title: Ivermectin selection on beta-tubulin: evidence in Onchocerca volvulus and Haemonchus contortus.
abstract: Ivermectin resistance is common in trichostrongylid nematodes of livestock, such as Haemonchus contortus. This anthelmintic is the only drug approved for mass administration to control onchocerciasis caused by the nematode parasite, Onchocerca volvulus. In parts of West Africa up to 18 rounds of ivermectin treatment have been administered to communities and there are reports of poor parasitological responses to treatment. Understanding ivermectin resistance and ivermectin selection is an important step to reduce selection pressure for resistance, and to develop molecular markers which can be used to monitor the development of resistance and its spread. Here we report evidence that ivermectin selection changes the frequency of beta-tubulin alleles in both the sheep parasite, H. contortus, and the human parasite, O. volvulus. In O. volvulus we have been able to look at the frequency of beta-tubulin alleles in O. volvulus obtained before any ivermectin was used in humans in Africa, and following its widespread use. In H. contortus, we have been able to look at the frequency of beta-tubulin alleles in a strain which has not seen any anthelmintic selection and in an ivermectin selected strain derived from the unselected strain. We have found ivermectin selects on beta-tubulin in both of these nematode species. In the case of O. volvulus, we had previously reported that ivermectin selects for specific single nucleotide polymorphisms in the O. volvulus beta-tubulin gene. This polymorphism results in three amino acid changes in the H3 helix of beta-tubulin, as well as deletions in an associated intron. We report a simple PCR assay to detect the amplicon length polymorphism, resulting from these intronic deletions, which can be used to monitor the frequency of the beta-tubulin allele selected for by ivermectin in O. volvulus.
authors.affiliation: Institute of Parasitology, McGill University, Sainte Anne-de-Bellevue, Quebec, Canada.
authors.forename: J K L
authors.initials: JK
authors.lastname: Eng
authors.forename: W J
authors.initials: WJ
authors.lastname: Blackhall
authors.forename: M Y
authors.initials: MY
authors.lastname: Osei-Atweneboana
authors.forename: C
authors.initials: C
authors.lastname: Bourguinat
authors.forename: D
authors.initials: D
authors.lastname: Galazzo
authors.forename: R N
authors.initials: RN
authors.lastname: Beech
authors.forename: T R
authors.initials: TR
authors.lastname: Unnasch
authors.forename: K
authors.initials: K
authors.lastname: Awadzi
authors.forename: G W
authors.initials: GW
authors.lastname: Lubega
authors.forename: R K
authors.initials: RK
authors.lastname: Prichard
mesh_terms: D000354:Africa, Western; D000818:Animals; D004351:Drug Resistance; D005369:Filaricides / Q000494:pharmacology / Q000627:therapeutic use*; D005787:Gene Frequency; D006188:Haemonchiasis / Q000188:drug therapy* / Q000469:parasitology; D006189:Haemonchus / Q000187:drug effects* / Q000235:genetics; D006801:Humans; D007559:Ivermectin / Q000494:pharmacology / Q000627:therapeutic use*; D008842:Microfilariae / Q000235:genetics / Q000302:isolation & purification; D017181:Onchocerca volvulus / Q000187:drug effects* / Q000235:genetics / Q000254:growth & development; D009855:Onchocerciasis / Q000188:drug therapy* / Q000469:parasitology; D012150:Polymorphism, Restriction Fragment Length; D018807:Polymorphism, Single-Stranded Conformational; D012756:Sheep / Q000469:parasitology; D012867:Skin / Q000473:pathology; D014404:Tubulin / Q000235:genetics*
chemical_list: D005369:Filaricides; D014404:Tubulin; D007559:Ivermectin