differed significantly between the two groups. Additionally, GBS reproductive tract colonization was positively correlated with the presence of other microorganisms within the microenvironment. Consistent with multifactor regression studies conducted in various countries and regions [12-14, 26], this study revealed a significant association between premature rupture of membranes and preterm birth and genital tract GBS colonization in the third trimester. Additionally, in line with recent findings from both animal models and maternal cohorts with gestational diabetes [10, 27, 28], this study revealed that gestational diabetes may be associated with an increased risk of GBS colonization. Some studies have also indicated that pregestational diabetes can increase the likelihood of GBS colonization during pregnancy [29]. All pregnant women should initiate and maintain regular blood glucose monitoring as early as possible and actively manage their blood glucose levels to prevent reproductive tract microecological alterations and reduce the incidence of GBS colonization. This study also revealed a correlation between reproductive tract GBS colonization and the white blood cell count as well as fungal infections in pregnant women in this region, which may be the cause of the higher degree of vaginal cleanliness among GBS culture-positive women compared with GBS culture-negative women in the third trimester (Grade III vs. Grade II, P0.05) and was significantly associated with concurrent fungal infection (p<0.05). GBS colonization is closely linked to vaginitis, which can disrupt the reproductive tract microbial ecosystem and result in severe complications [30, 31]. In summary, GBS colonization of the reproductive tract may lead to adverse pregnancy outcomes or serious neonatal complications by altering the host reproductive tract microecology. We conducted further investigations into the relationship between GBS colonization and specific microecological community compositions. GBS colonization is prevalent in the female gastrointestinal tract and lower genital tract [32]. However, the relationship between GBS colonization and the microecological community is complex, and few studies have focused on the associations between specific reproductive tract microbial compositions and GBS colonization during pregnancy [33]. A large-scale cross-sectional study conducted in the United States demonstrated significant differences in both the alpha and beta diversity of the gastrointestinal microbiota between GBS carriers and non-carriers [32]. However, the relationship between GBS colonization during pregnancy and the alpha and beta diversity indices of the reproductive tract microbiome remains controversial. Several studies have reported that there is no significant correlation between the alpha diversity index of the reproductive tract microbiota and GBS colonization status [20, 34, 35]. Conversely, other studies have indicated that in late pregnancy, the alpha diversity of the reproductive tract microbiota significantly differs according to GBS colonization status [24]. In the present study, the GBS culture-positive group exhibited greater species richness. However, we observed no statistically significant difference in alpha diversity, perhaps because of the reduced abundance of Lactobacillus during the third trimester of pregnancy and the substantial interindividual variation in the relative abundance of GBS bacteria. Nevertheless, GBS colonization compromised the stability of the microecological community, leading to an increased abundance of GBS and its emergence as the predominant strain. Previous studies have demonstrated that the beta diversity index of microflora components does not exhibit statistically significant differences during bacterial infections [36]. However, in our study, a significant difference in the beta diversity index was observed between the GBS culture-positive and GBS culture-negative groups. Additionally, there was a notable distinction in the composition of the reproductive tract microbiome between the two groups, which aligns with findings from a study conducted in Egypt [24]. Our study also demonstrated that the intergroup differences were more pronounced than the intragroup differences. While our findings suggest that GBS colonization alters the distribution of the reproductive tract microbiota, the sample size was limited, necessitating validation with larger datasets. Pregnant women should undergo routine monitoring for GBS reproductive tract colonization during pregnancy, and prompt intervention should be provided to those who test positive for GBS. Our study revealed that, at the phylum level, Firmicutes species predominated in the core reproductive tract microbiome of pregnant women, irrespective of GBS infection status. At the genus level, Lactobacillus was the most abundant, followed by Gardnerella . Notably, the first principal component accounted for a substantial proportion of the total variation between the groups, and significant differences were observed in the microbial composition between the groups. Specifically, the abundance of Lactobacillus was markedly reduced in the GBS-positive group. In the GBS-positive group, Firmicutes represented 72% of the total bacteria, and Actinomyces accounted for 8%. In contrast, in the GBS-negative group, Firmicutes represented 46%, and Actinomyces accounted fo31%. Additionally, within the GBS-negative culture group, Lactobacillus and Gardnerella accounted for 65% and 4%, respectively. In the GBS-positive culture group, the proportions of Lactobacillus and Gardnerella were 31% and 22%, respectively . The normal reproductive tract microbiota plays a crucial role in protecting the genital mucosa from colonization by potential pathogenic bacteria. The presence of Lactobacillus species, particularly Lactobacillus crispatus , is negatively correlated with GBS colonization [37]. Consistent with previous studies, our findings indicate that Lactobacillus remains the predominant bacterial genus in the reproductive tract flora during the third trimester of pregnancy [38, 39]. Notably, the abundance of Lactobacillus crispatus was significantly greater in the GBS-negative group [24], whereas a marked decrease in Lactobacillus abundance was observed in patients with positive GBS cultures, suggesting a potential association between reduced Lactobacillus levels and GBS colonization. Consistent with the results of a longitudinal study on persistent GBS infection during pregnancy [33], this study also revealed a reduction in the reproductive tract microbes of the Lactobacillus crispatus community in the GBS-positive culture group (GBS-positive culture group vs. GBS-negative culture group: 10% vs. 41%) and a relative increase in the ratio of Lactobacillus crispatus to Lactobacillus inerta (GBS-positive vs. GBS-negative: 50.0% vs. 36.6%). These findings suggest that an elevated proportion of Lactobacillus inerta may be associated with an increased risk of GBS colonization, warranting further investigation. Lactobacillus may inhibit GBS colonization by modulating the pH of the microenvironment of the reproductive tract [40] and decreasing the number of pathogen-induced neutrophils [41]. Several studies have demonstrated that Lactobacillus inoculation can prevent and protect against GBS colonization [41-43]. Consistent with the aforementioned findings of Shabayek et al. [24], this study revealed an increase in the Bifidobacterium content. However, other studies have explored Bifidobacterium as potential probiotics for preventing GBS infection [44], as its abundance in infants and young children often decreases following antibiotic treatment. Therefore, the role of Bifidobacterium in GBS colonization remains inconclusive. In this study, we observed a substantial increase in the prevalence of Gardnerella bacteria within the reproductive tract microbiome among pregnant women colonized by GBS compared with those without GBS (23% vs. 4%). This finding supports the conclusion of a 2021 study in a pregnant mouse model in the United States [45] that demonstrated that infection with Gardnerella significantly elevates the risk of GBS colonization. There are several limitations to our study that warrant acknowledgement. First, the sample size was relatively small, and these findings should be validated in future studies with larger cohorts. Second, the second-generation sequencing technology utilized in this study has inherent limitations; namely, it cannot accurately identify all bacterial species present in the reproductive tract, thereby constraining species-level analysis. Conclusion GBS shares intricate relationships and other bacterial taxa within the reproductive tract. This study demonstrates, for the first time, a positive correlation between GBS colonization during late pregnancy and the presence of other microbial communities in the microenvironment of the reproductive tract. This correlation may be a significant factor contributing to serious complications such as vaginitis, preterm birth, and premature rupture of membranes in late pregnancy. Understanding the composition and dynamics of the reproductive tract microflora in women during the third trimester of pregnancy can provide theoretical support and guidance for clinical prevention and treatmentof GBS colonization. Optimizing the composition of the reproductive tract microbiota can decrease the likelihood of GBS colonization during pregnancy, thereby reducing adverse pregnancy outcomes and neonatal infectious diseases. Abbreviations GBS: Group B Streptococcus, OTUs: operational taxonomic units, LDA: linear discriminant analysis, ANOVA: analysis of variance, CI: confidence interval. Declarations Clinical trial number Not applicable Consent for Publication statement Not applicable Ethics approval and consent to participate This study was approved by the ethics committee of the Medical Ethics Committee of Shenyang Women and Infants Hospital and in compliance with the Declaration of Helsinki. In addition, written informed consent was obtained from all participants. Consent for publication All the authors read, commented on, and approved the final manuscript. Availability of data and materials The datasets generated and/or analysed during the current study are available in the National Center for Biotechnology Information (NCBI) repository with the citation accession PRJNA1256138, [https://www.ncbi.nlm.nih.gov/sra/PRJNA1256138]”. Competing interests The authors declare that they have no competing interests. Funding Scientific research project of the Shenyang Health Commission (RC230916) Authors’ contributions Tong Zhang, Chenglong Tong, Jialin Wang, Shuang Gao, Kangyi Li and Xiaona Wang collected the data used in this project. Tong Zhang and Xiaona Wang designed the project. Tong Zhang and Xiaona Wang wrote and edited this first draft. All the authors read, commented on, and approved the final manuscript. Acknowledgements We are grateful for the collaboration of the patients, medical and nursery staff and data managers in this study. We would like to thank Springer Nature Editing Service (https://authorservices.springernature.com) for the English language editing. This study was funded by the Scientific Research Project of the Shenyang Health Commission (RC230916). None of the authors of this manuscript have conflicts of interest. References Pellegrini A, Motta C, Menegussi EB, Pierangelini A, Viglio S, Coppolino F, et al. The serine-rich repeat glycoprotein Srr2 mediates Streptococcus agalactiae interaction with host fibronectin. BMC Microbiol. 2024;24:221. Litman E, Young B, Spiel M. Novel insights on group B Streptococcus in pregnancy. Clin Obstet Gynecol. 2024;67:633–43. Bigus D, Lewandowska W, Biega E, Grela A, Siedlar A, Sosnowska M, et al. Ultra-fast impedimetric immunoassay for detection of Streptococcus agalactiae using carbon electrode with nanodiamonds film. Micromachines (Basel). 