Intraspecific mitochondrial DNA variation of Fasciola hepatica eggs from sheep with different level of anthelmintic resistance.

In the current study, Fasciola hepatica strains of sheep with different degrees of resistance to anthelmintics were analyzed by sequencing the cytochrome C oxidase (COX1) and the NADH dehydrogenase (NAD1) subunits. The strains were as follows: LS, susceptible to all drugs tested; CS, resistant to albendazole and triclabendazole; and SV, resistant to albendazole and clorsulon. The molecular characterization was done in eggs recovered from sheep infected by LS and CS. In relation to SV, eggs were recovered before (SV0) and after a treatment with albendazole (SVA) and clorsulon (SVC). Nested PCRs were carried out to amplify a fragment of 798 bp of the COX1 subunit and 870 bp of the NAD1 subunit. The pairwise sequence identity between eggs was analyzed for each strain. Population diversity indices, neutrality indices, and the degree of gene flow among the strains were evaluated. As a result, we have shown that there was homogeneity in the demographic expansion of the studied strains, and, according to the pairwise fixation index, these were not genetically differentiated. Although we found that the resistant strains had lower pairwise percentage similarities, higher haplotype diversity, and higher frequencies of specific SNPs, especially in the COX1 subunit, these differences were not very significant. Therefore, we conclude that the presence of adult flukes resistant to anthelmintics does not result in significant higher genetic diversity in the mtDNA of their eggs.