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      Sarcocystis spp. in domestic sheep in Kunming City, China: prevalence, morphology, and molecular characteristics Translated title: Sarcocystis spp. chez les moutons domestiques à Kunming, en Chine : prévalence, morphologie et caractéristiques moléculaires

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      1 , 2 , * , 1 , 1 , 3 , 1 , 1
      Parasite
      EDP Sciences
      Sheep, Sarcocystis, Prevalence, Morphology, Molecular characteristics, China

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          Abstract

          Sheep ( Ovis aries) are intermediate hosts for at least six named species of Sarcocystis: S. tenella, S. arieticanis, S. gigantea, S. medusiformis, S. mihoensis, and S. microps. Here, only two species, S. tenella and S. arieticanis, were found in 79 of 86 sheep (91.9%) in Kunming, China, based on their morphological characteristics. Four genetic markers, i.e., 18S rRNA gene, 28S rRNA gene, mitochondrial cox1 gene, and ITS-1 region, were sequenced and characterized for the two species of Sarcocystis. Sequences of the three former markers for S. tenella shared high identities with those of S. capracanis in goats, i.e., 99.0%, 98.3%, and 93.6%, respectively; the same three marker sequences of S. arieticanis shared high identities with those of S. hircicanis in goats, i.e., 98.5%, 96.5%, and 92.5%, respectively. No sequences in GenBank were found to significantly resemble the ITS-1 regions of S. tenella and S. arieticanis. Identities of the four genetic markers for S. tenella and S. arieticanis were 96.3%, 95.4%, 82.5%, and 66.2%, respectively.

          Translated abstract

          Les moutons ( Ovis aries) sont des hôtes intermédiaires pour au moins 6 espèces nommées de Sarcocystis : S. tenella, S. arieticanis, S. gigantea, S. medusiformis, S. mihoensis et S. microps. Ici, seules deux espèces, S. tenella et S. arieticanis ont été trouvées dans 79 (91.9 %) de 86 moutons à Kunming, en Chine, en fonction de leurs caractéristiques morphologiques. Quatre marqueurs génétiques, le gène de l’ARNr 18S, le gène de l’ARNr 28S, le gène mitochondrial cox1 et la région de l’ITS-1 ont été séquencés et caractérisés pour les deux espèces de Sarcocystis. Les séquences des trois premiers marqueurs de S. tenella partageaient des similarités élevées avec celles de S. capracanis chez les chèvres, soit 99.0 %, 98.3 % et 93.6 %, respectivement; les trois mêmes marqueurs de S. arieticanis partageaient des identités élevées avec ceux de S. hircicanis chez les chèvres, à savoir 98.5 %, 96.5 % et 92.5 %, respectivement. Aucune séquence de GenBank ne ressemble significativement aux régions ITS-1 de S. tenella et S. arieticanis. Les identités des quatre marqueurs génétiques de S. tenella et S. arieticanis étaient respectivement de 96.3 %, 95.4 %, 82.5 % et 66.2 %.

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          Most cited references14

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          The characterization of enzymatically amplified eukaryotic 16S-like rRNA-coding regions.

          Polymerase chain reaction conditions were established for the in vitro amplification of eukaryotic small subunit ribosomal (16S-like) rRNA genes. Coding regions from algae, fungi, and protozoa were amplified from nanogram quantities of genomic DNA or recombinant plasmids containing rDNA genes. Oligodeoxynucleotides that are complementary to conserved regions at the 5' and 3' termini of eukaryotic 16S-like rRNAs were used to prime DNA synthesis in repetitive cycles of denaturation, reannealing, and DNA synthesis. The fidelity of synthesis for the amplification products was evaluated by comparisons with sequences of previously reported rRNA genes or with primer extension analyses of rRNAs. Fewer than one error per 2000 positions were observed in the amplified rRNA coding region sequences. The primary structure of the 16S-like rRNA from the marine diatom, Skeletonema costatum, was inferred from the sequence of its in vitro amplified coding region.
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            Sarcocystis species in red deer revisited: with a re-description of two known species as Sarcocystis elongata n. sp. and Sarcocystis truncata n. sp. based on mitochondrial cox1 sequences.