2023;14:1076. Nakano S, Koide S, Hosaka Y, Hasegawa Y, Ishida-Kuroki K, Kawakami S, et al. Enrichment culture evaluation and characterization of Streptococcus agalactiae among pregnant women in Japan. J Med Microbiol. 2024;73:001849. Constantinou G, Ayers S, Mitchell EJ, Moore S, Jones AM, Downe S, et al. The acceptability of group B Streptococcal bacteria (GBS) testing to women, including self-swabbing procedures: a qualitative study. Midwifery. 2024;135:104063. Esmaon R, Lim BK, Gan F, Hamdan M, Tan PC. Sexual activity, vaginal symptoms, maternal perineal hygiene behavior, and constipation on ano-vaginal colonization of group B Streptococcus in near term pregnancy. BMC Pregnancy Childbirth. 2024;24:461. Gad A, Alkhdr M, Terkawi R, Alsharif H, Ibrahim M, Amin R, et al. Associations between maternal bacteremia during the peripartum period and early-onset neonatal sepsis: a retrospective cohort study. BMC Pediatr. 2024;24:526. Tesfaye A, Fekede M, Getu F, Mekuria S, Abebe T, Tollosa D, et al. Vertical transmission of group B Streptococcus , prevalence, associated factors, and antimicrobial susceptibility profile among newborns delivered at health facilities in Jigjiga city, Ethiopia. Int J Microbiol. 2024;2024:5673366. Armistead B, Oler E, Waldorf KA, Rajagopal L. The double life of group B Streptococcus : asymptomatic colonizer and potent pathogen. J Mol Biol. 2019;431:2914–31. Mercado-Evans V, Zulk JJ, Hameed ZA, Patras KA. Gestational diabetes as a risk factor for GBS maternal rectovaginal colonization: a systematic review and meta-analysis. BMC Pregnancy Childbirth. 2024;24:488. Afsari M, White A, Adhikari EH. Group B Streptococcus and intraamniotic inflammation and infection. Clin Obstet Gynecol. 2024;67:576–88. Korir ML, Doster RS, Lu J, Guevara MA, Spicer SK, Moore RE, et al. Streptococcus agalactiae cadD alleviates metal stress and promotes intracellular survival in macrophages and ascending infection during pregnancy. Nat Commun. 2022;13:5392. Galvez A, De Teran EDA, Espinosa JA, Perez-Pedregosa J, Bartha-Rasero JL, Del Valle JG, et al. Ligilactobacillus salivarius V4II-90 eradicates group B Streptococcus colonisation during pregnancy: a randomised, double-blind, placebo-controlled trial. Benef Microbes. 2024;15:387–96. Ren J, Qiang Z, Li YY, Zhang JN. Biomarkers for a histological chorioamnionitis diagnosis in pregnant women with or without group B Streptococcus infection: a case-control study. BMC Pregnancy Childbirth. 2021;21:250. Khan UB, Dyster V, Chaguza C, Van Sorge NM, Van De Beek D, Man WK, et al. Genetic markers associated with host status and clonal expansion of group B Streptococcus in the Netherlands. Front Microbiol. 2024;15:1410651. Coppolino F, De Gaetano GV, Claverie C, Sismeiro O, Varet H, Legendre R, et al. The SaeRS two-component system regulates virulence gene expression in group B Streptococcus during invasive infection. mBio. 2024;15:e0197524. Dominguez K, Pearah AN, Lindon AK, Worthington LA, Carter RR, Edwards NJL, et al. The impact of butyrate on group B Streptococcus -induced intestinal barrier disruption. Infect Immun. 2024;92:e0020024. Greenfield KG, Harlow OS, Witt LT, Dziekan EM, Tamar CR, Meier J, et al. Neonatal intestinal colonization of Streptococcus agalactiae and the multiple modes of protection limiting translocation. Gut Microbes. 2024;16:2379862. Job AM, Doran KS, Spencer BL. A group B Streptococcal type VII secreted LXG toxin mediates interbacterial competition and colonization of the female genital tract. bioRxiv 20240610598350. 2024. Akbari MS, Joyce LR, Spencer BL, Brady A, McIver KS, Doran KS. Identification of glyoxalase A in group B Streptococcus and its contribution to methylglyoxal tolerance and virulence. Infect Immun. 2025. 10.1128/iai.00540-24 . Aznar E, Strazielle N, Costa L, Poyart C, Tazi A, Ghersi-Egea JF, et al. The hypervirulent group B Streptococcus HvgA adhesin promotes central nervous system invasion through transcellular crossing of the choroid plexus. Fluids Barriers CNS. 2024;21:66. Khan K. Neurodevelopmental impairment associated with neonatal invasive group B Streptococcus disease: are animal models on track in understanding the mechanisms at play? Brain Behav Immun Health. 2024;40:100831. Marschalko M, Ambrus L. Characteristics and physiologic role of female lower genital microbiome. Orv Hetil. 2023;164:923–30. Shabayek S, Abdellah AM, Salah M, Ramadan M, Fahmy N. Alterations of the vaginal microbiome in healthy pregnant women positive for group B Streptococcus colonization during the third trimester. BMC Microbiol. 2022;22:313. Menichini D, Chiossi G, Monari F, De Seta F, Facchinetti F. Supplementation of probiotics in pregnant women targeting group B Streptococcus colonization: a systematic review and meta-analysis. Nutrients. 2022;14:4520. Dechen TC, Sumit K, Ranabir P. Correlates of vaginal colonization with group B Streptococci among pregnant women. J Glob Infect Dis. 2010;2:236–41. Edwards JM, Watson N, Focht C, Wynn C, Todd CA, Walter EB, et al. Group B Streptococcus (GBS) colonization and disease among pregnant women: a historical cohort study. Infect Dis Obstet Gynecol. 2019;2019:5430493. Pitts SI, Maruthur NM, Langley GE, Pondo T, Shutt KA, Hollick R, et al. Obesity, diabetes, and the risk of invasive group B Streptococcal disease in nonpregnant adults in the united states. Open Forum Infect Dis. 2018;5:ofy030. Field C, Bank TC, Spees CK, Germann K, Landon MB, Gabbe S, et al. Association between glycemic control and group B Streptococcus colonization among pregnant individuals with pregestational diabetes. Am J Reprod Immunol. 2023;90:e13779. Chee WJY, Chew SY, Than LTL. Vaginal microbiota and the potential of Lactobacillus derivatives in maintaining vaginal health. Microb Cell Fact. 2020;19:203. Nguyen ATC, Le Nguyen NT, Hoang TTA, Nguyen TT, Tran TTQ, Tran DNT, et al. Aerobic vaginitis in the third trimester and its impact on pregnancy outcomes. BMC Pregnancy Childbirth. 2022;22:432. Cowley ES, Chaves IZ, Osman F, Suen G, Anantharaman K, Hryckowian AJ. Determinants of gastrointestinal group B Streptococcus carriage in adults. bioRxiv 20230817553755. 2023. Maidment TI, Pelzer ES, Borg DJ, Cheung E, Begun J, Nitert MD, et al. Group B Streptococcus vaginal colonisation throughout pregnancy is associated with decreased Lactobacillus crispatus and increased Lactobacillus iners abundance in the vaginal microbial community. Front Cell Infect Microbiol. 2024;14:1435745. Rosen GH, Randis TM, Desai PV, Sapra KJ, Ma B, Gajer P, et al. Group B Streptococcus and the vaginal microbiota. J Infect Dis. 2017;216:744–51. Le TM, Choi Y, Nguyen HDT, Lee D, Lee OE, Chong GO, et al. Relationship between maternal group B Streptococcal colonization and gestational vaginal microbiome composition: a pilot study. Indian J Med Microbiol. 2023;46:100426. Sayres LC, Younge NE, Rikard B, Corcoran DL, Modliszewski JL, Hughes BL. The gestational membrane microbiome in the presence or absence of intraamniotic infection. Am J Obstet Gynecol MFM. 2023;5:100837. Starc M, Lucovnik M, Vrlic PE, Jeverica S. Protective effect of Lactobacillus crispatus against vaginal colonization with group B Streptococci in the third trimester of pregnancy. Pathogens. 2022;11:980. Mejia ME, Mercado-Evans V, Zulk JJ, Ottinger S, Ruiz K, Ballard MB, et al. Vaginal microbial dynamics and pathogen colonization in a humanized microbiota mouse model. NPJ Biofilms Microbiomes. 2023;9:87. Mejia ME, Robertson CM, Patras KA. Interspecies interactions within the host: the social network of group B Streptococcus . Infect Immun. 2023;91:e0044022. Marziali G, Foschi C, Parolin C, Vitali B, Marangoni A. In-vitro effect of vaginal Lactobacilli against group B Streptococcus . Microb Pathog. 2019;136:103692. De Gregorio PR, Tomas MSJ, Nader-Macias ME. Immunomodulation of Lactobacillus reuteri CRL1324 on group B Streptococcus vaginal colonization in a murine experimental model. Am J Reprod Immunol. 2016;75:23–35. De Gregorio PR, Tomas MSJ, Terraf MCL, Nader-Macias ME. Preventive effect of Lactobacillus reuteri CRL1324 on group B Streptococcus vaginal colonization in an experimental mouse model. J Appl Microbiol. 2015;118:1034–47. Liu Y, Huang Y, Cai W, Li D, Zheng W, Xiao Y, et al. Effect of oral Lactobacillus rhamnosus GR-1 and Lactobacillus reuteri RC-14 on vaginal group B Streptococcus colonization and vaginal microbiome in late pregnancy. Nan Fang Yi Ke Da Xue Xue Bao. 2020;40:1753–9. Aloisio I, Mazzola G, Corvaglia LT, Tonti G, Faldella G, Biavati B, et al. Influence of intrapartum antibiotic prophylaxis against group B Streptococcus on the early newborn gut composition and evaluation of the anti- Streptococcus activity of Bifidobacterium strains. Appl Microbiol Biotechnol. 2014;98:6051–60. Gilbert NM, Foster LR, Cao B, Yin Y, Mysorekar IU, Lewis AL. Gardnerella vaginalis promotes group B Streptococcus vaginal colonization, enabling ascending uteroplacental infection in pregnant mice. Am J Obstet Gynecol. 2021;224:e5301–17. Additional Declarations No competing interests reported. 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Hospital","correspondingAuthor":false,"prefix":"","firstName":"Chenglong","middleName":"","lastName":"Tong","suffix":""},{"id":462698535,"identity":"88ad59b6-9804-412b-825b-edccbe493826","order_by":2,"name":"Jialin Wang","email":"","orcid":"","institution":"Shenyang Women's and Children's Hospital","correspondingAuthor":false,"prefix":"","firstName":"Jialin","middleName":"","lastName":"Wang","suffix":""},{"id":462698536,"identity":"54510567-9392-4183-a840-fdd4fbfe834f","order_by":3,"name":"Shuang Gao","email":"","orcid":"","institution":"Shenyang Women's and Children's Hospital","correspondingAuthor":false,"prefix":"","firstName":"Shuang","middleName":"","lastName":"Gao","suffix":""},{"id":462698537,"identity":"033cee5b-6bd8-4b32-8791-f188e1ea5aae","order_by":4,"name":"Kangyi Li","email":"","orcid":"","institution":"Shenyang Women's and Children's 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2","display":"","copyAsset":false,"role":"figure","size":177012,"visible":true,"origin":"","legend":"\u003cp\u003eHistogram of species distributions\u003c/p\u003e","description":"","filename":"Figure2.Histogramofspeciesdistributions.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6487350/v1/27e1f29ffd054f6f413097f8.jpg"},{"id":83623546,"identity":"936763ae-f448-4acd-8ff3-58ee5163cc74","added_by":"auto","created_at":"2025-05-29 15:55:41","extension":"jpg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":105696,"visible":true,"origin":"","legend":"\u003cp\u003eBox plot of alpha diversity indexes\u003c/p\u003e","description":"","filename":"Figure3.Boxplotofalphadiversityindexes.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6487350/v1/8916d8dc27ca3b619ebde4c9.jpg"},{"id":83623538,"identity":"565ec0c3-4b24-448d-a964-9283727b1991","added_by":"auto","created_at":"2025-05-29 15:55:40","extension":"jpg","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":93542,"visible":true,"origin":"","legend":"\u003cp\u003ePCA diagram\u003c/p\u003e","description":"","filename":"Figure4.PCAdiagram.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6487350/v1/fcd613c2609a681e629213f1.jpg"},{"id":83623540,"identity":"856fa65e-11c1-47bf-9d5e-f65836fe9c40","added_by":"auto","created_at":"2025-05-29 15:55:41","extension":"jpg","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":68326,"visible":true,"origin":"","legend":"\u003cp\u003ePERMANOVA/ANOSIM analysis box diagram\u003c/p\u003e","description":"","filename":"Figure5.PERMANOVAANOSIManalysisboxdiagram.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6487350/v1/6cb0e1ad2873f49087e26ae8.jpg"},{"id":83623955,"identity":"10b6dceb-a6d7-46fd-be04-3180f665eb9e","added_by":"auto","created_at":"2025-05-29 16:03:41","extension":"jpg","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":119439,"visible":true,"origin":"","legend":"\u003cp\u003eHistogram of the LDA value distribution\u003c/p\u003e","description":"","filename":"Figure6.HistogramoftheLDAvaluedistribution.