Incidence of pyrethroid‐resistant oilseed rape pests in Germany

BACKGROUND: Failures in pollen beetle control using pyrethroids since 2005 indicated pyrethroid resistance in Germany. Therefore, resistance monitoring using bioassays was established in Germany for oilseed rape pest insects. RESULTS: The spread and intensity of pyrethroid resistance of Meligethes aeneus increased from 2005 onwards, with no sensitive samples left in any region of Germany in 2011. Sensitivity also declined for the newly introduced actives bifenthrin, etofenprox (both class‐I pyrethroids) and tau‐fluvalinate; all three claimed to be less affected by resistance, although there was no clear cross‐resistance to lambda‐cyhalothrin (class‐II pyrethroid). In the German region with the longest tradition and high intensity of oilseed rape production, pyrethroid resistance of Psylliodes chrysocephala and Ceutorhynchus obstrictus, with resistance factors of up to 81 and 140 respectively, was detected. CONCLUSION: The intensive use of only one mode of action for many years is risky, because even pest insects with a low intrinsic resistance risk may develop resistance. Therefore, resistance strategies need to include several control options for pest insects needing regular treatments. Copyright © 2012 Society of Chemical Industry     

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     

Insecticide-resistant pollen beetles (Meligethes aeneus F) found in Danish oilseed rape (Brassica napus L) fields

The pollen beetle is the most important pest in Danish oilseed rape fields. In 2001, we screened a broad range of pollen beetle populations for pyrethroid and dimethoate resistance. A standard dip-test was used to test insecticide resistance in 18 populations collected from oilseed winter and spring rape fields. The beetles were treated with four different insecticides: the pyrethroids tau-fluvalinate, lambda-cyhalothrin and esfenvalerate, and the organophosphate dimethoate. The results show that up to 99% of the pollen beetles survived Danish standard doses of pyrethroids and up to 36% of the beetles survived standard doses of dimethoate.     

Pyrethroid resistance and cross‐resistance in the German cockroach,Blattella germanica (L)

A German cockroach (Blatella germanica (L)) strain, Apyr‐R, was collected from Opelika, Alabama after control failures with pyrethroid insecticides. Levels of resistance to permethrin and deltamethrin in Apyr‐R (97‐ and 480‐fold, respectively, compared with a susceptible strain, ACY) were partially or mostly suppressed by piperonyl butoxide (PBO) and S,S,S,‐tributylphosphorotrithioate (DEF), suggesting that P450 monooxygenases and hydrolases are involved in resistance to these two pyrethroids in Apyr‐R. However, incomplete suppression of pyrethroid resistance with PBO and DEF implies that one or more additional mechanisms are involved in resistance. Injection, compared with topical application, resulted in 43‐ and 48‐fold increases in toxicity of permethrin in ACY and Apyr‐R, respectively. Similarly, injection increased the toxicity of deltamethrin 27‐fold in ACY and 28‐fold in Apyr‐R. These data indicate that cuticular penetration is one of the obstacles for the effectiveness of pyrethroids against German cockroaches. However, injection did not change the levels of resistance to either permethrin or deltamethrin, suggesting that a decrease in the rate of cuticular penetration may not play an important role in pyrethroid resistance in Apyr‐R. Apyr‐R showed cross‐resistance to imidacloprid, with a resistance ratio of 10. PBO treatment resulted in no significant change in the toxicity of imidacloprid, implying that P450 monooxygenase‐mediated detoxication is not the mechanism responsible for cross‐resistance. Apyr‐R showed no cross‐resistance to spinosad, although spinosad had relatively low toxicity to German cockroaches compared with other insecticides tested in this study. This result further confirmed that the mode of action of spinosad to insects is unique. Fipronil, a relatively new insecticide, was highly toxic to German cockroaches, and the multi‐resistance mechanisms in Apyr‐R did not confer significant cross‐resistance to this compound. Thus, we propose that fipronil could be a valuable tool in integrated resistance management of German cockroaches. © 2001 Society of Chemical Industry     

Impact of alpha-cypermethrin on honey bees foraging on spring oilseed rape (Brassica napus) flowers in field conditions

