Enantioselective mixture toxicity of the azole fungicide imazalil with the insecticide α-cypermethrin in Chironomus riparius: Investigating the importance of toxicokinetics and enzyme interactions

The fungicide imazalil is a chiral compound with one R- and one S-enantiomer. Enantiomers, while having the same chemical properties, can differ in their biological activity expressed as efficacy/toxicity as well as in their degradation kinetics and pathways. Azoles such as imazalil have been shown to synergize the effect of pyrethroid insecticides like alpha-cypermethrin through inhibition of cytochrome P450 monooxygenase responsible for pyrethroid detoxification. The aim of this study was to investigate, if the enantiomers of imazalil are selective in their synergistic potential in a mixture with a pyrethroid insecticide tested in Chironomus riparius. Potential enantioselectivity was studied on the level of uptake and elimination, inhibition of cytochrome P450 activity measured in vitro and in vivo and on synergistic potential of alpha-cypermethrin induced immobilization. Synergy was measured as an increase in alpha-cypermethrin toxicity after 144h applying a constant non-lethal imazalil concentration of 0.65 mu mol/L. The R- and S-imazalil enantiomers increased alpha-cypermethrin toxicity from an EC50 of 1580 +/- 980 pmol/L to an EC50 of 83 +/- 10 pmol/L and 53 +/- 8 pmol/L, respectively. The relatively small potency difference between imazalil enantiomers could not be explained by the in vitro cytochrome P450 inhibition, as the IC50 values were similar (0.11 +/- 0.01 and 0.09 +/- 0.01 mu mol/L for R- and S-imazalil). Measuring in vivo P450 inhibition and the toxicokinetic of imazalil did not show a clear trend of selectivity towards one or the other enantiomer. The study therefore suggests that cytochrome P450 enzymes involved in detoxification in C. riparius are not enantioselective for imazalil. (C) 2019 Elsevier Ltd. All rights reserved.