Alteration of the binding constants of malathion and carbaryl to cytochrome P-450 due to hydrolysis by rat hepatic microsomal esterases

During studies to determine spectral binding properties of malathion and carbaryl to rat liver microsomal cytochrome P-450, it was observed that final concentrations of 5 μM malathion and 10 μM carbaryl produced unstable binding spectra. Evidence is presented which demonstrates that a major reason for the unstable binding spectra of these insecticides is rapid degradation by hepatic microsomal esterases. In this respect, carbaryl and malathion, and possibly other compounds that contain ester linkages, appear to be unique, since most compounds are not metabolized under the conditions in which binding spectra are determined, i.e. in the absence of an NADPH generating system.     

Delayed action insecticides and their role in mosquito and malaria control

There is considerable interest in the management of insecticide resistance in mosquitoes. One possible approach to slowing down the evolution of resistance is to use late-life-acting (LLA) insecticides that selectively kill only the old mosquitoes that transmit malaria, thereby reducing selection pressure favoring resistance. In this paper we consider an age-structured compartmental model for malaria with two mosquito strains that differ in resistance to insecticide, using an SEI approach to model malaria in the mosquitoes and thereby incorporating the parasite developmental times for the two strains. The human population is modeled using an SEI approach. We consider both conventional insecticides that target all adult mosquitoes, and LLA insecticides that target only old mosquitoes. According to linearised theory the potency of the insecticide affects mainly the speed of evolution of resistance. Mutations that confer resistance can also affect other parameters such as mean adult life span and parasite developmental time. For both conventional and LLA insecticides the stability of the malaria-free equilibrium, with only the resistant mosquito strain present, depends mainly on these other parameters. This suggests that the main long term role of an insecticide could be to induce genetic changes that have a desirable effect on a vital parameter such as adult life span. However, when this equilibrium is unstable, numerical simulations suggest that a potent LLA insecticide can slow down the spread of malaria in humans but that the timing of its action is very important.     

Genetic diversity of the cotton aphid Aphis gossypii in the unstable environment of a cotton growing area

1 Spatial and temporal habitat heterogeneity represented by annual crops is a major factor influencing population dynamics of phytophagous insect pests such as the cotton aphid Aphis gossypii Glover. We studied the effects of instability of the cotton agroecosystem resulting from the temporary availability of the plant resource and the repeated use of insecticides on the genetic variability of the cotton aphids.
2 Samples of A. gossypii were collected in cotton plots, treated or not with insecticides and from vegetable crops (Malvaceae, Cucurbitaceae and Solanaceae) within the cotton growing area of northern Cameroon. The genetic structure of the samples was assessed using eight microsatellite markers. Insecticide resistance was estimated through the detection of two mutations in the ace -1 gene that are associated with insensitivity of acetylcholinesterase to carbamate and organophosphate insecticides.
3 The results obtained show that both host plants and insecticides act in genetic structuring of A. gossypii . Ninety-three percent of aphids collected on cotton were characterized by the same microsatellite multilocus genotype, Burk1 , which also displays the insecticide resistant alleles.
4 During the dry season, the cotton crop season after, the genotype Burk1 was principally found on two other malvaceous cultivated plants, rosella and okra, acting as suitable reservoir plants. The ability of the cotton aphid to move among asynchronous suitable habitats in response to changes in resource availability enables the pest to exploit unstable cropping systems. An understanding of the cotton aphid life system may aid to improve strategies for integrated resistance management.     

Genetics,Synergists, and Age Affect Insecticide Sensitivity of the Honey Bee,Apis mellifera