BACKGROUND: Cruciferous oil-bearing crops have gained in importance worldwide. The expansion of the growing area of these crops has caused a proliferation of pests. Exposure to organophosphate, carbamate and pyrethroid insecticides has been associated with bee poisoning in food crops. This study examines the repellent effect of alpha-cypermethrin on the number of foraging honey bees, Apis mellifera L., on fields of spring oilseed rape, Brassica napus L. var. oleifera. RESULTS: The first experiment was conducted on differently sprayed 10 m(2) experimental plots where alpha-cypermethrin was applied at different times. Another experiment was conducted on a 4 ha seed production field divided into two parts: one part was treated with alpha-cypermethrin and the other was not treated with this insecticide. The results show that there was no difference in the number of honey bees between alpha-cypermethrin-treated and untreated patches. The result persisted through three observation years, regardless of varying flower and honey bee densities. CONCLUSION: No repellent effect of the insecticide on honey bees was found even 24 h after spraying. The density of oilseed rape flowers most likely played a major role in choosing the foraging area.     

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.     

Experiences in controlling resistant pollen beetle by type I ether pyrethroid Trebon 30 EC in Germany

Oil seed rape (OSR) is an important crop in Germany with a long growing history. Due to the prevalence of various springtime pests an average of two pyrethroid applications are made. Pyrethroid resistance in Germany was found locally in 2003, widespread distribution of resistance was observed in 2006, a year of very strong pest pressure. Field trials in 2005 and the following years and laboratory analysis showed good control of resistant pollen beetle ( Meligethes aeneus ) by type I ether pyrethroid etofenprox, the active substance of Trebon 30 EC. Etofenprox differs significantly from common pyrethroids, which are usually ester pyrethroids including an α-amino-group and halogen substitutions. Trebon 30 EC has comparable efficacy on all important OSR pests but it has superior efficacy on resistant pollen beetle compared to common pyrethroids. To cope with pyrethroid resistance of pollen beetle, three different modes of action (MOA)s are postulated, which may be type I pyrethroids, neonicotinoids and organophosphates (OPs). Because type I pyrethroids, such as Trebon 30 EC, perform well at low temperatures, have a broad spectrum of activity, are considered as not to be toxic to bees, and give a high level of control of resistant pollen beetle, they will have an important role in the insecticide spraying regime of OSR.     

Monooxygenase mediated pyrethroid detoxification in sea lice (Lepeophtheirus salmonis)

The role of monooxygenases in detoxification of the pyrethroids cypermethrin and deltamethrin was examined. Four strains of sea lice (Lepeophtheirus salmonis Krøyer) with normal or moderately reduced sensitivity towards the pyrethroids were tested in bioassays by exposure to the pyrethroid alone and in combination with an oxygenase inhibitor, piperonyl butoxide (PBO). The normal (baseline) sensitivity was considered as the sensitivity range for the two most sensitive strains. Pre‐treatment with PBO elevated the sensitivity (P < 0.01) compared with groups exposed to the pyrethroid only. A positive, but not statistically significant, correlation between the activity of haem peroxidases and the pyrethroid concentration immobilizing 50% of the parasites was demonstrated (ρ = 0.500 for deltamethrin and ρ = 0.310 for cypermethrin). The results indicate that cytochrome P450 monooxygenases are involved in detoxification of pyrethroids in sea lice. ¹⁴C‐Deltamethrin was absorbed in a lesser amount in a group of sea lice exposed to a mixture of the compound and PBO than in a group exposed to ¹⁴C‐deltamethrin alone. A significant difference could be demonstrated both immediately after exposure (P < 0.01) and 24 h after exposure (P < 0.05). No significant differences were found between groups pre‐treated with PBO and groups exposed to ¹⁴C‐deltamethrin only. ¹⁴C‐Deltamethrin was taken up mainly through the cuticle, especially the cuticle on the extremities of the ventral surface, and subsequently distributed throughout the body of the parasite. Copyright © 2005 Society of Chemical Industry     

Effect of pretreatment with piperonyl butoxide on pyrethroid efficacy against insecticide-resistant Helicoverpa armigera (Lepidoptera: Noctuidae) and Bemisia tabaci (Sternorrhyncha: Aleyrodidae)

Pyrethroid resistance in B-type Bemisia tabaci Gennadius and Australian Helicoverpa armigera Hübner field populations is primarily conferred by esterase isoenzymes which metabolise and sequester pyrethroid insecticides. It has been shown previously that pyrethroid resistance-associated esterases in H. armigera are inhibited by the insecticide synergist piperonyl butoxide (PBO) over a 22-h period. It is demonstrated here that similar inhibition can be obtained against B-type B. tabaci. Small-scale field trials showed excellent levels of pyrethroid control when insects were pretreated with PBO and then dosed with pyrethroid during the time of maximum esterase inhibition. These results demonstrate that PBO can restore pyrethroid efficacy in the field against both B-type B. tabaci and resistant H. armigera.     

