Identification of Echinococcus granulosus Strains in Isolated Hydatid Cyst Specimens from Animals by PCR-RFLP Method in West Azerbaijan - Iran.

  • Haleh Hanifian Dept. of Medical Parasitology and Mycology, Faculty of medicine, Urmia University of Medical Sciences, Urmia, Iran.
  • Kambiz Diba Dept. of Medical Parasitology and Mycology, Faculty of medicine, Urmia University of Medical Sciences, Urmia, Iran.
  • Khosrow Hazrati Tappeh Cellular and Molecular Research center, Urmia University of Medical Seiences, Urmia-Iran.
  • Habib Mohammadzadeh Dept. of Medical Parasitology and Mycology, Faculty of medicine, Urmia University of Medical Sciences, Urmia, Iran.
  • Rahim Mahmoudlou Dept. of surgery, Faculty of medicine, Urmia University of Medical Sciences, Urmia-Iran.
Keywords: Hydatid cyst, ITS1, RFLP-PCR, Strain

Abstract

 Background: The aim of this study was DNA extraction from protosco-lecses of Echinococcus granulosus and identification of these strains in West- Azerbaijan Province, north western Iran. Methods: Thirty one livestock isolates from sheep and cattle were collected from abattoirs of the province. To investigate the genetic variation of the isolates, after DNA extraction by Glass beads-phenol chloroform method; PCR-RLFP analysis of rDNA-ITS1 was performed using three different re-striction enzymes of Taq 1, Rsa 1 and Alu 1. Result: Amplified PCR products for all isolates were 1000bp band which is expected band in sheep strains (G1-G3 complex). The results of RFLP analy-sis also were the same for all isolates. PCR-RFLP patterns restriction en-zymes were identical as follows, Rsa1 bands under UV showed two bands approximately 655bp and 345bp. Alu1 bands were as follows: two approx-imately 800bp and 200bp and Taq1 did not cut any region and bands were approximately 1000 bp in all samples. Conclusions: Based on PCR-RFLP patterns of ITS1 fragment produced with endonucleases enzyme digestion in animal isolates, it can be concluded that a single strain of E. granulosus (sheep strain or G1-G3 complex) is domi-nant genotype in this province.

References

Simsek S, Köroğlu E, Dumanli N, Aktaş M, Şaki Ce, Altay K, Ütük AE. Seroprevalance of cattle Hydatidosis in some districts in the east Anatolian region of Turkey.Turk J Vet Anim Sci. 2005; 29:1305–1310.

Parsa F, Haghpanah B, Pestechian N, Salehi M. Molecular epidemiology of Echinococcus granulo-sus strains in domestic herbivores of Lorestan, Iran. Jundishapur J Microbiol. 2011; 4(2):123-130.

Erdem A, Simsek S, Koroglub E, Donald P (2008). Molecular genetics characterization of different isolates of Echinococcus granulosus in east and southeast regions of Turkey. Acta Trop. 2008; 107:192–194.

Ahmadi N, Dalimi A. Characterization of Echinococcus granulosus isolates from human, sheep and camel in Iran. Infect Genet Evol. 2006; 6:85-90. 5. Sharbatkhori M, Mirhendi H, Jex AR, Pangasa A, Campbell BE, Kia EB, Eshraghian MR, Ha-randi MF, Gasser RB. Genetic categorization of Echinococcus granulosus from humans and her-bivorous hosts in Iran using an integrated mu-tation scanning phylogenetic approach. Elec-trophoresis. 2009; 30(15): 2648-55.

Zhang L, Eslami A, Hosseini SH, McManus DP. Indication of the presence of two distinct strains of Echinococcus granulosus in Iran by mito-chondrial DNA markers. Am J Trop Med Hyg. 1998; 59:171-174. 7. Keith Stanley McLachlan. The Boundaries of Mod-ern Iran, Published by UCL Press: 1994; pp.: 55.