The number of honey bee colonies in the United States has declined to half of its peak level in the 1940s, and colonies lost over the winter have reached levels that are becoming economically unstable. While the causes of these losses are numerous and the interaction between them is very complex, the role of insecticides has garnered much attention. As a result, there is a need to better understand the risk of insecticides to bees, leading to more studies on both toxicity and exposure. While much research has been conducted on insecticides and bees, there have been very limited studies to elucidate the role that bee genotype and age has on the toxicity of these insecticides. The goal of this study was to determine if there are differences in insecticide sensitivity between honey bees of different genetic backgrounds (Carniolan, Italian, and Russian stocks) and assess if insecticide sensitivity varies with age. We found that Italian bees were the most sensitive of these stocks to insecticides, but variation was largely dependent on the class of insecticide tested. There were almost no differences in organophosphate bioassays between honey bee stocks (<1-fold), moderate differences in pyrethroid bioassays (1.5 to 3-fold), and dramatic differences in neonicotinoid bioassays (3.4 to 33.3-fold). Synergism bioassays with piperonyl butoxide, amitraz, and coumaphos showed increased phenothrin sensitivity in all stocks and also demonstrated further physiological differences between stocks. In addition, as bees aged, the sensitivity to phenothrin significantly decreased, but the sensitivity to naled significantly increased. These results demonstrate the variation arising from the genetic background and physiological transitions in honey bees as they age. This information can be used to determine risk assessment, as well as establishing baseline data for future comparisons to explain the variation in toxicity differences for honey bees reported in the literature.     

Monitoring for resistance to organophosphorus and pyrethroid insecticides in Varroa mite populations

The occurrence of resistance in Varroa mite populations is a serious threat to the beekeeping industry and to crops that rely on the honey bee for pollination. Integrated pest management strategies for control of this pest include the judicious use of insecticides. To monitor field populations of Varroa mite for insecticide resistance, a glass vial bioassay procedure was developed to use in the development of a resistance management strategy. Diagnostic concentrations needed to separate susceptible genotypes from resistant individuals were determined for cypermethrin (0.1 microg per vial), fluvalinate (5.0 microg per vial), malathion (0.01 microg per vial), coumaphos (10.0 microg per vial), diazinon (5.0 microg per vial), methomyl (0.5 microg per vial), propoxur (0.1 microg per vial), and endosulfan (2.5 microg per vial). Resistance to organophosphorus insecticides (malathion, coumaphos) and pyrethroids (cypermetrhrin, fluvalinate) was widespread in both La Media Ranch, TX, and Wewahitchka, FL, from 2007 to 2009. There was no resistance to endosulfan, diazinon, methomyl, and propoxur in field populations of Varroa mite in the two locations where resistance was monitored. The seasonal patterns of resistance in Wewahitchka were different from those of La Media Ranch. In the former location, the frequency of resistance to all insecticides tested decreased significantly from 2007 to 2009, whereas it increased in the latter location. Resistance levels were unstable, suggesting that resistance could be successfully managed. The results validate use of the glass vial bioassay to monitor for resistance in Varroa mite and provide the basis for the development of a resistance management strategy designed to extend the efficacy of all classes of insecticides used for control of Varroa mite.     

Expression of the housefly acetylcholinesterase in a bioreactor and its potential application in the detection of pesticide residues

Acetylcholinesterase is a key enzyme of the animal nerve system. The enzyme is the primary target of organophosphorous (OP) and carbamate (CB) insecticides. The insect AChE is being extensively used in development of new insecticides or in vitro selection of the new designed insecticides, and in pharmacological and toxicological field. Rapid assays using AChE-based methods have been proposed as an efficient and rapid method for the detection of pesticides, especially in many Asian markets. In this study, the acetylcholinesterase gene was cloned from housefly (Musca domestica) susceptible to organophosphate (OP) and carbamate (CB) insecticides, and expressed in baculovirus-insect cells system using a bioreactor with oxygen supplementation. The recombinant housefly AChE was purified using ammonium sulfate precipitation and procainamide affinity chromatography, and approximately 0.42 mg of the purified AChE with high biological activity (118.9 U/mg) was obtained from 100 ml of culture solution. The purified AChE was highly sensitive to OP and CBs insecticides. In conclusion, an efficient expression and purification system has been developed for large-scale production of recombinant housefly AChE. The recombinant enzyme is potential to be used for the detection of pesticide residues.     