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6487350/v1/652268aadfa75d6163c2e25e.jpg"},{"id":87756673,"identity":"d443535a-2fee-4de3-a60f-66eaff138aec","added_by":"auto","created_at":"2025-07-28 16:07:29","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1431003,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6487350/v1/4544fd89-6263-40db-ad09-78263d9ff757.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"An analysis of the vaginal microflora in women positive for Group B Streptococcus during the third trimester of pregnancy","fulltext":[{"header":"Background","content":"\u003cp\u003eGroup B Streptococcus (GBS), or \u003cem\u003eStreptococcus agalactiae\u003c/em\u003e, is an opportunistic pathogen that can affect pregnant women and colonize the gastrointestinal and reproductive tracts [\u003cspan additionalcitationids=\"CR2\" citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. Studies have demonstrated that 20\u0026ndash;40% of pregnant women globally carry GBS, with variations observed across different racial backgrounds and regional populations. Generally, the prevalence of GBS infection is more pronounced in low- and middle-income countries. For example, while a whole-genome sequencing analysis conducted in Nagasaki Prefecture, Japan, in 2024 revealed a GBS carriage rate of 19% among pregnant women [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e], and a contemporaneous study in the UK indicated that 20\u0026ndash;25% of pregnant women in the region were carriers of GBS [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e], a prospective cross-sectional study in Malaysia reported GBS colonization in 41.0% (235 out of 573) of participants [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e], and a study in Qatar reported GBS infection rates as high as 39.2% among bacteraemia-positive women [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. Additionally, a study in Ethiopia reported a GBS colonization rate of 19.08% (162 out of 849) [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. However, there is limited research on GBS colonization in the reproductive tract during late pregnancy in China.\u003c/p\u003e \u003cp\u003eIn general, GBS colonization may present as mild and asymptomatic [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. However, emerging evidence suggests that even asymptomatic GBS colonization involves bacterial factors that can facilitate transmission and increase virulence [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. GBS colonization is closely associated with infectious disease in both mothers and newborns [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e, \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e] and increases the risk of preterm birth [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e], premature rupture of membranes [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e, \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e], neonatal infectious diseases [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e, \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e], neonatal sepsis [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e, \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e], neonatal pneumonia [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e, \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e, \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e, \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e], neonatal meningitis [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e, \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e], and neonatal neurodevelopmental disorders [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e]. Nevertheless, the pathogenesis of GBS colonization remains incompletely understood, particularly in terms of the mechanisms by which GBS transitions from a harmless commensal to a pathogenic organism. The genital tract microbiome plays a crucial role in modulating local immune responses, defending against pathogens, and maintaining immune tolerance to physiological changes [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e]. Moreover, the microbiome across different body sites defines host metabolic characteristics and significantly contributes to host immune homeostasis and resistance to invading pathogens [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e]. Research has demonstrated that probiotic supplementation during the third trimester of pregnancy can reduce rectal GBS colonization in the reproductive tract and increase perinatal safety [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e, \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e]. Further investigating changes in the reproductive tract microbiome can provide deeper insights into the complex interactions between GBS and other microorganisms, thereby facilitating a more comprehensive understanding of how GBS contributes to disease onset and progression.\u003c/p\u003e \u003cp\u003eIn summary, GBS infection may be associated with alterations in the microflora of the reproductive tract. However, the potential interaction between GBS colonization and the composition of the reproductive tract microbiome remains poorly understood [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e], and the current research exhibits several limitations, including unclear confounding factors, low sampling rates for GBS, limited data on GBS-infected populations in China, and the inability of classical culture-based methods to detect low-abundance or unculturable microorganisms. Consequently, there is no consensus regarding the relationship between reproductive tract microecological imbalance and GBS infection, necessitating further investigation. High-throughput sequencing technology offers a novel approach to explore the distribution and function of reproductive tract microflora. Therefore, in this study, we have carried out a retrospective case‒control analysis using 16S rRNA next-generation sequencing to examine the reproductive tract microbiome of third-trimester pregnant women. Specifically, we compared the microbiomes of 22 GBS culture-positive and 31 GBS culture-negative women with third-trimester pregnancies, analysed the microbial diversity of the microbiomes and investigated the functional pathways associated with GBS carriage to elucidate its clinical significance and potential impact.\u003c/p\u003e"},{"header":"Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eSample source and grouping\u003c/h2\u003e \u003cp\u003eThis retrospective case‒control study examined pregnant women at 35\u0026ndash;37 weeks of gestation who were treated at Shenyang Women and Infants Hospital between November 1, 2022, and July 1, 2024. The study included 31 patients in the GBS culture-positive group (hereafter referred to as the SY group) and 31 patients in the GBS culture-negative group (hereafter referred to as the DZ group). All participants provided informed consent, and this study was approved by the institutional ethics committee.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eInstruments and reagents\u003c/h3\u003e\n\u003cp\u003eThe NovaSeq 6000 sequencing platform (Illumina, USA), Agilent 2100 Bioanalyzer (Agilent, USA), Qubit 2.0 nucleic acid and protein quantifier (Thermo Field, USA), and Qsep400 high-throughput nucleic acid protein analysis system (Bioptic, Taiwan) were used. A TGuide S96 genomic DNA extraction kit (Tiangen Biochemical Technology, Beijing) and an MGISP-960 high-throughput automated sample preparation system (Huada Zhizao Technology, Shenzhen) were used. Data analysis was conducted using IBM SPSS Statistics 27 (International Business Machines Corporation, USA).\u003c/p\u003e\n\u003ch3\u003eInclusion criteria and exclusion criteria\u003c/h3\u003e\n\u003cp\u003eGBS culture was performed, and 31 patients who tested positive for GBS infection were categorized into the GBS-positive group. An additional 31 patients tested during the same period who did not have GBS infection were classified into the GBS-negative group. Using the Illumina NovaSeq sequencing platform, paired-end libraries were constructed and sequenced. Nine patients in the GBS culture-positive group whose sequencing data did not include streptococcus were excluded from the analysis.\u003c/p\u003e\n\u003ch3\u003eStatistical methods\u003c/h3\u003e\n\u003cp\u003eChi-square tests and multiple logistic regression analyses were employed to identify factors associated with GBS infection, with statistical significance set at P\u0026thinsp;\u0026lt;\u0026thinsp;0.05. Sociodemographic and clinical characteristics of the study participants were collected through a thorough review of medical records.\u003c/p\u003e\n\u003ch3\u003eBioinformatics analysis\u003c/h3\u003e\n\u003cp\u003eTrimmomatic (version 0.33) was utilized for quality filtering of the raw data, whereas Cutadapt (version 1.9.1) was employed for the identification and removal of primer sequences. The paired-end reads were subsequently assembled using USEARCH (version 10), followed by chimaera detection and removal using UCHIME (version 8.1). This process yielded high-quality sequences suitable for further analysis. Sequence characteristic analysis was conducted using USEARCH (version 10.0) and QIIME2 (version 2020.6). For all samples, operational taxonomic units (OTUs) were defined by clustering at a similarity threshold of 97%.\u003c/p\u003e"},{"header":"Results","content":"\u003cp\u003e\u003cstrong\u003e\u003cem\u003eDemographic data and 16S sequence characteristics\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe microorganisms present on reproductive tract swabs collected from 53 pregnant women were sequenced using the Illumina NovaSeq platform for 16S rRNA microbiome analysis. The cohort comprised 31 GBS culture-negative participants and 22 GBS culture-positive participants. The subjects ranged in age from 22 to 46 years, with a mean age of 29.96 \u0026plusmn; 4.84 years. The mean age of the GBS-positive participants was 29.45 \u0026plusmn; 2.61 years, and that of the GBS-negative participants was 30.32 \u0026plusmn; 5.96 years. There was no statistically significant difference in age between the two groups (P \u0026gt; 0.05). The average parity for the tested patients was 1.15 \u0026plusmn; 0.41, 1.23 \u0026plusmn; 0.43, and 1.10 \u0026plusmn; 0.40, respectively, with no statistically significant difference between the groups (P \u0026gt; 0.05). The mean gravidity for the subjects was 2.74 \u0026plusmn; 1.36, 2.97 \u0026plusmn; 1.66, and 2.41 \u0026plusmn; 0.67, respectively, with no statistically significant difference between the groups (P \u0026gt; 0.05). Additionally, the chi-square test revealed significant differences in vaginal cleanliness (P = 0.003) and fungal infection (P = 0.042) between the groups. As shown in Table 1, adverse pregnancy outcomes, such as premature rupture of membranes and preterm delivery, gestational diabetes, abnormal vaginal cleanliness (grade III or IV) in the third trimester, elevated leucorrhoeal white blood cell count, and concurrent fungal infection, were associated with an increased risk of GBS infection (P \u0026lt; 0.05). This study also revealed that, compared with lower uterine segment caesarean section, vaginal delivery was associated with a lower risk of GBS infection. Factors such as anaemia and twin pregnancy were observed to potentially increase the risk of GBS infection, although these differences did not reach statistical significance.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 1.