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.     

Pyrethroid resistance monitoring in European populations of pollen beetle (Meligethes spp.): a coordinated approach through the Insecticide Resistance Action Committee (IRAC)

BACKGROUND: Pollen beetle (Meligethes spp.) is a major pest of European oilseed rape crops. Its resistance to pyrethroid insecticides has been recorded in samples of beetles collected in Europe since at least 1999, and problems with the control of the beetle in the field have been widely reported. In 2007, a Pollen Beetle Working Group was formed through the Insecticide Resistance Action Committee (IRAC) in order to coordinate efforts for surveying pyrethroid resistance development. RESULTS: The results of the first 3 years of the pollen beetle pyrethroid susceptibility survey using a laboratory test are presented in this paper. Resistant beetle samples were collected from 20 of the 21 countries surveyed, with a general trend of increasing frequency and spread of resistant samples in European oilseed‐rape‐growing regions. CONCLUSION: Pyrethroid‐resistant beetles dominate in Western and Central Europe and are becoming established in the North and East, the main oilseed‐rape‐growing areas of Europe. The development and spread of pyrethroid‐resistant pollen beetles highlights the need for effective management strategies for oilseed rape insect pests. Copyright © 2011 Society of Chemical Industry     

Pyrethroid resistance and thiacloprid baseline susceptibility of European populations of Meligethes aeneus (Coleoptera: Nitidulidae) collected in winter oilseed rape

BACKGROUND: Pollen beetle, Meligethes aeneus F. (Coleoptera: Nitidulidae), is a major pest in European winter oilseed rape. Recently, control failures with pyrethroid insecticides commonly used to control this pest have been reported in many European countries. For resistance management purposes, the neonicotinoid insecticide thiacloprid was widely introduced as a new mode of action for pollen beetle control. RESULTS: A number of pollen beetle populations collected in Germany, France, Austria, Great Britain, Sweden, Denmark, Finland, Poland, Czech Republic and Ukraine were tested for pyrethroid resistance using lambda‐cyhalothrin‐coated glass vials (adult vial test). Most of the populations tested exhibited substantial levels of resistance to lambda‐cyhalothrin, and resistance ratios ranged from < 10 to > 2000. A similar resistance monitoring bioassay for the neonicotinoid insecticide thiacloprid was developed and validated by assessing baseline susceptibility data for 88 European pollen beetle populations. A variation of less than fivefold in response to thiacloprid was detected. The thiacloprid adult vial bioassay is based on glass vials coated with an oil‐dispersion‐based formulation of thiacloprid, resulting in a much better bioavailability compared with technical material. Analytical measurements revealed a > 56 and 28 day stability of thiacloprid and lambda‐cyhalothrin in coated glass vials at room temperature, respectively. No cross‐resistance between thiacloprid and lambda‐cyhalothrin based on log‐dose probit–mortality data was detected. CONCLUSION: Pyrethroid resistance in many European populations of M. aeneus was confirmed, whereas all populations are susceptible to thiacloprid when tested in a newly designed and validated monitoring bioassay based on glass vials coated with oil‐dispersion‐formulated thiacloprid. Based on the homogeneous results, it is concluded that thiacloprid could be an important chemical tool for pollen beetle resistance management strategies in European winter oilseed rape. Copyright © 2011 Society of Chemical Industry     

Incidence and spread of knockdown resistance (kdr) in German Colorado potato beetle (Leptinotarsa decemlineata Say) populations

The Colorado beetle, Leptinotarsa decemlineata (Say), is one of the most important pests in many potato‐producing regions. Colorado beetle infestations are normally kept under economic damage thresholds by applying insecticides such as pyrethroids. Pyrethroids are known to act on voltage‐gated sodium channels and have been used for several decades to control L. decemlineata. Their continuous and widespread use has resulted in the development of resistance, which is often linked to a L1014F target‐site mutation in the voltage‐gated sodium channel, known as knockdown resistance (kdr). Since pyrethroids are used in many potato‐growing regions in Germany, more than 140 L. decemlineata samples were collected and analysed for the presence of the kdr allele by pyrosequencing diagnostics. Results showed that kdr is present in many German L. decemlineata populations, but even its homozygous presence does not substantially compromise the efficacy of recommended label rates of pyrethroids, as demonstrated by bioassays and crossing experiments. The implications of these findings for resistance management are briefly discussed.     

