title: Introgression of ivermectin resistance genes into a susceptible Haemonchus contortus strain by multiple backcrossing.
abstract: Anthelmintic drug resistance in livestock parasites is already widespread and in recent years there has been an increasing level of anthelmintic drug selection pressure applied to parasitic nematode populations in humans leading to concerns regarding the emergence of resistance. However, most parasitic nematodes, particularly those of humans, are difficult experimental subjects making mechanistic studies of drug resistance extremely difficult. The small ruminant parasitic nematode Haemonchus contortus is a more amenable model system to study many aspects of parasite biology and investigate the basic mechanisms and genetics of anthelmintic drug resistance. Here we report the successful introgression of ivermectin resistance genes from two independent ivermectin resistant strains, MHco4(WRS) and MHco10(CAVR), into the susceptible genome reference strain MHco3(ISE) using a backcrossing approach. A panel of microsatellite markers were used to monitor the procedure. We demonstrated that after four rounds of backcrossing, worms that were phenotypically resistant to ivermectin had a similar genetic background to the susceptible reference strain based on the bulk genotyping with 18 microsatellite loci and individual genotyping with a sub-panel of 9 microsatellite loci. In addition, a single marker, Hcms8a20, showed evidence of genetic linkage to an ivermectin resistance-conferring locus providing a starting point for more detailed studies of this genomic region to identify the causal mutation(s). This work presents a novel genetic approach to study anthelmintic resistance and provides a "proof-of-concept" of the use of forward genetics in an important model strongylid parasite of relevance to human hookworms. The resulting strains provide valuable resources for candidate gene studies, whole genome approaches and for further genetic analysis to identify ivermectin resistance loci.
authors.affiliation: Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, United Kingdom.
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Redman
authors.forename: Neil
authors.initials: N
authors.lastname: Sargison
authors.forename: Fiona
authors.initials: F
authors.lastname: Whitelaw
authors.forename: Frank
authors.initials: F
authors.lastname: Jackson
authors.forename: Alison
authors.initials: A
authors.lastname: Morrison
authors.forename: David Jon
authors.initials: DJ
authors.lastname: Bartley
authors.forename: John Stuart
authors.initials: JS
authors.lastname: Gilleard
mesh_terms: D000818:Animals; D000871:Anthelmintics / Q000494:pharmacology*; D004351:Drug Resistance / Q000235:genetics*; D005818:Genetic Engineering / Q000379:methods*; D006189:Haemonchus / Q000235:genetics*; D007178:Inbreeding; D007559:Ivermectin / Q000494:pharmacology*; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D000871:Anthelmintics; D007559:Ivermectin