\u003c/strong\u003e Risk factors for GBS colonization\u003c/p\u003e\n\u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\" width=\"100%\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 36.7347%;\"\u003e\n \u003cp\u003eGroup\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.3878%;\"\u003e\n \u003cp\u003eNumber of cases\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 13.2653%;\"\u003e\n \u003cp\u003ePercentage\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.2041%;\"\u003e\n \u003cp\u003eP\u003cem\u003e\u0026nbsp;\u003c/em\u003evalue\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.4082%;\"\u003e\n \u003cp\u003eOR (95% CI)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 36.7347%;\"\u003e\n \u003cp\u003eNumber of foetuses\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.3878%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 13.2653%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.2041%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.4082%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 36.7347%;\"\u003e\n \u003cp\u003eSingle birth\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.3878%;\"\u003e\n \u003cp\u003e48\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 13.2653%;\"\u003e\n \u003cp\u003e90.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.2041%;\"\u003e\n \u003cp\u003e0.162\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.4082%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 36.7347%;\"\u003e\n \u003cp\u003eTwins\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.3878%;\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 13.2653%;\"\u003e\n \u003cp\u003e9.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.2041%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.4082%;\"\u003e\n \u003cp\u003e2.84 (0.34\u0026ndash;23.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 36.7347%;\"\u003e\n \u003cp\u003eDelivery mode\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.3878%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 13.2653%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.2041%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.4082%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 36.7347%;\"\u003e\n \u003cp\u003eLower uterine caesarean section\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.3878%;\"\u003e\n \u003cp\u003e30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 13.2653%;\"\u003e\n \u003cp\u003e56.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.2041%;\"\u003e\n \u003cp\u003e0.096\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.4082%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 36.7347%;\"\u003e\n \u003cp\u003eVaginal delivery\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.3878%;\"\u003e\n \u003cp\u003e14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 13.2653%;\"\u003e\n \u003cp\u003e26.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.2041%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.4082%;\"\u003e\n \u003cp\u003e1.8 (0.49\u0026ndash;6.65)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 36.7347%;\"\u003e\n \u003cp\u003eOthers\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.3878%;\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 13.2653%;\"\u003e\n \u003cp\u003e17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.2041%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.4082%;\"\u003e\n \u003cp\u003e3.5 (0.62\u0026ndash;19.68)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 36.7347%;\"\u003e\n \u003cp\u003ePremature rupture of membranes\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.3878%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 13.2653%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.2041%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.4082%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 36.7347%;\"\u003e\n \u003cp\u003eNo\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.3878%;\"\u003e\n \u003cp\u003e44\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 13.2653%;\"\u003e\n \u003cp\u003e83\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.2041%;\"\u003e\n \u003cp\u003e\u0026lt; 0.001\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.4082%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 36.7347%;\"\u003e\n \u003cp\u003eYes\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.3878%;\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 13.2653%;\"\u003e\n \u003cp\u003e17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.2041%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.4082%;\"\u003e\n \u003cp\u003e1.99 (0.47\u0026ndash;8.45)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 36.7347%;\"\u003e\n \u003cp\u003ePremature delivery\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.3878%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 13.2653%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.2041%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.4082%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 36.7347%;\"\u003e\n \u003cp\u003eNo\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.3878%;\"\u003e\n \u003cp\u003e44\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 13.2653%;\"\u003e\n \u003cp\u003e83\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.2041%;\"\u003e\n \u003cp\u003e\u0026lt; 0.001\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.4082%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 36.7347%;\"\u003e\n \u003cp\u003eYes\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.3878%;\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 13.2653%;\"\u003e\n \u003cp\u003e17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.2041%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.4082%;\"\u003e\n \u003cp\u003e1.16 (0.27\u0026ndash;4.9)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 36.7347%;\"\u003e\n \u003cp\u003eAnaemia\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.3878%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 13.2653%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.2041%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.4082%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 36.7347%;\"\u003e\n \u003cp\u003eNo\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.3878%;\"\u003e\n \u003cp\u003e29\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 13.2653%;\"\u003e\n \u003cp\u003e54.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.2041%;\"\u003e\n \u003cp\u003e0.25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.4082%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 36.7347%;\"\u003e\n \u003cp\u003eMild anaemia\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.3878%;\"\u003e\n \u003cp\u003e16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 13.2653%;\"\u003e\n \u003cp\u003e30.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.2041%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.4082%;\"\u003e\n \u003cp\u003e1.42 (0.29\u0026ndash;6.81)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 36.7347%;\"\u003e\n \u003cp\u003eModerate anaemia\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.3878%;\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 13.2653%;\"\u003e\n \u003cp\u003e15.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.2041%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.4082%;\"\u003e\n \u003cp\u003e1.67 (0.3\u0026ndash;9.27)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 36.7347%;\"\u003e\n \u003cp\u003eDiabetes\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.3878%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 13.2653%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.2041%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.4082%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 36.7347%;\"\u003e\n \u003cp\u003eNo\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.3878%;\"\u003e\n \u003cp\u003e39\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 13.2653%;\"\u003e\n \u003cp\u003e73.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.2041%;\"\u003e\n \u003cp\u003e\u0026lt; 0.001\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.4082%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 36.7347%;\"\u003e\n \u003cp\u003eYes\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.3878%;\"\u003e\n \u003cp\u003e14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 13.2653%;\"\u003e\n \u003cp\u003e26.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.2041%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.4082%;\"\u003e\n \u003cp\u003e2.38 (0.69\u0026ndash;8.26)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 36.7347%;\"\u003e\n \u003cp\u003eCleanliness of leucorrhoea\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.3878%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 13.2653%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.2041%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.4082%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 36.7347%;\"\u003e\n \u003cp\u003eⅠ/Ⅱ\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.3878%;\"\u003e\n \u003cp\u003e36\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 13.2653%;\"\u003e\n \u003cp\u003e67.9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.2041%;\"\u003e\n \u003cp\u003e\u0026lt; 0.001\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.4082%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 36.7347%;\"\u003e\n \u003cp\u003eⅢ/Ⅳ\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.3878%;\"\u003e\n \u003cp\u003e17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 13.2653%;\"\u003e\n \u003cp\u003e32.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.2041%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.4082%;\"\u003e\n \u003cp\u003e9.75 (2.53\u0026ndash;37.64)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 36.7347%;\"\u003e\n \u003cp\u003eNumber of white blood cells\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.3878%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 13.2653%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.2041%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.4082%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 36.7347%;\"\u003e\n \u003cp\u003e\u0026le; 5/HPF\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.3878%;\"\u003e\n \u003cp\u003e41\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 13.2653%;\"\u003e\n \u003cp\u003e77.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.2041%;\"\u003e\n \u003cp\u003e0.007\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.4082%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 36.7347%;\"\u003e\n \u003cp\u003e\u0026gt; 5/HPF\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.3878%;\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 13.2653%;\"\u003e\n \u003cp\u003e22.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.2041%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.4082%;\"\u003e\n \u003cp\u003e6.46 (1.5\u0026ndash;27.9)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 36.7347%;\"\u003e\n \u003cp\u003eFungal infection\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.3878%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 13.2653%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.2041%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.4082%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 36.7347%;\"\u003e\n \u003cp\u003eNo\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.3878%;\"\u003e\n \u003cp\u003e43\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 13.2653%;\"\u003e\n \u003cp\u003e81.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.2041%;\"\u003e\n \u003cp\u003e0.034\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.4082%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 36.7347%;\"\u003e\n \u003cp\u003eYes\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19.3878%;\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 13.2653%;\"\u003e\n \u003cp\u003e18.9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10.2041%;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.4082%;\"\u003e\n \u003cp\u003e4.36 (0.98\u0026ndash;19.34)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eAbbreviations: HPF: high-power field of view.