Molecular characterization of cystic echinococcosis cysts isolated from cattle in Shiraz, Iran

Abstract

Cystic echinococcosis (CE), is one of the common parasitic infections of domestic animals and humans caused by the larval stages of the tapeworm Echinococcus granulosus. This research aimed to identify of CE cysts in cattle isolates from Shiraz slaughterhouse using molecular technique. Thirty CE cysts isolates were collected and total DNA was extracted from protoscolices or the germinal layer of each cyst followed by polymerase chain reaction (PCR) using cytochrome c oxidase I (cox1) and NADH dehydrogenase subunit 1 (NADH-1) mitochondrial genes (~ 450 bp) and sequencing. The sequence results were compared with available reference sequences in GenBank by BLAST system. Phylogenetic analysis was performed using MEGA 5.0 software and the maximum likelihood method. The results showed that all 30 samples examined in this study were identified as E. granulosus sensu stricto (G1-G3 genotypes) with G1 dominant.

Keywords: Cystic echinococcosis cyst; Echinococcus granulosus sensu stricto; Iran; Slaughtered cattle.

Fig. 1

Electrophoresis of PCR products, using primers of JB3 and JB4.5 for the COX1 fragment on 1.5% agarose gel. M: molecular weight marker, 1–10: isolated samples, +: positive control for COX1, and −: negative control

Fig. 2

Electrophoresis of PCR products, using primers of MS1 and MS2 for the NADH1 fragment on 1.5% agarose gel. M: molecular weight marker, 27–30: isolated samples, +: positive control for NADH1, and −: negative control

Fig. 3

Comparison of the sequences obtained in this study based on the COX1 gene with the sequences available in the gene bank. Numbers above branches correspond to bootstrap valued based on 1000 replicates

Fig. 4

Comparison of the sequences obtained in this study based on the NADH1 gene with the sequences available in the gene bank Numbers above branches correspond to bootstrap valued based on 1000 replicates