Varcasia A, Canu S, Lightowlers MW, Scala A, Garippa G. Molecular characterization of Echinococcus granulosus strains in Sardinia. Parasi-tol Res. 2006; 98: 273-277.

Thompson RCA, Mc Manus DP (2001). Aeti-ology: parasites and life cycles. In: WHO/OIE Manual on Echinococcosis in Human and animals: A public health problem of global concern .Eds, Eckert J, Gemmell MA, Meslin FX, Pawlowski ZS. World Organization for Animal Health Paris. 2001; pp.: 1-19.

Bart JM, Abdukader M, Zhang YL et al. Geno-typing of human cystic echinococcosis in Xin-jiang, PR China. Parasitology. 2006; 133:571-579.

Li T, Ito A, Nakaya K, et al. Species identifi-cation of human echinococcosis using histo-pathology and genotyping in northwestern China. Trans R Soc Trop Med Hyg. 2008; 102(6): 585-590.

Thompson RCA. Biology and systematics of Echinococcus. In: Echinococcus and Hydatid Disease . Eds, Thompson RCA, Lymbery AJ. UK. 1995; pp.: 1-50.

Thompson RCA. Echinococcosis. In: Principles and Practice of Clinical Parasitology .Eds, Gillespie SH, Pearson, RD. UK. 2001; pp.595-612.

Thompson RCA, Mc Manus DP. Aetiology: parasites and life cycles. In: WHO/OIE Manual on Echinococcosis in Human and animals: A public health problem of global concern .Eds, Eckert J, Gemmell MA, Meslin FX, Pawlowski ZS. World Organization for Animal Health Paris. 2001; pp.: 19-29.

Bowles J, McManus DP. Rapid discrimination of Echinococcus species and strains using a poly-merase chain reaction-based RFLP method. Mol Biochem Parasitol.1993; 57:231-239.

Bhattacharya D, Bera AK, Bera BC, Pan D, Das SK. Molecular appraisal of Indian animal isolates of Echinococcus granulosus. Indian J Med Res. 2008; 127:383-287.

Villalobos N, et al. Molecular identification of Echinococcus granulosus genotypes (G1 and G7) isolated from pigs in Mexico. Vet Parasitol. 2007; 20(147): 185-9.

Jamali R, Ghazanchaei A, Asgharzadeh M. Identification and Characterization of Echinococ-cus granulosus by PCR-RFLP Technique. Tabriz District. Journal of Parasitic Diseases. 2008; 28(2): 69-72.

M'rad S, Filisetti D, Oudni M, et al. Molecu-lar evidence of ovine (G1) and camel (G6) strains of Echinococcus granulosus in Tunisia and putative role of cattle in human contamination. Vet Parasitol. 2005; 129: 267-272.

Mwambete KD, Ponce-Gordo F, Cuesta-Ban-dera C. Genetic identification and host range of the Spanish strains of Echinococcus granulosus. Acta Trop. 2004; 91: 87-93.

Schneider R, Gollackner B, Edel B et al. De-velopment of a new PCR protocol for the de-tection of species and genotypes (strains) of Echinococcus in formalin-fixed, parafin emb-edded tissues. Int J Parasitol. 2008; 38: 1065- 1071.

Rokni MB. Echinococcosis /hydatidosis in Iran. Iranian J Parasitol. 2009; 4(1):1-16.

Dousti M, Abdi J, Bakhtiyari S, Mohebali M, Mirhendi SH, Rokni MB. Genotyping of Hydatid Cyst Isolated from Human and Do-mestic Animals in Ilam Province, Western Iran Using PCR-RFLP. Iranian J Parasitol. 2013; 8(1):47-52.

How to Cite
1.
Hanifian H, Diba K, Tappeh KH, Mohammadzadeh H, Mahmoudlou R. Identification of Echinococcus granulosus Strains in Isolated Hydatid Cyst Specimens from Animals by PCR-RFLP Method in West Azerbaijan - Iran. IJPA. 8(3):376-81.
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