\u003c/p\u003e\n\u003cp\u003eNotes:\u003c/p\u003e\n\u003cp\u003e\u003csup\u003e*\u003c/sup\u003e P value \u0026lt; 0.05.\u003c/p\u003e\n\u003cp\u003eThe species composition of the samples was elucidated through a series of bioinformatics processes, including filtering, clustering or denoising, species annotation, and abundance analysis of the reads. A total of 3,877,318 paired-end reads were obtained from 53 samples. After quality control and splicing, a total of 3,502,966 clean reads were generated, with each sample yielding at least 43,459 clean reads, with an average of 66,094 clean reads per sample.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eDistribution of\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e\u003cem\u003ethe\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e\u003cem\u003emicroecological flora\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eA Venn diagram (Figure 1) was constructed to illustrate the presence of both shared and unique operational taxonomic units (OTUs) between the GBS culture-negative group and the GBS culture-positive group. A total of 16,495 OTUs were identified in the GBS culture-negative group, whereas 15,740 OTUs were found in the GBS culture-positive group. Specifically, 2,023 OTUs were common to both groups, leaving 14,472 unique OTUs in the GBS culture-negative group and 13,717 unique OTUs in the GBS culture-positive group. The specie diversity was lower in the GBS culture-positive group than in the GBS culture-negative group. However, in addition to these differences, the overall composition of the vaginal microflora in pregnant women from both groups also exhibited many similarities.\u003c/p\u003e\n\u003cp\u003eAs illustrated in the species distribution histogram (Figure 2), Firmicutes was the predominant phylum in the core reproductive tract microbiome, followed by \u003cem\u003eActinobacteria\u003c/em\u003e, \u003cem\u003eBacteroidetes\u003c/em\u003e, \u003cem\u003eProteobacteria\u003c/em\u003e, and \u003cem\u003eAcidobacteria\u003c/em\u003e, in both groups. At the genus level, \u003cem\u003eLactobacillus\u003c/em\u003e was the most abundant, followed by \u003cem\u003eGardnerella\u003c/em\u003e. However, significant variations were observed in the proportions of microecological flora components across the different groups. \u003cem\u003eFirmicutes\u003c/em\u003e constituted 72% and \u003cem\u003eActinomyces\u003c/em\u003e constituted 8% of the total species in the GBS-positive group, whereas \u003cem\u003eFirmicutes\u003c/em\u003e accounted for 46% and \u003cem\u003eActinomyces\u003c/em\u003e 31% in the GBS-negative group (P\u0026lt;0.01). Additionally, \u003cem\u003eLactobacillus\u003c/em\u003e and \u003cem\u003eGardnerella\u003c/em\u003e represented 65% and 4% of the bacteria in the GBS-negative group, respectively, whereas they represented 31% and 22% of the bacteria in the GBS-positive group, respectively (P\u0026lt;0.01). Furthermore, our analysis revealed greater species diversity in the GBS culture-positive group than in the GBS culture-negative group. In addition to the aforementioned species, the GBS culture-positive group presented greater abundances of \u003cem\u003eVerrucomicrobia\u003c/em\u003e, \u003cem\u003eChloroflexi\u003c/em\u003e, \u003cem\u003eMyxococcus\u003c/em\u003e, and \u003cem\u003eBlastomonas\u003c/em\u003e. Conversely, the GBS culture-negative group presented greater abundances of \u003cem\u003eChloroflexi\u003c/em\u003e, \u003cem\u003eVerrucomicrobia\u003c/em\u003e, \u003cem\u003eBlastomonas\u003c/em\u003e, and \u003cem\u003eMyxococcus\u003c/em\u003e. At the genus level, the GBS culture-positive group presented greater abundances of \u003cem\u003eStreptococcus\u003c/em\u003e, \u003cem\u003eBifidobacterium\u003c/em\u003e, \u003cem\u003eAtopobium\u003c/em\u003e, \u003cem\u003ePrevotella\u003c/em\u003e, \u003cem\u003eLactobacillus\u003c/em\u003e \u003cem\u003emucosus\u003c/em\u003e \u003cem\u003ereuteri\u003c/em\u003e, \u003cem\u003eCorynebacterium\u003c/em\u003e, and \u003cem\u003eBacteroides\u003c/em\u003e. In contrast, the GBS culture-negative group presented greater abundances of \u003cem\u003ePrevotella\u003c/em\u003e, \u003cem\u003eBacteroides\u003c/em\u003e, \u003cem\u003eAtopobium\u003c/em\u003e, \u003cem\u003eCorynebacterium\u003c/em\u003e, \u003cem\u003eLactobacillus\u003c/em\u003e \u003cem\u003ereuteri\u003c/em\u003e, \u003cem\u003eStreptococcus\u003c/em\u003e, and \u003cem\u003eBifidobacterium\u003c/em\u003e.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eAlpha and\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e\u003cem\u003ebeta diversity\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe microbial community in the reproductive tract in the GBS-positive group presented significantly greater diversity and unevenness than that in the GBS-negative group, with a notable reduction in the abundance of Lactobacillus. Therefore, we conducted further analysis of the differences in reproductive tract microbial composition across the different groups. Alpha diversity analysis further revealed that the microbiota of the GBS culture-positive group presented greater species richness (Figure 3); however, the difference between the groups was not statistically significant. For example, the Shannon index was not significantly different between the DZ group and the SY group (3.6 vs. 4.2, P = 0.47). Similarly, the differences in the Simpson index (0.6 vs. 0.7, P = 0.11), PD whole-tree index (20.0 vs. 19.4, P = 0.85), ACE index (631.7 vs. 812.1, P = 0.45), and Chao 1 index (635.1 vs. 815.0, P = 0.45) values also did not reach statistical significance. Principal component analysis (Figure 4) demonstrated that the first principal component accounted for a substantial proportion of the total variation (54.66%). We subsequently calculated various distance metrics, including binary Jaccard, Bray‒Curtis, and unweighted UniFrac, to quantify the dissimilarity between samples and derive the beta diversity values. Our analysis revealed significant differences in species composition. Specifically, PERMANOVA utilizing the Bray‒Curtis and weighted UniFrac algorithms indicated that the differences between the two groups were statistically significant (P \u0026lt; 0.01). This analysis revealed significant differences in the composition of the reproductive tract microbiomes between the third-trimester GBS culture-positive and GBS culture-negative groups (Figure 5). Additionally, intergroup differences were more pronounced than intragroup variations (Table 2). Together, these data clearly indicate a positive correlation between GBS reproductive tract colonization and the presence of other microorganisms within this microenvironment.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 2.\u003c/strong\u003e Beta diversity analysis\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"100%\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 23px;\"\u003e\n \u003cp\u003eIndex\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003eANOSIM\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"5\" valign=\"top\" style=\"width: 57px;\"\u003e\n \u003cp\u003ePERMANOVA\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 23px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 14px;\"\u003e\n \u003cp\u003eSumsOfSqs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12px;\"\u003e\n \u003cp\u003eMeanSqs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11px;\"\u003e\n \u003cp\u003eF.Model\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003eR\u003csup\u003e2\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003eP value\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 23px;\"\u003e\n \u003cp\u003ebinary_jaccard\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.05\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.09\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 14px;\"\u003e\n \u003cp\u003e0.49\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12px;\"\u003e\n \u003cp\u003e0.49\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11px;\"\u003e\n \u003cp\u003e1.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.02\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.27\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 23px;\"\u003e\n \u003cp\u003ebray_curtis\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.23\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e\u0026lt; 0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 14px;\"\u003e\n \u003cp\u003e1.33\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12px;\"\u003e\n \u003cp\u003e1.33\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11px;\"\u003e\n \u003cp\u003e3.29\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.06\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e\u0026lt; 0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 23px;\"\u003e\n \u003cp\u003eunweighted_unifrac\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.05\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 14px;\"\u003e\n \u003cp\u003e0.34\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12px;\"\u003e\n \u003cp\u003e0.34\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11px;\"\u003e\n \u003cp\u003e1.55\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.03\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.13\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 23px;\"\u003e\n \u003cp\u003eweighted_unifrac\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.18\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e\u0026lt; 0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 14px;\"\u003e\n \u003cp\u003e0.83\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12px;\"\u003e\n \u003cp\u003e0.83\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11px;\"\u003e\n \u003cp\u003e4.61\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.08\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e\u0026lt; 0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eAbbreviations: F.Model: F-test value; MeanSqs: mean square error; R: degree of difference; R\u003csup\u003e2\u003c/sup\u003e: The degree of explanation for differences between sample groups; SumsOfSqs: total variance.\u003c/p\u003e\n\u003cp\u003eNotes:\u003c/p\u003e\n\u003cp\u003e\u003csup\u003e*\u003c/sup\u003e P \u0026lt; 0.05;\u003c/p\u003e\n\u003cp\u003eR \u0026gt; 0: difference between groups.\u003c/p\u003e\n\u003cp\u003eR \u0026gt; 0.75: large difference.\u003c/p\u003e\n\u003cp\u003eR \u0026gt; 0.5: moderate difference.\u003c/p\u003e\n\u003cp\u003eR \u0026gt; 0.25: small difference.\u003c/p\u003e\n\u003cp\u003eIf R = 0 or is close to 0, there is no difference between groups.\u003c/p\u003e\n\u003cp\u003eIf R \u0026lt; 0, the within-group difference is significantly greater than the between-group difference.\u003c/p\u003e\n\u003cp\u003eA larger R\u003csup\u003e2\u003c/sup\u003e indicates a greater degree of explanation for between-group differences.\u003c/p\u003e\n\u003cp\u003eNext, we aimed to investigate which bacterial taxa contribute to the differences in reproductive tract microecological populations under various GBS infection states. Linear discriminant analysis (LDA) conducted on samples from different groups (Figure 6) revealed that \u003cem\u003eActinomycetaceae\u003c/em\u003e, \u003cem\u003eBifidobacteriaceae\u003c/em\u003e, and \u003cem\u003eStreptococcaceae\u003c/em\u003e, particularly \u003cem\u003eStreptococcus agalactiae\u003c/em\u003e, were significantly associated with GBS-positive status (LDA \u0026gt; 4). Conversely, \u003cem\u003eLactobacillus inertus\u003c/em\u003e and \u003cem\u003eFirmicutes\u003c/em\u003e (specifically \u003cem\u003eBacilli\u003c/em\u003e and \u003cem\u003eLactobacillaceae\u003c/em\u003e) were closely correlated with GBS-negative status (LDA \u0026gt; 4). Analysis of variance (ANOVA) revealed significant differences in the populations of \u003cem\u003eActinomycetaceae\u003c/em\u003e, \u003cem\u003eFirmicutes\u003c/em\u003e, \u003cem\u003eBacilli\u003c/em\u003e, \u003cem\u003eLactobacillaceae\u003c/em\u003e, \u003cem\u003eBifidobacteriaceae\u003c/em\u003e, and \u003cem\u003eGardnerella\u003c/em\u003e under different GBS infection states (P \u0026lt; 0.05). Additionally, G-TEST analysis corroborated these findings, revealing significant differences in the abundance of \u003cem\u003eLactobacillus\u003c/em\u003e, \u003cem\u003eGardnerella\u003c/em\u003e, \u003cem\u003eMicrobacterium\u003c/em\u003e, and \u003cem\u003eCorynebacterium\u003c/em\u003e among the different groups (P \u0026lt; 0.05).