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      Genetic Diversity of Giardia duodenalis: Multilocus Genotyping Reveals Zoonotic Potential between Clinical and Environmental Sources in a Metropolitan Region of Brazil

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          Abstract

          Background

          Giardia duodenalis is a flagellate protozoan that parasitizes humans and several other mammals. Protozoan contamination has been regularly documented at important environmental sites, although most of these studies were performed at the species level. There is a lack of studies that correlate environmental contamination and clinical infections in the same region. The aim of this study is to evaluate the genetic diversity of a set of clinical and environmental samples and to use the obtained data to characterize the genetic profile of the distribution of G. duodenalis and the potential for zoonotic transmission in a metropolitan region of Brazil.

          Methodology/Principal Findings

          The genetic assemblages and subtypes of G. duodenalis isolates obtained from hospitals, a veterinary clinic, a day-care center and important environmental sites were determined via multilocus sequence-based genotyping using three unlinked gene loci. Cysts of Giardia were detected at all of the environmental sites. Mixed assemblages were detected in 25% of the total samples, and an elevated number of haplotypes was identified. The main haplotypes were shared among the groups, and new subtypes were identified at all loci. Ten multilocus genotypes were identified: 7 for assemblage A and 3 for assemblage B.

          Conclusions/Significance

          There is persistent G. duodenalis contamination at important environmental sites in the city. The identified mixed assemblages likely represent mixed infections, suggesting high endemicity of Giardia in these hosts. Most Giardia isolates obtained in this study displayed zoonotic potential. The high degree of genetic diversity in the isolates obtained from both clinical and environmental samples suggests that multiple sources of infection are likely responsible for the detected contamination events. The finding that many multilocus genotypes (MLGs) and haplotypes are shared by different groups suggests that these sources of infection may be related and indicates that there is a notable risk of human infection caused by Giardia in this region.

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          Most cited references58

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              DnaSP, DNA polymorphism analyses by the coalescent and other methods.

              DnaSP is a software package for the analysis of DNA polymorphism data. Present version introduces several new modules and features which, among other options allow: (1) handling big data sets (approximately 5 Mb per sequence); (2) conducting a large number of coalescent-based tests by Monte Carlo computer simulations; (3) extensive analyses of the genetic differentiation and gene flow among populations; (4) analysing the evolutionary pattern of preferred and unpreferred codons; (5) generating graphical outputs for an easy visualization of results. The software package, including complete documentation and examples, is freely available to academic users from: http://www.ub.es/dnasp
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, USA )
                1932-6203
                2014
                23 December 2014
                : 9
                : 12
                : e115489
                Affiliations
                [1 ]Molecular Biology and Genetic Engineering Center (CBMEG), University of Campinas (UNICAMP), Campinas, SP, Brazil
                [2 ]APTA - São Paulo Agency for Agribusiness Technology, Piracicaba, SP, Brazil
                [3 ]Department of Animal Biology, Biology Institute, University of Campinas (UNICAMP), Campinas, SP, Brazil
                [4 ]Department of Plant Biology, Biology Institute, University of Campinas (UNICAMP), Campinas, SP, Brazil
                Food and Drug Administration, United States of America
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Conceived and designed the experiments: MD APS. Performed the experiments: MD. Analyzed the data: MD AGA MIZ RMBF. Contributed reagents/materials/analysis tools: MD APS. Wrote the paper: MD AGA MIZ RMBF APS.

                Article
                PONE-D-14-11526
                10.1371/journal.pone.0115489
                4275228
                25536055
                de9c5acc-f5ac-4929-9013-0d98bf0b18eb
                Copyright @ 2014

                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
                : 19 March 2014
                : 24 November 2014
                Page count
                Pages: 27
                Funding
                This work was supported by the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) through a research grant and a research fellowship awarded to APS (number 577032/2008-9 and 304914/2010-0), by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES - BioComputacional Program) and by the Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) through a graduate fellowship to MD (MS 2008/52197-4 and DD 2011/50413-4). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Biology and Life Sciences
                Evolutionary Biology
                Population Genetics
                Genetic Polymorphism
                Haplotypes
                Genetics
                Microbial Genetics
                Molecular Genetics
                Microbiology
                Protozoology
                Molecular Biology
                Parasitology
                Intestinal Parasites
                Veterinary Science
                Veterinary Diseases
                Zoonoses
                Medicine and Health Sciences
                Epidemiology
                Genetic Epidemiology
                Molecular Epidemiology
                Infectious Diseases
                Tropical Diseases
                Neglected Tropical Diseases
                Custom metadata
                The authors confirm that all data underlying the findings are fully available without restriction. All sequence files are available from the GenBank database (accession number(s) JN116442–JN116504, KM495697-KM495727 and KF922892-KF923021).

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                Uncategorized

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