Binding of bis(p-chlorophenyl) acetic acid with bovine serum albumin

The binding of bis(p-chlorophenyl) acetic acid with bovine serum albumin has been monitored by observing the changes in the relaxation times of bis(p-chlorophenyl) acetic acid proton magnetic resonance peaks. The addition of bovine serum albumin to an aqueous solution of bis(p-chlorophenyl) acetic acid increased linewidths in a manner suggesting rapid, reversible binding. Line broadening was found to be greater for the ring protons than for the benzylic proton, indicating that the ring presents the stronger binding site. The equilibrium constant has been approximated. The binding results are discussed in terms of environmental and toxicological considerations.     

Chemical structure and physical properties of candidate insecticides in relation to residual toxicity to adult mosquitos

Data are presented on changes in physical properties and insecticidal activity resulting from modifications in chemical structure in two groups of compounds. The first group consists of propoxur and three closely related N-methylcarbamates and the corresponding N-acetyl compounds The parent compounds are all highly toxic to adult mosquitos by topical application in solution and deposits from water-dispersible powder formulations on plywood and plaster of Paris panels have long residual activity. N-acetylation is not accompanied by excessive loss of toxicity to mosquitos, but increases volatility to the extent that none of the four N-acetyl derivatives can be considered as potential residual insecticides. The second group consists of tetrachlorvinphos and related vinyl phosphates and vinyl phosphorothionates which show a wide range of toxicity both to adult mosquitos and to mammals.     

Plant growth substance action on lecithin and lecithin/cholesterol vesicles

The adsorption of some plant growth regulating compounds onto lecithin and equimolar lecithin/cholesterol vesicles and the effect of these substances on the rate of chloride/nitrate and sodium/potassium ion exchange across the vesicular membranes have been examined. On comparing the binding of 2,4-D, 2,6-D, 2,4,5-T and IAA to lecithin vesicles in solution at various pH values it is found that the unionised form of these acids is bound very much more strongly than the ionised form. Compounds having relatively low oil/water partition coefficients such as 2,4-D, 2,6-D and 2,4,5-T are adsorbed onto vesicles, from solutions containing equimolar equilibrium concentrations of the unionised molecules, to markedly different extents (depending on the structure of the lipophilic portion of these molecules). Where the oil/water partition coefficient is higher, as for 2,4-dichlorophenol and 2-(2,4-dichlorophenoxy)ethanol, binding to vesicles may also arise due to non-specific solution within the hydrocarbon region of the lipid bilayers. The affinity of these compounds for lecithin has also been assessed by noting the extent to which the inclusion of lecithin in the oil phase increases the oil/water partition coefficient. This has shown that affinity for lecithin depends on the structure of both the lipophilic and hydrophilic portions of the molecule. Only the unionised form of the compounds examined had any large effect on ion flux across the vesicle membranes. Significant increases in the rate of chloride/nitrate exchange were obtained on introducing quite high concentrations (0.015 to 1.5 mM) of the compounds 2,4-D, 2,6-D, 2,4,5-T, IAA, and 2,4-dichlorophenol, but not 2,4-dichlorophenoxyethanol, to suspensions of lecithin vesicles. Similar flux increases were observed with lecithin/cholesterol vesicles except that more pronounced effects on flux were obtained on adding 2,4-dichlorophenol and 2,4-dichlorophenoxyethanol. In contrast 2,4-D, 2,6-D, 2,4-dichlorophenol and IAA had no significant effect on the rate of Na⁺/K⁺ ion exchange across lecithin vesicles although increases were observed with equimolar lecithin/cholesterol vesicles. Interpretations of these phenomena are suggested and their relevance to plant growth substance effects at the plasmalemma membrane discussed.     

