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Journal of Clinical Microbiology Jun 1988New and selective Rlk and SA media, combined with cold enrichment at 4 to 5 degrees C, allowed isolation of Mobiluncus species from patients with bacterial vaginosis at...
New and selective Rlk and SA media, combined with cold enrichment at 4 to 5 degrees C, allowed isolation of Mobiluncus species from patients with bacterial vaginosis at higher rates than with conventional cultivation methods. Rlk medium consists of Columbia CNA agar supplemented with peptone, yeast extract, 5% laked rabbit or sheep blood, nalidixic acid, and tinidazole. SA medium consists of Columbia CNA agar supplemented with 2% rabbit serum, 1.6% laked rabbit or sheep blood, nalidixic acid, and tinidazole. Use of these selective media plus the cold enrichment technique permitted Mobiluncus species to propagate at rates similar to those of other anaerobic members of the vaginal flora.
Topics: Bacteria, Anaerobic; Cold Temperature; Culture Media; Female; Humans
PubMed: 3384924
DOI: 10.1128/jcm.26.6.1134-1137.1988 -
PeerJ 2016Background. It is widely assumed that the uterine cavity in non-pregnant women is physiologically sterile, also as a premise to the long-held view that human infants...
Background. It is widely assumed that the uterine cavity in non-pregnant women is physiologically sterile, also as a premise to the long-held view that human infants develop in a sterile uterine environment, though likely reflecting under-appraisal of the extent of the human bacterial metacommunity. In an exploratory study, we aimed to investigate the putative presence of a uterine microbiome in a selected series of non-pregnant women through deep sequencing of the V1-2 hypervariable region of the 16S ribosomal RNA (rRNA) gene. Methods. Nineteen women with various reproductive conditions, including subfertility, scheduled for hysteroscopy and not showing uterine anomalies were recruited. Subjects were highly diverse with regard to demographic and medical history and included nulliparous and parous women. Endometrial tissue and mucus harvesting was performed by use of a transcervical device designed to obtain endometrial biopsy, while avoiding cervicovaginal contamination. Bacteria were targeted by use of a barcoded Illumina MiSeq paired-end sequencing method targeting the 16S rRNA gene V1-2 region, yielding an average of 41,194 reads per sample after quality filtering. Taxonomic annotation was pursued by comparison with sequences available through the Ribosomal Database Project and the NCBI database. Results. Out of 183 unique 16S rRNA gene amplicon sequences, 15 phylotypes were present in all samples. In some 90% of the women included, community architecture was fairly similar inasmuch B. xylanisolvens, B. thetaiotaomicron, B. fragilis and an undetermined Pelomonas taxon constituted over one third of the endometrial bacterial community. On the singular phylotype level, six women showed predominance of L. crispatus or L. iners in the presence of the Bacteroides core. Two endometrial communities were highly dissimilar, largely lacking the Bacteroides core, one dominated by L. crispatus and another consisting of a highly diverse community, including Prevotella spp., Atopobium vaginae, and Mobiluncus curtisii. Discussion. Our findings are, albeit not necessarily generalizable, consistent with the presence of a unique microbiota dominated by Bacteroides residing on the endometrium of the human non-pregnant uterus. The transcervical sampling approach may be influenced to an unknown extent by endocervical microbiota, which remain uncharacterised, and therefore warrants further validation. Nonetheless, consistent with our understanding of the human microbiome, the uterine microbiota are likely to have a previously unrecognized role in uterine physiology and human reproduction. Further study is therefore warranted to document community ecology and dynamics of the uterine microbiota, as well as the role of the uterine microbiome in health and disease.
PubMed: 26823997
DOI: 10.7717/peerj.1602 -
Microbiology Insights 2023The eradication of neonatal Group B (GBS) infections, considered as a major public health priority, necessarily requires a mastery of the data on vaginal carriage in...