Biotechnological production of plant-based insecticides

The demand for natural and nonpersistent insecticides is increasing day by day. Plant cell cultures could be an alternative to conventional methods of production of insecticides from field-grown plants. In vitro cultured plant cells produce a wide array of insecticides as a part of their secondary metabolism. Their ability to synthesize key enzymes and the manipulation of these could lead to the enhanced production of many insecticides of industrial importance. The development of a high-yielding hairy root culture system for thiophenes, nicotine, and phytoecdysones is of considerable interest. In this article, the current literature on various factors that influence the growth, production, and secretion of six insecticidal compounds, namely, pyrethrins, azadirachtin, thiophenes, nicotine, rotenoids, and phytoecdysones which have been prospects for the scale-up of cell cultures, genetic engineering to obtain transgenic plants, and metabolically engineered plants for increased production of bio-molecules, has been discussed. Environmental safety clearance and the future prospects of application of biomolecules for plant-derived insecticides are presented.     

Development and stability of insecticide resistance in the leafminer Liriomyza trifolii (Diptera: Agromyzidae) to cyromazine, abamectin, and spinosad

Three populations of the leafminer, Liriomyza trifolii (Burgess), were collected from commercial ornamental production greenhouses in the United States and tested for susceptibility to three commercial insecticides. A leaf dip bioassay of leaves containing young (1-2-d-old) larvae was used. Based on larval mortality and compared with a susceptible laboratory reference colony, the three strains varied in spectrum and level of resistance to the insecticides. CA-1, collected from Gerbera daisy, was moderately resistant to cyromazine (18.1-fold) and abamectin (22.0-fold), but highly resistant to spinosad (> 188-fold). CA-2, collected from chrysanthemums, was not resistant to abamectin, had a low level of resistance to cyromazine (8.2-fold), but was extremely resistant to spinosad (1,192-fold). GA-1, collected from chrysanthemums, had very low levels of resistance to cyromazine (5.4-fold) and spinosad (1.9-fold) but was moderately resistant to abamectin (30.6-fold). When reared in the absence of insecticide selection pressure, all three strains reverted to approximately the level of the reference strain. The CA-1 strain reverted in nine generations to cyromazine; however, the lowest levels of abamectin and spinosad resistance reverted to was 3.1-fold at F8 and 3.2 at the F10, respectively. The CA-2 strain reverted in five generations to both cyromazine and spinosad. GA-1 reverted in five generations to abamectin. Based on the results, resistance to these three insecticides was unstable. Additionally, there was no cross-resistance among these three insecticides.     

Automated phenotyping of mosquito larvae enables high-throughput screening for novel larvicides and offers potential for smartphone-based detection of larval insecticide resistance

Pyrethroid-impregnated nets have contributed significantly to halving the burden of malaria but resistance threatens their future efficacy and the pipeline of new insecticides is short. Here we report that an invertebrate automated phenotyping platform (INVAPP), combined with the algorithm Paragon, provides a robust system for measuring larval motility in Anopheles gambiae (and An. coluzzi) as well as Aedes aegypti with the capacity for high-throughput screening for new larvicides. By this means, we reliably quantified both time- and concentration-dependent actions of chemical insecticides faster than using the WHO standard larval assay. We illustrate the effectiveness of the system using an established larvicide (temephos) and demonstrate its capacity for library-scale chemical screening using the Medicines for Malaria Venture (MMV) Pathogen Box library. As a proof-of-principle, this library screen identified a compound, subsequently confirmed to be tolfenpyrad, as an effective larvicide. We have also used the INVAPP / Paragon system to compare responses in larvae derived from WHO classified deltamethrin resistant and sensitive mosquitoes. We show how this approach to monitoring larval response to insecticides can be adapted for use with a smartphone camera application and therefore has potential for further development as a simple portable field-assay with associated real-time, geo-located information to identify hotspots.     

Biotechnological Production of Plant-Based Insecticides

ABSTRACT:?