The status and development of insecticide resistance in Danish populations of the housefly Musca domestica L

Samples of housefly (Musca domestica) field populations were collected from Danish livestock farms in 1997. The tolerance of the first‐generation offspring was determined for a number of insecticides. Dose‐response values were obtained by topical application for the pyrethroids bioresmethrin and pyrethrum, both synergised with piperonyl butoxide, and the organophosphate dimethoate. The organophosphates azamethiphos and propetamphos and the carbamate methomyl were tested in discriminating dose feeding bioassays. Resistance was low to moderate in most of the populations for most of the compounds tested, but this study also revealed the existence of high resistance to pyrethroid, organophosphate and carbamate insecticides in some populations. The resistance factors at LD₅₀ for bioresmethrin/piperonyl butoxide ranged between 2 and 98, and for pyrethrum/piperonyl butoxide between 2 and 29. Our results indicate that pyrethroid resistance in Denmark is increasing, since four of the 21 farms showed more than 100‐fold resistance at LD₉₅, a level of resistance only observed once before. Resistance factors at LD₅₀ for dimethoate ranged from 9 to 100, and showed two distinct trends: populations with either decreasing or increasing resistance. Resistance to azamethiphos was found to be widespread and high. Although two strains with high methomyl and propetamphos resistance were observed, methomyl and propetamphos resistance is moderate and appears not to be increasing. © 2001 Society of Chemical Industry     

Insecticidal activity against Aedes aegypti of m-pentadecadienyl-phenol isolated from Myracrodruon urundeuva seeds

BACKGROUND: Myracrodruon urundeuva Fr. Allemao is a common tree in the Caatinga that has been widely used for various medical purposes. Previous studies showed that the ethanol seed extract of M. urundeuva has potent activity against the larval stage of the dengue vector Aedes aegypti. Given this potential insecticidal activity, bioguided separation steps were performed in order to isolate the active compound(s). RESULTS: The isolation process resulted in only one active chemical compound, identified by infrared spectroscopy and mass spectrometry as m-pentadecadienyl-phenol. This compound presented potent larvicidal and pupicidal activity (LC₅₀ 10.16 and 99.06 µg mL⁻¹ respectively) and great egg hatching inhibitory activity (IC₅₀ 49.79 µg mL⁻¹). The mode of action was investigated through observations of behavioural and morphological changes performed in third-instar larvae treated with m-pentadecadienyl-phenol solution after 1, 6, 12, 16 and 20 h of exposure. Some changes were observed as flooding of the tracheal system, alterations in siphonal valves and anal gills and lethargy, probably caused by the strong anticholinesterasic activity reported previously. CONCLUSION: The compound isolated from M. urundeuva seeds, m-pentadecadienyl-phenol, showed potent activity against immature stages of dengue vector, Ae. aegypti, being considered the main larvicidal principle. Copyright © 2012 Society of Chemical Industry     

Chronic chlorpyrifos-induced oxidative changes in the testes and pituitary gland of Wistar rats: Ameliorative effects of vitamin C

Chlorpyrifos (CPF), a chlorinated organophosphate insecticide that is widely used in agriculture and public health, has been implicated in male reproductive toxicity. Apart from acetylcholinesterase inhibition, CPF has been shown to induce changes characteristic of oxidative stress. Therefore, the aim of the present study was to evaluate the effects of vitamin C on oxidative changes in the testes and pituitary gland of rats chronically exposed to CPF. Twenty adult male Wistar rats were divided into four groups of five animals each: Group I (S/oil) received soya oil (2 ml/kg); Group II (VC) was administered with vitamin C (100 mg/kg); Group III (CPF) was given CPF (10.6 mg/kg; ∼1/8th LD₅₀); Group IV (VC + CPF) was pretreated with vitamin C (100 mg/kg) and then given CPF (10.6 mg/kg), 30 min later. The regimens were administered orally by gavage once daily for 15 weeks. Thereafter, the rats were sacrificed and the testes and pituitary glands were evaluated for the concentration of malonaldehyde (MDA) and activities of superoxide dismutase (SOD) and catalase (CAT). The result shows that CPF increased MDA concentration and reduced activities of SOD and CAT, which were ameliorated by pretreatment with vitamin C.     