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eFunctional changes\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e\u003cem\u003ein the\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e\u003cem\u003e\u0026nbsp;reproductive tract microbiota\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eTo investigate the functional capabilities of the reproductive tract microbiota in women during the third trimester of pregnancy, we conducted a comprehensive analysis of the dataset involving various gene function predictions. These predictions included KEGG pathway analysis, COG function classification, BugBase phenotype prediction, and FAPROTAX ecological function prediction. Our findings revealed significant differences in the relative abundance percentages of metabolic pathways between the GBS culture-positive group and the GBS culture-negative group (p=0.0056). Notably, the RNA processing and modification metabolic pathway differed significantly between the two groups (p=0.0019). Additionally, there were statistically significant differences (p\u0026lt;0.01) in the chemical heterotrophy, animal parasitism/symbiosis, fermentation, mammalian intestinal microbial functions, and human intestinal microbial functions pathways. Furthermore, the GBS culture-positive group presented a marked increase in aerobic capacity and biofilm formation ability, whereas the ability to harbour mobile elements decreased.\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eIn this study, no significant differences were observed between the GBS-positive and GBS-negative groups in terms of age, maternal history, or other fundamental sociodemographic characteristics. In addition to known correlates such as premature rupture of membranes, preterm delivery, and gestational diabetes, this study identified additional factors associated with an increased risk of genital GBS colonization in pregnant women during the third trimester. These factors include the cleanliness of vaginal discharge, the leukocyte count in vaginal secretions, and the presence of concurrent fungal infections. Through an in-depth analysis of the distribution of reproductive tract microbial populations in pregnant women with different GBS infection statuses, this study identified both common and distinct characteristics between the GBS culture-negative and GBS culture-positive groups. The GBS culture-positive group presented greater species diversity, reduced homogeneity, and significantly decreased Lactobacillus abundance. The reproductive tract microbiome composition differed significantly between the two groups. Additionally, GBS reproductive tract colonization was positively correlated with the presence of other microorganisms within the microenvironment.\u003c/p\u003e\n\u003cp\u003eConsistent with multifactor regression studies conducted in various countries and regions [12-14, 26], this study revealed a significant association between premature rupture of membranes and preterm birth and genital tract GBS colonization in the third trimester. Additionally, in line with recent findings from both animal models and maternal cohorts with gestational diabetes [10, 27, 28], this study revealed that gestational diabetes may be associated with an increased risk of GBS colonization. Some studies have also indicated that pregestational diabetes can increase the likelihood of GBS colonization during pregnancy [29]. All pregnant women should initiate and maintain regular blood glucose monitoring as early as possible and actively manage their blood glucose levels to prevent reproductive tract microecological alterations and reduce the incidence of GBS colonization. This study also revealed a correlation between reproductive tract GBS colonization and the white blood cell count as well as fungal infections in pregnant women in this region, which may be the cause of the higher degree of vaginal cleanliness among GBS culture-positive women compared with GBS culture-negative women in the third trimester (Grade III vs. Grade II, P\u0026lt;0.05). The leukocyte count was greater in the GBS culture-positive group than in the GBS culture-negative group (2+ vs. 1+, p\u0026gt;0.05) and was significantly associated with concurrent fungal infection (p\u0026lt;0.05). GBS colonization is closely linked to vaginitis, which can disrupt the reproductive tract microbial ecosystem and result in severe complications [30, 31]. In summary, GBS colonization of the reproductive tract may lead to adverse pregnancy outcomes or serious neonatal complications by altering the host reproductive tract microecology. We conducted further investigations into the relationship between GBS colonization and specific microecological community compositions.\u003c/p\u003e\n\u003cp\u003eGBS colonization is prevalent in the female gastrointestinal tract and lower genital tract [32]. However, the relationship between GBS colonization and the microecological community is complex, and few studies have focused on the associations between specific reproductive tract microbial compositions and GBS colonization during pregnancy [33]. A large-scale cross-sectional study conducted in the United States demonstrated significant differences in both the alpha and beta diversity of the gastrointestinal microbiota between GBS carriers and non-carriers [32]. However, the relationship between GBS colonization during pregnancy and the alpha and beta diversity indices of the reproductive tract microbiome remains controversial. Several studies have reported that there is no significant correlation between the alpha diversity index of the reproductive tract microbiota and GBS colonization status [20, 34, 35]. Conversely, other studies have indicated that in late pregnancy, the alpha diversity of the reproductive tract microbiota significantly differs according to GBS colonization status [24]. In the present study, the GBS culture-positive group exhibited greater species richness. However, we observed no statistically significant difference in alpha diversity, perhaps because of the reduced abundance of Lactobacillus during the third trimester of pregnancy and the substantial interindividual variation in the relative abundance of GBS bacteria. Nevertheless, GBS colonization compromised the stability of the microecological community, leading to an increased abundance of GBS and its emergence as the predominant strain. Previous studies have demonstrated that the beta diversity index of microflora components does not exhibit statistically significant differences during bacterial infections [36]. However, in our study, a significant difference in the beta diversity index was observed between the GBS culture-positive and GBS culture-negative groups. Additionally, there was a notable distinction in the composition of the reproductive tract microbiome between the two groups, which aligns with findings from a study conducted in Egypt [24]. Our study also demonstrated that the intergroup differences were more pronounced than the intragroup differences. While our findings suggest that GBS colonization alters the distribution of the reproductive tract microbiota, the sample size was limited, necessitating validation with larger datasets. Pregnant women should undergo routine monitoring for GBS reproductive tract colonization during pregnancy, and prompt intervention should be provided to those who test positive for GBS.\u003c/p\u003e\n\u003cp\u003eOur study revealed that, at the phylum level, \u003cem\u003eFirmicutes\u003c/em\u003e species predominated in the core reproductive tract microbiome of pregnant women, irrespective of GBS infection status. At the genus level, \u003cem\u003eLactobacillus\u003c/em\u003e was the most abundant, followed by \u003cem\u003eGardnerella\u003c/em\u003e. Notably, the first principal component accounted for a substantial proportion of the total variation between the groups, and significant differences were observed in the microbial composition between the groups. Specifically, the abundance of \u003cem\u003eLactobacillus\u003c/em\u003e was markedly reduced in the GBS-positive group. In the GBS-positive group, \u003cem\u003eFirmicutes\u003c/em\u003e represented 72% of the total bacteria, and \u003cem\u003eActinomyces\u003c/em\u003e accounted for 8%. In contrast, in the GBS-negative group, \u003cem\u003eFirmicutes\u003c/em\u003e represented 46%, and \u003cem\u003eActinomyces\u003c/em\u003e accounted fo31%. Additionally, within the GBS-negative culture group, \u003cem\u003eLactobacillus\u003c/em\u003e and \u003cem\u003eGardnerella\u003c/em\u003e accounted for 65% and 4%, respectively. In the GBS-positive culture group, the proportions\u0026nbsp;\u003cem\u003eof Lactobacillus and Gardnerella were 31% and 22%, respectively\u003c/em\u003e.\u003c/p\u003e\n\u003cp\u003eThe normal reproductive tract microbiota plays a crucial role in protecting the genital mucosa from colonization by potential pathogenic bacteria. The presence of \u003cem\u003eLactobacillus\u003c/em\u003e species, particularly\u003cem\u003e\u0026nbsp;Lactobacillus crispatus\u003c/em\u003e, is negatively correlated with GBS colonization [37]. Consistent with previous studies, our findings indicate that \u003cem\u003eLactobacillus\u003c/em\u003e remains the predominant bacterial genus in the reproductive tract flora during the third trimester of pregnancy [38, 39]. Notably, the abundance of \u003cem\u003eLactobacillus\u003c/em\u003e \u003cem\u003ecrispatus\u003c/em\u003e was significantly greater in the GBS-negative group [24], whereas a marked decrease in \u003cem\u003eLactobacillus\u003c/em\u003e abundance was observed in patients with positive GBS cultures, suggesting a potential association between reduced Lactobacillus levels and GBS colonization. Consistent with the results of a longitudinal study on persistent GBS infection during pregnancy [33], this study also revealed a reduction in the reproductive tract microbes of the\u0026nbsp;\u003cem\u003eLactobacillus crispatus\u0026nbsp;\u003c/em\u003ecommunity in the GBS-positive culture group (GBS-positive culture group vs. GBS-negative culture group: 10% vs. 41%) and a relative increase in the ratio of \u003cem\u003eLactobacillus crispatus\u003c/em\u003e to \u003cem\u003eLactobacillus inerta\u003c/em\u003e (GBS-positive vs. GBS-negative: 50.0% vs. 36.6%). These findings suggest that an elevated proportion of \u003cem\u003eLactobacillus inerta\u003c/em\u003e may be associated with an increased risk of GBS colonization, warranting further investigation. \u003cem\u003eLactobacillus\u003c/em\u003e may inhibit GBS colonization by modulating the pH of the microenvironment of the reproductive tract\u0026nbsp;[40]\u0026nbsp;and decreasing the number of pathogen-induced neutrophils\u0026nbsp;[41]. Several studies have demonstrated that Lactobacillus inoculation can prevent and protect against GBS colonization\u0026nbsp;[41-43]. Consistent with the aforementioned findings of Shabayek et al.