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      The association between ethnicity and vaginal microbiota composition in Amsterdam, the Netherlands

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          Abstract

          Objective

          To evaluate whether ethnicity is independently associated with vaginal microbiota (VMB) composition in women living in Amsterdam, the Netherlands, as has been shown for American women.

          Methods

          Women (18–34 years, non-pregnant, N = 610) representing the six largest ethnic groups (Dutch, African Surinamese, South-Asian Surinamese, Turkish, Moroccan, and Ghanaian) were sampled from the population-based HELIUS study. Sampling was performed irrespective of health status or healthcare seeking behavior. DNA was extracted from self-sampled vaginal swabs and sequenced by Illumina MiSeq (16S rRNA gene V3-V4 region).

          Results

          The overall prevalence of VMBs not dominated by lactobacilli was 38.5%: 32.2% had a VMB resembling bacterial vaginosis and another 6.2% had a VMB dominated by Bifidobacteriaceae (not including Gardnerella vaginalis), Corynebacterium, or pathobionts (streptococci, staphylococci, Proteus or Enterobacteriaceae). The most prevalent VMB in ethnically Dutch women was a Lactobacillus crispatus-dominated VMB, in African Surinamese and Ghanaian women a polybacterial G. vaginalis-containing VMB, and in the other ethnic groups a L. iners-dominated VMB. After adjustment for sociodemographic, behavioral and clinical factors, African Surinamese ethnicity (adjusted odds ratio (aOR) 5.1, 95% confidence interval (CI) 2.1–12.0) and Ghanaian ethnicity (aOR 4.8, 95% CI 1.8–12.6) were associated with having a polybacterial G. vaginalis-containing VMB, and African Surinamese ethnicity with a L. iners-dominated VMB (aOR 2.8, 95% CI 1.2–6.2). Shorter steady relationship duration, inconsistent condom use with casual partners, and not using hormonal contraception were also associated with having a polybacterial G. vaginalis-containing VMB, but human papillomavirus infection was not. Other sexually transmitted infections were uncommon.

          Conclusions

          The overall prevalence of having a VMB not dominated by lactobacilli in this population-based cohort of women aged 18–34 years in Amsterdam was high (38.5%), and women of sub-Saharan African descent were significantly more likely to have a polybacterial G. vaginalis-containing VMB than Dutch women independent of modifiable behaviors.

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          Most cited references25

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          Temporal dynamics of the human vaginal microbiota.

          Elucidating the factors that impinge on the stability of bacterial communities in the vagina may help in predicting the risk of diseases that affect women's health. Here, we describe the temporal dynamics of the composition of vaginal bacterial communities in 32 reproductive-age women over a 16-week period. The analysis revealed the dynamics of five major classes of bacterial communities and showed that some communities change markedly over short time periods, whereas others are relatively stable. Modeling community stability using new quantitative measures indicates that deviation from stability correlates with time in the menstrual cycle, bacterial community composition, and sexual activity. The women studied are healthy; thus, it appears that neither variation in community composition per se nor higher levels of observed diversity (co-dominance) are necessarily indicative of dysbiosis.
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            Bacterial Communities in Women with Bacterial Vaginosis: High Resolution Phylogenetic Analyses Reveal Relationships of Microbiota to Clinical Criteria

            Background Bacterial vaginosis (BV) is a common condition that is associated with numerous adverse health outcomes and is characterized by poorly understood changes in the vaginal microbiota. We sought to describe the composition and diversity of the vaginal bacterial biota in women with BV using deep sequencing of the 16S rRNA gene coupled with species-level taxonomic identification. We investigated the associations between the presence of individual bacterial species and clinical diagnostic characteristics of BV. Methodology/Principal Findings Broad-range 16S rRNA gene PCR and pyrosequencing were performed on vaginal swabs from 220 women with and without BV. BV was assessed by Amsel’s clinical criteria and confirmed by Gram stain. Taxonomic classification was performed using phylogenetic placement tools that assigned 99% of query sequence reads to the species level. Women with BV had heterogeneous vaginal bacterial communities that were usually not dominated by a single taxon. In the absence of BV, vaginal bacterial communities were dominated by either Lactobacillus crispatus or Lactobacillus iners. Leptotrichia amnionii and Eggerthella sp. were the only two BV-associated bacteria (BVABs) significantly associated with each of the four Amsel’s criteria. Co-occurrence analysis revealed the presence of several sub-groups of BVABs suggesting metabolic co-dependencies. Greater abundance of several BVABs was observed in Black women without BV. Conclusions/Significance The human vaginal bacterial biota is heterogeneous and marked by greater species richness and diversity in women with BV; no species is universally present. Different bacterial species have different associations with the four clinical criteria, which may account for discrepancies often observed between Amsel and Nugent (Gram stain) diagnostic criteria. Several BVABs exhibited race-dependent prevalence when analyzed in separate groups by BV status which may contribute to increased incidence of BV in Black women. Tools developed in this project can be used to study microbial ecology in diverse settings at high resolution.
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              Differences in vaginal microbiome in African American women versus women of European ancestry.