            In a previous investigation, five Sarcocystis species were described from Norwegian red deer and believed to be conspecific with species occurring in either reindeer or moose based on sarcocyst morphology and nucleotide sequences of the nuclear ribosomal DNA unit. The aim of the present study was to characterize numerous isolates of these sarcocyst types at the mitochondrial cytochrome c oxidase subunit I gene (cox1) in order to corroborate or refute previous species designations of Sarcocystis in red deer. The Sarcocystis tarandi- and Sarcocystis rangiferi-like taxa in red deer and reindeer, respectively, were thoroughly compared by sequencing 14-27 isolates of each type. Sequence comparisons revealed four distinct sequence types, which by phylogenetic analyses were placed in four monophyletic groups according to host origin, and they were therefore considered to represent four separate species. The two taxa of this type in red deer were named Sarcocystis elongata and Sarcocystis truncata, respectively. Sequencing of many isolates of Sarcocystis hjorti and Sarcocystis ovalis from red deer and moose confirmed that these species occur in both hosts. A revised description of the two new species is given and the current knowledge concerning all six Sarcocystis species in red deer is reviewed.
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              Molecular characterisation of Sarcocystis rileyi from a common eider (Somateria mollissima) in Norway.

              The breast and leg muscles of a common eider (Somateria mollissima; Anatidae: Anseriformes) from northern Norway contained numerous grossly visible cigar-shaped sarcocysts measuring about 5 × 1 mm. Light microscopic examination of isolated sarcocysts revealed that they were encapsulated by a thin fibrous layer, underneath which there was a thin and fairly smooth cyst wall with no visible protrusions. The cystozoites were straight, spindle-shaped and about 13 μm long. Genomic DNA was extracted from 12 excised sarcocysts and each DNA isolate was subjected to PCR amplification of one to four loci: the 18S and 28S ribosomal RNA genes (four isolates), the internal transcribed spacer 1 (ITS1) region (six isolates) and the mitochondrial cytochrome c oxidase subunit 1 gene (cox1) (12 isolates). At the three nuclear loci, the new sequences showed 99.9-100% sequence identity with previous sequences of Sarcocystis rileyi from the mallard duck in Lithuania and USA, and they could therefore be assigned to this species. At cox1, the new sequences of S. rileyi were most similar to Sarcocystis arctica and Sarcocystis neurona, but the most closely related Sarcocystis spp. in birds have not been sequenced at this locus. There was no sequence variation at any locus between the 4-12 examined isolates of S. rileyi. This is the first genetically verified record of S. rileyi in the common eider, as well as in any bird species in Norway. The phylogenetic placement of S. rileyi was inferred separately from 28S rRNA gene and cox1 sequences, and similar results were obtained in both analyses.
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                Author and article information

                Journal
                Parasite
                Parasite
                parasite
                Parasite
                EDP Sciences
                1252-607X
                1776-1042
                2017
                02 August 2017
                : 24
                : ( publisher-idID: parasite/2017/01 )
                : 30
                Affiliations
                [1 ] School of Biological Sciences, Yunnan University Kunming 650091 PR China
                [2 ] Southeast Asia Biodiversity Research Institute, Chinese Academy of Sciences, Yezin Nay Pyi Taw 05282 Myanmar
                [3 ] Department of Biology, Wake Forest University Winston-Salem NC 27106 USA
                Author notes
                [* ]Corresponding author: jjhu@ 123456ynu.edu.cn
                Article
                parasite170057 10.1051/parasite/2017025
                10.1051/parasite/2017025
                5539603
                28766501
                01e92cd2-c679-408a-a6f1-acd51a98549d
                © J.-J. Hu et al., published by EDP Sciences, 2017

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

                History
                : 06 May 2017
                : 09 June 2017
                Page count
                Figures: 5, Tables: 1, Equations: 0, References: 18, Pages: 8
                Categories
                Research Article

                sheep,sarcocystis,prevalence,morphology,molecular characteristics,china

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