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MSphere Jan 2021Preterm birth (PTB) is the largest contributor to infant death in sub-Saharan Africa and globally. With a global estimate of 773,600, Nigeria has the third highest rate...
Preterm birth (PTB) is the largest contributor to infant death in sub-Saharan Africa and globally. With a global estimate of 773,600, Nigeria has the third highest rate of PTB worldwide. There have been a number of microbiome profiling studies to identify vaginal microbiomes suggestive of preterm and healthy birth outcome. However, studies on the pregnancy vaginal microbiome in Africa are sparse with none performed in Nigeria. Moreover, few studies have considered the concurrent impact of steroid hormones and the vaginal microbiome on pregnancy outcome. We assessed two key determinants of pregnancy progression to gain a deeper understanding of the interactions between vaginal microbiome composition, steroid hormone concentrations, and pregnancy outcome. Vaginal swabs and blood samples were prospectively collected from healthy midtrimester pregnant women. Vaginal microbiome compositions were assessed by analysis of the V3-V5 region of 16S rRNA genes, and potential functional metabolic traits of identified vaginal microbiomes were imputed by PICRUSt (phylogenetic investigation of communities by reconstruction of unobserved states) analysis, while plasma estradiol (E2) and progesterone (P1) levels were quantified by the competitive enzyme-linked immunosorbent assay (ELISA). PTB vaginal samples were characterized by increased microbial richness, high diversity, and depletion of lactobacilli compared to term delivery samples. Women who delivered preterm were characterized by an -dominated vagitype. High relative abundance of at the midtrimester was highly predictive of PTB (area under the receiving operator characteristics [AUROC] of 0.983). There was a marked overlap in the range of plasma E2 and P1 values between term and PTB groups. Giving birth too soon accounts for half of all newborn deaths worldwide. Clinical symptoms alone are not sufficient to identify women at risk of giving birth too early, as such a pragmatic approach to reducing the incidence of preterm birth entails developing early strategies for intervention before it materializes. In view of the role played by the vaginal microbiome and maternal steroid hormones in determining obstetric outcome, we assessed the vaginal microbiome composition and steroid hormone during pregnancy and examined their relationship in predicting preterm birth risk in Nigerian women. This study highlights a potential early-driver microbial marker for prediction of preterm birth risk and supports the notion that vaginal microbiome composition varies across populations. A knowledge of relevant preterm birth microbial markers specific to populations would enhance the development of personalized therapeutic interventions toward restoring a microbiome that optimizes reproductive health fitness, therefore reducing the incidence of preterm birth.
Topics: Actinobacteria; Adult; Estradiol; Female; Humans; Microbiota; Pregnancy; Pregnancy Trimester, Second; Premature Birth; Progesterone; Vagina
PubMed: 33504666
DOI: 10.1128/mSphere.01261-20 -
Pathogens (Basel, Switzerland) Jan 2021Healthy women of reproductive age have a vaginal pH around 4.5, whereas little is known about pH in the upper genital tract. A shift in the vaginal microbiota may result...
INTRODUCTION
Healthy women of reproductive age have a vaginal pH around 4.5, whereas little is known about pH in the upper genital tract. A shift in the vaginal microbiota may result in an elevated pH in the upper genital tract. This might contribute to decreased fertility and increased risk of preterm birth. Therefore, we aimed to measure pH in different compartments of the female genital tract in both nonpregnant and pregnant women, stratifying into a normal and abnormal vaginal microbiota.
MATERIAL AND METHODS
In this descriptive study, we included 6 nonpregnant, 12 early-pregnant, and 8 term-pregnant women. A pH gradient was recorded with a flexible pH probe. An abnormal vaginal microbiota was diagnosed by a quantitative polymerase chain reaction technique for ; ; ; bacterial vaginosis-associated bacterium 1, 2, 3, and TM7; and spp. among others.
RESULTS
In all participants we found the pH gradient in the lower reproductive canal to be most acidic in the lower vagina and most alkaline in the upper uterine cavity. Women with an abnormal vaginal microbiota had an increased pH in the lower vagina compared to the other groups.
