title: Phenobarbital induction and chemical synergism demonstrate the role of UDP-glucuronosyltransferases in detoxification of naphthalophos by Haemonchus contortus larvae.
abstract: We used an enzyme induction approach to study the role of detoxification enzymes in the interaction of the anthelmintic compound naphthalophos with Haemonchus contortus larvae. Larvae were treated with the barbiturate phenobarbital, which is known to induce the activity of a number of detoxification enzymes in mammals and insects, including cytochromes P450 (CYPs), UDP-glucuronosyltransferases (UDPGTs), and glutathione (GSH) S-transferases (GSTs). Cotreatment of larvae with phenobarbital and naphthalophos resulted in a significant increase in the naphthalophos 50% inhibitory concentration (IC50) compared to treatment of larvae with the anthelmintic alone (up to a 28-fold increase). The phenobarbital-induced drug tolerance was reversed by cotreatment with the UDPGT inhibitors 5-nitrouracil, 4,6-dihydroxy-5-nitropyrimidine, probenecid, and sulfinpyrazone. Isobologram analysis of the interaction of 5-nitrouracil with naphthalophos in phenobarbital-treated larvae clearly showed the presence of strong synergism. The UDPGT inhibitors 5-nitrouracil, 4,6-dihydroxy-5-nitropyrimidine, and probenecid also showed synergistic effects with non-phenobarbital-treated worms (synergism ratio up to 3.2-fold). This study indicates that H. contortus larvae possess one or more UDPGT enzymes able to detoxify naphthalophos. In highlighting the protective role of this enzyme group, this study reveals the potential for UDPGT enzymes to act as a resistance mechanism that may develop under drug selection pressure in field isolates of this species. In addition, the data indicate the potential for a chemotherapeutic approach utilizing inhibitors of UDPGT enzymes as synergists to increase the activity of naphthalophos against parasitic worms and to combat detoxification-mediated drug resistance if it arises in the field.
authors.affiliation: CSIRO Agriculture Flagship, St. Lucia, Queensland, Australia andrew.kotze@csiro.au.
authors.forename: Andrew C
authors.initials: AC
authors.lastname: Kotze
authors.affiliation: CSIRO Agriculture Flagship, St. Lucia, Queensland, Australia.
authors.forename: Angela P
authors.initials: AP
authors.lastname: Ruffell
authors.affiliation: CSIRO Agriculture Flagship, St. Lucia, Queensland, Australia.
authors.forename: Aaron B
authors.initials: AB
authors.lastname: Ingham
mesh_terms: D000818:Animals; D000871:Anthelmintics / Q000378:metabolism / Q000494:pharmacology*; D004351:Drug Resistance / Q000187:drug effects; D004357:Drug Synergism; D004790:Enzyme Induction / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005786:Gene Expression Regulation; D014453:Glucuronosyltransferase / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D006189:Haemonchus / Q000187:drug effects* / Q000201:enzymology / Q000235:genetics; D015801:Helminth Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D008658:Inactivation, Metabolic / Q000187:drug effects; D007814:Larva / Q000187:drug effects* / Q000201:enzymology / Q000235:genetics; D009943:Organophosphorus Compounds / Q000378:metabolism / Q000494:pharmacology*; D010634:Phenobarbital / Q000494:pharmacology*; D011339:Probenecid / Q000494:pharmacology; D013442:Sulfinpyrazone / Q000494:pharmacology; D014498:Uracil / Q000031:analogs & derivatives / Q000494:pharmacology
chemical_list: D000871:Anthelmintics; D004791:Enzyme Inhibitors; D015801:Helminth Proteins; D009943:Organophosphorus Compounds; D014498:Uracil; C057449:5-nitrouracil; D014453:Glucuronosyltransferase; C008897:naftalofos; D011339:Probenecid; D013442:Sulfinpyrazone; D010634:Phenobarbital