Limits...
Molecular Analysis of Sarcocystis Spp. Isolated from Sheep (Ovis aries) in Babol Area, Mazandaran Province, Northern Iran.

Kalantari N, Khaksar M, Ghaffari S, Hamidekish SM - Iran J Parasitol (2016 Jan-Mar)

Bottom Line: Eight and thirty-four variations in nucleotide position were seen in partial sequence of the18S rRNA gene of S. gigantea and S. moulei.Sheep can be considered as an alternative intermediate host for S. moulei.Furthermore, multiple alignments showed some variations in the consensus sequences of the isolates obtained in the current study compared with previously published isolates.

View Article: PubMed Central - PubMed

Affiliation: Health Research Institute, Cellular and Molecular Biology Research Center, Babol University of Medical Sciences, Babol, Iran.

ABSTRACT

Background: To differentiate Sarcocystis macro-cyst-forming species in slaughtered sheep in Babol area, Mazandaran Province, sequence analysis of 18S rRNA gene was performed.

Methods: Overall, 150 slaughtered sheep were examined macroscopically in slaughterhouse, Babol and intra-abdominal and diaphragm muscles tissues infected with macro-cyst of Sarcocystis spp. were collected in 2013. One macro-cyst was isolated from the infected muscles of each sheep. The partial 18S rRNA gene was amplified by PCR and sequenced afterward.

Results: The rate of infection with macro-cyst producing Sarcocystis spp. was 33.3% (50 / 150). The partial 18S rRNA gene of Sarcocystis species was amplified at the expected PCR product size (∼1100 bp) from all 50 macroscopic cysts samples. From 30 sequences DNA samples, 20 samples (66.7%), six (20%) and four (13.3%) isolates were identified as S. gigantea, S. moulei and Sarcocystis spp., respectively. Eight and thirty-four variations in nucleotide position were seen in partial sequence of the18S rRNA gene of S. gigantea and S. moulei.

Conclusion: Sheep can be considered as an alternative intermediate host for S. moulei. Furthermore, multiple alignments showed some variations in the consensus sequences of the isolates obtained in the current study compared with previously published isolates. To understand better the genetic diversity among Sarcocystis species complete sequences of the18S rRNA gene or sequence analysis of other genetic loci would be beneficial.

No MeSH data available.


Related in: MedlinePlus

Phylogeny of Sarcocystis spp. isolates by the programme Mega v.6. using maximum likelihood method and bootstrap of 1000 replicates based on 18S rRNA gene. The reference sequences accession numbers are included
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC4835472&req=5

Figure 3: Phylogeny of Sarcocystis spp. isolates by the programme Mega v.6. using maximum likelihood method and bootstrap of 1000 replicates based on 18S rRNA gene. The reference sequences accession numbers are included

Mentions: Thirty PCR products of the 18S rRNA gene were successfully sequenced. Each PCR product yielded a fragment containing 850 to 1021 consensus nucleotides. Overall, 20 out of 30 samples (66.7%) and six/thirty (20%) isolates had a similarity of more than 98% and 100% coverage to S. gigantea accession number KC209733 or L24384 and S. moulei accession number L76473 or KC 508513, respectively (NCBI GenBank). 13.3% isolates showed a similarity of more than 89% and 50% coverage to Sarcocystis spp. accession number GQ131808. Multiple alignments showed some variation in the consensus sequences of the isolates obtained in the current study compared with previously published S. gigantea isolates and with each other (Fig. 2). The same results were observed for S. moulei (Fig. 2). A phylogenetic tree of three S. gigantea, two S. moulei and some published sequences (NCBI GenBank) is shown in Fig. 3.


Molecular Analysis of Sarcocystis Spp. Isolated from Sheep (Ovis aries) in Babol Area, Mazandaran Province, Northern Iran.

Kalantari N, Khaksar M, Ghaffari S, Hamidekish SM - Iran J Parasitol (2016 Jan-Mar)

Phylogeny of Sarcocystis spp. isolates by the programme Mega v.6. using maximum likelihood method and bootstrap of 1000 replicates based on 18S rRNA gene. The reference sequences accession numbers are included
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4835472&req=5

Figure 3: Phylogeny of Sarcocystis spp. isolates by the programme Mega v.6. using maximum likelihood method and bootstrap of 1000 replicates based on 18S rRNA gene. The reference sequences accession numbers are included
Mentions: Thirty PCR products of the 18S rRNA gene were successfully sequenced. Each PCR product yielded a fragment containing 850 to 1021 consensus nucleotides. Overall, 20 out of 30 samples (66.7%) and six/thirty (20%) isolates had a similarity of more than 98% and 100% coverage to S. gigantea accession number KC209733 or L24384 and S. moulei accession number L76473 or KC 508513, respectively (NCBI GenBank). 13.3% isolates showed a similarity of more than 89% and 50% coverage to Sarcocystis spp. accession number GQ131808. Multiple alignments showed some variation in the consensus sequences of the isolates obtained in the current study compared with previously published S. gigantea isolates and with each other (Fig. 2). The same results were observed for S. moulei (Fig. 2). A phylogenetic tree of three S. gigantea, two S. moulei and some published sequences (NCBI GenBank) is shown in Fig. 3.

Bottom Line: Eight and thirty-four variations in nucleotide position were seen in partial sequence of the18S rRNA gene of S. gigantea and S. moulei.Sheep can be considered as an alternative intermediate host for S. moulei.Furthermore, multiple alignments showed some variations in the consensus sequences of the isolates obtained in the current study compared with previously published isolates.

View Article: PubMed Central - PubMed

Affiliation: Health Research Institute, Cellular and Molecular Biology Research Center, Babol University of Medical Sciences, Babol, Iran.

ABSTRACT

Background: To differentiate Sarcocystis macro-cyst-forming species in slaughtered sheep in Babol area, Mazandaran Province, sequence analysis of 18S rRNA gene was performed.

Methods: Overall, 150 slaughtered sheep were examined macroscopically in slaughterhouse, Babol and intra-abdominal and diaphragm muscles tissues infected with macro-cyst of Sarcocystis spp. were collected in 2013. One macro-cyst was isolated from the infected muscles of each sheep. The partial 18S rRNA gene was amplified by PCR and sequenced afterward.

Results: The rate of infection with macro-cyst producing Sarcocystis spp. was 33.3% (50 / 150). The partial 18S rRNA gene of Sarcocystis species was amplified at the expected PCR product size (∼1100 bp) from all 50 macroscopic cysts samples. From 30 sequences DNA samples, 20 samples (66.7%), six (20%) and four (13.3%) isolates were identified as S. gigantea, S. moulei and Sarcocystis spp., respectively. Eight and thirty-four variations in nucleotide position were seen in partial sequence of the18S rRNA gene of S. gigantea and S. moulei.

Conclusion: Sheep can be considered as an alternative intermediate host for S. moulei. Furthermore, multiple alignments showed some variations in the consensus sequences of the isolates obtained in the current study compared with previously published isolates. To understand better the genetic diversity among Sarcocystis species complete sequences of the18S rRNA gene or sequence analysis of other genetic loci would be beneficial.

No MeSH data available.


Related in: MedlinePlus