              Women of European ancestry are more likely to harbour a Lactobacillus-dominated microbiome, whereas African American women are more likely to exhibit a diverse microbial profile. African American women are also twice as likely to be diagnosed with bacterial vaginosis and are twice as likely to experience preterm birth. The objective of this study was to further characterize and contrast the vaginal microbial profiles in African American versus European ancestry women. Through the Vaginal Human Microbiome Project at Virginia Commonwealth University, 16S rRNA gene sequence analysis was used to compare the microbiomes of vaginal samples from 1268 African American women and 416 women of European ancestry. The results confirmed significant differences in the vaginal microbiomes of the two groups and identified several taxa relevant to these differences. Major community types were dominated by Gardnerella vaginalis and the uncultivated bacterial vaginosis-associated bacterium-1 (BVAB1) that were common among African Americans. Moreover, the prevalence of multiple bacterial taxa that are associated with microbial invasion of the amniotic cavity and preterm birth, including Mycoplasma, Gardnerella, Prevotella and Sneathia, differed between the two ethnic groups. We investigated the contributions of intrinsic and extrinsic factors, including pregnancy, body mass index, diet, smoking and alcohol use, number of sexual partners, and household income, to vaginal community composition. Ethnicity, pregnancy and alcohol use correlated significantly with the relative abundance of bacterial vaginosis-associated species. Trends between microbial profiles and smoking and number of sexual partners were observed; however, these associations were not statistically significant. These results support and extend previous findings that there are significant differences in the vaginal microbiome related to ethnicity and demonstrate that these differences are pronounced even in healthy women.
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                Author and article information

                Contributors
                Role: Data curationRole: Formal analysisRole: InvestigationRole: Project administrationRole: VisualizationRole: Writing – original draftRole: Writing – review & editing
                Role: InvestigationRole: Writing – review & editing
                Role: InvestigationRole: ResourcesRole: SupervisionRole: Writing – review & editing
                Role: Data curationRole: Project administrationRole: Writing – review & editing
                Role: ConceptualizationRole: Writing – review & editing
                Role: InvestigationRole: ResourcesRole: SupervisionRole: Writing – review & editing
                Role: Data curationRole: Project administrationRole: ResourcesRole: Writing – review & editing
                Role: Funding acquisitionRole: ResourcesRole: Writing – review & editing
                Role: ConceptualizationRole: SupervisionRole: Writing – review & editing
                Role: ConceptualizationRole: SupervisionRole: Writing – review & editing
                Role: ConceptualizationRole: Formal analysisRole: Funding acquisitionRole: MethodologyRole: SupervisionRole: Writing – original draftRole: Writing – review & editing
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                11 July 2017
                2017
                : 12
                : 7
                : e0181135
                Affiliations
                [1 ] Amsterdam Institute for Global Health and Development, Amsterdam, The Netherlands
                [2 ] Center for Infection and Immunity Amsterdam, Academic Medical Center, Amsterdam, The Netherlands
                [3 ] Public Health Laboratory, Public Health Service of Amsterdam (GGD), Amsterdam, The Netherlands
                [4 ] Department of Medical Microbiology and Infection Control, VU University Medical Center, Amsterdam, The Netherlands
                [5 ] Department of Infectious Diseases, Public Health Service of Amsterdam (GGD), Amsterdam, the Netherlands
                [6 ] Department of Dermatology, Academic Medical Center, Amsterdam, The Netherlands
                [7 ] Department of Public Health, Academic Medical Center, Amsterdam, The Netherlands
                [8 ] Department of Clinical Epidemiology, Biostatistics and Bioinformatics, Academic Medical Center, Amsterdam, The Netherlands
                [9 ] Department of Internal Medicine, Division of Infectious Diseases, Academic Medical Center, Amsterdam, The Netherlands
                [10 ] Department of Clinical Infection, Microbiology and Immunology, Institute of Infection and Global Health, University of Liverpool, Liverpool, United Kingdom
                Fred Hutchinson Cancer Research Center, UNITED STATES
                Author notes