Genetics of Pyrethroid Resistance in a Strain (ALHF) of House Flies (Diptera: Muscidae)

Five house fly lines were derived from crosses of the pyrethroid-resistant ALHF (wildtype) and the susceptible aabys (bearing recessive morphological markers on each of five autosomes) strains. Each line was homozygous for one mutant-type marker from aabys. The level of resistance to permethrin was measured for each line to determine the genetic linkage of pyrethroid resistance in ALHF. Permethrin resistance in ALHF was 6600-fold compared with that in aabys. Resistance in flies bearing a mutant-type marker on autosome 4 was similar to that in ALHF. Flies with mutant-type markers on autosomes 1 and 2 had relatively lower resistance than ALHF; flies with mutant-type markers on autosomes 3 and 5 had much lower levels of resistance. These results demonstrated that factors on autosomes 3 and 5 play very important roles in pyrethroid resistance, whereas factors on autosomes 1 and 2 may have relatively small roles in resistance. Piperonyl butoxide (PBO) increased toxicity of permethrin in strains with mutant-type markers on autosomes 3 and 4 similar to that in ALHF. Slightly decreased synergism ratios in strains with autosomes 1 and 2 mutant-type markers compared with ALHF indicated that factors on autosomes 1 and 2 might make a small contribution in P450 monooxygenase-mediated resistance. However, when the autosome 5 mutant-type marker was present, PBO did not substantially decrease resistance, suggesting that the factor(s) on autosome 5 plays the most important role in P450 monooxygenase-mediated resistance. The resistance ratios of permethrin + PBO in strains with mutant-type markers on autosomes 1, 2, and 5 were significantly lower than those in ALHF, suggesting that factors on autosomes 1, 2, and 5 might be involved in pyrethroid resistance mechanisms other than P450-mediated detoxication. Injection did not change levels of resistance in the house flies tested, revealing that decreased rate of cuticular penetration (pen) probably does not play an important role in pyrethroid resistance in ALHF. The interaction and regulation of different mechanisms and/or factors involved in pyrethroid resistance in house flies are discussed.     

Differential efficacy of three commonly used pyrethroids against laboratory and field‐collected larvae and adults of Helicoverpa zea (Lepidoptera: Noctuidae) and significance for pyrethroid resistance management

BACKGROUND: Helicoverpa zea (Boddie) pyrethroid resistance monitoring programs typically utilize cypermethrin in the adult vial test. Here we investigated if differences in insect growth stage and pyrethroid structure affect resistance ratios and discuss implications for pyrethroid resistance management. RESULTS: Vial bioassays with cypermethrin, esfenvalerate and bifenthrin were conducted on H. zea third instars and male moths from a susceptible laboratory colony and the F1 generation of a pyrethroid‐resistant field population. In the susceptible population, both growth stages were most sensitive to bifenthrin and adults were more sensitive to esfenvalerate than cypermethrin. LC₅₀ resistance ratios for the larvae and adults of the resistant population were approximately two times higher for bifenthrin than cypermethrin or esfenvalerate. CONCLUSION: For the resistant population, vial assays using either growth stage gave similar resistance ratios for each of the three pyrethroids, respectively, proving the adult vial test accurately reflects larval resistance. However, as resistance ratios varied considerably depending on the pyrethroid used, resistance ratio values obtained with one pyrethroid may not be predictive of resistance ratios for other pyrethroids. Our results suggest that carefully chosen pyrethroid structures diagnostic for specific mechanisms of resistance could improve regional monitoring programs. Copyright © 2009 Society of Chemical Industry     

Target-site resistance to pyrethroids in European populations of pollen beetle, Meligethes aeneus F.

