Exploring the diversity of Gardnerella vaginalis in the genitourinary tract microbiota of monogamous couples through subtle nucleotide variation

© The Author(s), 2011. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in PLoS One 6 (2011): e26732, doi:10.1371/journal.pone.0026732.Bacterial vaginosis (BV) is an enigmatic disease of unknown origin that affects a large percentage of women. The vaginal microbiota of women with BV is associated with serious sequelae, including abnormal pregnancies. The etiology of BV is not fully understood, however, it has been suggested that it is transmissible, and that G. vaginalis may be an etiological agent. Studies using enzymatic assays to define G. vaginalis biotypes, as well as more recent genomic comparisons of G. vaginalis isolates from symptomatic and asymptomatic women, suggest that particular G. vaginalis strains may play a key role in the pathogenesis of BV. To explore G. vaginalis diversity, distribution and sexual transmission, we developed a Shannon entropy-based method to analyze low-level sequence variation in 65,710 G. vaginalis 16S rRNA gene segments that were PCR-amplified from vaginal samples of 53 monogamous women and from urethral and penile skin samples of their male partners. We observed a high degree of low-level diversity among G. vaginalis sequences with a total of 46 unique sequence variants (oligotypes), and also found strong correlations of these oligotypes between sexual partners. Even though Gram stain-defined normal and some Gram stain-defined intermediate oligotype profiles clustered together in UniFrac analysis, no single G. vaginalis oligotype was found to be specific to BV or normal vaginal samples. This study describes a novel method for investigating G. vaginalis diversity at a low level of taxonomic discrimination. The findings support cultivation-based studies that indicate sexual partners harbor the same strains of G. vaginalis. This study also highlights the fact that a few, reproducible nucleotide variations within the 16S rRNA gene can reveal clinical or epidemiological associations that would be missed by genus-level or species-level categorization of 16S rRNA data.This work is supported by funding from the Research Institute for Children in New Orleans and NIH grant 5RO1AI79071-2

Bacterial communities in penile skin, male urethra, and vaginas of heterosexual couples with and without bacterial vaginosis

© The Author(s), 2016. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Microbiome 4 (2016): 16, doi:10.1186/s40168-016-0161-6.The epidemiology of bacterial vaginosis (BV) suggests it is sexually transmissible, yet no transmissible agent has been identified. It is probable that BV-associated bacterial communities are transferred from male to female partners during intercourse; however, the microbiota of sexual partners has not been well-studied. Pyrosequencing analysis of PCR-amplified 16S rDNA was used to examine BV-associated bacteria in monogamous couples with and without BV using vaginal, male urethral, and penile skin specimens. The penile skin and urethral microbiota of male partners of women with BV was significantly more similar to the vaginal microbiota of their female partner compared to the vaginal microbiota of non-partner women with BV. This was not the case for male partners of women with normal vaginal microbiota. Specific BV-associated species were concordant in women with BV and their male partners. In monogamous heterosexual couples in which the woman has BV, the significantly higher similarity between the vaginal microbiota and the penile skin and urethral microbiota of the male partner, supports the hypothesis that sexual exchange of BV-associated bacterial taxa is common.This work was supported by National Institute of Health Grant R01 AI079071-01A1

Prevalence and Abundance of Uncultivated Megasphaera-Like Bacteria in the Human Vaginal Environment▿

Cultivation-independent analysis of 16S rRNA gene sequences in vaginal samples revealed two previously unrecognized, uncultivated Megasphaera-like phylotypes. Phylogenetic analysis and environmental distribution suggest that these Megasphaera types may be unique to the vaginal environment. Quantitative PCR suggests that both phylotypes are present in higher concentrations in women with bacterial vaginosis

Cultivation-Independent Analysis of Changes in Bacterial Vaginosis Flora Following Metronidazole Treatment

PCR was used to survey bacterial vaginosis flora before and after metronidazole treatment. The species composition for pretreatment patients was variable. Lactobacillus iners was prominent in all patients posttreatment. Atopobium vaginae concentrations were highest for patients who failed or responded incompletely to treatment and lowest for patients who were cured

Quantitative PCR Assessments of Bacterial Species in Women with and without Bacterial Vaginosis▿

Knowledge of the abundance of bacterial species in vaginal communities will help us to better understand their role in health and disease. However, progress in this field has been limited because quantifying bacteria in natural specimens is an arduous process. We developed quantitative real-time PCR (qPCR) assays to facilitate assessments of bacterial abundance in vaginal specimens and evaluated the utility of these assays by measuring species abundance in patients whose vaginal floras were clinically described as normal, intermediate, or bacterial vaginosis (BV) as defined by Nugent's criteria. The qPCR measurements showed that Lactobacillus species were predominant in normal vaginal specimens and that high Lactobacillus crispatus and Lactobacillus jensenii abundance was specific to normal specimens, while Lactobacillus iners abundance was high in all categories including BV. The abundances of all non-Lactobacillus species were higher in BV specimens than in normal specimens. Prevotella species were prevalent in all specimens and represented a high percentage of total species in BV specimens. qPCR assays can be a useful tool for describing the structure of vaginal communities and elucidating their role in health and disease

Psytrance as a Genre of Paralel Reality

S psytrancem jsem se poprvé náhodně setkala v pražském klubu Futurum v roce 2001. Do té doby jsem se pravidelně setkávala s různými žánry ETH (techno, house, drum'n'bas, jungle apod.), ale žánr psytrance pro mne byl v mnohém atypický a do té doby z běžných "náhodných zdrojů" nezaregistrován. Jeho hudební a vizuální stránka se od doposud poznané ETH znatelně odlišovala. Pestrost a netradiční výraznost zvuků vybízely k podstatně pozornějšímu poslechu a stimulovaly fantazii s doposud nepoznanou intenzitou. Všechny stěny klubu byly dekorovány fluorescenčními obrazy s fantaskními motivy, s kterými byli svým oděvem sladěni i někteří účastníci. Celý jinak poměrně fádní klub v tomto podání připomínal spíše jakousi futuristickou svatyni. Součástí celého prostoru byla také obdobným způsobem pojatá čajovna, což na ostatních akcích nebylo zvykem. Mnohem častěji podobné místo s relaxační hudbou však na jiných akcích úplně chybělo

Additional file 1: Table S1. of Bacterial communities in penile skin, male urethra, and vaginas of heterosexual couples with and without bacterial vaginosis

Metadata from female patients. (XLS 72 kb

Additional file 2: Table S2. of Bacterial communities in penile skin, male urethra, and vaginas of heterosexual couples with and without bacterial vaginosis

Metadata from male sexual partners. (XLS 60 kb

Bacterial communities in heterosexual couples

<p>Bacterial tag-encoded FLX amplicon pyrosequencing analysis of the V4 -V6 region of the 16S rRNA gene using primers 530F: GTGCCAGCMGCNGCGG and 1100R: GGGTTNCGNTCGTTG. Sequence files are in FASTA format.</p> <p>DNA was extracted from penile skin (PS), urethral (UT) and vaginal (VG) specimens from normal (N) women and women with bacterial vaginosis (BV) and their sexual partners.</p