Molecular Epidemiology of Blastocystis sp. in Various Animal Groups from Two French Zoos and Evaluation of Potential Zoonotic Risk

RESEARCH ARTICLE Molecular Epidemiology of Blastocystis sp. in Various Animal Groups from Two French Zoos and Evaluation of Potential Zoonotic Risk Amandine Cian1☯, Dima El Safadi1,2☯, Marwan Osman1,2☯, Romain Moriniere3, Nausicaa Gantois1, Sadia Benamrouz-Vanneste1,4, Pilar Delgado-Viscogliosi1, Karine Guyot1, Luen-Luen Li5, Sébastien Monchy5, Christophe Noël6, Philippe Poirier7, Céline Nourrisson7, Ivan Wawrzyniak7, Frédéric Delbac7, Stéphanie Bosc3, Magali Chabé1, Thierry Petit8, Gabriela Certad1,9, Eric Viscogliosi1* a1111111111 a1111111111 a1111111111 a1111111111 a1111111111 OPEN ACCESS Citation: Cian A, El Safadi D, Osman M, Moriniere R, Gantois N, Benamrouz-Vanneste S, et al. (2017) Molecular Epidemiology of Blastocystis sp. in Various Animal Groups from Two French Zoos and Evaluation of Potential Zoonotic Risk. PLoS ONE 12 (1): e0169659. doi:10.1371/journal.pone.0169659 Editor: Vyacheslav Yurchenko, University of Ostrava, CZECH REPUBLIC Received: October 17, 2016 Accepted: December 20, 2016 Published: January 6, 2017 Copyright: © 2017 Cian 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. Data Availability Statement: All relevant data are within the paper and its Supporting Information files. Funding: This work was supported by grants from the Programme Orientations Stratégiques from the University of Lille 2, the Fonds Hospitalier d’Aide à l’Emergence from the CHRU of Lille, the Centre National de la Recherche Scientifique and the Institut Pasteur of Lille. DES and MO were supported by PhD fellowships from the Conseil National de la Recherche Scientifique and the AZM 1 Université de Lille, CNRS, Inserm, CHU Lille, Institut Pasteur de Lille, U1019 –UMR 8204 –CIIL–Centre d’Infection et d’Immunité de Lille, Lille, France, 2 Laboratoire Microbiologie Santé et Environnement (LMSE), Ecole Doctorale des Sciences et de Technologie, Faculté de Santé Publique, Université Libanaise, Tripoli, Lebanon, 3 Parc Zoologique de Lille, Lille, France, 4 Laboratoire Ecologie et Biodiversité, Faculté Libre des Sciences et Technologies de Lille, Université Catholique de Lille, Lille, France, 5 Laboratoire d’Océanologie et de Géosciences, CNRS UMR 8187, Université du Littoral Côte d’Opale, Wimereux, France, 6 Geneius Laboratories Ltd., INEX Business Centre, Newcastle-upon-Tyne, United Kingdom, 7 Clermont Université, Université Blaise Pascal-Université d’Auvergne—CNRS UMR 6023 Laboratoire Microorganismes: Génome et Environnement, Clermont-Ferrand, France, 8 Parc Zoologique de La Palmyre, Les Mathes, France, 9 Département de la Recherche Médicale, Groupement des Hôpitaux de l’Institut Catholique de Lille, Faculté de Médecine et Maïeutique, Université Catholique de Lille, France ☯ These authors contributed equally to this work. * Abstract Blastocystis sp. is a common intestinal parasite infecting humans and a wide range of animals worldwide. It exhibits an extensive genetic diversity and 17 subtypes (STs) have thus far been identified in mammalian and avian hosts. Since several STs are common to humans and animals, it was proposed that a proportion of human infections may result from zoonotic transmission. However, the contribution of each animal source to human infection remains to be clarified. Therefore, the aim of this study was to expand our knowledge of the epidemiology and host specificity of this parasite by performing the largest epidemiological survey ever conducted in animal groups in terms of numbers of species screened. A total of 307 stool samples from 161 mammalian and non-mammalian species in two French zoos were screened by real-time PCR for the presence of Blastocystis sp. Overall, 32.2% of the animal samples and 37.9% of the species tested were shown to be infected with the parasite. A total of 111 animal Blastocystis sp. isolates were subtyped, and 11 of the 17 mammalian and avian STs as well as additional STs previously identified in reptiles and insects were found with a varying prevalence according to animal groups. These data were combined with those obtained from previous surveys to evaluate the potential risk of zoonotic transmission of Blastocystis sp. through the comparison of ST distribution between human and animal hosts. This suggests that non-human primates, artiodactyls and birds may serve as reservoirs for human infection, especially in animal handlers. In contrast, other mammals such as carnivores, and non-mammalian groups including reptiles and insects, do not seem PLOS ONE | DOI:10.1371/journal.pone.0169659 January 6, 2017 1 / 29 Molecular Epidemiology of Blastocystis sp. in Zoo Animals & Saade Association and AC by a PhD fellowship from the University of Lille 2 and the Institut Pasteur of Lille. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Geneius Laboratories Ltd. and the zoos of La Palmyre and Lille provided support in the form of salaries for authors (C. Noël, RM, SB, TP), but did not have any additional role in the study design, data collection and analysis, or decision to publish. to represent significant sources of Blastocystis sp. infection in humans. In further studies, more intensive sampling and screening of potential new animal hosts will reinforce these statements and expand our understanding of the circulation of Blastocystis sp. in animal and human populations. Competing Interests: C. Noël is employed by Geneius Laboratories Ltd., RM and SB by the zoo of Lille and TP by the zoo of La Palmyre. There are no patents, products in development or marketed products to declare in relation with our study. This does not alter our adherence to all the PLOS ONE policies on sharing data and materials, as detailed online in the guide for authors. Blastocystis sp. is an enteric protist with a worldwide distribution belonging to the group Stramenopiles and currently identified as one of the most common single-celled eukaryotes found in human stool samples [1–3]. Indeed, its prevalence can reach an average of 20% in industrialized countries [4,5] and can largely exceed 50% in developing countries [6]. A recent study even showed a prevalence of 100% in a cohort of children living in a rural area of Senegal [7]. Such a high prevalence of Blastocystis sp. clearly raises the question of the impact of this parasite in human health. Since asymptomatic carriage by Blastocystis sp. is very common, its role in human health and disease remains uncertain [8,9]. However, recent genomic data [10] coupled with in-vitro and in-vivo studies [11,12] allowed the identification of putative virulence factors and demonstrated the damaging effects of the parasite on the intestinal barrier, leading to plausible models of pathogenesis [8,13]. In addition, Bla (...truncated)