Cargando…
Morphological and molecular characterization of Sarcocystis wenzeli in chickens (Gallus gallus) in China
BACKGROUND: There has been considerable confusion concerning the number and classification of Sarcocystis spp. in chickens. Scarce nucleotide data of Sarcocystis spp. from chickens are provided in GenBank. The study aimed to investigate the morphological and molecular characteristics of Sarcocystis...
Autores principales: | , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2020
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7557040/ https://www.ncbi.nlm.nih.gov/pubmed/33054838 http://dx.doi.org/10.1186/s13071-020-04390-x |
_version_ | 1783594334576181248 |
---|---|
author | Pan, Jing Ma, Chunli Huang, Zhumei Ye, Yulong Zeng, Hongxia Deng, Shuangsheng Hu, Junjie Tao, Jianping |
author_facet | Pan, Jing Ma, Chunli Huang, Zhumei Ye, Yulong Zeng, Hongxia Deng, Shuangsheng Hu, Junjie Tao, Jianping |
author_sort | Pan, Jing |
collection | PubMed |
description | BACKGROUND: There has been considerable confusion concerning the number and classification of Sarcocystis spp. in chickens. Scarce nucleotide data of Sarcocystis spp. from chickens are provided in GenBank. The study aimed to investigate the morphological and molecular characteristics of Sarcocystis spp. found in chickens in China. METHODS: Tissues from 33 chickens were collected in 2019. Sarcocysts were observed using light (LM) and transmission electron microscopy (TEM). Individual sarcocysts from different chickens were selected for DNA extraction, and five loci, 18S rDNA, 28S rDNA, ITS1 region, the mitochondrial cox1 gene and the apicoplastic rpoB gene, were amplified from each sarcocyst, sequenced and analyzed. RESULTS: Only S. wenzeli was found in 14 of 33 (42.4%) chickens. Under LM, the sarcocysts were microscopic and exhibited palisade-like villar protrusions measuring 1.5–2.8 μm. Ultrastructurally, the sarcocyst wall contained numerous stubby hill-like villar protrusions. The protrusions included scattered microtubules, which extended from the tips of the protrusions into the ground substance. The five loci were successfully sequenced and the sequences deposited in GenBank. At 18S rDNA, ITS1 and cox1, the most similar sequences in GenBank were those of Sarcocystis sp. obtained from the brains of chickens, i.e. 99.9–100%, 98.1–98.5% and 99.3% identity, respectively. The five loci (18S rDNA, 28S rDNA, ITS1, cox1 and rpoB) showed different levels of interspecific sequence similarity with other closely related species of Sarcocystis (e.g. 99.8%, 99.0–99.2%, 89.3–89.7%, 98.5%, and 97.5%, respectively, with S. anasi). Phylogenetic analysis based on four of the loci (18S rDNA, cox1, rpoB and ITS1) revealed that S. wenzeli formed an independent clade with Sarcocystis spp. that utilize geese or ducks as intermediate hosts and canines as the known or presumed definitive host. CONCLUSIONS: To our knowledge, the sequences of 28S rDNA and rpoB reported here constitute the first records of genetic markers of Sarcocystis spp. in chickens. Based on molecular analysis, S. wenzeli might be responsible for the neurological disease in chickens, and ITS1 and rpoB are more suitable for discriminating it from closely related Sarcocystis spp. Phylogenetic analysis revealed that S. wenzeli presents a close relationship with Sarcocystis spp. in geese or ducks. [Image: see text] |
format | Online Article Text |
id | pubmed-7557040 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-75570402020-10-15 Morphological and molecular characterization of Sarcocystis wenzeli in chickens (Gallus gallus) in China Pan, Jing Ma, Chunli Huang, Zhumei Ye, Yulong Zeng, Hongxia Deng, Shuangsheng Hu, Junjie Tao, Jianping Parasit Vectors Research BACKGROUND: There has been considerable confusion concerning the number and classification of Sarcocystis spp. in chickens. Scarce nucleotide data of Sarcocystis spp. from chickens are provided in GenBank. The study aimed to investigate the morphological and molecular characteristics of Sarcocystis spp. found in chickens in China. METHODS: Tissues from 33 chickens were collected in 2019. Sarcocysts were observed