The demand for natural and nonpersistent insecticides is increasing day by day. Plant cell cultures could be an alternative to conventional methods of production of insecticides from field-grown plants. In vitro cultured plant cells produce a wide array of insecticides as a part of their secondary metabolism. Their ability to synthesize key enzymes and the manipulation of these could lead to the enhanced production of many insecticides of industrial importance. The development of a high-yielding hairy root culture system for thiophenes, nicotine, and phytoecdysones is of considerable interest. In this article, the current literature on various factors that influence the growth, production, and secretion of six insecticidal compounds, namely, pyrethrins, azadirachtin, thiophenes, nicotine, rotenoids, and phytoecdysones which have been prospects for the scale-up of cell cultures, genetic engineering to obtain transgenic plants, and metabolically engineered plants for increased production of bio-molecules, has been discussed. Environmental safety clearance and the future prospects of application of bio-molecules for plant-derived insecticides are presented.     

Insecticide susceptibility of Cnaphalocrocis medinalis (Lepidoptera: Pyralidae) in China

Insecticide control is the major measure for suppression of Cnaphalocrocis medinalis (Guenée) (Lepidoptera: Pyralidae) damage, and a few insecticides used for long time have proved to fail to control this pest in China. Several new chemicals have been introduced for control of C. medinalis. However, there was no baseline susceptibility data of C. medinalis to insecticides used or will be in use. In this study, a seedling dipping method was developed for bioassay of insecticide susceptibility of C. medinalis. Dose responses of C. medinalis to 11 insecticides were tested. Interpopulation sensitivity to insecticides was compared. Based on LC50 values, C. medinalis was most susceptible to antibiotic insecticides (abamectin, emamectin benzoate, and spinosad) and least sensitive to monosultap and a Bacillus thuringiensis (Bt) product. Chlorantraniliprole and insect growth regulator (IGR) insecticides (tebufenozide and hexaflumuron) exhibited great efficacy against C. medinalis. No susceptibility difference was observed for antibiotic insecticide and IGR insecticides among three populations. Narrow variation in tolerant level was detected for organophosphates insecticides, chlorantraniliprole, monosultap, and Bt. The results in this study provided baseline susceptibility data of C. medinalis to 11 insecticides and also offered useful information for choice of alternative insecticide and for integrated resistance management of C. medinalis.     

杀虫剂使用失当是甜菜夜蛾大发生的重要原因

近年来,甜菜夜蛾Spodoptera exigua(H櫣bner)在我国的危害面积越来越广,造成的损失越来越大。为了更好地了解甜菜夜蛾的发生和成灾规律,改善和提高预测与防治技术,文章就杀虫剂对甜菜夜蛾及其天敌的影响进行了分析。结果表明,杀虫剂使用失当是甜菜夜蛾大发生的重要原因。甜菜夜蛾对杀虫剂的抗药性水平超过天敌,而天敌的飞行、搜索、攻击和产卵等行为,以及生长、发育、生殖和寿命等又容易受杀虫剂的干扰,产生负面影响,从而降低天敌对甜菜夜蛾的控制能力。因此,如何合理使用杀虫剂,在有效控制甜菜夜蛾种群的同时,达到保护天敌的目的,将是我们面临的新课题。     

昆虫毒蕈碱型乙酰胆碱受体研究进展

毒蕈碱型乙酰胆碱受体（muscarinic acetylcholine receptor,mAchR）是昆虫神经系统中一种十分重要的神经递质受体,属于G蛋白偶联受体超家族,与配体结合后激活G蛋白并通过胞内第二信使产生生物学效应,参与昆虫多种神经生理功能,是农药开发的一个潜在靶标。昆虫毒蕈碱型乙酰胆碱受体的研究,有助于提高昆虫神经生理及行为调节机制等方面的认识,并为以此为靶标的新型杀虫剂研制提供新思路。本文综述了国内外对昆虫毒蕈碱型乙酰胆碱受体的研究概况,并对GenBank中已上传的昆虫mAchR序列进行了分子进化分析,最后对昆虫mAchR研究中存在的问题及前景进行了分析和展望。     

