Cytochrome P450 mediated pyrethroid resistance in European populations of Meligethes aeneus (Coleoptera: Nitidulidae)

Pollen beetle, Meligethes aeneus (Coleoptera: Nitidulidae) is a major pest on several million hectares in European winter oilseed rape cultivation. Synthetic pyrethroids have been successfully used for many years to keep them under economic damage thresholds. Recently wide-spread resistance development to pyrethroids in pollen beetle populations was described in many European countries, including Germany, France, Poland, Denmark and others. Resistance monitoring is conducted by incubating beetles for 24 h in glass vials coated with different concentrations of lambda-cyhalothrin. Using such an assay format we were able to show cross-resistance to other pyrethroids, such as deltamethrin, cypermethrin, and to a somewhat lower extent bifenthrin, etofenprox and tau-fluvalinate. Here we also investigated in more detail in 27 different populations the biochemical mechanism of pyrethroid resistance. Synergism experiments revealed a high synergistic potential for piperonyl butoxide in vivo, whereas other compounds such as S,S,S-tributylphosphorotrithioate and diethylmaleimide failed to suppress pyrethroid resistance. Incubating microsomal fractions of pollen beetle with deltamethrin and subsequent LC–MS/MS analysis revealed 4-OH-deltamethrin as the major metabolite. Metabolite formation in vitro and pyrethroid resistance in vivo is correlated and inhibition trials with piperonyl butoxide, tebuconazole and aminobenzotriazole suggest the involvement of cytochrome P450′s. Furthermore we were able to show cross-resistance to tau-fluvalinate which is supported by the competitive inhibition of 4-OH-deltamethrin formation by increasing concentrations of tau-fluvalinate in microsomal hydroxylation assays. Although we provided clear experimental evidence for an oxidative mechanism of resistance in numerous populations, other mechanisms might be involved based on the data discussed.