Pollen beetle, Meligethes aeneus F. (Coleoptera: Nitidulidae) is a major univoltine pest of oilseed rape in many European countries. Winter oilseed rape is cultivated on several million hectares in Europe and the continuous use of pyrethroid insecticides to control pollen beetle populations has resulted in high selection pressure and subsequent development of resistance. Resistance to pyrethroid insecticides in this pest is now widespread and the levels of resistance are often sufficient to result in field control failures at recommended application rates. Recently, metabolic resistance mediated by cytochrome P450 monooxygenases was implicated in the resistance of several pollen beetle populations from different European regions. Here, we have also investigated the possible occurrence of a target-site mechanism caused by modification of the pollen beetle para-type voltage-gated sodium channel gene. We detected a single nucleotide change that results in an amino acid substitution (L1014F) within the domain IIS6 region of the channel protein. The L1014F mutation, often termed kdr, has been found in several other insect pests and is known to confer moderate levels of resistance to pyrethroids. We developed a pyrosequencing-based diagnostic assay that can detect the L1014F mutation in individual beetles and tested more than 350 populations collected between 2006 and 2010 in 13 European countries. In the majority of populations tested the mutation was absent, and only samples from two countries, Denmark and Sweden, contained pollen beetles heterozygous or homozygous for the L1014F mutation. The mutation was first detected in a sample from Denmark collected in 2007 after reports of field failure using tau-fluvalinate, and has since been detected in 7 out of 11 samples from Denmark and 25 of 33 samples from Sweden. No super-kdr mutations (e.g. M918T) known to cause resistance to pyrethroids were detected. The implications of these results for resistance management strategies of pollen beetle populations in oilseed rape crops are discussed.     

Risk posed to honeybees (Apis mellifera L,Hymenoptera) by an imidacloprid seed dressing of sunflowers

In a greenhouse metabolism study, sunflowers were seed‐treated with radiolabelled imidacloprid in a 700 g kg^?1 WS formulation (Gaucho® WS 70) at 0.7 mg AI per seed, and the nature of the resulting residues in nectar and pollen was determined. Only the parent compound and no metabolites were detected in nectar and pollen of these seed‐treated sunflower plants (limit of detection <0.001 mg kg^?1). In standard LD₅₀ laboratory tests, imidacloprid showed high oral toxicity to honeybees (Apis mellifera), with LD₅₀ values between 3.7 and 40.9 ng per bee, corresponding to a lethal food concentration between 0.14 and 1.57 mg kg^?1. The residue level of imidacloprid in nectar and pollen of seed‐treated sunflower plants in the field was negligible. Under field‐growing conditions no residues were detected (limit of detection: 0.0015 mg kg^?1) in either nectar or pollen. There were also no detectable residues in nectar and pollen of sunflowers planted as a succeeding crop in soils which previously had been cropped with imidacloprid seed‐treated plants. Chronic feeding experiments with sunflower honey fortified with 0.002, 0.005, 0.010 and 0.020 mg kg^?1 imidacloprid were conducted to assess potential long‐term adverse effects on honeybee colonies. Testing end‐points in this 39‐day feeding study were mortality, feeding activity, wax/comb production, breeding performance and colony vitality. Even at the highest test concentration, imidacloprid showed no adverse effects on the development of the exposed bee colonies. This no‐adverse‐effect concentration of 0.020 mg kg^?1 compares with a field residue level of less than 0.0015 mg kg^?1 ( = limit of detection in the field residue studies) which clearly shows that a sunflower seed dressing with imidacloprid poses no risk to honeybees. This conclusion is confirmed by observations made in more than 10 field studies and several tunnel tests. © 2001 Society of Chemical Industry     

Detection of organophosphate and pyrethroid resistance alleles in Czech Leptinotarsa decemlineata (Coleoptera: Chrysomelidae) populations by molecular methods