\u0026nbsp;[24], this study\u0026nbsp;revealed\u0026nbsp;an increase in\u0026nbsp;the\u003cem\u003e\u0026nbsp;Bifidobacterium\u003c/em\u003e content. However, other studies have explored \u003cem\u003eBifidobacterium\u0026nbsp;\u003c/em\u003eas potential probiotics for preventing GBS infection\u0026nbsp;[44],\u0026nbsp;as\u0026nbsp;its\u0026nbsp;abundance\u0026nbsp;in infants and young children often\u0026nbsp;decreases\u0026nbsp;following antibiotic treatment. Therefore, the role of\u0026nbsp;\u003cem\u003eBifidobacterium\u003c/em\u003e in GBS colonization remains inconclusive. In this study, we observed a substantial increase in the prevalence of \u003cem\u003eGardnerella\u003c/em\u003e bacteria within the reproductive tract microbiome among pregnant women colonized by GBS compared\u0026nbsp;with\u0026nbsp;those without\u0026nbsp;GBS\u0026nbsp;(23% vs.\u0026nbsp;4%). This finding supports the conclusion of a 2021 study in a pregnant mouse model in the United States\u0026nbsp;[45]\u0026nbsp;that demonstrated that infection with Gardnerella significantly elevates the risk of GBS colonization.\u003c/p\u003e\n\u003cp\u003eThere are several limitations to our study that warrant acknowledgement. First, the sample size was relatively small, and these findings should be validated in future studies with larger cohorts. Second, the second-generation sequencing technology utilized in this study has inherent limitations; namely, it cannot accurately identify all bacterial species present in the reproductive tract, thereby constraining species-level analysis.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eGBS shares intricate relationships and other bacterial taxa within the reproductive tract. This study demonstrates, for the first time, a positive correlation between GBS colonization during late pregnancy and the presence of other microbial communities in the microenvironment of the reproductive tract. This correlation may be a significant factor contributing to serious complications such as vaginitis, preterm birth, and premature rupture of membranes in late pregnancy. Understanding the composition and dynamics of the reproductive tract microflora in women during the third trimester of pregnancy can provide theoretical support and guidance for clinical prevention and treatmentof GBS colonization. Optimizing the composition of the reproductive tract microbiota can decrease the likelihood of GBS colonization during pregnancy, thereby reducing adverse pregnancy outcomes and neonatal infectious diseases.\u003c/p\u003e"},{"header":"Abbreviations","content":"GBS: Group B Streptococcus, OTUs: operational taxonomic units, LDA: linear discriminant analysis, ANOVA: analysis of variance, CI: confidence interval."},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003e\u003cem\u003eClinical trial number\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eConsent for Publication statement\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eEthics approval and consent to participate\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis study was approved by the ethics committee of the Medical Ethics Committee of Shenyang Women and Infants Hospital and in compliance with the Declaration of Helsinki. In addition, written informed consent was obtained from all participants.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eConsent for publication\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll the authors read, commented on, and approved the final manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eAvailability of data and materials\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe datasets generated and/or analysed during the current study are available in the National Center for Biotechnology Information (NCBI) repository with the citation accession PRJNA1256138, [https://www.ncbi.nlm.nih.gov/sra/PRJNA1256138]”.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eCompeting\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e\u003cem\u003einterests\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that they have no competing interests.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eFunding\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eScientific research project of the Shenyang Health Commission (RC230916)\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eAuthors’ contributions\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eTong Zhang, Chenglong Tong, Jialin Wang, Shuang Gao, Kangyi Li and Xiaona Wang collected the data used in this project. Tong Zhang and Xiaona Wang designed the project. Tong Zhang and Xiaona Wang wrote and edited this first draft. All the authors read, commented on, and approved the final manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eAcknowledgements\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe are grateful for the collaboration of the patients, medical and nursery staff and data managers in this study. We would like to thank Springer Nature Editing Service \u0026nbsp;(https://authorservices.springernature.com) for the English language editing. This study was funded by the Scientific Research Project of the Shenyang Health Commission (RC230916). None of the authors of this manuscript have conflicts of interest.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003ePellegrini A, Motta C, Menegussi EB, Pierangelini A, Viglio S, Coppolino F, et al. The serine-rich repeat glycoprotein Srr2 mediates \u003cem\u003eStreptococcus agalactiae\u003c/em\u003e interaction with host fibronectin. BMC Microbiol. 2024;24:221.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLitman E, Young B, Spiel M. Novel insights on group B \u003cem\u003eStreptococcus\u003c/em\u003e in pregnancy. Clin Obstet Gynecol. 2024;67:633\u0026ndash;43.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBigus D, Lewandowska W, Biega E, Grela A, Siedlar A, Sosnowska M, et al. Ultra-fast impedimetric immunoassay for detection of \u003cem\u003eStreptococcus agalactiae\u003c/em\u003e using carbon electrode with nanodiamonds film. Micromachines (Basel). 2023;14:1076.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNakano S, Koide S, Hosaka Y, Hasegawa Y, Ishida-Kuroki K, Kawakami S, et al. Enrichment culture evaluation and characterization of \u003cem\u003eStreptococcus agalactiae\u003c/em\u003e among pregnant women in Japan. J Med Microbiol. 2024;73:001849.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eConstantinou G, Ayers S, Mitchell EJ, Moore S, Jones AM, Downe S, et al. The acceptability of group B Streptococcal bacteria (GBS) testing to women, including self-swabbing procedures: a qualitative study. Midwifery. 2024;135:104063.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEsmaon R, Lim BK, Gan F, Hamdan M, Tan PC. Sexual activity, vaginal symptoms, maternal perineal hygiene behavior, and constipation on ano-vaginal colonization of group B \u003cem\u003eStreptococcus\u003c/em\u003e in near term pregnancy. BMC Pregnancy Childbirth. 2024;24:461.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGad A, Alkhdr M, Terkawi R, Alsharif H, Ibrahim M, Amin R, et al. Associations between maternal bacteremia during the peripartum period and early-onset neonatal sepsis: a retrospective cohort study. BMC Pediatr. 2024;24:526.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTesfaye A, Fekede M, Getu F, Mekuria S, Abebe T, Tollosa D, et al. Vertical transmission of group B \u003cem\u003eStreptococcus\u003c/em\u003e, prevalence, associated factors, and antimicrobial susceptibility profile among newborns delivered at health facilities in Jigjiga city, Ethiopia. Int J Microbiol. 2024;2024:5673366.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eArmistead B, Oler E, Waldorf KA, Rajagopal L. The double life of group B \u003cem\u003eStreptococcus\u003c/em\u003e: asymptomatic colonizer and potent pathogen. J Mol Biol. 2019;431:2914\u0026ndash;31.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMercado-Evans V, Zulk JJ, Hameed ZA, Patras KA. Gestational diabetes as a risk factor for GBS maternal rectovaginal colonization: a systematic review and meta-analysis. BMC Pregnancy Childbirth. 2024;24:488.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAfsari M, White A, Adhikari EH. Group B \u003cem\u003eStreptococcus\u003c/em\u003e and intraamniotic inflammation and infection. Clin Obstet Gynecol. 2024;67:576\u0026ndash;88.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKorir ML, Doster RS, Lu J, Guevara MA, Spicer SK, Moore RE, et al. \u003cem\u003eStreptococcus agalactiae\u003c/em\u003e cadD alleviates metal stress and promotes intracellular survival in macrophages and ascending infection during pregnancy. Nat Commun. 2022;13:5392.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGalvez A, De Teran EDA, Espinosa JA, Perez-Pedregosa J, Bartha-Rasero JL, Del Valle JG, et al. \u003cem\u003eLigilactobacillus salivarius\u003c/em\u003e V4II-90 eradicates group B \u003cem\u003eStreptococcus\u003c/em\u003e colonisation during pregnancy: a randomised, double-blind, placebo-controlled trial. Benef Microbes. 2024;15:387\u0026ndash;96.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRen J, Qiang Z, Li YY, Zhang JN. Biomarkers for a histological chorioamnionitis diagnosis in pregnant women with or without group B \u003cem\u003eStreptococcus\u003c/em\u003e infection: a case-control study. BMC Pregnancy Childbirth. 2021;21:250.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKhan UB, Dyster V, Chaguza C, Van Sorge NM, Van De Beek D, Man WK, et al. Genetic markers associated with host status and clonal expansion of group B \u003cem\u003eStreptococcus\u003c/em\u003e in the Netherlands. Front Microbiol. 2024;15:1410651.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCoppolino F, De Gaetano GV, Claverie C, Sismeiro O, Varet H, Legendre R, et al. The SaeRS two-component system regulates virulence gene expression in group B \u003cem\u003eStreptococcus\u003c/em\u003e during invasive infection. mBio. 2024;15:e0197524.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDominguez K, Pearah AN, Lindon AK, Worthington LA, Carter RR, Edwards NJL, et al. The impact of butyrate on group B \u003cem\u003eStreptococcus\u003c/em\u003e-induced intestinal barrier disruption. Infect Immun. 2024;92:e0020024.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGreenfield KG, Harlow OS, Witt LT, Dziekan EM, Tamar CR, Meier J, et al. Neonatal intestinal colonization of \u003cem\u003eStreptococcus agalactiae\u003c/em\u003e and the multiple modes of protection limiting translocation. Gut Microbes. 2024;16:2379862.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJob AM, Doran KS, Spencer BL. A group B Streptococcal type VII secreted LXG toxin mediates interbacterial competition and colonization of the female genital tract. bioRxiv 20240610598350. 2024.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAkbari MS, Joyce LR, Spencer BL, Brady A, McIver KS, Doran KS. Identification of glyoxalase A in group B \u003cem\u003eStreptococcus\u003c/em\u003e and its contribution to methylglyoxal tolerance and virulence. Infect Immun. 2025. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1128/iai.00540-24\u003c/span\u003e\u003cspan address=\"10.1128/iai.00540-24\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAznar E, Strazielle N, Costa L, Poyart C, Tazi A, Ghersi-Egea JF, et al. The hypervirulent group B \u003cem\u003eStreptococcus\u003c/em\u003e HvgA adhesin promotes central nervous system invasion through transcellular crossing of the choroid plexus. Fluids Barriers CNS. 2024;21:66.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKhan K. Neurodevelopmental impairment associated with neonatal invasive group B \u003cem\u003eStreptococcus\u003c/em\u003e disease: are animal models on track in understanding the mechanisms at play? Brain Behav Immun Health. 2024;40:100831.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMarschalko M, Ambrus L. Characteristics and physiologic role of female lower genital microbiome. Orv Hetil. 2023;164:923\u0026ndash;30.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eShabayek S, Abdellah AM, Salah M, Ramadan M, Fahmy N. Alterations of the vaginal microbiome in healthy pregnant women positive for group B \u003cem\u003eStreptococcus\u003c/em\u003e colonization during the third trimester. BMC Microbiol. 