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      The Effects of Circumcision on the Penis Microbiome

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          Abstract

          Background

          Circumcision is associated with significant reductions in HIV, HSV-2 and HPV infections among men and significant reductions in bacterial vaginosis among their female partners.

          Methodology/Principal Findings

          We assessed the penile (coronal sulci) microbiota in 12 HIV-negative Ugandan men before and after circumcision. Microbiota were characterized using sequence-tagged 16S rRNA gene pyrosequencing targeting the V3–V4 hypervariable regions. Taxonomic classification was performed using the RDP Naïve Bayesian Classifier. Among the 42 unique bacterial families identified, Pseudomonadaceae and Oxalobactericeae were the most abundant irrespective of circumcision status. Circumcision was associated with a significant change in the overall microbiota (PerMANOVA p = 0.007) and with a significant decrease in putative anaerobic bacterial families (Wilcoxon Signed-Rank test p = 0.014). Specifically, two families—Clostridiales Family XI ( p = 0.006) and Prevotellaceae ( p = 0.006)—were uniquely abundant before circumcision. Within these families we identified a number of anaerobic genera previously associated with bacterial vaginosis including: Anaerococcus spp., Finegoldia spp., Peptoniphilus spp., and Prevotella spp.

          Conclusions/Significance

          The anoxic microenvironment of the subpreputial space may support pro-inflammatory anaerobes that can activate Langerhans cells to present HIV to CD4 cells in draining lymph nodes. Thus, the reduction in putative anaerobic bacteria after circumcision may play a role in protection from HIV and other sexually transmitted diseases.

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          Most cited references37

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          Database resources of the National Center for Biotechnology Information

          In addition to maintaining the GenBank® nucleic acid sequence database, the National Center for Biotechnology Information (NCBI) provides analysis and retrieval resources for the data in GenBank and other biological data made available through the NCBI web site. NCBI resources include Entrez, the Entrez Programming Utilities, MyNCBI, PubMed, PubMed Central, Entrez Gene, the NCBI Taxonomy Browser, BLAST, BLAST Link (BLink), Electronic PCR, OrfFinder, Spidey, Splign, RefSeq, UniGene, HomoloGene, ProtEST, dbMHC, dbSNP, Cancer Chromosomes, Entrez Genomes and related tools, the Map Viewer, Model Maker, Evidence Viewer, Clusters of Orthologous Groups (COGs), Retroviral Genotyping Tools, HIV-1/Human Protein Interaction Database, Gene Expression Omnibus (GEO), Entrez Probe, GENSAT, Online Mendelian Inheritance in Man (OMIM), Online Mendelian Inheritance in Animals (OMIA), the Molecular Modeling Database (MMDB), the Conserved Domain Database (CDD), the Conserved Domain Architecture Retrieval Tool (CDART) and the PubChem suite of small molecule databases. Augmenting many of the web applications is custom implementation of the BLAST program optimized to search specialized data sets. All of the resources can be accessed through the NCBI home page at www.ncbi.nlm.nih.gov.
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            GenBank® is a comprehensive database that contains publicly available nucleotide sequences for more than 300 000 organisms named at the genus level or lower, obtained primarily through submissions from individual laboratories and batch submissions from large-scale sequencing projects. Most submissions are made using the web-based BankIt or standalone Sequin programs, and accession numbers are assigned by GenBank® staff upon receipt. Daily data exchange with the European Molecular Biology Laboratory Nucleotide Sequence Database in Europe and the DNA Data Bank of Japan ensures worldwide coverage. GenBank is accessible through the National Center for Biotechnology Information (NCBI) Entrez retrieval system, which integrates data from the major DNA and protein sequence databases along with taxonomy, genome, mapping, protein structure and domain information, and the biomedical journal literature via PubMed. BLAST provides sequence similarity searches of GenBank and other sequence databases. Complete bimonthly releases and daily updates of the GenBank database are available by FTP. To access GenBank and its related retrieval and analysis services, begin at the NCBI Homepage: www.ncbi.nlm.nih.gov.
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              Determination of bacterial load by real-time PCR using a broad-range (universal) probe and primers set.

              The design and evaluation of a set of universal primers and probe for the amplification of 16S rDNA from the Domain Bacteria to estimate total bacterial load by real-time PCR is reported. Broad specificity of the universal detection system was confirmed by testing DNA isolated from 34 bacterial species encompassing most of the groups of bacteria outlined in Bergey's Manual of Determinative Bacteriology. However, the nature of the chromosomal DNA used as a standard was critical. A DNA standard representing those bacteria most likely to predominate in a given habitat was important for a more accurate determination of total bacterial load due to variations in 16S rDNA copy number and the effect of generation time of the bacteria on this number, since rapid growth could result in multiple replication forks and hence, in effect, more than one copy of portions of the chromosome. The validity of applying these caveats to estimating bacterial load was confirmed by enumerating the number of bacteria in an artificial sample mixed in vitro and in clinical carious dentine samples. Taking these parameters into account, the number of anaerobic bacteria estimated by the universal probe and primers set in carious dentine was 40-fold greater than the total bacterial load detected by culture methods, demonstrating the utility of real-time PCR in the analysis of this environment.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, USA )
                1932-6203
                2010
                6 January 2010
                : 5
                : 1
                : e8422
                Affiliations
                [1 ]Translational Genomics Research Institute (TGen), Flagstaff, Arizona, United States of America
                [2 ]Northern Arizona University, Flagstaff, Arizona, United States of America
                [3 ]Johns Hopkins Medical Institutions, Baltimore, Maryland, United States of America
                [4 ]University of Maryland School of Medicine, Baltimore, Maryland, United States of America
                [5 ]School of Public Health, Makerere University, Kampala, Uganda
                [6 ]Bloomberg School of Public Health, Johns Hopkins University, Baltimore, Maryland, United States of America
                Charité-Universitätsmedizin Berlin, Germany
                Author notes

                Conceived and designed the experiments: LBP CML KEJ MA MKL JR PK DS MJW RHG. Performed the experiments: LBP CML MA MKL JRB. Analyzed the data: LBP CML KEJ MA MKL JRB JR PK MJW RHG. Contributed reagents/materials/analysis tools: LBP CML KEJ MA JR DS MJW RHG. Wrote the paper: LBP CML KEJ MA MKL JRB JR PK MJW RHG.

                Article
                09-PONE-RA-13954
                10.1371/journal.pone.0008422
                2798966
                20066050
                d175e92d-529b-4ef6-b169-32408cfd3df9
                Price 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
                : 24 October 2009
                : 9 November 2009
                Page count
                Pages: 12
                Categories
                Research Article
                Infectious Diseases
                Microbiology
                Public Health and Epidemiology
                Computational Biology/Metagenomics
                Ecology/Community Ecology and Biodiversity
                Microbiology/Medical Microbiology
                Dermatology/Skin Infections
                Infectious Diseases/Bacterial Infections
                Infectious Diseases/Gynecologic Infections
                Infectious Diseases/HIV Infection and AIDS
                Infectious Diseases/Sexually Transmitted Diseases
                Infectious Diseases/Urological Infections
                Public Health and Epidemiology/Infectious Diseases

                Uncategorized
                Uncategorized

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