Biochemical evidence that an S431F mutation in acetylcholinesterase-1 of Aphis gossypii mediates resistance to pirimicarb and omethoate

Molecular changes in acetylcholinesterase (AChE) leading to target-site resistance to carbamate and organophosphate insecticides have recently been identified. Of particular interest is the S431F mutation in ace2 and its orthologue ace1 of Myzus persicae Sulzer and Aphis gossypii Glover, respectively. This mutation has been correlated with resistance to pirimicarb, but biochemical evidence has not yet been provided. Here, we describe for the first time that recombinantly expressed AChE1 from A gossypii carrying the S431F mutation is insensitive to pirimicarb and omethoate, but sensitive to demeton-S-methyl and hypersensitive to carbofuran. Furthermore, site-directed mutagenesis of the serine residue at position 431 in ace1 from a pirimicarb-susceptible clone of A gossypii conferred insensitivity to pirimicarb. We conclude that AChE1 of A gossypii is the target of toxicological relevance of carbamates and organophosphates.     

Mode of action of etoxazole

The mode of action of the 2,4-diphenyl-1,3-oxazoline acaricide/insecticide etoxazole has been argued to be moulting inhibition, but experimental results supporting this hypothesis are lacking. This study investigated the effect of etoxazole on chitin biosynthesis in the fall armyworm, Spodoptera frugiperda (Smith) (Lepidoptera: Noctuidae). Etoxazole induced moulting defects in fall armyworm larvae similar, if not identical, to those caused by benzoylphenylureas, a well-known class of insecticidal chitin biosynthesis inhibitors. Furthermore, in contrast to untreated larvae, the chitin content in the integuments of larvae several days after treatment did not differ from that in freshly ecdysed individuals, thus suggesting strong chitin biosynthesis inhibition in vivo. A more detailed investigation of the inhibitory potential by incubating cultured integument pieces from larvae of S. frugiperda with [14C]N-acetyl-D-glucosamine, a radiolabelled chitin precursor, revealed I50 values of 2.95 and 0.071 microM for etoxazole and triflumuron respectively. The incorporation of radiolabel into potassium hydroxide-resistant material was inhibited by etoxazole in a dose-dependent manner. Based on these results, it is concluded that the acaricidal and insecticidal mode of action of etoxazole is chitin biosynthesis inhibition.     

Monitoring of spirodiclofen susceptibility in field populations of European red mites, Panonychus ulmi (Koch) (Acari: Tetranychidae), and the cross-resistance pattern of a laboratory-selected strain

BACKGROUND: Phytophagous mites such as the European red mite, Panonychus ulmi (Koch), are serious pests in European fruit tree orchards, and a number of acaricides are frequently used to control them. Spirodiclofen (Envidor^®) has been a commonly used acaricide for several years. In the present study, European field populations collected in 2009 and 2010 were checked for their susceptibility to spirodiclofen by using discriminating dose and full dose response bioassays. RESULTS: In 2009 and 2010, a total of 63 field populations (including winter eggs) of European red mites were collected in different European countries, and in several populations from south‐western Germany a shifting in susceptibility against spirodiclofen was observed. Full dose response bioassays on different developmental stages of field‐collected strains suggested an age‐dependent expression of resistance because eggs remain fully susceptible to spirodiclofen. Artificial selection with spirodiclofen of one of the field strains resulted in resistance ratios of > 7000. Synergism studies suggest a possible role of cytochrome‐P450‐dependent monooxygenases in spirodiclofen detoxification. Most of the other acaricides from different chemical classes displayed no or low cross‐resistance in a spirodiclofen‐selected strain. CONCLUSION: In order to preserve spirodiclofen as an important tool in spider mite resistance management, the efficacy situation should be continuously monitored, and it is suggested that spirodiclofen be alternated with acaricides coming from different mode‐of‐action classes. An observed age‐specific expression of resistance revealed full susceptibility of eggs, so targeting spirodiclofen particularly against eggs is likely to reduce the selection pressures imposed on other life stages. Copyright © 2011 Society of Chemical Industry