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First molecular characterization of Sarcocystis tenella in Tatra chamois (Rupicapra rupicapra tatrica) in Poland.

Kolenda R, Schierack P, Zieba F, Zwijacz-Kozica T, Bednarski M - Parasitol. Res. (2015)

Bottom Line: When compared at a haplotype level, all the sheep sequences of cox1 differed from those isolated from Tatra chamois.In conclusion, we showed that cox1 and ssu rRNA genes can be used as genetic markers for identification of the S. tenella, with cox1 gene providing better resolution during phylogenetic analyses.However, both genetic population analysis and phylogenetic inference with cox1 and ssu rRNA genes demonstrated that they do not constitute good markers for spatial differentiation of S. tenella.

View Article: PubMed Central - PubMed

Affiliation: Faculty of Natural Sciences, Brandenburg University of Technology Cottbus-Senftenberg, Großenhainer Str. 57, D-01968, Senftenberg, Germany, rkolenda@B-TU.de.

ABSTRACT
In this study, sarcocysts from three Polish Tatra chamois were isolated and identified using morphological and molecular methods for the first time. Six cysts were found in the latissimus dorsi muscle and another two in the diaphragm. No sarcocysts were detected in the myocardium, tongue, and esophagus. The isolated cysts were long with rounded ends, 0.35-0.61 mm in length, and 0.02-0.06 mm in width. All the sarcocysts were identified as Sarcocystis tenella on the basis of light microscopy and sequencing of cytochrome C oxidase subunit I (cox1) and small-subunit rRNA (ssu rRNA) genes. Comparative analysis showed a 99.23% identity of the cox1 gene sequences from Tatra chamois and sheep sarcocysts, and an even higher degree of sequence identity (99.88%) was documented in the case of the ssu rRNA gene. When compared at a haplotype level, all the sheep sequences of cox1 differed from those isolated from Tatra chamois. In contrast, one out of the two ssu rRNA haplotypes from the sheep isolates was identical with the haplotype from Tatra chamois. In conclusion, we showed that cox1 and ssu rRNA genes can be used as genetic markers for identification of the S. tenella, with cox1 gene providing better resolution during phylogenetic analyses. However, both genetic population analysis and phylogenetic inference with cox1 and ssu rRNA genes demonstrated that they do not constitute good markers for spatial differentiation of S. tenella.

No MeSH data available.


Related in: MedlinePlus

Maximum likelihood for DNA phylograms of selected Sarcocystidae and Coccidia. a The cox1 tree was constructed based on an alignment of partial cox1 gene sequences from eight Polish Sarcocystis tenella isolates and available cox1 gene sequences of related species deposited in the GenBank (a total of 375 sequences). b The ssu rRNA tree was constructed based on an alignment of nearly complete ssu rRNA gene sequences from eight Polish S. tenella isolates and available ssu rRNA gene sequences of related species deposited in the GenBank (a total 184 sequences). The trees were rooted with Eimeria spp. The values between the branches represent percent bootstrap value per 1000 replicates. Bootstrap values below 50 % are not shown. Polish isolates are marked in red. The GenBank accession numbers of all sequences used for construction of the trees are given in Table S1
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Related In: Results  -  Collection


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Fig1: Maximum likelihood for DNA phylograms of selected Sarcocystidae and Coccidia. a The cox1 tree was constructed based on an alignment of partial cox1 gene sequences from eight Polish Sarcocystis tenella isolates and available cox1 gene sequences of related species deposited in the GenBank (a total of 375 sequences). b The ssu rRNA tree was constructed based on an alignment of nearly complete ssu rRNA gene sequences from eight Polish S. tenella isolates and available ssu rRNA gene sequences of related species deposited in the GenBank (a total 184 sequences). The trees were rooted with Eimeria spp. The values between the branches represent percent bootstrap value per 1000 replicates. Bootstrap values below 50 % are not shown. Polish isolates are marked in red. The GenBank accession numbers of all sequences used for construction of the trees are given in Table S1

Mentions: The phylogenetic analyses of the cox1 gene sequences from Sarcocystis spp. isolated from cattle, cervids, and sheep were conducted with three methods: ML, MP, and Bayesian inference. As the trees with highly similar clade arrangements were obtained irrespective of the method used, we present only the ML tree (Fig. 1). All the Polish cox1 sequences from S. tenella are placed alongside the sequences from sheep in one cluster. All the sequences from Tatra chamois can be found in separate nodes of this cluster, but they do not tend to group in a single subclade. These findings are further supported by low pairwise distance values between the groups, as shown in Table 3. The Norwegian isolates showed higher intragroup pairwise distance (0.009 ± 0.002) as compared to the intergroup pairwise distance between the Polish and Norwegian isolates (0.008 ± 0.002).Fig. 1


First molecular characterization of Sarcocystis tenella in Tatra chamois (Rupicapra rupicapra tatrica) in Poland.