CONCLUSIONS
There is a pronounced pH gradient within the female genital tract. This gradient is not disrupted in women with an abnormal vaginal microbiota.
PubMed: 33498288
DOI: 10.3390/pathogens10020090 -
Scientific Reports Jan 2021Persistent HPV infection associated with immune modulation may result in high-grade squamous intraepithelial lesions (CIN)2/3. Currently, there is little information on...
Persistent HPV infection associated with immune modulation may result in high-grade squamous intraepithelial lesions (CIN)2/3. Currently, there is little information on the cervicovaginal microbiome, local cytokine levels and HPV infection related to CIN. Follow-up of patients after local surgery provides an opportunity to monitor changes in the cervicovaginal environment. Accordingly, we undertook this longitudinal retrospective study to determine associations between HPV genotypes, cervicovaginal microbiome and local cytokine profiles in 41 Japanese patients with CIN. Cervicovaginal microbiota were identified using universal 16S rRNA gene (rDNA) bacterial primers for the V3/4 region by PCR of genomic DNA, followed by MiSeq sequencing. We found that Atopobium vaginae was significantly decreased (p < 0.047), whereas A. ureaplasma (p < 0.022) increased after surgery. Cytokine levels in cervical mucus were measured by multiplexed bead-based immunoassays, revealing that IL-1β (p < 0.006), TNF-α (p < 0.004), MIP-1α (p < 0.045) and eotaxin (p < 0.003) were significantly decreased after surgery. Notably, the level of eotaxin decreased in parallel with HPV clearance after surgery (p < 0.028). Thus, local surgery affected the cervicovaginal microbiome, status of HPV infection and immune response. Changes to the cervicovaginal microbiota and cervical cytokine profile following surgery for cervical intraepithelial neoplasia may be important for understanding the pathogenesis of CIN in future.
Topics: Adult; Cervix Uteri; Cytokines; Female; Genotype; Humans; Microbiota; Papillomaviridae; Phylogeny; Time Factors; Uterine Cervical Neoplasms; Vagina; Uterine Cervical Dysplasia
PubMed: 33495564
DOI: 10.1038/s41598-020-80176-6 -
Archives of Microbiology Apr 2021The exploration of vaginal microbiota by using next-generation sequencing (NGS) of 16S ribosomal RNA (rRNA) gene is widely used. Up to now, different hypervariable...
The exploration of vaginal microbiota by using next-generation sequencing (NGS) of 16S ribosomal RNA (rRNA) gene is widely used. Up to now, different hypervariable regions have been selected to study vaginal microbiota by NGS and there is no standard method for analysis. The study aimed to characterize vaginal microbiota from clinical samples using NGS targeting the 16S rRNA gene and to determine the performance of individual and concatenated hypervariable region sequences to generate the taxonomic profiles of the vaginal microbiota. Fifty-one vaginal DNA samples were subjected to 16S rRNA gene NGS based on the Ion Torrent PGM platform with the use of two primer sets spanning seven hypervariable regions of the 16S rRNA gene. Our analysis revealed that the predominant bacterial genera were Lactobacillus, Gardnerella and Atopobium, which accounted for 78%, 14% and 2%, respectively, of sequences from all vaginal bacterial genera. At the species level, Lactobacillus iners, Gardnerella vaginalis and Atopobium vaginae accounted for 72%, 10% and 6%, respectively, of the bacterial cells present. Analyses using the V3 region generally indicated the highest bacterial diversity followed by the V6-V7 and V4 regions, while the V9 region gave the lowest bacterial resolution. NGS based on the 16S rRNA gene can give comprehensive estimates of the diversity of vaginal bacterial communities. Selection of sequences from appropriate hypervariable regions is necessary to provide reliable information on bacterial community diversity.
Topics: Bacteria; DNA, Bacterial; Female; Genetic Variation; High-Throughput Nucleotide Sequencing; Humans; Microbiota; RNA, Ribosomal, 16S; Vagina
PubMed: 33221964
DOI: 10.1007/s00203-020-02114-4 -
Microorganisms Nov 2020The physiological microbiota of the vagina is responsible for providing a protective barrier, but Some factors can disturb the balance in its composition. At that time,...
