Morphological and molecular characterizations of Sarcocystis miescheriana and Sarcocystis suihominis in domestic pigs (Sus scrofa) in China

Huang, Zhumei; Ye, Yulong; Zhang, Hengzhen; Deng, Shuangsheng; Tao, Jianping; Hu, Junjie; Yang, Yurong

doi:10.1007/s00436-019-06521-5

Cited by 28 publications

(21 citation statements)

References 13 publications

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“…Nucleotide sequence analysis has proven to be a useful tool for delineating or identifying species of Sarcocystis from the same or different hosts, and different genetic markers have revealed different levels of intra-or interspeci c sequence diversity [7,11]. There are currently no nucleotide data for Sarcocystis spp.…”

Section: Discussionmentioning

confidence: 99%

“…The ultrastructure of sarcocysts is traditionally a reliable characteristic for identifying different Sarcocystis species in a given host. Currently, PCR assays and sequencing procedures are considered much more practical, accurate, and reliable methods for the delineation and identi cation of Sarcocystis species than traditional methods based on morphological characteristics [6,7]. However, there are currently no records of the nucleotide sequences of Sarcocystis spp.…”

Section: Introductionmentioning

confidence: 99%

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Morphological and Molecular Characterization of Sarcocystis Wenzeli in Chickens (Gallus gallus) in China

Pan

Huang

et al. 2020

Preprint

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Background: There has been considerable confusion concerning the number and classification of Sarcocystis spp. found in chickens. To date, no nucleotide sequences of Sarcocystis spp. from chickens have been provided in GenBank. The present study aimed to investigate the morphological and molecular characteristics of Sarcocystis spp. 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, the ITS-1 region (ITS-1), the mitochondrial cox1 gene (cox1), and the apicoplastic rpoB gene (rpoB), were amplified from each sarcocyst, cloned, 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. The bradyzoites were lancet-shaped, measuring 9.2–12.6 × 1.5–3.5 μm. Ultrastructurally, the sarcocyst wall contained numerous stubby hill-like villar protrusions that were up to 1.2 μm long and 1.0 μm wide. The protrusions included scattered microtubules, which extended from the tips of the protrusions into the ground substance. The intraspecific similarity of the five loci (18S rDNA, 28S rDNA, ITS-1, cox1, and rpoB) among the newly obtained sequences ranged from 99.8–100%, 99.7–100%, 99.0–99.9%, 100% and 98.9–100%, respectively. The five loci showed different levels of interspecific sequence similarity with 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 three of the loci (18S rDNA, cox1, and rpoB) revealed that S. wenzeli formed an independent clade with Sarcocystis spp. that employ geese or ducks as intermediate hosts and canines as the known or presumed definitive host.Conclusions: The sequences of 18S rDNA, 28S rDNA, ITS-1, cox1 and rpoB from S. wenzeli reported here constitute the first records of genetic markers of Sarcocystis from chickens made available in GenBank. Based on sequence analysis, ITS-1 and rpoB are more suitable for discriminating S. wenzeli from closely related species of Sarcocystis. Phylogenetic analysis revealed that S. wenzeli presents a close relationship with Sarcocystis spp. from geese or ducks.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Morphological and Molecular Characterization of Sarcocystis Wenzeli in Chickens (Gallus gallus) in China

Pan

Huang

et al. 2020

Preprint

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“…Nucleotide sequence analysis has proven to be a useful tool for delineating or identifying species of Sarcocystis from the same or different hosts, and different genetic markers have revealed different levels of intra-or interspeci c sequence diversity [6,7,11]. There are only one 18S rDNA sequence, one ITS-1 sequence and one cox1 sequence of Sarcocystis sp.…”

Section: Discussionmentioning

confidence: 99%

“…The ultrastructure of sarcocysts is traditionally a reliable characteristic for identifying different Sarcocystis species in a given host. Currently, PCR assays and sequencing procedures are considered much more practical, accurate, and reliable methods for the delineation and identi cation of Sarcocystis species than traditional methods based on morphological characteristics [6,7]. However, there are currently only one 18S rDNA sequence (783 bp), one ITS-1 region (ITS-1) sequence (923 bp) and one mitochondrial cox1 gene (cox1) sequence (547 bp) of an undescribed and unnamed species of Sarcocystis obtained from chickens deposited in GenBank.…”

Section: Introductionmentioning

confidence: 99%

Morphological and molecular characterization of Sarcocystis wenzeli in chickens (Gallus gallus) in China

Pan

Huang

et al. 2020

Preprint

Self Cite

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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, the ITS-1 region ( ITS-1 ), the mitochondrial cox1 gene ( cox1 ), and the apicoplastic rpoB gene ( rpoB ), 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 deposited in GenBank. At 18S rDNA, ITS-1 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, ITS-1 , 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 three of the loci (18S rDNA, cox1 , and rpoB ) revealed that S . wenzeli formed an independent clade with Sarcocystis spp. that employ geese or ducks as intermediate hosts and canines as the known or presumed definitive host. Conclusions: 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 ITS-1 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.

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“…2017; Huang et al . 2019). These approaches, however, are not appropriate for identifying certain Sarcocystis taxa that display very few morphological differences or for host range studies.…”

Section: Introductionmentioning

confidence: 99%

Molecular phylogeny of Sarcocystis fayeri (Apicomplexa: Sarcocystidae) from the domestic horse Equus caballus based on 18S rRNA gene sequences and its prevalence

Abdel‐Gaber

Quraishy

Dkhil

et al. 2020

Lett Appl Microbiol

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Sarcocystosis is a parasitic disease caused by an intracellular protozoan parasite Sarcocystis belonging to the phylum Apicomplexa. These parasites have a requisite two‐host life cycle. Recently, there are many Sarcocystis species that identified morphologically. In the present study, diaphragmatic muscle samples from the domestic horse (Equus caballus) were examined for Sarcocystis infection. The natural infection with sarcocysts was recorded to be 62·5% for only microcysts in the infected muscles. Molecular analysis using the 18S rRNA gene was conducted to swiftly and accurately identify the recovered species. Studies on the expression of the 18S rRNA gene have confirmed that the present parasite isolates belong to the Sarcocystis genus. The sequence data showed significant identities (>80%) with archived gene sequences from species within the Sarcocystidae family, and a dendrogram showing the phylogenetic relationship was constructed. The most closely related species were the previously described Sarcocystis fayeri and Sarcocystis bertrami. The current data showed that the present species was identified as S. fayeri and deposited in GenBank (accession number MF614956.1). This study highlights the importance of the genetic data in the exact taxonomy within sarcocystid species.

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Morphological and molecular characterizations of Sarcocystis miescheriana and Sarcocystis suihominis in domestic pigs (Sus scrofa) in China

Cited by 28 publications

References 13 publications

Morphological and Molecular Characterization of Sarcocystis Wenzeli in Chickens (Gallus gallus) in China

Morphological and Molecular Characterization of Sarcocystis Wenzeli in Chickens (Gallus gallus) in China

Morphological and molecular characterization of Sarcocystis wenzeli in chickens (Gallus gallus) in China

Molecular phylogeny of Sarcocystis fayeri (Apicomplexa: Sarcocystidae) from the domestic horse Equus caballus based on 18S rRNA gene sequences and its prevalence

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