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. 2017 Oct-Dec;12(4):614-621.

Genotyping Echinococcus granulosus from Canine Isolates in Ilam Province, West of Iran

Affiliations

Genotyping Echinococcus granulosus from Canine Isolates in Ilam Province, West of Iran

Abdolhossein Dalimi et al. Iran J Parasitol. 2017 Oct-Dec.

Abstract

Background: Cystic Echinococcosis (CE) is one of the most common parasitic zoonosis caused by Echinococcus granulosus worldwide. This study investigated genotype diversity of Echinococcus granulosus isolated from stray dogs and golden jackals in Ilam province, West of Iran.

Methods: Adult worms were collected from the small intestine of the stray dogs and golden jackals from Ilam Province roads during 2012-2014. DNA was extracted from the adult worms and the partial mitochondrial NADH dehydrogenase subunit1 (nad1) was amplified by PCR then the products were digested by using HpaII, Rsa1, and Alu1 restriction enzymes. In order to confirm RFLP results, a number of PCR products were bi-directionally sequenced.

Results: Totally, 20 stray dogs out of 75 (26.66%) and two out of 73 (2.74%) golden jackal showed infection with E. granulosus. Amplified PCR product for all isolates was a band of approximately 550bp. Alu1 digested the product into two bands of approximately 160bp and 390bp fragments, while the Rsa1 cut the product into 320bp and 230bp fragments and the HpaII had no effect on the PCR product for both dog and jackal samples. The isolate sequences of mtDNAnad1 gene indicated 100% homology with references G1, G2 and G3 sequences in the GenBank database.

Conclusion: The genotype of adult E. granulosus was similar to larval stage genotypes of parasite (G1-G3 complex).

Keywords: Echinococcus granulosus; Genotyping; Golden jackal; Iran; Stray dog.

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Conflict of interest statement

Conflict of Interest All authors declare that they have no conflict of interest.

Figures

Fig. 1:
Fig. 1:
The collection sites of canine examined from Ilam Province between 2012 and 2014
Fig. 2:
Fig. 2:
PCR products of mtDNA-nad1 of adult E. granulosuson agar gel: M: marker with 50bp molecular weight, Co.n: control negative, 1: dog, 2: jackal samples
Fig. 3:
Fig. 3:
Digestion pattern of 550bp PCR products of mtDNAnad-1 fragment with AluI enzyme on agar gel M: marker with 50bp molecular weight, Co.n: negative control, Pp: PCR products without enzyme, 1: dog, 2: jackal samples
Fig. 4:
Fig. 4:
Digestion pattern of 550bp PCR products of mtDNA-nad1 fragment with Rsa1 enzyme on agar gel M: marker with 50bp molecular weight, Co.n: negative control, Pp: PCR products without enzyme, 1: dog, 2: jackal samples
Fig. 5:
Fig. 5:
Digestion pattern of 550bp PCR products of mtDNA-nad1 fragment with HpaII enzyme on agar gel M: marker with 50bp molecular weight, Co.n: negative control, Pp: PCR products without enzyme, 1: dog, 2: jackal samples
Fig. 6:
Fig. 6:
Phylogenetic relationships among E. granulosus isolated from dogs and jackals based on mtDNA nad1 gene sequence. The evolutionary history was inferred using the neighbor-joining method, supported by 1000 bootstrap replicates

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