Effect of temperature on the morbidity and recovery of the Pacific Coast wireworm, Limonius canus, following contact with tefluthrin-treated wheat seeds

Late instars of the Pacific Coast wireworm, Limonius canus (LeConte) (Coleoptera: Elateridae), were exposed to wheat seeds treated with tefluthrin at 10 g active ingredient/100 kg wheat seed, for 2 min, at 10, 15, 20, and 25 °C. All wireworms were moribund within 20 min of first exposure and recovered fully within 7 h. Morbidity induction time (i) decreased considerably as the temperature (t) was increased from 10 to 15 °C, but did not decrease further when the temperature was increased from 15 to 25 °C. For each temperature, induction time decreased as wireworm weight (w) increased, so that . A strong negative temperature coefficient was observed in wireworm recovery, the time required for recovery (r) after exposure decreasing as both temperature (t) and wireworm weight (w) increased, so that r = 1102.47(w)^?0.1848 * e^?0.1012t. The temperature at which wireworms contact insecticides in the field may significantly affect the induction and duration of morbidity, and determine whether wireworms will become moribund before they are repelled by tefluthrin. The ability of wireworms to recover from tefluthrin‐induced morbidity may seriously limit the efficacy of this insecticide in reducing wireworm populations in the field.     

Possible aversion learning in the Pacific Coast wireworm

The effects of carbon dioxide and the induction of morbidity on aversion learning in larvae of the Pacific Coast wireworm Limonius canus LeConte (Coleoptera: Elateridae) are discussed. Wireworms preconditioned by exposing them one or four times to odour of Tefluthrin 20SC and Dividend XLRTA [Syngenta Crop Protection (Canada), Inc., Canada] during the induction of temporary morbidity subsequently contact tefluthrin‐treated wheat seeds in soil bioassays for as long as naïve (i.e. not preconditioned) larvae but are repelled four to five‐fold more frequently by Dividend‐treated seeds in soil bioassays than naïve wireworms, suggesting that wireworms are capable of associating a novel odour (i.e. Dividend) with morbidity but require a minimum of 10–15 min subsequent contact time with treated seeds before being repelled. Wireworms preconditioned by exposure to peppermint odour during the induction of morbidity are not subsequently repelled by peppermint odour in soil bioassays, suggesting that wireworms are either not capable of aversion learning or that the presence of a CO₂ source and/or a suitable host plant may override a negative cue (i.e. peppermint odour). In studies conducted in the absence of soil, a host plant and CO₂ production, wireworms are repelled slightly by droplets of 1.0% but not 0.1% peppermint oil. Previous exposure to peppermint odour or contact with peppermint oil‐treated filter paper during one induction of morbidity does not increase the repellency of wireworms to 1.0% peppermint oil significantly. Repellency to 1.0% peppermint oil is almost eliminated when morbidity is induced five times in the absence of peppermint odour but is restored when peppermint odour is present during preconditioning. These findings suggest that wireworm sensitivity to repellent compounds decreases when repeatedly made moribund, although the results are not sufficient to conclude that wireworms are capable of associative learning.     

Varietal susceptibility of potatoes to wireworm herbivory

1 Wireworms, the soil dwelling larvae of click beetles, Agriotes spp., have recently become a more prevalent pest of potatoes. The present study investigated whether potato varieties showed variable susceptibility to wireworm herbivory, and also tested whether increased susceptibility was associated with lower concentrations of glycoalkaloids. Twelve varieties were originally screened across a range of experimental scales, including laboratory and tunnel experiments and a large‐scale field trial involving over 2000 tubers. 2 In laboratory no‐choice tests, Maris Peer, Marfona and Rooster varieties were significantly more susceptible to wireworm attack, with 63% of tubers showing damage, compared with just 15% of the less susceptible varieties of King Edward, Nadine and Maris Piper. There was also greater tissue consumption and weight gain when wireworms were reared on the most susceptible varieties. 3 In choice tests, wireworms showed a significant preference for those varieties previously identified as being the most susceptible to wireworm herbivory (4.2 holes per tuber) compared with the least susceptible (1.2 holes per tuber). Similar patterns of susceptibility were seen in the field trial, although there was generally more variation in susceptibility. 4 In a tunnel experiment, Marfona and Maris Peer were significantly more susceptibile to wireworm attack (47% of tubers showing damage) compared with Nadine, King Edward and Maris Piper (27% of tubers showing damage). Although Nadine, in particular, had the highest glycoalkaloid concentrations (309.33 mg/kg) and lowest amounts of wireworm herbivory, the relationship between susceptibility and glycoalkaloid concentrations was weak, suggesting that this is unlikely to be the sole mechanism underpinning varietal susceptibility.     

