High paraoxon-hydrolyzing activity in organophosphorus insecticide-resistant mosquitoes.

We found a strong paraoxon-hydrolyzing activity (23.4 +/- 8.50 nmol/h/individual and 137 +/- 86.2 nmol/h/mg protein) in the crude extract from larvae of Culex tritaeniorhynchus Toyama 89, which is markedly resistant to organophosphorous insecticides. The activity was higher than those from Cx. tritaeniorhynchus re-e-ae (0.175 +/- 0.0336 and 1.83 +/- 0.651), Anopheles omorii (0.112 +/- 0.0301 and 1.86 +/- 0.746) and An. stephensi (0.0651 +/- 0.0713 and 0.789 +/- 0.910), which are susceptible to organophosphorous insecticides. These facts suggest that the high paraoxon-hydrolyzing activity plays a role in the development of organophosphorous resistance in Cx. tritaeniorhynchus. The enzyme preparation obtained from Toyama 89 showed higher activity in the alkaline pH range and its Km values to paraoxon were 0.67 mM in larvae and 0.50 mM in adults. A calcium ion was strictly required for the hydrolysis of paraoxon. Fenitroxon was also hydrolyzed, in addition to paraoxon. However, it did not degradate parathion and fenitrothion at all. Dichlorvos and phenyl acetate competitively inhibited the enzyme. The phenyl acetate-hydrolyzing activity in the preparation of Toyama 89 was significantly (p less than 0.01) lower than those in susceptible strains, and was irreversibly inhibited by paraoxon. Therefore, the paraoxon-hydrolyzing activity belongs to the class of organophosphate compound hydrolases; it must be thus distinguished from bacterial phosphotriesterase.