The physiological microbiota of the vagina is responsible for providing a protective barrier, but Some factors can disturb the balance in its composition. At that time, the amounts of the genus decrease, which may lead to the development of infection and severe complications during pregnancy. The aim of the study was the analysis of the bacterial composition of the vagina in 32 Caucasian women at each trimester of pregnancy using the next-generation sequencing method and primers targeting V3-V4 regions. In the studied group, the dominant species were and . Statistically significant differences in the quantitative composition between trimesters were observed in relation to , , spp. Out of the 32 patients, 20 demonstrated fluctuations within the genus and 9 of them, at different stages of pregnancy, exhibited the presence of potentially pathogenic microbiota, among others: spp., , and The composition of the vaginal microbiota during pregnancy was subject to partial changes over trimesters. Although in one-third of the studied patients, both the qualitative and quantitative composition of microbiota was relatively constant, in the remaining patients, physiological and potentially pathogenic fluctuations were distinguished.
PubMed: 33217908
DOI: 10.3390/microorganisms8111813 -
Frontiers in Public Health 2020Abnormal vaginal microbiota (AVM), including bacterial vaginosis (BV), is caused by a microbiota imbalance. Nugent scoring is the gold standard for the laboratory...
Abnormal vaginal microbiota (AVM), including bacterial vaginosis (BV), is caused by a microbiota imbalance. Nugent scoring is the gold standard for the laboratory diagnosis of BV; however, it is somewhat subjective to interpret, and challenging to distinguish bacteria. Hence, there is a need for improved technologies for the accurate diagnosis of AVM. To this end, next-generation sequencing (NGS) technology has been shown to yield comprehensive information on the pathophysiology of AVM. Hence, to evaluate the relationship between microbiota composition and the pathophysiology of AVM and its clinical significance, we characterized vaginal swab samples from 212 pregnant Korean women using both Nugent scoring and NGS analysis. Of these, the Nugent scoring identified 175 subjects (82.5%; 175/212) with normal flora (NF), 20 (9.4%; 20/212) with intermediate flora (IF), and 17 (8.0%; 17/212) with BV. NGS analysis followed by the characterization of vaginal microbiota composition, as represented by alpha and beta diversity, revealed the relative abundance of specific bacterial taxa at the genus and species level. Moreover, we identified all five predominant community state types (CSTs) along with three smaller CSTs. Analysis of the vaginal microbiota revealed the dominance of one or two spp. in the NF group. Meanwhile, the IF and BV groups were dominated by the genera , and . These two groups also showed higher alpha diversity than the NF group ( < 0.05). Principal coordinate analysis (PCoA) indicated that the NF group was significantly different from the AVM groups ( < 0.05), whereas no significant difference was observed between IF and BV groups ( = 0.25). Lastly, to investigate the characteristics of vaginal microbiota based on taxonomic composition, the IF and BV groups (AVM groups) were reclassified using the unweighted pair group method with arithmetic mean (UPGMA) clustering. Consequently, they were reclassified into BV1 -dominated), BV2-1 (-dominated), BV2-2 ( s1 or s2 and -dominated), and BV3 [mixed population of , and other bacteria ( < 0.05)]. Collectively, these findings could serve to advance the current understanding regarding AVM pathophysiology.
Topics: Actinobacteria; Female; Humans; Lactobacillus; Microbiota; Pregnancy; Republic of Korea
PubMed: 33194936
DOI: 10.3389/fpubh.2020.507024 -
Scientific Reports Oct 2020Vaginal infection is a gynecological problem in women of reproductive age with multiple health outcomes. The most common forms of infection include bacterial vaginosis...
