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     

Degradation of Cry1Ab protein from genetically modified maize in the bovine gastrointestinal tract

Immunoblotting assays using commercial antibodies were established to investigate the unexpected persistence of the immunoactive Cry1Ab protein in the bovine gastrointestinal tract (GIT) previously suggested by enzyme-linked immunosorbent assay (ELISA). Samples of two different feeding experiments in cattle were analyzed with both ELISA and immunoblotting methods. Whereas results obtained by ELISA suggested that the concentration of the Cry1Ab protein increased during the GIT passage, the immunoblotting assays revealed a significant degradation of the protein in the bovine GIT. Samples showing a positive signal in the ELISA consisted of fragmented Cry1Ab protein of approximately 17 and 34 kDa size. Two independent sets of gastrointestinal samples revealed the apparent discrepancy between the results obtained by ELISA and immunoblotting, suggesting that the antibody used in the ELISA reacts with fragmented yet immunoactive epitopes of the Cry1Ab protein. It was concluded that Cry1Ab protein is degraded during digestion in cattle. To avoid misinterpretation, samples tested positive for Cry1Ab protein by ELISA should be reassessed by another technique.     

Kinetics and electron transport of soil hydrogenases catalyzing the oxidation of atmospheric hydrogen

The soil hydrogenases of chernozem and eolian sand were different with respect to their kinetic properties. Increase of soil moisture above optimum moisture content or prior incubation of the soils under very high H₂-mixing ratios (i.e. 1%) resulted in a decrease of V_max or in an increase of the K_m of the H₂ oxidation reaction. Under anaerobic conditions, the K_m for H₂ was higher and V_max was lower than under aerobic conditions. The anaerobic H₂-oxidation activity of both soils was stimulated by the addition of artificial electron acceptors with redox potentials of at least 80 mV. Ferricyanide as the most efficient stimulator did not function as a final electron acceptor for anaerobic H₂-oxidation, but acted as a catalyst by bypassing a rate-limiting electron transport step. In eolian sand, the aerobic as well as the anaerobic activity for atmospheric H₂ oxidation decreased upon exposure to very high H₂-mixing ratios (i.e. 1%). A similar effect was observed after incubation with ferricyanide which enabled the inflow of excess electrons from soil reductants or added NADH into the electron transport system of the soil hydrogenase with anaerobic activity. The activity for atmospheric H₂ oxidation was regenerated during incubation in H₂-free atmospheres, especially in the presence of oxygen. Inhibition and regeneration were probably due to alterations in components of the soil hydrogenases caused by the extent of a maximal electron flow through the electron transport system of the soil hydrogenases. Two classes of hydrogenase activities were discerned in eolian sand: one predominantly active under aerobic and the other under anaerobic conditions.     

Effects of chronic ingestion of ochratoxin a on blood levels and excretion of the mycotoxin in sheep

Ruminants are relatively resistant to the acutely toxic effects of ochratoxin A, due to extensive degradation of ochratoxin A to its less toxic metabolite ochratoxin alpha by rumen microorganisms. However, most estimates of the degradation capacity for ochratoxin A in ruminants are based on in vitro studies. In the current study, the metabolism of ochratoxin A was investigated over a period of 29 days, feeding various doses of the mycotoxin (0, 9.5, 19.0, and 28.5 mug ochratoxin A/kg body weight) to sheep. Animals were fed diets consisting of 70% concentrates and 30% grass silage. Significant concentrations of undegraded ochratoxin A were detected in serum of sheep at all levels of ochratoxin A tested. Serum concentrations of ochratoxin A slightly accumulated with time of exposure and were linearly dependent on the administered dose of ochratoxin A. Furthermore, a constant proportion (6-8%) of the dose was excreted in the urine. The results of this study indicate that even at moderate to low levels of ochratoxin A in the diet, considerable amounts of the mycotoxin are absorbed by ruminants and may accumulate in tissues. Therefore, feeding of ochratoxin A-contaminated feedstuffs to ruminants does not seem to be a reliable means for using these feedstuffs.     

Nitrification activity in tropical rain forest soils of the Coastal Lowlands and Atherton Tablelands,Queensland, Australia

Intact soil cores from a montane tropical rain forest site in the Atherton Tablelands (Kauri Creek) and from a lowland tropical rain forest site in the Coastal Lowlands (Bellenden Ker), Queensland, Australia were investigated during different hygric seasons for the magnitude of gross nitrification rates using the Barometric Process Separation technique (BaPS). Pronounced seasonal variations of gross nitrification rates were found at both sites with highest values during the transition period between dry and wet season (montane site: 24.0 mg N (kg SDW)^—1 d^—1; lowland site: 13.1 mg N (kg SDW)^—1 d^—1) and significantly lower rates of gross nitrification during the dry and wet season. Rates of gross nitrification were always higher at the montane site than at the lowland site, but the opposite was found for N₂O emissions. The results indicated that the high losses of N₂O at the lowland tropical rain forest site may be contributed largely by high denitrification activity due to its wetter and warmer climate as compared to the dryer and colder climate at the montane tropical rain forest site. This conclusion was supported by analysis of cell numbers of microbes involved in N‐cycling. Higher numbers of denitrifiers were present at the lowland site, whereas higher numbers of nitrifiers were found at the montane site.     

Sediment toxicity in a connected oxbow lake of the Upper Rhine (Germany): EROD induction in fish cells

Purpose  

Although the water quality of the Rhine River in Europe has improved, contaminated sediments may still threaten fish populations. This is particularly likely in Upper Rhine oxbow lakes, which are connected to the main river channel only at one end, except for extreme flood events. The impact of such extreme flood events on the sediment toxicity has, however, not yet been investigated, although numerous oxbow lakes exist with varying degrees of connection to their main river channels.     

Validation of CADESI-03, a severity scale for clinical trials enrolling dogs with atopic dermatitis

In dogs, atopic dermatitis (AD) is a common and chronic allergic skin disease that often necessitates treatment with pharmacological interventions. In the last 30 years, numerous clinical trials testing the efficacy of anti-inflammatory drugs have been reported, but there has been a lack of consistency in the assessment of outcome measures. Several clinical scales have been employed over time, but none of these scoring systems were ever tested for validity and reliability. A committee of the International Task Force on Canine Atopic Dermatitis evaluated the currently available scales used to assess disease morbidity in humans and dogs with AD, and a third version of the Canine Atopic Dermatitis Extent and Severity Index (CADESI-03) was designed. This version was expanded from previous ones by redistribution and increase in body sites tested, the use of an additional lesion reflecting underlying pruritus (e.g. self-induced alopecia) and an increase in the numerical range of severity for each lesion. The CADESI-03 scale was tested for validity and reliability in a cohort of 38 dogs with AD. Overall, this revised version of the CADESI was found to exhibit acceptable content, construct, criterion, and inter- and intra-observer reliability and sensitivity to change. As a result, this scale is recommended as a validated tool for assessment of disease severity in clinical trials testing the efficacy of interventions in dogs with AD.