BACKGROUND: Pyrethroids and organophosphates are the most frequently used insecticides for Colorado potato beetle, Leptinotarsa decemlineata (Say), control in the Czech Republic. Based on molecular methods, organophosphate and pyrethroid resistance alleles have been detected in samples from three sites. The accuracy of restriction fragment length polymorphism (RFLP) and bidirectional polymerase chain reaction amplification of specific alleles (Bi‐PASA) for detection of resistance alleles is compared. RESULTS: Leptinotarsa decemlineata from three sites showed higher frequencies of resistance alleles to organophosphates than to pyrethroids. The rates of occurrence of individuals homozygous resistant (RR) to pyrethroids ranged from 20.0 to 22.9%, while the rates of occurrence of individuals RR to organophosphates ranged from 52.9 to 66.7%. The incidences of individuals with resistance alleles to both organophosphates and pyrethroids ranged from 8.6 to 13.6%. No relationship was found between incidence of the pyrethroid resistance allele and site, while incidence of the organophosphate resistance allele differed significantly according to site. CONCLUSION: Both RFLP and Bi‐PASA were suitable for detecting resistance alleles to pyrethroids, and in most cases also for detecting resistance alleles to organophosphates. In contrast to Bi‐PASA, RFLP was also suitable for samples with lower DNA quality when testing for the resistance allele to pyrethroids. On the other hand, RFLP was not as accurate as Bi‐PASA in detection of the organophosphate resistance allele. Copyright © 2010 Society of Chemical Industry     

Pyrethroid tolerance in Culex pipiens pipiens var molestus from Marin County, California

In May 2001 a sample of Culex pipiens pipiens variety molestus Forskål from Marin County, California, collected as larvae and reared to adults, was found to show reduced resmethrin and permethrin knock‐down responses in bottle bioassays relative to a standard susceptible Cx pipiens quinquefasciatus Say colony (CQ1). Larval susceptibility tests, using CQ1 as standard susceptible, indicated that the Marin mosquitoes had LC₅₀ resistance ratios of 18.3 for permethrin, 12 for deltamethrin and 3.3 for pyrethrum. A colony of Marin was established and rapidly developed higher levels of resistance in a few generations after exposure to permethrin as larvae. These selected larvae were shown to cross‐resist to lambda‐cyhalothrin as well as to DDT. However, adult knock‐down time in the presence of permethrin, resmethrin and pyrethrum was not increased after increase in tolerance to pyrethroids as larvae. Partial and almost complete reversion to susceptibility as larvae was achieved with S, S, S‐tributylphosphorotrithioate and piperonyl butoxide (PBO), respectively, suggesting the presence of carboxylesterase and P450 monooxygenase mediated resistance. Insensitive target site resistance (kdr) was also detected in some Marin mosquitoes by use of an existing PCR‐based diagnostic assay designed for Cx p pipiens L mosquitoes. Carboxylesterase mediated resistance was supported by use of newly synthesized novel pyrethroid‐selective substrates in activity assays. Bottle bioassays gave underestimates of the levels of tolerance to pyrethroids of Marin mosquitoes when compared with mortality rates in field trials using registered pyrethroid adulticides with and without PBO. This study represents the first report of resistance to pyrethroids in a feral population of a mosquito species in the USA. Copyright © 2003 Society of Chemical Industry     

Investigating the potential of selected natural compounds to increase the potency of pyrethrum against houseflies Muscadomestica (Diptera: Muscidae)

BACKGROUND: A study was undertaken to determine the efficacy of seven natural compounds compared with piperonyl butoxide (PBO) in synergising pyrethrum, with the intention of formulating an effective natural synergist with pyrethrum for use in the organic crop market. RESULTS: Discriminating dose bioassays showed PBO to be significantly more effective at synergising pyrethrum in houseflies than the seven natural compounds tested, causing 100% mortality in insecticide‐susceptible WHO and resistant 381zb strains of housefly. The most effective natural synergists against WHO houseflies were dillapiole oil, grapefruit oil and parsley seed oil, with 59, 50 and 41% mortality respectively, compared with 18% mortality with unsynergised pyrethrum. Against 381zb houseflies, the most effective natural synergists were parsley seed oil and dillapiole oil. Esterase inhibition by the natural compounds and PBO in vitro showed no correlation with pyrethrum synergism in vivo, whereas the inhibition of oxidases in vitro more closely correlated with pyrethrum synergism in vivo. CONCLUSION: Dillapiole oil and parsley seed oil showed the greatest potential as pyrethrum synergists. PBO remained the most effective synergist, possibly owing to its surfactant properties, enhancing penetration of pyrethrins. The results suggest the involvement of oxidases in pyrethroid resistance in houseflies, with the efficacy of synergists showing a high correlation with inhibition of oxidases. Copyright © 2011 Society of Chemical Industry     