Interaction of substerilizing Gamma radiation and thiodicarb treatment for management of the Tobacco caterpillarSpodoptera litura

The F₁ sterility technique (inherited sterility using substerile insects) has been proposed for reproductive suppression of lepidopteran pests. Management of lepidopteran pests using F₁ sterility may be greatly influenced by other possible strategies, including chemical control measures. Within this context, a laboratory investigation was undertaken of the tobacco caterpillarSpodoptera litura (Fabricius) (Lepidoptera: Noctuidae) to assess the bioefficacy of a substerilizing dose (70 Gy) of gamma radiation on bio-characteristics(viz., adult survival, fecundity, egg hatch, F₁ larval weight, F₁ pupation, F₁ sex ratio); to establish the median lethal dose (LD₅₀) of an oxime carbamate insecticide, thiodicarb (Larvin®75 WP), toS. litura; and determine the interaction of thiodicarb with the F₁ insects derived from a substerilizing gamma dose. A dose of 70 Gy administered to 7-day-old male pupae reduced the the eclosion by ~ 10%. When the eclosed males (derived from treated pupae) were mated with normal females, the fecundity and fertility were decreased as compared with control (0 Gy). Analysis of body weight of the F₁ larvae showed no significant influence of radiation,i.e., growth was comparable to that of normal progeny, which is a positive factor in the sterile-insect technique. Bioassay on newly molted 6th instar larvae with thiodicarb (Larvin®75 WP) showed that the LD₅₀ of the insecticide was 6.76µg g^-1 in the combined treatment (radiation to P₁ male pupae + thiodicarb to F₁ larvae), as compared with 28.67 µg g_-1 in the chemical treatment (thiodicarb), indicating that the toxicity of the insecticide to the F₁ progeny was increased 4.24 times by the radiation treatment, which is a markedly pronounced effect. Further studies of the combined effect of radiation and insecticide on pupation and sex ratio of F₁ progeny showed that pupation was affected more by the combined treatment than by the insecticide alone, whereas the sex ratio was skewed in favor of males in both the combined treatment and the chemical treatment, indicating that thiodicarb was selectively more toxic to females. The possible interaction and potential and the feasible approaches of integrating F₁ sterility and thiodicarb treatment are discussed for management of the lepidopteran pestS. litura.     

Role of oxidative stress in organophosphate insecticide toxicity - Short review

The objective of this paper is to present a short review of the state of knowledge regarding oxidative stress and its role in toxicity of organophosphate insecticides. The information has been obtained by searching the relevant literature using chemical abstracts, PubMed, scopus, medline and other data bases. The significance of the problem has been elucidated. Organophosphate insecticides (OP), apart from inhibition of cholinesterase and presence of cholinergic effects, oxidative stress and hyperglycemia has been reported by many authors as one of the adverse effects in poisoning by OP in both humans and animals. Oxidative stress induced by organophosphate leads to disturbances in the function of different organs and tissues. In subchronic or chronic OP exposition induction of oxidative stress has been reported, by many authors, as the main mechanism of its toxicity. Data were categorized according to animal studies (in vitro and in vivo) and clinical studies. On the basis of relevant literature it is concluded, that determination of oxidative stress parameters can be useful for monitoring people exposed to OP professionally. Supplementation with natural or synthetic antioxidant may be beneficial in OP poisoning, however the rat models of OP poisoning used in those studies do not completely reflect clinical situation. For this reason the clinical trials are needed to explore effectiveness of these antioxidants in protection against toxicity of OP.     

A new model membrane system for the investigation of penetrative properties of plant growth substances

A new model membrane system has been developed which simulates in many ways the behaviour of a plant cell membrane. It is very suitable for studying the penetration and associated effects of plant growth substances because of its large size (4.9±0.3 cm²), its ease of preparation and its reproducible properties. It consists of a Millipore filter disc impregnated with egg yolk lecithin, n-octanol and n-tetradecane. The effect of a number of plant growth and allied compounds on membrane conductivity at 25±0.5°c has been assessed by the addition of various concentrations of these substances to the solutions (0.01 M-NaCl) on each side of the membrane. Certain compounds present in the solutions (pH 3.0) in the concentration range 10^?4 M to 10^?3 M considerably reduced the membrane conductivity. This effect was seen to be related to the structure of the hydrophobic portion of the molecule. The flux through the membrane at 25±0.5°c of various organic substances (2,4-D, benzoic acid, benzyl alcohol, benzamide and benzaldehyde) was measured for membranes containing no lecithin and compared with the flux obtained in comparable experiments, with lecithin present in the membrane. The presence of lecithin increased the flux, and the percentage increase was used to assess in each case the part played by lecithin in transport. The flux of 2,4-D was much greater at low pH values. A possible interpretation of these phenomena is suggested.     

