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. 2014 Sep;9(3):358-64.

PCR-RFLP Analysis of 28 SrDNA for Specification of Fasciola gigantica (Cobbold, 1855) in the Infected Lymnaea auricularia (Linnaeus, 1785) Snails from Northwestern Iran

Mohammad Yakhchali  1 , Reza Malekzadeh-Viayeh  2 , Abass Imani-Baran  3
Affiliations

PCR-RFLP Analysis of 28 SrDNA for Specification of Fasciola gigantica (Cobbold, 1855) in the Infected Lymnaea auricularia (Linnaeus, 1785) Snails from Northwestern Iran

Mohammad Yakhchali et al. Iran J Parasitol. 2014 Sep.

Abstract

Background: Fasciolosis in livestock is a crucial concern in the globe, mainly due to its impact on human health. The aim of this study was to determine the rate of infection with Fasciola gigantica (Cobbold, 1855) larvae in the field-collected snails of Lymnaea auricularia (Linnaeus, 1785) from northwestern Iran using a molecular approach.

Methods: A total of 6,759 pond snails were collected from 28 freshwater bodies in West Azarbaijan. PCR was performed to amplify a 618-bp fragment of the nuclear 28 SrRNA gene of Fasciola. The PCR products were digested by AvaII restriction enzyme to create restriction fragment length polymorphism (RFLP) patterns specific for the detection of F. gigantica.

Results: Of the total collected snails 496 (7.34 %) were L. auricularia, among which 4.64% (23 out of 496) were infected with a Fasciola species according to the PCR analysis. Only 2.22% (11 out of 496) of the infected snails were from the mountainous areas. The highest Fasciola infection rate recorded in the snails of a single site was 1.81% (9 out of 496 snails). Based on the RFLP pattern, F. gigantica accounted for 2.42% of the infection rates in the study sites.

Conclusion: Application of PCR-RFLP was proven to be a useful approach for valid and robust detection of the infection with F. gigantica in its intermediate host snails. These findings may therefore be applicable for establishment of the control programs against dissemination of the infection in different regions.

Keywords: 28SrDNA; Fasciola gigantica; Iran; Lymnaea auricularia.

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Figures

Fig. 1
Fig. 1
Position of sampling sites and distribution of Lymnaea auricularia in West Azarbaijan Province (WAP), North West Iran. Go: Golestaneh; Is: Ismjondi: Zr: Zarineh-Roud
Fig. 2
Fig. 2
PCR-amplified 618-bp-long 28SrRNA of Fasciola sp. in Lymnaea auricularia (Lanes 1-14). N: negative control; P: positive control; M: 250bp DNA size marker
Fig. 3
Fig. 3
RFLP patterns of the PCR products digested by AvaII restriction enzyme. Lanes 1-5: two restricted fragments of 269 and 322bp-long specific for Fasciola gigantica in the infected Lymnaea auricularia snails. Lane 6: a Fasciola isolate other than F. gigantica. M: 250bp DNA size marker
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