杀虫药剂的神经毒理学研究进展

大多数杀虫药剂都具有较强的神经毒性,它们对神经系统的作用靶标不同。有机磷类杀虫剂不仅抑制乙酰胆碱酯酶活性和乙酰胆碱受体功能,影响乙酰胆碱的释放,而且还具有非胆碱能毒性,有些有机磷杀虫剂还能引发迟发性神经毒性。新烟碱类杀虫剂作为烟碱型乙酰胆碱受体（nAChR）的激动剂,作用于该类受体的α亚基;它对昆虫的毒性比对哺乳动物的毒性大得多,乃是因为它对昆虫和哺乳动物nAChR的作用位点不同。拟除虫菊酯类杀虫剂主要作用于神经细胞钠通道,引起持续开放,导致传导阻滞;该类杀虫剂也可抑制钙通道。另外,这类杀虫剂还干扰谷氨酸递质和多巴胺神经元递质的释放。拟除虫菊酯类杀虫剂对昆虫的选择毒性很可能是因为昆虫神经元的钠通道结构与哺乳动物的不同。阿维菌素类杀虫剂主要作用于γ-氨基丁酸（GABA）受体,它能促进GABA的释放,增强GABA与GABA受体的结合,使氯离子内流增加,导致突触后膜超级化。由于这类杀虫剂难以穿透脊椎动物的血脑屏障而与中枢神经系统的GABA受体结合,故该类杀虫剂对脊椎动物的毒性远低于对昆虫的毒性。多杀菌素类杀虫剂可与中枢神经系统的nAChR作用,引起Ach长时间释放,此外,这类杀虫剂还可作用于昆虫的GABA受体,改变GABA门控氯通道的功能。     

Acetylcholinesterase. Two types of modifications confer resistance to insecticide.

Quantitative and qualitative changes in acetylcholinesterase confer resistance to insecticides. We have constructed several Drosophila melanogaster strains producing various amounts of enzyme by P-mediated transformation. Toxicological analysis of these strains demonstrates that resistance to organophosphorus insecticides is correlated with the amount of acetylcholinesterase in the central nervous system. Resistance may also be qualitatively determined. Comparison of the Drosophila acetylcholinesterase gene between a resistant strain caught in the wild and a wild type susceptible strain only revealed one nucleotide transition resulting in the replacement of a phenylalanine by a tyrosine. Flies mutant for acetylcholinesterase and rescued with a minigene mutagenized for this same transition produced an altered enzyme which renders flies resistant to pesticides.     

An unstable giant satellite associated with chromosomes 21 and 22 in the same individual.

The short arms of the acrocentric chromosomes are among the most common sites in which to find human chromosomal heteromorphisms. Heteromorphic chromosomes are noted for their variability between individuals and populations; however, they generally are consistent within an individual. Contrary to this general rule, a normal female was found to have a giant satellite on the short arm of a chromosome 22 in most lymphocytes and fibroblasts, but in other cells, it was attached to a chromosome 21. Furthermore, in some cells, it was found on multiple chromosomes, that is, on both 22's or on a 21 and a 22. The familial nature of this heteromorphism was established when it was found in the woman's mother, where it was confined exclusively to chromosome 22. These results suggest an unstable giant satellite associated with both G-group chromosomes of a normal individual. Results are discussed in the light of the patient's occupational exposure to insecticides at a mushroom farm.     

褐飞虱对吡虫啉的抗性机理和靶标分子毒理学

褐飞虱Nilaparvata lugens是水稻最重要的害虫之一,长期依赖化学防治导致了该害虫对不同类型杀虫剂抗性的产生,对新烟碱类杀虫剂吡虫啉高水平抗性的产生更是造成了巨大的粮食生产损失。近年来在褐飞虱对吡虫啉抗性机理,以及在抗药性机理研究推动下吡虫啉作用靶标褐飞虱神经系统烟碱型乙酰胆碱受体（nicotinic acetylcholine receptors, nAChRs）毒理学等方面取得了许多研究进展。nAChRs是昆虫神经系统中最重要的神经递质受体,是几类重要杀虫剂的作用靶标,其中以新烟碱类杀虫剂为代表。通过对比敏感品系和室内连续筛选获得的高抗吡虫啉品系,在褐飞虱两个nAChRs亚基Nlα1和Nlα3中均发现了抗性相关点突变Y151S,该突变导致了受体与吡虫啉结合亲和力的显著下降,而对内源神经递质乙酰胆碱的亲和力影响很小。Nlα1与褐飞虱另外两个亚基Nlα2和Nlβ1共聚成一个受体,构成吡虫啉低亲和力结合位点;Nlα3与褐飞虱另外两个亚基Nlα8和Nlβ1共聚成一个受体,构成吡虫啉高亲和力结合位点。不仅褐飞虱nAChRs与吡虫啉抗性相关,某些nAChRs附属蛋白也直接影响褐飞虱对吡虫啉的抗性,如Lynx蛋白。关于褐飞虱nAChRs组成、抗药性相关变异、受体附属蛋白对抗药性的影响等方面的研究,均为国内外前沿报道,不仅有助于对新烟碱类杀虫剂抗性机理的理解,对昆虫nAChRs毒理学同样具有很大的推动作用。     