The eradication of neonatal Group B (GBS) infections, considered as a major public health priority, necessarily requires a mastery of the data on vaginal carriage in pregnant women. The aims of this study were to determine the prevalence of vaginal carriage of GBS in pregnant women, antibiotic susceptibility, and associated risk factors. This was a cross-sectional, descriptive study conducted over a period of 9 months (July 2020 to March 2021) in pregnant women between 34 and 38 weeks of gestation (WG) followed at the Nabil Choucair health center in Dakar. Identification and antibiotic susceptibility of GBS isolates were performed on the Vitek 2 from vaginal swabs cultured on Granada medium. Demographic and obstetric interview data were collected and analyzed on SPSS (version 25). The level of significance for all statistical tests was set at < .05. The search of GBS vaginal carriage had involved 279 women aged 16 to 46 years, with a median pregnancy age of 34 (34-37) weeks' gestation. GBS was found in 43 women, for a vaginal carriage rate of 15.4%. In 27.9% (12/43) of volunteers screened, this carriage was monomicrobial, while in 72.1% (31/43) of women, GBS was associated with other pathogens such as spp. (60.5%), (2.3%), (34.9%) and/or spp. (11.6%). The level of resistance was 27.9% (12/43) for penicillin G, 53.5% (23/43) for erythromycin, 25.6% (11/43) for clindamycin and 100% for tetracycline. However, the strains had retained fully susceptible to vancomycin and teicoplanin. The main risk factor associated with maternal GBS carriage were ectocervical inflammation associated with contact bleeding (OR = 3.55; = .005). The high rate of maternal vaginal GBS carriage and the levels of resistance to the various antibiotics tested confirm the importance of continuous GBS surveillance in our resource-limited countries.
PubMed: 37275206
DOI: 10.1177/11786361231174419 -
American Journal of Perinatology Mar 2016This study aims to evaluate vaginal microbiota differences by bacterial vaginosis (BV), birth timing, and race, and to estimate parameters to power future vaginal...
OBJECTIVE
This study aims to evaluate vaginal microbiota differences by bacterial vaginosis (BV), birth timing, and race, and to estimate parameters to power future vaginal microbiome studies.
METHODS
Previously, vaginal swabs were collected at 21 to 25 weeks (stored at -80°C), and vaginal smears evaluated for BV (Nugent criteria). In a blinded fashion, 40 samples were selected, creating 8 equal-sized groups stratified by race (black/white), BV (present/absent), and birth timing (preterm/term). Samples were thawed, DNA extracted, and prepared. Polymerase chain reaction (PCR) with primers targeting the 16S rDNA V4 region was used to prepare an amplicon library. PCR products were sequenced and analyzed using quantitative insight into microbial ecology; taxonomy was assigned using ribosomal database program classifier (threshold 0.8) against the modified Greengenes database.
RESULTS
After quality control, 97,720 sequences (mean) per sample, single-end 250 base-reads, were analyzed. BV samples had greater microbiota diversity (p < 0.05)-with BVAB1, Prevotella, and unclassified genus, Bifidobacteriaceae family (all p < 0.001) more abundant; there was minimal content of Gardnerella or Mobiluncus. Microbiota did not differ by race or birth timing, but there was an association between certain microbial clusters and preterm birth (p = 0.07). To evaluate this difference, 159 patients per group are needed.
CONCLUSIONS
There are differences in the vaginal microbiota between patients with and without BV. Larger studies should assess the relationship between microbiota composition and preterm birth.
Topics: Adolescent; Adult; Alabama; DNA, Ribosomal; Female; Humans; Microbiota; Polymerase Chain Reaction; Pregnancy; Sequence Analysis, DNA; Vagina; Vaginosis, Bacterial; Young Adult
PubMed: 26479170
DOI: 10.1055/s-0035-1565919 -
PloS One 2011The spectrum of bacteria associated with bacterial vaginosis (BV) has recently expanded through taxonomic changes and the use of molecular methods. These methods have...
BACKGROUND
The spectrum of bacteria associated with bacterial vaginosis (BV) has recently expanded through taxonomic changes and the use of molecular methods. These methods have yet to be used in large-scale epidemiological studies in Africa where BV is highly prevalent.
METHODS
An analysis of samples obtained during a clinical trial of the management of vaginal discharge in four West African countries. Samples were available from 1555 participants; 843 (54%) had BV. Nucleic acids of 13 bacterial genera or species potentially associated with BV were detected through the polymerase chain reaction.
RESULTS
The associations between various components of the vaginal flora were complex. Excluding Lactobacillus, the other 12 micro-organisms were all associated with each other at the p≤0.001 level. The prevalence of various bacterial genera or species varied according to age, sexual activity and HIV status. In multivariate analysis, the presence of Gardnerella vaginalis, Bifidobacterium, Megasphaera elsdenii, Dialister, Mycoplasma hominis, Leptotrichia, and Prevotella were independently associated with BV as was the absence of Lactobacillus and Peptoniphilus. However, Mobiluncus, Atopobium vaginae, Anaerococcus, and Eggerthella were not independently associated with BV. Unexpectedly, after treatment with a regimen that included either metronidazole or tinidazole, the proportion of patients with a complete resolution of symptoms by day 14 increased with the number of bacterial genera or species present at enrolment.