2022;22:313.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMenichini D, Chiossi G, Monari F, De Seta F, Facchinetti F. Supplementation of probiotics in pregnant women targeting group B \u003cem\u003eStreptococcus\u003c/em\u003e colonization: a systematic review and meta-analysis. Nutrients. 2022;14:4520.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDechen TC, Sumit K, Ranabir P. Correlates of vaginal colonization with group B \u003cem\u003eStreptococci\u003c/em\u003e among pregnant women. J Glob Infect Dis. 2010;2:236\u0026ndash;41.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEdwards JM, Watson N, Focht C, Wynn C, Todd CA, Walter EB, et al. Group B \u003cem\u003eStreptococcus\u003c/em\u003e (GBS) colonization and disease among pregnant women: a historical cohort study. Infect Dis Obstet Gynecol. 2019;2019:5430493.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePitts SI, Maruthur NM, Langley GE, Pondo T, Shutt KA, Hollick R, et al. Obesity, diabetes, and the risk of invasive group B Streptococcal disease in nonpregnant adults in the united states. Open Forum Infect Dis. 2018;5:ofy030.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eField C, Bank TC, Spees CK, Germann K, Landon MB, Gabbe S, et al. Association between glycemic control and group B \u003cem\u003eStreptococcus\u003c/em\u003e colonization among pregnant individuals with pregestational diabetes. Am J Reprod Immunol. 2023;90:e13779.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChee WJY, Chew SY, Than LTL. Vaginal microbiota and the potential of \u003cem\u003eLactobacillus\u003c/em\u003e derivatives in maintaining vaginal health. Microb Cell Fact. 2020;19:203.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNguyen ATC, Le Nguyen NT, Hoang TTA, Nguyen TT, Tran TTQ, Tran DNT, et al. Aerobic vaginitis in the third trimester and its impact on pregnancy outcomes. BMC Pregnancy Childbirth. 2022;22:432.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCowley ES, Chaves IZ, Osman F, Suen G, Anantharaman K, Hryckowian AJ. Determinants of gastrointestinal group B \u003cem\u003eStreptococcus\u003c/em\u003e carriage in adults. bioRxiv 20230817553755. 2023.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMaidment TI, Pelzer ES, Borg DJ, Cheung E, Begun J, Nitert MD, et al. Group B \u003cem\u003eStreptococcus\u003c/em\u003e vaginal colonisation throughout pregnancy is associated with decreased \u003cem\u003eLactobacillus crispatus\u003c/em\u003e and increased \u003cem\u003eLactobacillus iners\u003c/em\u003e abundance in the vaginal microbial community. Front Cell Infect Microbiol. 2024;14:1435745.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRosen GH, Randis TM, Desai PV, Sapra KJ, Ma B, Gajer P, et al. Group B \u003cem\u003eStreptococcus\u003c/em\u003e and the vaginal microbiota. J Infect Dis. 2017;216:744\u0026ndash;51.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLe TM, Choi Y, Nguyen HDT, Lee D, Lee OE, Chong GO, et al. Relationship between maternal group B \u003cem\u003eStreptococcal\u003c/em\u003e colonization and gestational vaginal microbiome composition: a pilot study. Indian J Med Microbiol. 2023;46:100426.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSayres LC, Younge NE, Rikard B, Corcoran DL, Modliszewski JL, Hughes BL. The gestational membrane microbiome in the presence or absence of intraamniotic infection. Am J Obstet Gynecol MFM. 2023;5:100837.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eStarc M, Lucovnik M, Vrlic PE, Jeverica S. Protective effect of \u003cem\u003eLactobacillus crispatus\u003c/em\u003e against vaginal colonization with group B \u003cem\u003eStreptococci\u003c/em\u003e in the third trimester of pregnancy. Pathogens. 2022;11:980.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMejia ME, Mercado-Evans V, Zulk JJ, Ottinger S, Ruiz K, Ballard MB, et al. Vaginal microbial dynamics and pathogen colonization in a humanized microbiota mouse model. NPJ Biofilms Microbiomes. 2023;9:87.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMejia ME, Robertson CM, Patras KA. Interspecies interactions within the host: the social network of group B \u003cem\u003eStreptococcus\u003c/em\u003e. Infect Immun. 2023;91:e0044022.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMarziali G, Foschi C, Parolin C, Vitali B, Marangoni A. In-vitro effect of vaginal \u003cem\u003eLactobacilli\u003c/em\u003e against group B \u003cem\u003eStreptococcus\u003c/em\u003e. Microb Pathog. 2019;136:103692.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDe Gregorio PR, Tomas MSJ, Nader-Macias ME. Immunomodulation of \u003cem\u003eLactobacillus reuteri\u003c/em\u003e CRL1324 on group B \u003cem\u003eStreptococcus\u003c/em\u003e vaginal colonization in a murine experimental model. Am J Reprod Immunol. 2016;75:23\u0026ndash;35.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDe Gregorio PR, Tomas MSJ, Terraf MCL, Nader-Macias ME. Preventive effect of \u003cem\u003eLactobacillus reuteri\u003c/em\u003e CRL1324 on group B \u003cem\u003eStreptococcus\u003c/em\u003e vaginal colonization in an experimental mouse model. J Appl Microbiol. 2015;118:1034\u0026ndash;47.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLiu Y, Huang Y, Cai W, Li D, Zheng W, Xiao Y, et al. Effect of oral \u003cem\u003eLactobacillus rhamnosus\u003c/em\u003e GR-1 and \u003cem\u003eLactobacillus reuteri\u003c/em\u003e RC-14 on vaginal group B \u003cem\u003eStreptococcus\u003c/em\u003e colonization and vaginal microbiome in late pregnancy. Nan Fang Yi Ke Da Xue Xue Bao. 2020;40:1753\u0026ndash;9.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAloisio I, Mazzola G, Corvaglia LT, Tonti G, Faldella G, Biavati B, et al. Influence of intrapartum antibiotic prophylaxis against group B \u003cem\u003eStreptococcus\u003c/em\u003e on the early newborn gut composition and evaluation of the anti-\u003cem\u003eStreptococcus\u003c/em\u003e activity of \u003cem\u003eBifidobacterium\u003c/em\u003e strains. Appl Microbiol Biotechnol. 2014;98:6051\u0026ndash;60.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGilbert NM, Foster LR, Cao B, Yin Y, Mysorekar IU, Lewis AL. Gardnerella vaginalis promotes group B \u003cem\u003eStreptococcus\u003c/em\u003e vaginal colonization, enabling ascending uteroplacental infection in pregnant mice. Am J Obstet Gynecol. 2021;224:e5301\u0026ndash;17.\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"bmc-microbiology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"mcro","sideBox":"Learn more about [BMC Microbiology](http://bmcmicrobiol.biomedcentral.com/)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/mcro","title":"BMC Microbiology","twitterHandle":"#bmcmicrobiology","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"em","reportingPortfolio":"BMC Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"group B Streptococcus, pregnant, vaginal microflora, vagina","lastPublishedDoi":"10.21203/rs.3.rs-6487350/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6487350/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003eBackground\u003c/h2\u003e \u003cp\u003ePresently, 20\u0026ndash;40% of pregnant women are colonized with \u003cem\u003eStreptococcus agalactiae\u003c/em\u003e, which is commonly referred to as Group B Streptococcus (GBS). Numerous studies have demonstrated the association of GBS infection with adverse pregnancy outcomes and neonatal infectious diseases. However, few studies have explored the complex interactions between GBS and other reproductive tract microbes.\u003c/p\u003e\u003ch2\u003eMethod\u003c/h2\u003e \u003cp\u003eThis study employed a retrospective case‒control design. The research subjects included 62 pregnant women at 35\u0026ndash;37 weeks of gestation who received treatment at Shenyang Women and Infants Hospital between November 1, 2022, and July 1, 2024. Chi-square tests and multiple logistic regression analyses were performed to identify factors associated with genital tract colonization in GBS patients. Additionally, reproductive tract swabs from 53 pregnant women were subjected to 16S rRNA microbiome analysis using the Illumina NovaSeq platform.\u003c/p\u003e\u003ch2\u003eResults\u003c/h2\u003e \u003cp\u003eOur analysis revealed that factors such as premature rupture of membranes, preterm delivery, diabetes mellitus, vaginal cleanliness, elevated leukocyte count in the vaginal discharge, and fungal infection were associated with an increased risk of GBS colonization. Significant differences in the composition of the reproductive microflora were observed across different GBS infection statuses, with notably greater species diversity in the GBS culture-positive group. At the genus level, Lactobacillus was the predominant genus in the GBS culture-positive group, followed by Gardnerella, Streptococcus, and Bifidobacterium. In contrast, in the GBS culture-negative group, Lactobacillus and Gardnerella remained the most abundant genera but were followed by Prevotella, Bacteroides, and others. Furthermore, GBS reproductive tract colonization was positively correlated with the presence of other microorganisms within the microenvironment.\u003c/p\u003e\u003ch2\u003eConclusion\u003c/h2\u003e \u003cp\u003eGBS colonization is positively correlated with the presence of other microorganisms in the reproductive tract microenvironment during late pregnancy. This association may contribute significantly to severe complications, including reproductive tract inflammation, preterm delivery, and premature rupture of membranes in the later stages of pregnancy.\u003c/p\u003e","manuscriptTitle":"An analysis of the vaginal microflora in women positive for Group B Streptococcus during the third trimester of pregnancy","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-05-29 15:55:36","doi":"10.21203/rs.3.rs-6487350/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-05-28T07:02:53+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-05-27T19:20:37+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-05-25T14:09:13+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"29870710311173079970651455006919512984","date":"2025-05-25T12:34:25+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"289199684155938745555580655607423585542","date":"2025-05-23T13:30:53+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-05-23T11:08:17+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-05-22T16:56:54+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2025-05-05T05:59:05+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-05-02T13:53:03+00:00","index":"","fulltext":""},{"type":"submitted","content":"BMC Microbiology","date":"2025-05-02T13:51:55+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"bmc-microbiology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"mcro","sideBox":"Learn more about [BMC Microbiology](http://bmcmicrobiol.biomedcentral.com/)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/mcro","title":"BMC Microbiology","twitterHandle":"#bmcmicrobiology","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"em","reportingPortfolio":"BMC Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"27af7482-b45d-496d-96cc-b19afd21f176","owner":[],"postedDate":"May 29th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2025-07-28T16:00:12+00:00","versionOfRecord":{"articleIdentity":"rs-6487350","link":"https://doi.org/10.1186/s12866-025-04184-0","journal":{"identity":"bmc-microbiology","isVorOnly":false,"title":"BMC Microbiology"},"publishedOn":"2025-07-26 15:57:15","publishedOnDateReadable":"July 26th, 2025"},"versionCreatedAt":"2025-05-29 15:55:36","video":"","vorDoi":"10.1186/s12866-025-04184-0","vorDoiUrl":"https://doi.org/10.1186/s12866-025-04184-0","workflowStages":[]},"version":"v1","identity":"rs-6487350","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6487350","identity":"rs-6487350","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}