Molecular Analysis of Sarcocystis Spp. Isolated from Sheep (Ovis aries) in Babol Area, Mazandaran Province, Northern Iran
AbstractBackground: 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.
Chhabra MB, Samantaray S. Sarcocystis and sarcocystosis in India: status and emerging perspectives. J Parasi Dis. 2013; 37:1-10.
Fayer R. Sarcocystis spp. in human infections. Clin Microbiol Rev. 2004; 17:894-902.
Heckeroth AR, Tenter AM. Development and validation of species-specific nested PCRs for diagnosis of acute sarcocystiosis in sheep. Int J Parasitol. 1999; 29:1331-49.
Heckeroth AR, Tenter AM. Comparison of immunological and molecular methods for the diagnosis of infections with pathogenic Sarcocystis species in sheep. Tokai J Exp Clin Med. 1998; 23: 293-302.
Oryan A, Moghaddar N, Gaur SN. The distribution pattern of Sarcocystis species, their transmission and pathogenesis in sheep in Fars province of Iran. Vet Res Commun. 1996; 20 :243-53.
Rassouli M, Ahmadpanahi J, Alvandi A. Prevalence of Sarcocystis spp. and Hammondia spp. microcysts in esophagus tissue of sheep and cattle, emphasized on their morphological differences. Parasitol Res. 2014; 113:3801-5.
Bahari P, Salehi M, Seyedabadi M, Mohammadi A. Molecular identification of macroscopic and microscopic cysts of Sarcocystis in sheep in North Khorasan province, Iran. Int J Mol Cell Med. 2014; 3:51-6.
Dalimi AH, Paikari HA, Esmaeilzadeh M, Valizadeh M, Karimi GR, Motamedi GR, Godarzi MA. Identification of Sarcocystis species of slaughtered sheep in a slaughterhouse Ziyaran Qazvin by PCR-RFLP. Modares J Med Sci. 2008; 11:65-72.
Farhang-Pajuh F, Yakhchali M, Mardani K. Molecular determination of abundance of infection with Sarcocystis species in slaughtered sheep of Urmia, Iran.Vet Res Forum. 2014 ; 5:181-6.
Nourollahi-Fard SR, Kheirandish R, Sattari S. Prevalence and histopathological finding of thin-walled and thick-walled Sarcocysts in slaughtered cattle of Karaj abattoir, Iran. J Parasit Dis. 2015; 39: 272-5.
Fallah M, Matini M, Kia EB, Mobedi I. Study of Zoonotic Tissue Parasites (Hydatid Cyst, Fasciola, Dicrocoelium and Sarcocystis) in Hamadan Abattoir, 2009. Scientific J Hamadan University of Medical Sciences & Health Services. 2010; 17: 5-12.
Sharbatkhori M, Kia EB, Harandi MF, Jalalizand N, Zahabiun F, Mirhendi H. Comparison of five simple methods for DNA extraction from Echinococcus granulosus protoscoleces for PCR-amplification of ribosomal DNA. Iran J Parasitol. 2009; 4: 54-60.
Yang ZQ, Zuo YX, Yao YG, Chen XW, Yang GC, Zhang YP. Analysis of the 18S rRNA genes of Sarcocysti species suggests that the morphologically similar organisms from cattle and water buffalo should be considered the same species. Mol Bioch Parasitol. 2001; 115:283-8.
Kalantari N, Bayani M, Ghaffari S. Sarcocystis cruzi: First molecular identification from cattle in Iran. Int J Mol Cell Med. 2013; 2:125-30.
Tamura K, Stecher G,Peterson D, Filipski A, Kumar S. MEGA6: molecular evolutionary genetics analysis version 6.0. Mol Biol Evol. 2013; 30: 2725-9.
Gajadhar AA, Marquardt WC, Blair CD. Development of a model ribosomal RNA hybridization assay for the detection of Sarcocystis and other coccidia. Can J Vet Res. 1992; 56:208-13.
Al-Hoot AS, Al-Qureishy SA, Al-Rashid K, Bashtar AR. Microscopic study on Sarcocystis moulei from sheep and goats in Saudi Arabia. J Egypt Soc Parasitol. 2005; 35:295-312.
Ozkayhan MA, Karaer Z, Ilkme A, Karaer Z, Ilkme AN, Atmaca HT. The prevalence of Sarcocystis species in sheep slaughtered in municipality slaughterhouse in Kirikkale. Turkiye Parazitol Derg. 2007; 31:272-6.
Rejmanek D, Vanwormer E, Miller MA, Mazet JA, Nichelason AE, Melli AC, Packham AE, Jessup DA, Conrad PA. Prevalence and risk factors associated with Sarcocystis neurona infections in opossums (Didelphis virginiana) from central California. Vet Parasitol. 2009; 166:8-14.
Neefs J-M, Van de Peer Y, Hendriks L, De Wachter R. Compilation of small ribosomal subunit RNA sequences. Nucleic Acids Res. 1990; 18(Suppl):2237-317.
Marsh A, Barr BC, Madigan J, Lakritz J, Conrad PA. Sequence analysis and polymerase chain reaction amplification of small subunit ribosomal DNA from Sarcocystis neurona. Am J Vet Res. 1996; 57:975-81.
Hamidinejat H, Moetamedi H, Alborzi A, Hatami A. Molecular detection of Sarcocystis species in slaughtered sheep by PCR–RFLP from south-western of Iran. J Parasit Dis. 2014; 38:233-7.
Gjerde B. Phylogenetic relationships among Sarcocystis species in cervids, cattle and sheep inferred from the mitochondrial cytochrome c oxidase subunit I gene. Int J Parasitol. 2013; 43:579-91.
Jehle C, Dinkel A, Sander A, Morent M, Romig T, Luc PV, De TV, Thai VV, Mackenstedt U. Diagnosis of Sarcocystis spp. in cattle ( Bos taurus) and water buffalo ( Bubalus bubalis) in Northern Vietnam. Vet Parasitol. 2009; 166:314-20.
Kolenda R, Ugorski M, Bednarski M. Molecular characterization of Sarcocystis species from Polish roe deer based on ssu rRNA and cox1 sequence analysis. Parasitol Res. 2014; 113:3029-39.
Shekarforoush S, Razavi S, Dehghan S, Sarihi K. Prevalence of Sarcocystis species in slaughtered goats in Shiraz, Iran. Vet Record. 2005; 156:418-20.