Spontaneous electrical activity recorded from the aphid central nervous system

Whilst many classes of insecticides target the insect central nervous system (CNS), their effects in the CNS of pest aphids have not been demonstrated. In this report, we describe an electrophysiological method for recording spontaneous neuronal activity from the giant willow aphid (Tuberolachnus salignus). Using extracellular recording electrodes and two analysis methods (threshold and template search), spontaneous spike activity was shown to exhibit sensitivity to the neuroexcitatory insecticide imidacloprid. This method allows changes in the frequency of action-potentials to be monitored during direct bath exposure to chemical agents, enabling a means of assessing and comparing neurotoxic effects of insecticides in a previously inaccessible superfamily of pest insects.     

Assessment of Potential Sublethal Effects of Various Insecticides on Key Biological Traits of The Tobacco Whitefly,Bemisia tabaci

The tobacco whitefly Bemisia tabaci is one of the most devastating pests worldwide. Current management of B. tabaci relies upon the frequent applications of insecticides. In addition to direct mortality by typical acute toxicity (lethal effect), insecticides may also impair various key biological traits of the exposed insects through physiological and behavioral sublethal effects. Identifying and characterizing such effects could be crucial for understanding the global effects of insecticides on the pest and therefore for optimizing its management in the crops. We assessed the effects of sublethal and low-lethal concentrations of four widely used insecticides on the fecundity, honeydew excretion and feeding behavior of B. tabaci adults. The probing activity of the whiteflies feeding on treated cotton seedlings was recorded by an Electrical Penetration Graph (EPG). The results showed that imidacloprid and bifenthrin caused a reduction in phloem feeding even at sublethal concentrations. In addition, the honeydew excretions and fecundity levels of adults feeding on leaf discs treated with these concentrations were significantly lower than the untreated ones. While, sublethal concentrations of chlorpyrifos and carbosulfan did not affect feeding behavior, honeydew excretion and fecundity of the whitefly. We demonstrated an antifeedant effect of the imidacloprid and bifenthrin on B. tabaci, whereas behavioral changes in adults feeding on leaves treated with chlorpyrifos and carbosulfan were more likely caused by the direct effects of the insecticides on the insects'' nervous system itself. Our results show that aside from the lethal effect, the sublethal concentration of imidacloprid and bifenthrin impairs the phloem feeding, i.e. the most important feeding trait in a plant protection perspective. Indeed, this antifeedant property would give these insecticides potential to control insect pests indirectly. Therefore, the behavioral effects of sublethal concentrations of imidacloprid and bifenthrin may play an important role in the control of whitefly pests by increasing the toxicity persistence in treated crops.     

A Study of the Chronic Effects of Insecticides in Man

Signs of possible chronic intoxication due to insecticides were sought among 441 apple-growers. A group of 170 persons living in the same environment and 162 other persons having no contact with insecticides were used as controls. It was the first attempt to determine signs of chronic intoxication by organic phosphate and chlorinated hydrocarbon insecticides. The greater incidence of leukopenia and of neurologic manifestations found among the subjects suggest that insecticides may have some chronic effects in man. This assumption is reinforced by the similarities in the findings between the subjects and the environments, and by the parallelism between the duration of exposure to insecticides and the number of affected subjects. The repetition of this study at five-year intervals seems desirable until a sufficient number of apple-growers have been studied who have been exposed to large doses of insecticides for 10, 15 or 20 years.