title: Evidence from two independent backcross experiments supports genetic linkage of microsatellite Hcms8a20, but not other candidate loci, to a major ivermectin resistance locus in Haemonchus contortus.
abstract: Haemonchus contortus is the leading parasitic nematode species used to study anthelmintic drug resistance. A variety of candidate loci have been implicated as being associated with ivermectin resistance in this parasite but definitive evidence of their importance is still lacking. We have previously performed two independent serial backcross experiments to introgress ivermectin resistance loci from two H. contortus ivermectin-resistant strains - MHco4(WRS) and MHco10(CAVR) - into the genetic background of the ivermectin-susceptible genome reference strain MHco3(ISE). We have interrogated a number of candidate ivermectin resistance loci in the resulting backcross populations and assessed the evidence for their genetic linkage to an ivermectin resistance locus. These include the microsatellite marker Hcms8a20 and six candidate genes Hco-glc-5, Hco-avr-14, Hco-lgc-37 (previously designated Hco-hg-1), Hco-pgp-9 (previously designated Hco-pgp-1), Hco-pgp-2 and Hco-dyf-7. We have sampled the haplotype diversity of amplicon markers within, or adjacent to, each of these loci in the parental strains and fourth generation backcross populations to assess the evidence for haplotype introgression from the resistant parental strain into the genomic background of the susceptible parental strain in each backcross. The microsatellite Hcms8a20 locus showed strong evidence of such introgression in both independent backcrosses, suggesting it is linked to an important ivermectin resistance mutation in both the MHco4(WRS) and MHco10(CAVR) strains. In contrast, Hco-glc-5, Hco-avr-14, Hco-pgp-9 and Hco-dyf-7 showed no evidence of introgression in either backcross. Hco-lgc-37 and Hco-pgp-2 showed only weak evidence of introgression in the MHco3/4 backcross but not in the MHco3/10 backcross. Overall, these results suggest that microsatellite marker Hcms8a20, but not the other candidate genes tested, is linked to a major ivermectin resistance locus in the MHco4(WRS) and MHco10(CAVR) strains. This work also emphasises the need for genome-wide approaches to identify mutations responsible for the ivermectin resistance in this parasite.
authors.affiliation: Department of Comparative Biology and Experimental Medicine, Faculty of Veterinary Medicine, University of Calgary, Alberta, Canada.
authors.forename: Andrew M
authors.initials: AM
authors.lastname: Rezansoff
authors.affiliation: Institute of Biodiversity, Animal Health and Comparative Medicine, College of Medical, Veterinary and Life Sciences, University of Glasgow, Scotland, UK.
authors.forename: Roz
authors.initials: R
authors.lastname: Laing
authors.affiliation: Department of Comparative Biology and Experimental Medicine, Faculty of Veterinary Medicine, University of Calgary, Alberta, Canada. Electronic address: jsgillea@ucalgary.ca.
authors.forename: John S
authors.initials: JS
authors.lastname: Gilleard
mesh_terms: D000818:Animals; D000977:Antiparasitic Agents / Q000494:pharmacology*; D003001:Cloning, Molecular; D018509:DNA, Helminth / Q000378:metabolism; D004351:Drug Resistance / Q000235:genetics*; D005260:Female; D008040:Genetic Linkage; D006189:Haemonchus / Q000187:drug effects* / Q000235:genetics*; D006239:Haplotypes; D007559:Ivermectin / Q000494:pharmacology*; D008297:Male; D018895:Microsatellite Repeats; D009154:Mutation; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
chemical_list: D000977:Antiparasitic Agents; D018509:DNA, Helminth; D007559:Ivermectin
keywords: Anthelmintic; Drug resistance; Haemonchus contortus; Ivermectin; Nematode