CONCLUSIONS
Numerous bacterial genera or species were strongly associated with each other in a pattern that suggested a symbiotic relationship. BV cases with a simpler flora were less likely to respond to treatment. Overall, the vaginal flora of West African women with BV was reminiscent of that of their counterparts in industrialized countries.
Topics: Adult; Africa; Bacteria; DNA, Viral; Epidemiologic Studies; Female; Humans; Polymerase Chain Reaction; Randomized Controlled Trials as Topic; Vagina; Vaginosis, Bacterial; Young Adult
PubMed: 21949860
DOI: 10.1371/journal.pone.0025082 -
Diagnostic Microbiology and Infectious... Dec 2014As part of a larger study using 454 pyrosequencing to investigate the vaginal microbiota of women with bacterial vaginosis (BV), we found an association between a novel...
As part of a larger study using 454 pyrosequencing to investigate the vaginal microbiota of women with bacterial vaginosis (BV), we found an association between a novel BV-associated bacterium (BVAB1) and high Nugent scores and propose that BVAB1 is the curved Gram-negative rod traditionally identified as Mobiluncus spp. in vaginal Gram stains.
Topics: Adult; Colony Count, Microbial; Female; Gram-Negative Bacteria; Humans; Mobiluncus; RNA, Ribosomal, 16S; Vagina; Vaginosis, Bacterial
PubMed: 25262105
DOI: 10.1016/j.diagmicrobio.2014.09.008 -
Journal of Clinical Microbiology Feb 2003Fifteen strains of an anaerobic, catalase-negative, gram-positive diphtheroid-shaped bacterium recovered from human sources were characterized by phenotypic and...
Fifteen strains of an anaerobic, catalase-negative, gram-positive diphtheroid-shaped bacterium recovered from human sources were characterized by phenotypic and molecular chemical and molecular genetic methods. The unidentified bacterium showed some resemblance to Actinomyces species and related taxa, but biochemical testing, polyacrylamide gel electrophoresis analysis of whole-cell proteins, and amplified 16S ribosomal DNA restriction analysis indicated the strains were distinct from all currently named Actinomyces species and related taxa. Comparative 16S rRNA gene sequencing studies showed that the bacterium represents a hitherto-unknown phylogenetic line that is related to but distinct from Actinomyces, Actinobaculum, Arcanobacterium, and Mobiluncus: We propose, on the basis of phenotypic and phylogenetic evidence, that the unknown bacterium from human clinical specimens should be classified as a new genus and species, Varibaculum cambriensis gen. nov., sp. nov. The type strain of Varibaculum cambriensis sp. nov. is CCUG 44998(T) = CIP 107344(T).
Topics: Actinomycetaceae; Humans; Phylogeny; RNA, Ribosomal, 16S
PubMed: 12574260
DOI: 10.1128/JCM.41.2.640-644.2003 -
American Journal of Obstetrics and... Mar 2014Genital tract infection accounts for approximately 25-40% of all preterm births. We sought to assess the relationship between preterm birth and selected vaginal...
OBJECTIVE
Genital tract infection accounts for approximately 25-40% of all preterm births. We sought to assess the relationship between preterm birth and selected vaginal bacterial taxa associated with preterm birth either directly or through their association with bacterial vaginosis (BV).
STUDY DESIGN
Vaginal fluid for Gram stain was collected between 17 and 22 weeks' gestation as part of a randomized trial of ultrasound-indicated cerclage for preterm birth prevention in women at high risk for recurrent spontaneous preterm birth. Bacterial deoxyribonucleic acid was extracted from the Gram stain slides and analyzed using quantitative polymerase chain reaction.
RESULTS
Among the 499 participants, Mycoplasma was positively correlated with increased risk of preterm (risk ratio [RR], 1.83; 95% confidence interval [CI], 1.52-2.22) as was Mobiluncus (RR, 1.36; 95% CI, 1.07-1.73) and Atopobium (RR, 1.44; 95% CI, 1.1-1.87). However, there were strong interactions between the race/ethnic group and the presence of these and other individual taxa on risk of preterm birth. By contrast, bacterial vaginosis-associated bacteria (BVAB)-3 was consistently associated with a reduction in the risk of preterm birth for all racial/ethnic groups (0.55; 95% CI, 0.39-0.78).