Vaginal infection is a gynecological problem in women of reproductive age with multiple health outcomes. The most common forms of infection include bacterial vaginosis (BV), vulvovaginal candidiasis (VC), and aerobic vaginitis (AV). Our main goals were to evaluate different types of vaginal infections in Ecuadorian women in a large urban area (Quito) and to characterize the vaginal microbiota colonization by opportunistic species. We collected vaginal swabs and epidemiological surveys from 414 women from June 2016 to July of 2017. We analyzed vaginal samples for the presence of any vaginal infection. The microbiological examination was done through Gram-stain, wet mount smears, and polymerase chain reaction (PCR) assays using primers for target genes, such as 16S rRNA (Atopobium vaginae, Mobiluncus mulieris, and Gardnerella species), ddl (Enterococcus faecalis), adk (Escherichia coli) and KER1 (Candida albicans) genes. Most women showed a healthy vaginal microbiota (66.7%). Nearly one-tenth (10.4%) of the participants had intermediate microbiota, and the remaining women (22.9%) had a single vaginal infection (BV, AV, or VC) or coinfections. From the 95 participants that had an infection, AV was the main diagnosed vaginal infection (51.6%), followed by BV (24.2%) and finally VC (7.4%). The remaining women (16.8%) showed coinfections, being BV and AV the most common coinfection. Using univariable logistic regression analyses we found an increased odds of healthy microbiota in women with a sexual partner (P = 0.02, OR = 1.64). Also, women in a free union relationship (P = 0.000, OR = 16.65) had an increased odds of having coinfections. On the other hand, the use of birth control (condom OR = 0.388 or other contraceptive method OR = 0.363) was associated with significantly lower odds of intermediate microbiota (P ≤ 0.05). We found no statistically significant differences between women with infection and a particular group age. Using multivariate logistic regression analyses we initially found an increased odds of having BV in women with M. mulieris (P = 0.020, OR = 4.98) and Gardnerella species (P = 0.010, OR = 4.16). Women with E. coli showed an increased odds of having AV (P = 0.009, OR = 2.81). The presence of C. albicans in women showed an increased odds of having VC (P = 0.007, OR = 17.94). Finally, women with M. mulieris showed a reverse odds of having healthy microbiota (P = 0.008, OR = 0.06). We found no statistically significant differences between women with symptomatic and asymptomatic infections or the presence of Enterococcus faecalis. We found using logistic regression analyses that M. mulieris was the most prevalent opportunistic pathogen among women with vaginal infection. Further studies should evaluate the possibility to use M. mulieris as a potential key predictor for vaginal infections.
Topics: Adult; Age Factors; Candidiasis, Vulvovaginal; Ecuador; Female; Humans; Microbiota; Middle Aged; Prevalence; Risk Factors; Vagina; Vaginal Diseases; Vaginosis, Bacterial; Young Adult
PubMed: 33110095
DOI: 10.1038/s41598-020-74655-z -
American Journal of Obstetrics and... Mar 2021Bacterial vaginosis, pelvic inflammatory disease, and endometritis are infections of the genital tract that can lead to many adverse health outcomes, including... (Review)
Review
Bacterial vaginosis, pelvic inflammatory disease, and endometritis are infections of the genital tract that can lead to many adverse health outcomes, including infertility. Bacterial vaginosis is characterized by a lower prevalence of lactobacilli and a higher prevalence of anaerobic bacteria, including Gardnerella vaginalis, Megasphaera spp., and Atopobium vaginae. Endometritis and pelvic inflammatory disease are caused by the ascension of pathogenic bacteria to the uterus, although the mechanisms by which they do so are unclear. Bacterial vaginosis, chronic endometritis, and pelvic inflammatory disease have been linked to infertility in retrospective and prospective trials. Similarly, the causes of bacterial vaginosis and endometritis-related infertility are likely multifactorial and stem from inflammation, immune targeting of sperm antigens, the presence of bacterial toxins, and increased risk of sexually transmitted infections. Diagnosis and treatment of bacterial vaginosis, chronic endometritis, and pelvic inflammatory disease before attempting conception may be important components of preconceptional care for symptomatic women to improve outcomes of natural and assisted reproduction.
Topics: Endometritis; Female; Humans; Infertility, Female; Pelvic Inflammatory Disease; Vaginosis, Bacterial
PubMed: 33091407
DOI: 10.1016/j.ajog.2020.10.019 -
BMJ Open Oct 2020Recent studies in in vitro fertilisation (IVF) patients have associated abnormal vaginal microbiota (AVM) with poor clinical pregnancy rates of 6%-9% per embryo...