Effect of Juglone on Active Oxygen Species and Antioxidant Enzymes in Susceptible and Partially Resistant Banana Cultivars to Black Leaf Streak Disease

The black leaf streak disease (BLSD), caused by Mycosphaerella fijiensis, is the most destructive disease of bananas and plantains around the world. Breeding for resistance is the most promising strategy to fight this disease especially in small farmer plantations. Mycosphaerella fijiensis produces many phytotoxins such as juglone, which can be used, jointly with field and inoculations under controlled conditions, for screening banana cultivars for BLSD-resistance. This non-host specific phytotoxin has been shown to act on chloroplasts and disturbs the proton electrochemical gradient across the plasmalemma membrane. Moreover, an involvement of the oxidative burst during the interaction has been suggested. The present study was carried out using two cultivars that differed for either their juglone-responses or their resistance to BLSD (cv. Grande Naine susceptible to BLSD and juglone and cv. Fougamou partially resistant to BLSD and highly tolerant to juglone). The production of active oxygen species (AOS) and the enhancement of the enzymatic and/or non-enzymatic AOS-scavenging systems were investigated after treatment of the two cultivars with juglone. The time-course of AOS-production and AOS-scavenging was shown to be the key difference between these two tested cultivars after treatment with juglone. Thus, an early release of AOS (O₂⁻ radical and H₂O₂) and a quick stimulation of a preferment anti-oxidant system (superoxide dismutases, catalases, and peroxidases) was observed for cv. Fougamou as compared to cv. Grande Naine for which a late and weak generation of AOS accompanied by a late stimulation of the anti-oxidant systems were detected.     

Esterase-mediated tolerance to a formulation of the organophosphate insecticide monocrotophos in the entomopathogenic fungus, Beauveria bassiana (Balsamo) Vuill: a promising biopesticide

The use of biopesticides compatible with chemical pesticides is recommended in pest management as an effective and ecologically sound strategy. The entomopathogenic fungus Beauveria bassiana (Balsamo) Vuill, which is used as a biopesticide, was found to exhibit and lose tolerance to two organophosphorus insecticides widely used in Indian fields. The pattern of response is similar to the esterase-mediated organophosphate tolerance in aphids achieved through esterase gene duplication. Therefore the role of esterase in the tolerance exhibited by isolates of B. bassiana to the organophosphate, monocrotophos, was studied. Both the total soluble protein content and esterase activity were found to increase significantly in B. bassiana cultures that were able to grow in the presence of monocrotophos. With the hitherto established linkage between esterase overproduction and organophosphate tolerance in insects and the observed similarity in response of the insects and B. bassiana to the chemical, it is concluded that tolerance to monocrotophos in B. bassiana may be due to a mechanism similar to that operating in insects. Habituation of the fungus to monocrotophos to initiate expression of esterase gene may help in achieving compatibility between the two.     

4种喷雾助剂对解淀粉芽胞杆菌Lx-11的增效作用及安全性评价

为提高解淀粉芽胞杆菌Lx-11悬浮剂对水稻白叶枯病的防治效果，通过研究添加4种喷雾助剂 (有机硅、TM-10、卵磷脂和SY-6535) 对解淀粉芽胞杆菌Lx-11含菌量的影响、药液的表面张力和接触角的变化、助剂对水稻生长的安全性影响等，并测定了添加助剂生防菌对白叶枯病的盆栽防治效果以及在水稻叶表定殖动态，最终筛选出适合的喷雾助剂。结果表明:添加卵磷脂的解淀粉芽胞杆菌Lx-11悬浮剂的含菌量显著降低；添加有机硅、TM-10和SY-6535均能有效降低悬浮剂的表面张力及其在水稻叶片上的接触角。安全性评价显示，有机硅、TM-10和SY-6535对水稻幼苗生长无影响。盆栽试验结果表明，添加TM-10和SY-6535后，解淀粉芽胞杆菌Lx-11悬浮剂 (4.0 × 109 cfu/mL) 对水稻白叶枯病的防治效果达到57.70%和56.56%，显著高于未添加助剂的悬浮剂的防治效果；定殖试验结果表明，添加助剂后，解淀粉芽胞杆菌Lx-11在水稻叶表的定殖量也显著高于对照处理。     

