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      Molecular identification and subtyping of Blastocystis sp. in laboratory rats in China Translated title: Identification moléculaire et sous-typage de Blastocystis sp. chez des rats de laboratoire en Chine

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          Abstract

          Blastocystis sp. is a ubiquitous protist that has been frequently reported in humans and animals worldwide. A total of 355 fecal samples of experimental rats were collected from four laboratory rearing facilities in China, and Blastocystis sp. was detected by PCR amplification of the partial small subunit ribosomal (SSU) rRNA gene. Twenty-nine (8.2%, 29/355) samples were positive for Blastocystis sp., with the highest infection rate (20.7%, 24/116) in rats of the Zhengzhou1, followed by that in the Zhengzhou2 (5.0%, 2/40), Shenyang (3.0%, 3/100) and Wuhan (0) rearing facilities. Among the three rat strains, Sprague–Dawley (SD) rats had higher infection rates (11.3%, 17/151) compared to Wistar rats (8.7%, 9/104) and spontaneously hypertensive (SH) rats (3.0%, 3/100). Two Blastocystis sp. subtypes (ST4 and ST7) were identified. ST4 was the predominant subtype detected in 26 samples (89.7%). A phylogenetic analysis demonstrated that the sequences of ST4 and ST7 obtained in this study were clustered with their reference subtypes. To our knowledge, this is the first report of Blastocystis sp. in experimental rats in China. Pathogen infections in laboratory animals need to be monitored due to fecal-oral transmission.

          Translated abstract

          Blastocystis sp. est un protiste omniprésent qui a été fréquemment signalé chez l’homme et les animaux du monde entier. Un total de 355 échantillons fécaux de rats de laboratoire ont été prélevés dans quatre installations d’élevage en laboratoire en Chine et Blastocystis sp. a été détectée par amplification par PCR du gène partiel de la petite sous-unité de l’ARNr ribosomique (SSU). Vingt-neuf échantillons (8,2 %, 29/355) étaient positifs pour Blastocystis sp., avec le taux d’infection le plus élevé (20,7 %, 24/116) chez les rats des élevages de Zhengzhou1, suivi de ceux de Zhengzhou2 (5,0 %, 2/40), Shenyang (3,0 %, 3/100) et Wuhan (0). Parmi les trois souches de rats, les rats Sprague-Dawley (SD) avaient un taux d’infection plus élevé (11,3 %, 17/151) que les rats Wistar (8,7 %, 9/104) et les rats spontanément hypertendus (SH) (3,0 %, 3/100). Deux sous-types de Blastocystis sp. ont été identifiés, ST4 et ST7. ST4 était le sous-type prédominant, détecté dans 26 échantillons (89,7 %). Une analyse phylogénétique a démontré que les séquences de ST4 et ST7 obtenues dans cette étude étaient groupées avec leurs sous-types de référence. À notre connaissance, il s’agit du premier signalement de Blastocystis sp. chez des rats de laboratoire en Chine. Les infections pathogènes chez les animaux de laboratoire doivent être surveillées en raison de la transmission fécale-orale.

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          Most cited references 24

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          Subtype distribution of Blastocystis isolates from synanthropic and zoo animals and identification of a new subtype.

          Blastocystis isolates from 56 Danish synanthropic and zoo animals, 62 primates primarily from United Kingdom (UK) collections and 16 UK primate handlers were subtyped by PCR, sequencing and phylogenetic analysis. A new subtype (ST) from primates and artiodactyls was identified and designated as Blastocystis sp. ST10. STs isolated from non-human primates (n=70) included ST3 (33%), ST8 (21%), ST2 (16%), ST5 (13%), ST1 (10%), ST4 (4%) and ST10 (3%). A high prevalence of ST8 was seen among primate handlers (25%). This ST is normally very rare in humans, suggesting that acquisition of Blastocystis ST8 infections from primates by their handlers had occurred in these cases. Data from published studies of non-human primates, other mammals and birds were collected and interpreted to generate a comprehensive overview on the ST distribution in such animals. On the basis of information on 438 samples, it was found that Blastocystis from primates belong mainly to ST1, ST2, ST3, ST5 and ST8, ungulates and dogs mainly ST1, ST2, ST3, ST5 and ST10, rodents ST4 and birds mainly ST6 and ST7. The data indicate moderate host specificity, most clearly exemplified by the fact that STs isolated from avian and non-avian hosts rarely overlap.
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            Development of a new PCR protocol to detect and subtype Blastocystis spp. from humans and animals.

            Blastocystis spp. is commonly found in the feces of humans worldwide. Infection has been reported as asymptomatic, acute symptomatic, and chronic symptomatic. This wide range of responses to infection could be related to the genetic diversity of morphologically indistinguishable specimens obtained from infected hosts. The former name Blastocystis hominis is now reported as Blastocystis spp. because of its genetic diversity. Blastocystis is recognized as a complex of subtypes that have not been fully characterized as independent species. The finding of Blastocystis spp. in feces from several animal species suggests a zoonotic potential. Based on conserved regions of published nucleotide SSU rDNA sequences from all Blastocystis subtypes found in GenBank, a PCR and sequencing protocol was developed. The ~500 bp SSU rDNA gene fragment amplified by this PCR is highly sensitive compared with published primers and contains highly variable regions that allow phylogenetic analysis of Blastocystis. These primers were used to detect and subtype Blastocystis spp. specimens from naturally infected humans, primates, cattle, pigs, and chickens. Based on these findings, application of this method can elucidate the complexity of this heterogeneous genus and its role in human and animal disease, as well as its zoonotic potential.
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              Molecular Epidemiology of Blastocystis sp. in Various Animal Groups from Two French Zoos and Evaluation of Potential Zoonotic Risk

              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 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.
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                Author and article information

                Journal
                Parasite
                Parasite
                parasite
                Parasite
                EDP Sciences
                1252-607X
                1776-1042
                2020
                15 May 2020
                : 27
                : ( publisher-idID: parasite/2020/01 )
                Affiliations
                [1 ] Academy of Chinese Medical Sciences, Henan University of Chinese Medicine 450046 Zhengzhou PR China
                [2 ] College of Animal Science, Tarim University Alar 843300 Xinjiang PR China
                [3 ] Faculty of Veterinary Medicine and Animal Science, Bangabandhu Sheikh Mujibur Rahman Agricultural University 1706 Gazipur Bangladesh
                Author notes
                [* ]Corresponding author: qimengdz@ 123456163.com
                Article
                parasite200026 10.1051/parasite/2020035
                10.1051/parasite/2020035
                7227369
                32410727
                © J. Li et al., published by EDP Sciences, 2020

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( https://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                Page count
                Figures: 1, Tables: 1, Equations: 0, References: 25, Pages: 5
                Categories
                Short Note

                laboratory rats, subtype, identification, blastocystis sp.

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