The role of vector control in stopping the transmission of malaria: threats and opportunities

Malaria control, and that of other insect borne diseases such as dengue, is heavily dependent on our ability to control the mosquito populations that transmit these diseases. The major push over the last decade to reduce the global burden of malaria has been driven by the distribution of pyrethroid insecticide-treated bednets and an increase in coverage of indoor residual spraying (IRS). This has reduced malaria deaths by a third. Progress towards the goal of reducing this further is threatened by lack of funding and the selection of drug and insecticide resistance. When malaria control was initially scaled up, there was little pyrethroid resistance in the major vectors, today there is no country in Africa where the vectors remain fully susceptible to pyrethroids. The first pyrethroid resistance mechanisms to be selected produced low-level resistance which had little or no operational significance. More recently, metabolically based resistance has been selected, primarily in West Africa, which in some mosquito populations produces more than 1000-fold resistance. As this spreads the effectiveness of pyrethroid-based bednets and IRS will be compromised. New public health insecticides are not readily available. The pipeline of agrochemical insecticides that can be re-purposed for public health dried up 30 years ago when the target product profile for agricultural insecticides shifted from broad spectrum, stable, contact-acting insecticides to narrow spectrum stomach poisons that could be delivered through the plant. A public–private partnership, the Innovative Vector Control Consortium, was established in 2005 to stimulate the development of new public health pesticides. Nine potential new classes of chemistry are in the pipeline, with the intention of developing three into new insecticides. While this has been successfully achieved, it will still take 6–9 years for new insecticides to reach the market. Careful management of the resistance situation in the interim will be needed if current gains in malaria control are not to be reversed.     

Chronology of sodium channel mutations associated with pyrethroid resistance in Aedes aegypti

Aedes aegypti is the primary mosquito vector of dengue, yellow fever, Zika and chikungunya. Current strategies to control Ae. aegypti rely heavily on insecticide interventions. Pyrethroids are a major class of insecticides used for mosquito control because of their fast acting, highly insecticidal activities and low mammalian toxicity. However, Ae. aegypti populations around the world have begun to develop resistance to pyrethroids. So far, more than a dozen mutations in the sodium channel gene have been reported to be associated with pyrethroid resistance in Ae. aegypti. Co-occurrence of resistance-associated mutations is common in pyrethroid-resistant Ae. aegypti populations. As global use of pyrethroids in mosquito control continues, new pyrethroid-resistant mutations keep emerging. In this microreview, we compile pyrethroid resistance-associated mutations in Ae. aegypti in a chronological order, as they were reported, and summarize findings from functional evaluation of these mutations in an in vitro sodium channel expression system. We hope that the information will be useful for tracing possible evolution of pyrethroid resistance in this important human disease vector, in addition to the development of methods for global monitoring and management of pyrethroid resistance in Ae. aegypti.     

Stability of the resistance to lambda-cyhalothrin in the ladybird beetle Eriopis connexa

Natural enemies resistant to insecticides are expected to help control the remaining arthropod pests after insecticide application and, hence, prevent crop damage, pest resurgence, and resistance selection. Field-evolved resistance to lambda-cyhalothrin and to other pyrethroids exhibited by the neotropical ladybird beetle Eriopis connexa (Germar) (Coleoptera: Coccinellidae) has been characterized and enhanced under laboratory selection. In this study, we investigated the stability of the resistance and its relationship with detoxification enzyme activity and biological performance, which are important for a biocontrol agent. One subgroup of the resistant population of E. connexa was established without selection pressure (R-UNSEL) during eight generations, and compared to either a susceptible (SUS) or its parental resistant (R-SEL) population. The resistance ratio in R-UNSEL was reduced by 50% compared to R-SEL between the first and fourth generation without selection pressure, but stayed stable afterwards, from the fifth to the eighth generation. Despite eight generations without selection pressure and reduction in the resistance level, the resistance ratio in R-UNSEL was still 39× greater than in the SUS population. The reduced resistance in R-UNSEL correlated to reduction in esterase activity, but the R-UNSEL maintained greater activity than the SUS group. The absence of selection pressure and reduction in enzyme activity in R-UNSEL did not mitigate the adaptive costs, with 2.7× lower egg production compared to SUS females. These findings indicate that resistance to lambda-cyhalothrin in R-UNSEL has already stabilized in the population. Although the resistance ratio and detoxifying enzymes were reduced in R-UNSEL, the impact on fecundity was maintained. Furthermore, the absence of crossing with wild SUS individuals will allow the R-UNSEL offspring to retain the resistance allowing survival to lambda-cyhalothrin even when used at the highest recommended field rate.     