Kolenda R, Schierack P, Zieba F, Zwijacz-Kozica T, Bednarski M - Parasitol. Res. (2015)

Maximum likelihood for DNA phylograms of selected Sarcocystidae and Coccidia. a The cox1 tree was constructed based on an alignment of partial cox1 gene sequences from eight Polish Sarcocystis tenella isolates and available cox1 gene sequences of related species deposited in the GenBank (a total of 375 sequences). b The ssu rRNA tree was constructed based on an alignment of nearly complete ssu rRNA gene sequences from eight Polish S. tenella isolates and available ssu rRNA gene sequences of related species deposited in the GenBank (a total 184 sequences). The trees were rooted with Eimeria spp. The values between the branches represent percent bootstrap value per 1000 replicates. Bootstrap values below 50 % are not shown. Polish isolates are marked in red. The GenBank accession numbers of all sequences used for construction of the trees are given in Table S1
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4561999&req=5

Fig1: Maximum likelihood for DNA phylograms of selected Sarcocystidae and Coccidia. a The cox1 tree was constructed based on an alignment of partial cox1 gene sequences from eight Polish Sarcocystis tenella isolates and available cox1 gene sequences of related species deposited in the GenBank (a total of 375 sequences). b The ssu rRNA tree was constructed based on an alignment of nearly complete ssu rRNA gene sequences from eight Polish S. tenella isolates and available ssu rRNA gene sequences of related species deposited in the GenBank (a total 184 sequences). The trees were rooted with Eimeria spp. The values between the branches represent percent bootstrap value per 1000 replicates. Bootstrap values below 50 % are not shown. Polish isolates are marked in red. The GenBank accession numbers of all sequences used for construction of the trees are given in Table S1
Mentions: The phylogenetic analyses of the cox1 gene sequences from Sarcocystis spp. isolated from cattle, cervids, and sheep were conducted with three methods: ML, MP, and Bayesian inference. As the trees with highly similar clade arrangements were obtained irrespective of the method used, we present only the ML tree (Fig. 1). All the Polish cox1 sequences from S. tenella are placed alongside the sequences from sheep in one cluster. All the sequences from Tatra chamois can be found in separate nodes of this cluster, but they do not tend to group in a single subclade. These findings are further supported by low pairwise distance values between the groups, as shown in Table 3. The Norwegian isolates showed higher intragroup pairwise distance (0.009 ± 0.002) as compared to the intergroup pairwise distance between the Polish and Norwegian isolates (0.008 ± 0.002).Fig. 1

Bottom Line: When compared at a haplotype level, all the sheep sequences of cox1 differed from those isolated from Tatra chamois.In conclusion, we showed that cox1 and ssu rRNA genes can be used as genetic markers for identification of the S. tenella, with cox1 gene providing better resolution during phylogenetic analyses.However, both genetic population analysis and phylogenetic inference with cox1 and ssu rRNA genes demonstrated that they do not constitute good markers for spatial differentiation of S. tenella.

View Article: PubMed Central - PubMed

Affiliation: Faculty of Natural Sciences, Brandenburg University of Technology Cottbus-Senftenberg, Großenhainer Str. 57, D-01968, Senftenberg, Germany, rkolenda@B-TU.de.

ABSTRACT
In this study, sarcocysts from three Polish Tatra chamois were isolated and identified using morphological and molecular methods for the first time. Six cysts were found in the latissimus dorsi muscle and another two in the diaphragm. No sarcocysts were detected in the myocardium, tongue, and esophagus. The isolated cysts were long with rounded ends, 0.35-0.61 mm in length, and 0.02-0.06 mm in width. All the sarcocysts were identified as Sarcocystis tenella on the basis of light microscopy and sequencing of cytochrome C oxidase subunit I (cox1) and small-subunit rRNA (ssu rRNA) genes. Comparative analysis showed a 99.23% identity of the cox1 gene sequences from Tatra chamois and sheep sarcocysts, and an even higher degree of sequence identity (99.88%) was documented in the case of the ssu rRNA gene. When compared at a haplotype level, all the sheep sequences of cox1 differed from those isolated from Tatra chamois. In contrast, one out of the two ssu rRNA haplotypes from the sheep isolates was identical with the haplotype from Tatra chamois. In conclusion, we showed that cox1 and ssu rRNA genes can be used as genetic markers for identification of the S. tenella, with cox1 gene providing better resolution during phylogenetic analyses. However, both genetic population analysis and phylogenetic inference with cox1 and ssu rRNA genes demonstrated that they do not constitute good markers for spatial differentiation of S. tenella.

No MeSH data available.


Related in: MedlinePlus