                Competing Interests: M. F. Schim van der Loeff received HPV research funding from Sanofi Pasteur MSD; he is a co-investigator in a Merck-funded investigator-initiated study on Gardasil; he is an investigator on a Sanofi Pasteur MSD sponsored HPV vaccine trial; he served on a HPV vaccine advisory board of GSK; he received in-kind contribution for another HPV study from Stichting Pathologie Onderzoek en Ontwikkeling (SPOO); and his institution receives HPV research funding from Janssen Infectious Diseases and Vaccines. This does not alter our adherence to PLOS ONE policies on sharing data and materials. The other authors report no competing interests.

                Author information
                http://orcid.org/0000-0003-2728-4560
                Article
                PONE-D-17-08648
                10.1371/journal.pone.0181135
                5507447
                28700747
                8b625b78-032e-4591-86c9-54d3ed6ed5c2
                © 2017 Borgdorff et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 4 March 2017
                : 27 June 2017
                Page count
                Figures: 2, Tables: 3, Pages: 17
                Funding
                Funded by: funder-id http://dx.doi.org/10.13039/100007553, Aids Fonds;
                Award ID: 2011027
                Award Recipient :
                This work was supported by the Aids Fonds Netherlands [grant number 201102; https://aidsfonds.nl/], the Academic Medical Center Amsterdam [institutional funding; https://www.amc.nl/web/Zorg.htm], the Public Health Service of Amsterdam [institutional funding; http://www.ggd.amsterdam.nl/english/], the Dutch Heart Foundation [grant number 2010T084; https://www.hartstichting.nl/], the Netherlands Organization for Health Research and Development (ZonMw) [grant numbers 200500003, 7115 0001, and 204005002; https://www.zonmw.nl/en/], and the European Union (FP-7) [grant number 278901; https://ec.europa.eu/research/fp7/index_en.cfm]. Some analyses were carried out on the Dutch national e-infrastructure with the support of SURF Foundation [ https://www.surf.nl/en]. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                People and Places
                Population Groupings
                Ethnicities
                Africans
                People and Places
                Population Groupings
                Ethnicities
                Dutch People
                Biology and Life Sciences
                Organisms
                Bacteria
                Gut Bacteria
                Bifidobacterium
                Medicine and Health Sciences
                Epidemiology
                Ethnic Epidemiology
                Biology and Life Sciences
                Organisms
                Bacteria
                Corynebacteria
                People and Places
                Population Groupings
                Ethnicities
                Biology and Life Sciences
                Microbiology
                Medical Microbiology
                Microbiome
                Biology and Life Sciences
                Genetics
                Genomics
                Microbial Genomics
                Microbiome
                Biology and Life Sciences
                Microbiology
                Microbial Genomics
                Microbiome
                Biology and Life Sciences
                Behavior
                Custom metadata
                The HELIUS data are owned by the Academic Medical Center (AMC) in Amsterdam, The Netherlands. Any researcher can request the data by submitting a proposal to the HELIUS Executive Board as outlined at http://www.heliusstudy.nl/en/researchers/collaboration. Requests for further information and proposals can be submitted to Dr. Marieke Snijder, Scientific Coordinator and Data Manager of HELIUS, at m.b.snijder@ 123456amc.uva.nl , or to info@ 123456heliusstudie.nl . The “heliusstudy” website and generic email address will continue to be actively managed in the event Dr. Snijder should leave her post. The HELIUS Executive Board will check proposals for compatibility with the general objectives, ethical approvals and informed consent forms of the HELIUS study. There are no other restrictions to obtaining the data and all data requests will be processed in the same manner. Of the authors of this vaginal microbiota paper, Dr. Snijder (see above) and Professor Prins (Theme Leader Infectious Diseases) are affiliated with the HELIUS cohort. They were actively involved in this subproject of the HELIUS study as outlined in the authors' contributions section. HELIUS endorses the international ICMJE guidelines for co-authorship.

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