Sequence and transcript expression of the super-kdr locus of the horn fly,Haematobia irritans

In horn flies, Haematobia irritans irritans (Diptera: Muscidae) (Linnaeus, 1758), target site resistance to pyrethroids can be diagnosed by an allele-specific PCR that genotypes individual flies at both the super-kdr (skdr) and the knock down resistance (kdr) associated loci. When this technique uses genomic DNA as template, modifications, such as alternative RNA splicing and RNA editing are not specifically detected. Alternative splicing at the skdr locus has been reported in Dipterans; thus, the genomic DNA-based allele-specific PCR may not accurately reflect the frequency of the skdr mutation in horn fly field populations. To investigate if alternative splicing occurs at the skdr locus of horn flies, genomic DNA and cDNA sequences isolated from two wild populations and two laboratory-reared colonies with varying degrees of pyrethroid resistance were compared. There was no indication of alternative splicing at the super-kdr locus neither in the wild populations nor in the laboratory-reared colonies.     

Multiplicative interaction between the two major mechanisms of permethrin resistance,kdr and cytochrome P450‐monooxygenase detoxification,in mosquitoes

Epistasis is the nonadditive interaction between different loci which contribute to a phenotype. Epistasis between independent loci conferring insecticide resistance is important to investigate as this phenomenon can shape the rate that resistance evolves and can dictate the level of resistance in the field. The evolution of insecticide resistance in mosquitoes is a growing and world‐wide problem. The two major mechanisms that confer resistance to permethrin in Culex mosquitoes are target site insensitivity (i.e. kdr) and enhanced detoxification by cytochrome P450 monooxygenases. Using three strains of mosquitoes, and crosses between these strains, we assessed the relative contribution of the two independent loci conferring permethrin resistance, individually and when present together. We found that for all genotype combinations tested, Culex pipiens quinquefasciatus exhibited multiplicative interactions between kdr and P450 detoxification, whether the resistance alleles were homozygous or heterozygous. These results provide a basis for further analysis of the evolution and maintenance of insecticide resistance in mosquitoes.     

Mating behaviour and reproductive output in insecticide‐resistant and ‐susceptible strains of the maize weevil (Sitophilus zeamais)

Insecticide resistance is a broadly recognised and well‐studied management problem resulting from intensive insecticide use, which also provides useful evolutionary models of newly adapted phenotypes to changing environments. Two common assumptions in such models are the existence of fitness costs associated with insecticide resistance, which will place the resistant individuals at a disadvantage in insecticide‐free environments, and the prevalence of random mating among insecticide‐resistant and ‐susceptible individuals. However, cases of insecticide resistance lacking apparent fitness disadvantages do exist impacting the evolution and management of insecticide resistance. Assortative mating, although rarely considered, may also favour the evolution and spread of insecticide resistance. Thus, the possible existence of both conditions in the maize weevil (Sitophilus zeamais), a key pest of stored cereals, led to the assessment of the mating behaviour and reproductive fitness of insecticide‐resistant and ‐susceptible weevil strains and their reciprocal crosses. The patterns of female and male mating choice also were assessed. Although mating behaviour within and between weevil strains was similar without mate choice, mating within the resistant strain led to higher reproductive output than within the susceptible strain; inter‐strain matings led to even higher fertility. Thus, no apparent fitness cost associated with resistance seems to exist in these weevils, favouring the evolution of this phenotype that is further aided by the higher fertility of inter‐strain matings. Mate choice reduced latency to mate and no inter‐strain preference was detected, but female weevils were consistent in their mate selection between 1st and 2nd matings indicating existence of female mating preference among maize weevils. Therefore, if female mate selection comes to favour trait(s) associated with insecticide resistance, higher reproductive fitness will be the outcome of such matings favouring the evolution and spread of insecticide resistance among maize weevil populations reverting into a management concern.     