Effect of clindamycin and a live biotherapeutic on the reproductive outcomes of IVF patients with abnormal vaginal microbiota: protocol for a double-blind, placebo-controlled multicentre trial.
INTRODUCTION
Recent studies in in vitro fertilisation (IVF) patients have associated abnormal vaginal microbiota (AVM) with poor clinical pregnancy rates of 6%-9% per embryo transfer. The biological plausibility for this finding is hypothesised to be ascending infection to the endometrium which in turn hampers embryo implantation. New molecular based diagnosis may offer advantages compared to microscopical diagnosis of AVM which has huge inter-study variability ranging from 4 to 38%; however, the important question is whether screening and treatment of AVM would improve reproductive outcomes in IVF patients. Herein, we describe a protocol for an ongoing double-blind, placebo-controlled multicentre trial of IVF patients diagnosed with AVM and randomised in three parallel groups 1:1:1.
METHODS AND ANALYSIS
This is a drug intervention study where IVF patients will be screened for AVM, using a qPCR assay targeting and . If positive, patients will be randomised to one of the three study arms. The first arm consists of clindamycin 300 mg ×2 daily for 7 days followed by vaginal CTV-05 until clinical pregnancy scan week 7-9. The second arm consists of clindamycin and placebo CTV-05, whereas patients in the third arm will be treated with placebo/placebo. We used a superiority design to estimate that active treatment in both arms will increase the primary outcome, clinical pregnancy rate per embryo transfer, from 20% to 40%. A potential difference between the two active arms was considered exploratory. With a power of 80% and an alpha at 5%, the sample size is estimated to be 333 patients randomised. A pre-planned interim analysis is scheduled at 167 patients randomised.
ETHICS AND DISSEMINATION
All patients have to give informed consent. Dissemination of results is ensured in clinical trial agreements whether they be positive or not. Ethics committee, Central Denmark Region approved this protocol.
TRIAL REGISTRATION NUMBER
ICH-GCP monitored trial, EudraCT 2016-002385-31; Pre-results.
Topics: Actinobacteria; Clindamycin; Female; Fertilization in Vitro; Humans; Microbiota; Multicenter Studies as Topic; Pregnancy; Randomized Controlled Trials as Topic
PubMed: 33051228
DOI: 10.1136/bmjopen-2019-035866 -
Viruses Oct 2020Bacterial vaginosis (BV) is characterized by a reduction in spp. abundance and increased abundance of facultative anaerobes, such as spp. BV aetiology is not fully...
Bacterial vaginosis (BV) is characterized by a reduction in spp. abundance and increased abundance of facultative anaerobes, such as spp. BV aetiology is not fully understood; however, bacteriophages could play a pivotal role in the perturbation of the vaginal bacterial community. We investigated the vaginal viral community, including bacteriophages and the association to the bacterial community and BV-status. Vaginal samples from 48 patients undergoing IVF treatment for non-female factor infertility were subjected to metagenomic sequencing of purified virus-like particles. The vaginal viral community was characterized and correlated with the BV-status by Nugent score, bacterial community, structure, and the presence of key vaginal bacterial species. The majority of identified vaginal viruses belonged to the class of double-stranded DNA bacteriophages, with eukaryotic viruses constituting 4% of the total reads. Clear links between the viral community composition and BV ( = 0.006, = 0.26) as well as the presence of ( = 0.001, = 0.43), , spp., and were found ( < 0.002, > 0.15). The eukaryotic viral community also correlated with BV-status ( = 0.018, = 0.20). In conclusion, the vaginal virome was clearly linked with bacterial community structure and BV-status.
Topics: Adult; Bacteria; Bacteriophages; Cross-Sectional Studies; Dysbiosis; Female; Humans; Lactobacillus; Vagina; Vaginosis, Bacterial; Virome; Young Adult
PubMed: 33050261
DOI: 10.3390/v12101143