Increased Incidence of Erwinia Soft-rot on Calla Lilies in the Presence of Phosphorous

Erwinia soft-rot is an important disease of many ornamental potted crops and is one of the most limiting factors in greenhouse calla lily (Zantedeschia spp.) production. Experiments were conducted to test the effect of phosphorous added to soil-less mixes or to nutrient solutions used for irrigation on soft-rot caused by Erwinia carotovora subsp. carotovora (Ecc). Soft-rot incidence increased to 51% when soil-less mix was amended with superphosphate in comparison to regular soil-less mix (no superphosphate added) (31%). In contrast, addition of phosphorous in the nutrient solution met the phosphorous needs of the plant without enhancing soft-rot. Plant height, fresh mass, and number of flowers per plant were greater in calla lilies irrigated with nutrient solution containing phosphorous than no phosphorous treatments. Similar results were obtained in tests conducted in a commercial greenhouse with larger sample size. No statistical differences were found between tubers sprayed with water (control) or with a 0.5 mM solution of KH₂PO₄ in laboratory experiments to determine the effect of phosphorous on tuber root development. In other experiments, tubers were sprayed with either water, a bacterial cell suspension 1 × 10² cfu ml^–1, a solution of 0.5 mM KH₂PO₄, or a suspension of bacteria in KH₂PO₄. The results from these tests showed a significant increase of soft-rot development in tubers treated with the suspension of Ecc prepared in a solution of KH₂PO₄ relative to other treatments. Further laboratory tests indicated that enzymatic activity (polygalacturonase and pectate lyase) of Ecc increased when grown in the presence of phosphorous. These experiments suggest that increased soft-rot in the presence of phosphorous is due to increased virulence of Ecc.     

A novel immobilized cholinesterase for on-site screening of organophosphate and carbamate compounds

Acetylcholinesterase was physically adsorbed on polyvinylpyrrolidone K30 to prepare a novel immobilized cholinesterase, which was described for the first time. The immobilized enzyme was tested for its reactivity, enzyme activity stability, the solubility, the influence of pH and buffer, and characteristic reactivity with inhibitors, e.g., organophosphate and carbamate compounds. The result revealed that the immobilized cholinesterase held its original activity in solution, stable enough, easy to dissolve for convenient operation, and this immobilized cholinesterase could be well used for rapid, on-site screening of organophosphate and carbamate compounds, together with a portable pesticide analyzer.     

用正交试验法优选绿盲蝽若虫人工饲料配方

采用10因子3水平正交试验,得出影响绿盲蝽若虫总历期的人工饲料各因子顺序为:鸡蛋啤酒酵母小麦胚大豆卵磷脂、复合维生素卵黄利马豆粉蜂蜜水蔗糖大豆粉;影响绿盲蝽若虫存活率的因子顺序为:鸡蛋啤酒酵母利马豆粉大豆卵磷脂、复合维生素大豆粉蔗糖蜂蜜水小麦胚卵黄。综合两种结果,优选出绿盲蝽若虫人工饲料配方,其成分如下:蔗糖2.8 g、啤酒酵母粉0.25 g、50%蜂蜜水2.5 g、鸡蛋22.5 g、小麦胚10 g、利马豆粉30 g、大豆粉2.5 g、卵黄30 g、大豆卵磷脂1.5 g、复合维生素1.2 g、水164 g。此优化配方饲养绿盲蝽若虫存活率为80.57%,与四季豆饲养的若虫存活率71.13%没有显著差异;若虫总历期为11.80 d,较四季豆饲养的总历期10.4 d延长约1 d。     