Carlota,a new genus of Agrypnini from the Valdivian Forests of Chile (Elateridae,Agrypninae, Agrypnini)

Carlota gen. n., with one included species C. coigue sp. n., is described and illustrated from the Valdivian forests of Chile. The relationships of this genus to other Agrypnini from Chile are discussed and generic key for Chilean Agrypninae genera is provided.     

Resistance to carbosulfan in Anopheles gambiae from Ivory Coast,based on reduced sensitivity of acetylcholinesterase

Resistance to carbosulfan, a carbamate insecticide, was detected in field populations of the malaria vector mosquito Anopheles gambiae Giles (Diptera: Culicidae) from two ecologically contrasted localities near Bouaké, Ivory Coast: rural M'bé with predominantly M form of An. gambiae susceptible to pyrethroids; suburban Yaokoffikro with predominantly S form of An. gambiae highly resistant to pyrethroids (96% kdr). The discriminating concentration of 0.4% carbosulfan (i.e. double the LC100) was determined from bioassays with the susceptible An. gambiae Kisumu strain. Following exposure to the diagnostic dosage (0.4% carbosulfan for 1 h), mortality rates of female An. gambiae adults (reared from larvae collected from ricefields) were 62% and 29% of those from M'bé and Yaokoffikro, respectively, 24 h post-exposure. Exposure for 3 min to netting impregnated with the operational dosage of carbosulfan 200 mg/m2 gave mortality rates of 88% of those from M'bé and only 12.2% for Yaokoffikro. In each case the control untreated mortality rate was insignificant. Biochemical assays to detect possible resistance mechanism(s) revealed the presence of insensitive AChE in populations of An. gambiae at both localities, more prevalent in the S form at Yaokoffikro than in M form at M'bé, as expected from bioassays results. Our study demonstrates the need to monitor carbamate resistance among populations of the An. gambiae complex in Africa, to determine its spread and anticipate vector control failure if these insecticides are employed.     

The re-emergence of the bed bug as a nuisance pest: implications of resistance to the pyrethroid insecticides

A global resurgence of bed bugs (Hemiptera: Cimicidae) has led to renewed scientific interest in these insects. The current bed bug upsurge appears to have started almost synchronously in the late 1990 s in Europe, the U.S.A. and Australia. Several factors have led to this situation, with resistance to applied insecticides making a significant contribution. With a growing number of insecticides (DDT, carbamates, organophosphates etc.) being no longer available as a result of regulatory restrictions, the mainstay chemistry used for bed bug control over the past few decades has been the pyrethroid insecticides. With reports of increasing tolerance to pyrethroids leading to control failures on the rise, containing and eradicating bed bugs is proving to be a difficult task. Consequently, several recent studies have focused on determining the mode of action of pyrethroid resistance in bed bug populations sourced from different locations. Correct identification of the factor(s) responsible for the increasing resistance is critical to the development of effective management strategies, which need to be based, wherever possible, on firm scientific evidence. Here we review the literature on this topic, highlighting the mechanisms thought to be involved and the problems currently faced by pest control professionals in dealing with a developing pandemic.     

Activity of conventional and reduced-risk insecticides for protection of grapevines against the rose chafer, Macrodactylus subspinosus (Coleoptera: Scarabaeidae)

Abstract:  Bioassays were conducted to compare the residual toxicity and leaf protection activity of conventional broad-spectrum and reduced-risk insecticides against the rose chafer, Macrodactylus subspinosus . Insecticides were applied to a Vitis labrusca (F.) vineyard and residues were aged for 1, 3 or 7 days before leaves were collected and exposed to beetles in no-choice tests. Azinphosmethyl caused rapid knockdown and mortality for up to 1 week after application, with 1-day-old residues providing 95.6% protection against feeding, dropping to 51.6% when residues were a week old. Fenpropathrin caused mortality and knockdown after beetles had been exposed to fresh residues for 72 h. Although these effects diminished as residues aged, this compound provided the best protection of leaves against beetle feeding, with 77.9% reduction in feeding compared with the control after 7 days of aging in the vineyard. Of the reduced-risk insecticides, imidacloprid caused the greatest initial mortality and knockdown of beetles, providing protection against feeding that was equivalent to azinphosmethyl. Exposure to azadirachtin caused a low level of knockdown and mortality when residues were 1- and 3-days old. Protection against feeding was also low, lasting for only 3 days. Beetles were minimally affected by capsaicin and kaolin, with mortality and knockdown seen only when beetles were exposed to 1-day-old residues for 72 h. Foliage protection from these compounds was minimal, with between 10 and 15% reduction in feeding injury. Results are discussed in relation to development of semi-field bioassay methods for evaluating reduced-risk insecticides, and the management of M. subspinosus within grape pest management programs.     

