Molecular identification of Fasciola and Dicrocoelium species isolates in ruminants livestock from Kashan and Arak in center of Iran.

Abstract

Fascioliasis and Dicrocoeliasis are important trematode infections that affect humans and ruminants worldwide. Molecular techniques have a conclusive role in detection of liver flukes. The purpose of the current study was to find outthe genotypic diversity of Fasciola and Dicrocoelium spp. isolated from different hosts in Iran. Totally, 160 and 200 adult Fasciola and Dicrocoelium spp. isolates were collected from infected cattle, sheep, and goatsfrom two abattoirs in the center of Iran.PCR-RFLP, and DNA sequences nuclear markers (18 S, 28 S, ITS) and the mitochondrial marker (ND1, CO1) were applied. PCR products of Dicrocoelium and Fasciola samples were subjected to digestion by BfaI, Tru1I, BsrBI, ECO881, and Hind III enzymes. DNA from 60 isolates of Fasciola and Dicrocoelium of different hosts were sequenced and evaluated. The PCR reaction showed the length of 18 S, 28 S, ND1, CO1 of Fasciola at 260 bp, 618 bp, 700 bp, and 500 bp, and the length of the ITS2 and 28 S of Dicrocoelium was 236 bp and 963 bp respectively. D. dendriticum has an RFLP pattern of 110, and 126 bp (ITS2), and 116, 293, 409 bp (28 S) using, BfaI and Tru1I restriction enzymes. F. gigantica has a profile of 333, and 285 bp (28 S) using BsrBI enzyme. The RFLP pattern of genotype F. hepatica was 73, 120, and 507 bp (ND1) and 119 and 381 bp (CO1) in size using Hind III and ECO881 enzymes. Using the PCR-RFLP, two species of Fasciola (F. hepatica and F. gigantica), and one species of Dicrocoelium (D. dendriticum) were identified. To uncover the genetic population structure of liver flukes across the country, future studies are still required.