Molecular Identification of Fasciola and Dicrocoelium species isolates in ruminants live stock from Kashan and Arak in center of Iran

Fascioliasis and Dicrocoeliasis are important trematode infections that affect humans and ruminants worldwide. Molecular techniques have a conclusive role in detection liver flukes. The purpose of the current study was to find out the genotypic diversity of Fasciola and Dicrocoelium spp. isolated from different hosts in Iran. Totally, 160 and 200 adult Fasciola and Dicrocoelium spp. isolates collected from infected cattle, sheep, and goats from two abattoirs in the center of Iran. PCR-RFLP, and DNA sequences nuclear markers (18S, 28S, ITS) and the mitochondrial marker (ND1, CO1) were applied. PCR products of Dicrocoelium and Fasciola samples, were subjected to digestion by Bfa1, TruiI, BsrB1, 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 18S, 28S, ND1, CO1 of Fasciola at 260bp, 618bp, 700bp, and 500bp, and the length of the ITs2 and 28S of Dicrocoelium was 236bp and 963bp respectively. D. dendriticum has an RFLP pattern of 110, and 126bp (ITS2), and 116, 293, 409bp (28s) using, Bfa1 and Tru1I restriction enzymes. F. gigantica has a profile of 333, and 285bp (28s) using Bsrb1 enzyme. The RFLP pattern of genotype F . hepatica was 73, 120, and 507bp (ND1) and 119 and 381bp (CO1) in size using Hind III and ECO881 enzymes. Using the PCR-RFLP, three species of F. hepatica , F. gigantica , and D.dendriticum were identified. To uncover the genetic population structure of liver flukes across the country, future studies are still required.