Mortality and knockdown effects of imidacloprid and methomyl in house fly (Musca domestica L., Diptera: Muscidae) populations

In this study, the knockdown and mortality effects of imidacloprid and methomyl were investigated. The residual surface applications were carried out to determine the knockdown effects (KDt₅₀ and KDt₉₅) and mortality (LD₅₀ and LD₉₅) induced by each insecticide. For mortality comparisons, the susceptible house fly (Musca domestica L., Diptera: Muscidae) of a WHO population and three natural field‐collected M. domestica populations from Turkey were used. In conclusion, it was found that the resistance to imidacloprid and methomyl was significantly higher in the field populations when compared to the susceptible population from WHO. The results showed that applicators and pest management decision‐makers should control and conduct an integrated pest management strategy by including biological agents to prevent the development of high levels of resistance in the field populations.     

Reproductive status and flight activity of the overwintered Colorado potato beetle

Mating behavior of post-diapause Colorado potato beetles, Leptinotarsa decemlineata (Say), was observed within an overwintering site, a rotated potato field, newly colonized potato plants, and under laboratory conditions. The influence of spring mating on beetle flight in the presence and in the absence of host plants was investigated using a computer-linked flight mill system. Diapause was terminated simultaneously in male and female beetles, and the first matings were observed as early as within the first 24 h after the beetles emerged from the soil (60–90 DD accumulated). The beetles mated within the overwintering site, the potato field, and the fields rotated out of potatoes. Mating status did not affect flight behavior of overwintered beetles; however, unfed beetles displayed higher flight activity than fed beetles. Most flight activity took place soon after flight muscle regeneration, and then declined sharply by the 5th day after flight initiation. Mating in or near overwintering sites soon after diapause termination might be an important factor in providing gene flow between insecticide-resistant and insecticide-susceptible Colorado potato beetle populations, and should be considered in designing resistance management plans.     

Serological identification of wireworm and staphylinid predators of the Australian soldier fly (Inopus rubriceps) and wireworm feeding on plant and animal food

Pasture wireworm, Conoderus exsul (Sharp) fed more on larvae of the Australian soldier fly (Inopus rubriceps (Macquart)) than on germinating wheat seed when given a choice in the laboratory. Conversely, Agrypnus variabilis (Candeze), another wireworm found in New Zealand pastures, fed more on wheat seed. As determined by a serological technique (immuno-osmophoresis), 19% of C. exsul larvae, 20% of A. variabilis larvae and 80% of the adults of a staphylinid, Thyreocephalus oxthodoxus, (Olliff) collected in a pasture, had fed on I. rubriceps larvae. A predation rate of 12 soldier fly larvae/m²/day was estimated for C. exsul larvae.

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Résumé Pour déterminer la valeur de Conoderus exsul Sharp comme prédateur d'Inopus rubriceps Macquart, ravageur des prairies en Nouvelle Zélande, cet élatéride a été sur nourri des végétaux et animaux. En 72 heures, il s'est alimenté plus abondamment sur I. rubriceps que sur grains de blé. Par contre, un autre élatéride des prairies, Agrypnus variabilis Candeze, a consommé plus de grains de blé que de I. rubriceps.Une technique sérologique, l'immuno-osmophorèse, a montré que 19% des larves de C. exsul, 20% de celles de A. variabilis et 80% des adultes du staphylin, Thyreocephalus orthodoxus Olliff récoltés dans la nature, avaient consommé des larves de I. rubriceps.En se basant sur la densité en Elatérides, sur le pourcentage de réactions positives, sur les taux de consommation quotidiens, et la période d'observation de proies dans le tube digestif des prédateurs, on peut estimer que 12 larves/m²/jour de I. rubriceps ont été détruites par les larves de C. exsul pendant les 7 semaines de la période utilisée.