CONCLUSION
BV is characterized by a reduction of Lactobacillus, and lactic acid-producing bacteria and the presence of Mobiluncus; we found these factors and the presence of Mycoplasma to be associated with an increased risk of preterm birth. By contrast, the presence of a recently identified organism sufficient to cause BV, BVAB3, decreased the risk of preterm birth. These findings give insight into why treating BV has mixed impact on risk of preterm birth.
Topics: Adult; Black or African American; DNA, Bacterial; Female; Hispanic or Latino; Humans; Infant, Newborn; Mobiluncus; Mycoplasma; Obstetric Labor, Premature; Pregnancy; Pregnancy Complications, Infectious; Premature Birth; Risk Factors; Vagina; Vaginosis, Bacterial; White People
PubMed: 24096128
DOI: 10.1016/j.ajog.2013.10.003 -
Clinical and Diagnostic Laboratory... Sep 1996While bacterial vaginosis (BV) is a polymicrobial syndrome, Mobiluncus spp. are the organisms most highly associated with this condition. It is possible that serum...
While bacterial vaginosis (BV) is a polymicrobial syndrome, Mobiluncus spp. are the organisms most highly associated with this condition. It is possible that serum antibody to Mobiluncus spp. could be used as a serological marker for BV. Using immunofluorescence techniques, we studied the prevalence of antibody to M. curtisii among three cohorts-pregnant women, pediatric patients, and sexually inexperienced women. The prevalence of antibody in each of these three groups was 75, 6, and 0%, respectively. Of the three pediatric patients with antibody to Mobiluncus curtisii, two were neonates, and the only class of antibody detected was immunoglobulin G. Among the cohort of pregnant women, the presence of antibody could not be correlated with a clinical history of BV. Serum antibody to M. curtisii could be a useful serological marker for BV. The lack of correlation of antibody positivity to historical information regarding BV suggests that unrecognized or undiagnosed episodes of BV may be common.
Topics: Adolescent; Adult; Antibodies, Bacterial; Biomarkers; Child; Child, Preschool; Female; Humans; Immunoglobulin G; Infant, Newborn; Mobiluncus; Pregnancy; Prevalence; Sexual Behavior; Vaginosis, Bacterial
PubMed: 8877136
DOI: 10.1128/cdli.3.5.567-569.1996 -
PloS One 2013Bacterial vaginosis (BV) is the most common vaginal disorder of reproductive-age women. Yet the cause of BV has not been established. To uncover key determinants of BV,...
BACKGROUND
Bacterial vaginosis (BV) is the most common vaginal disorder of reproductive-age women. Yet the cause of BV has not been established. To uncover key determinants of BV, we employed a multi-omic, systems-biology approach, including both deep 16S rRNA gene-based sequencing and metabolomics of lavage samples from 36 women. These women varied demographically, behaviorally, and in terms of health status and symptoms.
PRINCIPAL FINDINGS
16S rRNA gene-based community composition profiles reflected Nugent scores, but not Amsel criteria. In contrast, metabolomic profiles were markedly more concordant with Amsel criteria. Metabolomic profiles revealed two distinct symptomatic BV types (SBVI and SBVII) with similar characteristics that indicated disruption of epithelial integrity, but each type was correlated to the presence of different microbial taxa and metabolites, as well as to different host behaviors. The characteristic odor associated with BV was linked to increases in putrescine and cadaverine, which were both linked to Dialister spp. Additional correlations were seen with the presence of discharge, 2-methyl-2-hydroxybutanoic acid, and Mobiluncus spp., and with pain, diethylene glycol and Gardnerella spp.
CONCLUSIONS
The results not only provide useful diagnostic biomarkers, but also may ultimately provide much needed insight into the determinants of BV.
Topics: Actinomycetales Infections; Adult; DNA, Bacterial; Female; Gene Regulatory Networks; Gram-Positive Bacterial Infections; Humans; Hydroxybutyrates; Lactobacillus; Metabolomics; Middle Aged; Mobiluncus; Polymerase Chain Reaction; RNA, Ribosomal, 16S; Vagina; Vaginal Diseases; Vaginosis, Bacterial; Young Adult
PubMed: 23405259
DOI: 10.1371/journal.pone.0056111