Cell death localization in situ in laboratory reared honey bee (Apis mellifera L.) larvae treated with pesticides

In this study, cell death detected by DNA fragmentation labeling and phosphatidylserine (PS) localization was investigated in the honey bee (Apis mellifera L.) midgut, salivary glands and ovaries after treating larvae with different pesticides offered via an artificial diet. To do this, honey bee larvae reared in an incubator were exposed to one of nine pesticides: chlorpyrifos, imidacloprid, amitraz, fluvalinate, coumaphos, myclobutanil, chlorothalonil, glyphosate and simazine. Following this, larvae were fixed and prepared for immunohistologically detected cellular death using two TUNEL techniques for DNA fragmentation labeling and Annexin V to detect the localization of exposed PS specific in situ binding to apoptotic cells. Untreated larvae experienced ∼10% midgut apoptotic cell death under controlled conditions. All applied pesticides triggered an increase in apoptosis in treated compared to untreated larvae. The level of cell death in the midgut of simazine-treated larvae was highest at 77% mortality and statistically similar to the level of cell death for chlorpyrifos (65%), imidacloprid (61%), myclobutanil (69%), and glyphosate (69%) treated larvae. Larvae exposed to fluvalinate had the lowest midgut columnar apoptotic cell death (30%) of any pesticide-treated larvae. Indications of elevated apoptotic cell death in salivary glands and ovaries after pesticide application were detected. Annexin V localization, indicative of apoptotic cell deletion, had an extensive distribution in the midgut, salivary glands and ovaries of pesticide-treated larvae. The data suggest that the tested pesticides induced apoptosis in tissues of honey bee larvae at the tested concentrations. Cell death localization as a tool for a monitoring the subclinical and sub-lethal effects of external influences on honey bee larval tissues is discussed.     

Occurrence of insecticide resistant pollen beetles (Meligethes aeneus F.) in Danish oilseed rape (Brassica napus L.) crops

The pollen beetle is the most important pest in Danish oilseed rape crops and it is essential that farmers are able to control this pest, especially in years when the economic damage threshold will be exceeded. About seven years ago, Danish oilseed rape growers began reporting that the pyrethroids seemed to be losing effectiveness towards pollen beetles. In 2001 18 populations collected from winter and spring oilseed rape fields were tested and very high level of insecticide resistance was found. In 2003 100 populations of pollen beetles were tested, using a FAO dip-test, for resistance to fenitrothion, lambda-cyhalothrin and tau-fluvalinate. It can be concluded that many Danish populations of pollen beetles are resistant to the pyrethroid lambda-cyhalothrin and to a lesser extent to the pyrethroid tau-fluvalinate. No resistance to the organophosphate fenitrothion was found.     

Buprofezin susceptibility survey, resistance selection and preliminary determination of the resistance mechanism in Nilaparvata lugens (Homoptera: Delphacidae)

BACKGROUND: Buprofezin has been used for many years to control Nilaparvata lugens (Stål). Assessment of susceptibility change in the insect is essential for maintaining control efficiency and resistance management. RESULTS: Eleven‐year surveys showed that most field populations were susceptible before 2004. However, substantially higher levels of resistance (up to 28‐fold) were found in most of the rice fields in China after 2004. A field population was collected and periodically selected for buprofezin resistance in the laboratory. After 65 generations (56 were selected), the colony successfully obtained 3599‐fold resistance to buprofezin. Synergism tests showed that O,O‐diethyl‐O‐phenyl phosphorothioate (SV1), piperonyl butoxide (PBO) and diethyl maleate (DEM) increased buprofezin toxicity in the resistant strain by only 1.5–1.6 fold, suggesting that esterases, P450‐monooxygenases and glutathione S‐transferases had no substantial effect on buprofezin resistance development. CONCLUSION: The results from this study indicate that N. lugens has the potential to develop high resistance to buprofezin. A resistance management program with rotation of buprofezin and other pesticides may efficiently delay or slow down resistance development in the insect. Further investigation is also necessary to understand the resistance mechanisms in N. lugens. Copyright © 2008 Society of Chemical Industry