     

Effects of permethrin at different temperatures on pyrethroid-resistant and susceptible strains of Anopheles

The influence of temperature (16, 22, 28, 37 degrees C) on effects of permethrin was investigated for susceptible and pyrethroid-resistant strains of the mosquitoes Anopheles gambiae and An. stephensi (Diptera: Culicidae). Young unfed female adult mosquitoes were exposed to 0.25% permethrin test papers or to polyester netting treated with permethrin 500mg a.i./m2. The time to 50% knockdown (KT50) declined as temperature increased, i.e. there was a positive temperature coefficient of this effect of the pyrethroid. Resistance ratios (comparing KT50 values) between resistant and susceptible An. stephensi ranged between 2.5 and 4.4 at the different temperatures. Comparative tests of pyrethroid tolerance of different strains would be valid over the 22-28 degrees C range but, when using a discriminating dose to detect resistance, more precise temperature control is desirable. Mortality 24h after exposure to 0.25% permethrin of both susceptible and resistant strains of An. stephensi showed a negative correlation with temperature between 16 and 22 degrees C and a positive correlation at higher temperatures. In An. gambiae, however, the correlation was positive over the whole range. Irritancy of permethrin-treated netting to Anopheles females (measured as time lapse until first flight take-off, and the number of take-offs during 7.5 min exposure) was positively correlated with temperature in all four strains and was much greater for the susceptible than the resistant strains.     

Pathological effects of the parasite Malameba locustae in males of the migratory grasshopper Melanoplus sanguinipes and its interaction with the insecticide, cypermethrin

Gross pathological changes in Malpighian tubules of adult male Melanoplus sanguinipes (Fab.) infected by the protozoan Malameba locustae (King & Taylor) included hypertrophy, rupture of the wall, loss of pigment and presence of melanotic lesions. Infected grasshoppers also had depleted fat bodies and were hyperactive and more resistant to the insecticide, cypermethrin.

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Résumé Des Melanoplus sanguinipes adultes mâles, infectés par le protozoaire parasite, Malameba locustae mûrissent au même taux et avec le même poids que les sains. Cependant, les acridiens infectés, une semaine et deux semaines après l'aptitude à l'envol, étaient hyperactifs et significativement plus résistants à la prostration provoquée par la cyperméthrine (insecticide pyrèthroïde). Les modifications pathologiques brutes observées sur les tubes de Malpighi des acridiens infectés comprenaient l'hypertrophie, la rupture de la paroi, la perte de la pigmentation et des lésions mélaniques. Les insectes infectés ont aussi un épuisement du corps gras. Ces modifications pathologiques ne sont apparemment pas responsables de l'augmentation de la résistance à la cyperméthrine des individus infectés puisque la plus grande résistance à la prostration a eu lieu chez ceux qui ont été exposés à M. locustae et traités simultanément au Triple-Sulpha-G pour empêcher le développment du parasite.

     

Glutathione S-transferase detoxification as a potential pyrethroid resistance mechanism in the maize weevil, Sitophilus zeamais

Insecticide resistance patterns among 16 Brazilian populations of the maize weevil, Sitophilus zeamais Motschulsky (Coleoptera: Curculionidae), were recognized by surveying resistance to three organophosphates (chlorpyrifos‐methyl, malathion, and pirimiphos‐methyl) and three pyrethroids (cypermethrin, deltamethrin, and permethrin). Two population clusters were obtained: one with three populations (Bragança Paulista, Cristalina, and Nova Andradina) showing low frequency of cypermethrin resistance (13–36%) and negligible frequency of deltamethrin resistance (2–9%); and another with six populations (Campos dos Goytacazes, Ivinhema, Patos de Minas, Penápolis, Uberlândia, and Venda Nova) showing low to negligible levels of pyrethroid resistance (0–23%). The remaining seven populations, including a susceptible, and a DDT‐ and pyrethroid‐resistant reference populations (Sete Lagoas and Jacarezinho, respectively), were significantly different from each other and from the two recognized clusters. In contrast with pyrethroid resistance, organophosphate resistance was negligible except for chlorpyrifos‐methyl in two populations (Fátima do Sul and Penápolis). There was no correlation between geographic distance and the Mahalanobis distance estimated from the resistance pattern ordination of the populations by canonical variate analysis, suggesting local selection and/or broad dispersal of resistant populations by grain trade. The results of biochemical in vitro studies measuring the activity of detoxification enzymes (esterases and glutathion S‐transferases) in conjunction with canonical correlation analysis suggest a major involvement of enhanced conjugation by glutathione S‐transferases (> 2‐fold increase) in pyrethroid resistance and, in the case of cypermethrin resistance, enhanced phosphotriesterase activity.