using light (LM) and transmission electron microscopy (TEM). Individual sarcocysts from different chickens were selected for DNA extraction, and five loci, 18S rDNA, 28S rDNA, ITS1 region, the mitochondrial cox1 gene and the apicoplastic rpoB gene, were amplified from each sarcocyst, sequenced and analyzed. RESULTS: Only S. wenzeli was found in 14 of 33 (42.4%) chickens. Under LM, the sarcocysts were microscopic and exhibited palisade-like villar protrusions measuring 1.5–2.8 μm. Ultrastructurally, the sarcocyst wall contained numerous stubby hill-like villar protrusions. The protrusions included scattered microtubules, which extended from the tips of the protrusions into the ground substance. The five loci were successfully sequenced and the sequences deposited in GenBank. At 18S rDNA, ITS1 and cox1, the most similar sequences in GenBank were those of Sarcocystis sp. obtained from the brains of chickens, i.e. 99.9–100%, 98.1–98.5% and 99.3% identity, respectively. The five loci (18S rDNA, 28S rDNA, ITS1, cox1 and rpoB) showed different levels of interspecific sequence similarity with other closely related species of Sarcocystis (e.g. 99.8%, 99.0–99.2%, 89.3–89.7%, 98.5%, and 97.5%, respectively, with S. anasi). Phylogenetic analysis based on four of the loci (18S rDNA, cox1, rpoB and ITS1) revealed that S. wenzeli formed an independent clade with Sarcocystis spp. that utilize geese or ducks as intermediate hosts and canines as the known or presumed definitive host. CONCLUSIONS: To our knowledge, the sequences of 28S rDNA and rpoB reported here constitute the first records of genetic markers of Sarcocystis spp. in chickens. Based on molecular analysis, S. wenzeli might be responsible for the neurological disease in chickens, and ITS1 and rpoB are more suitable for discriminating it from closely related Sarcocystis spp. Phylogenetic analysis revealed that S. wenzeli presents a close relationship with Sarcocystis spp. in geese or ducks. [Image: see text] BioMed Central 2020-10-14 /pmc/articles/PMC7557040/ /pubmed/33054838 http://dx.doi.org/10.1186/s13071-020-04390-x Text en © The Author(s) 2020 Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data. |
spellingShingle | Research Pan, Jing Ma, Chunli Huang, Zhumei Ye, Yulong Zeng, Hongxia Deng, Shuangsheng Hu, Junjie Tao, Jianping Morphological and molecular characterization of Sarcocystis wenzeli in chickens (Gallus gallus) in China |
title | Morphological and molecular characterization of Sarcocystis wenzeli in chickens (Gallus gallus) in China |
title_full | Morphological and molecular characterization of Sarcocystis wenzeli in chickens (Gallus gallus) in China |
title_fullStr | Morphological and molecular characterization of Sarcocystis wenzeli in chickens (Gallus gallus) in China |
title_full_unstemmed | Morphological and molecular characterization of Sarcocystis wenzeli in chickens (Gallus gallus) in China |
title_short | Morphological and molecular characterization of Sarcocystis wenzeli in chickens (Gallus gallus) in China |
title_sort | morphological and molecular characterization of sarcocystis wenzeli in chickens (gallus gallus) in china |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7557040/ https://www.ncbi.nlm.nih.gov/pubmed/33054838 http://dx.doi.org/10.1186/s13071-020-04390-x |
work_keys_str_mv | AT panjing morphologicalandmolecularcharacterizationofsarcocystiswenzeliinchickensgallusgallusinchina AT machunli morphologicalandmolecularcharacterizationofsarcocystiswenzeliinchickensgallusgallusinchina AT huangzhumei morphologicalandmolecularcharacterizationofsarcocystiswenzeliinchickensgallusgallusinchina AT yeyulong morphologicalandmolecularcharacterizationofsarcocystiswenzeliinchickensgallusgallusinchina AT zenghongxia morphologicalandmolecularcharacterizationofsarcocystiswenzeliinchickensgallusgallusinchina AT dengshuangsheng morphologicalandmolecularcharacterizationofsarcocystiswenzeliinchickensgallusgallusinchina AT hujunjie morphologicalandmolecularcharacterizationofsarcocystiswenzeliinchickensgallusgallusinchina AT taojianping morphologicalandmolecularcharacterizationofsarcocystiswenzeliinchickensgallusgallusinchina |