Changes in rat brain regions serotoninergic system following acephate poisoning

Acephate (O,S-dimethyl acetylphosphoramidothioic acid) is an organophosphate pesticide used throughout the world extensively. Anticholinesterase action of acephate produces cholinotoxicity and may produce compensatory changes in other neurotransmitter levels. The objective of the present investigation was (1) to find brain region specific changes and (2) duration related treatment effects of the organophosphate acephate exposure on serotoninergic system. Reports of repeated exposure to organophosphate pesticide induced changes in rat brain serotoninergic system are limited. The present study analyses changes in 5-hydroxytryptamine (5-HT) level and its metabolite 5-hydroxy indoleacetic acid (5-HIAA) level at 0, 4, 14, and 60 days of oral acephate exposure (178 mg/kg) in the rat brain regions: olfactory lobe, cerebellum, pons, medulla oblongata, spinal cord, hypothalamus, hippocampus, striatum, midbrain, and cortex. Acephate exposure decreased 5-HT level in cortex, hippocampus, striatum, hypothalamus, midbrain, and spinal cord brain regions at the 4, 14, and 60 days durations studied. Duration specific decrease in 5-HT level was observed in olfactory lobe at 4 and 14 days and in pons, medulla oblongata, and cerebellum brain regions at 14 and 60 days of treatment. 5-HIAA levels were decreased in the brain regions cortex, pons, medulla oblongata, and spinal cord at all the durations of treatment studied and in hypothalamus (4 days) and cerebellum (60 days). A short-term increase in 5-HIAA levels was observed in olfactory lobe and midbrain after 14 days of treatment. The results reveal that acephate exposure produced activation of the serotoninergic system and the increased activity of serotoninergic neurons results in decrease of the monoamine levels and its metabolite in brain regions.     

Ace and ace-like genes of invasive redlegged earth mite: copy number variation,target-site mutations,and their associations with organophosphate insensitivity

Background

Invasive Australian populations of redlegged earth mite, Halotydeus destructor (Tucker), are evolving increasing organophosphate resistance. In addition to the canonical ace gene, the target gene of organophosphates, the H. destructor genome contains many radiated ace-like genes that vary in copy number and amino acid sequence. In this work, we characterise copy number and target-site mutation variation at the canonical ace and ace-like genes and test for potential associations with organophosphate insensitivity. This was achieved through comparisons of whole-genome pool-seq data from alive and dead mites following organophosphate exposure.

Results

A combination of increased copy number and target-site mutations at the canonical ace was associated with organophosphate insensitivity in H. destructor. Resistant populations were segregating for G119S, A201S, F331Y at the canonical ace. A subset of populations also had copy numbers of canonical ace > 2, which potentially helps overexpress proteins carrying these target-site mutations. Haplotypes possessing different copy numbers and target-site mutations of the canonical ace gene may be under selection across H. destructor populations. We also detected some evidence that increases in copy number of radiated ace-like genes are associated with organophosphate insensitivity, which might suggest potential roles in sequestration or breakdown of organophosphates.

Conclusion

Different combinations of target-site mutations and (or) copy number variation in the canonical ace and ace-like genes may provide non-convergent ways for H. destructor to respond to organophosphate selection. However, these changes may only play a partial role in organophosphate insensitivity, which appears to have a polygenic architecture. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.     

A technique for studying the biological efficiency of small droplets of pesticide solutions and a consideration of the implications

A simple bioassay technique is described for small, uniformly-sized droplets of pesticide solutions obtained by use of a microtip spray nozzle. A soluble fluorescent tracer was added to the spraying solution; droplets of the solution were sprayed on leaf discs and counted under a microscope with the aid of an ultraviolet lamp. The technique was used to study the relationships between the number of drops, their size and the dicofol concentration, on the mortality of Tetranychus urticae eggs. LD₅₀ values decreased as the drop size was reduced; the concentration of the pesticide also affected the LD₅₀ values. The technique can be used to study the biological efficiency of ultra-low volume pesticide formulations in the laboratory.     

斑蝥素对菜蛾消化酶及酯酶的影响

测定斑蟊素饲喂对菜蛾Ｐｌｕｔｅｌｌａ ｘｙｌｏｓｔｅｌｌａ Ｌ．幼虫体主要消化酶和酯酶的影响。结果表明,斑蝥素处理后试虫的主要消化酶类平均活性变化不大,与对照组相比,蛋白酶、脂肪酶平均活性分别被抑制了８．７１％和１５．６９％,ａ－淀粉酶活性较对照组增加了６．７５％,均未达到显著差异水平;酯酶中的磷酸酯酶活性平均降低２５．３３％,未达显著差异水平,乙酰胆碱酯酶和羧酸酯酶活性显著提高,３６ｈ内平均比活     

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