Knockdown resistance to DDT and pyrethroids: from target-site mutations to molecular modelling

Naturally derived insecticides such as pyrethrum and man-made insecticides such as DDT and the synthetic pyrethroids act on the voltage-gated sodium channel proteins found in insect nerve-cell membranes. The correct functioning of these channels is essential for the normal transmission of nerve impulses, and this process is disrupted by binding of the insecticides, leading to paralysis and eventual death. Some insect pest populations have evolved modifications of the sodium channel protein that inhibit the binding of the insecticide and result in the insect developing resistance. This perspective outlines the current understanding of the molecular processes underlying target-site resistance to these insecticides (termed kdr and super-kdr), and how this knowledge may in future contribute to the design of novel insecticidal compounds.     

The mode of entry of contact insecticides

Experiments have been carried out to test the generally accepted view that contact insecticides enter into the central nervous system via the haemolymph. So far, they have only produced evidence to the contrary As an alternative route, it is suggested that these insecticides reach the site of action via the integumental tissues of the body wall and the tracheal system. The evidence for this is discussed.     

Insecticide Resistance in the Western Flower Thrips, Frankliniella occidentalis

The western flower thrips, Frankliniella occidentalis Pergande (Thysanoptera: Thripidae) is a serious pest on a wide range of crops throughout the world. F. occidentalis is difficult to control with insecticides because of its thigmokinetic behaviour and resistance to insecticides. Pesticide resistance can have a negative impact on integrated pest management programmes with chemical control as one of the components. Resistance to a number of different insecticides has been shown in many populations of F. occidentalis. This flower thrips has the potential of fast development of resistance owing to the short generation time, high fecundity, and a haplodiploid breeding system. The mechanisms conferring insecticide resistance in insects can be divided into four levels. First, an altered behaviour can aid the insect to avoid coming into contact with the insecticide. Second, a delayed penetration through the integument will reduce the effect of the insecticide at the target site. Third, inside the insect, detoxification enzymes may metabolise and thereby inactivate the insecticide. Fourth, the last level of resistance mechanisms is alterations at the target site for the insecticide. Knowledge of resistance mechanisms can give information and tools to be used in management of the resistance problem. Recently, studies have been carried out to investigate the underlying mechanisms conferring resistance in F. occidentalis. It appears that resistance in F. occidentalis is polyfactorial; different mechanisms can confer resistance in different populations and different mechanisms may coexist in the same population. Possible resistance mechanisms in F. occidentalis include: reduced penetration, detoxification by P450-monooxygenases, esterases and glutathione S-transferases, and alterations of acetylcholinesterase, the target site for organophosphate and carbamate insecticides. Target site resistance to pyrethroids (knockdown resistance) may also be a resistance mechanism in F. occidentalis.     

新疆有毒植物资源及其植物源杀虫剂开发应用前景

植物源杀虫剂(botanical insecticides)就是利用植物的某些部位或提取成分,加工成有杀虫作用的杀虫剂。在新疆,有毒植物资源种类繁多。通过介绍新疆有毒植物的资源概况,讨论有毒植物作为农业杀虫剂的应用现状和存在问题,提出其种质资源作为植物源杀虫剂的开发利用前景。     

6种杀虫剂对甜菜夜蛾的田间药效评价

通过6种不同杀虫剂对甜菜夜蛾的田间药效试验,结果表明:0.2%甲胺基阿维菌素苯甲酸盐微乳剂4000~6000倍稀释液对甜菜夜蛾有较好的防治效果,其防效在90.4%以上;2.02%甲胺基阿维·高氯乳油1000倍稀释液的第7d防效为91.9%;0.9%阿维菌素乳油1500倍稀释液在第3d的防效为89.5%,这些药剂均是当前防治甜菜夜蛾的优良药剂。     

Insect hormones and growth regulators

The endocrine system that controls development and reproduction in insects has many unique features that could provide targets for insecticides with novel modes of action. Such insecticides, which act by disrupting the growth and development, are often called insect growth regulators. Three components of the endocrine system are discussed in this context, and an example of the specific chemical disruption of one of them, which could form the basis for the development of such insecticides, is described.     

A simple method for monitoring the respiratory rhythm in intact insects and assessing the neurotoxicity of insecticides

A simple, non-invasive method, for the monitoring of the respiratory rhythm of insects was developed. The insect was restrained and a force displacement transducer attached to one abdominal segment of the dorsal region was used to monitor the respiratory rhythm. Although the method was applied to four species of insects, the main study focused on the yellow mealworm beetle, Tenebrio molitor, due to the simplicity of the recorded respiratory rhythm. In this case, the recorded rhythmic activity represents the synchronised contraction of the dorsal abdominal muscles, driven by the respiratory central pattern generator (CPG). This non-invasive method allows prolonged and stable recording of the respiratory rhythm. The records showed a decrease in the amplitude and the duration in the rhythmic contractions to about 25 and 31% of the initial values, respectively, in the first 7 h of continuous recording. The respiratory rhythm was strongly influenced by the presence of the CO₂, creating the CO₂-reflex. In the beginning of the application of CO₂, there was total inhibition of the respiratory rhythm and then there was a gradual increase in the tension developed by the dorsal abdominal muscles. Using this recording method, it was possible to quantify the function of the respiratory CPG, measuring the amplitude of the respiratory contractions, under normal conditions and in the presence of insecticides. The insecticides imidacloprid and deltamethrin of certain quantities were diluted in the appropriate solvent to make 50, 100 and 500 ng per insect, and were applied to the cuticle, topical application, of the immobilized insect, while recordings of the respiratory rhythm were made continuously.     

Steady-state and transient analysis of integument penetration by insecticides

Insecticide penetration studies are usually done by applying a finite dose of material to a finite area of the integument in a solvent, such as acetone, that evaporates very quickly. The time-course of penetration is then measured either as the buildup of material inside the insect or as the decline of material recoverable in a surface wash. Pharmacokinetic models are often used to describe such experiments, but identical models can be formulated from diffusion theory. Diffusion theory was applied to obtain equations that describe the steady-state behavior of a single-layer integument with partitioning between integument and solutions, and a double-layer integument with partitioning between layers and between each layer and the adjacent solution. The steady-state flux equations for both models were first-order, and it was shown that this would be the case for steady-state flux through any number of layers. The steady-state analysis has two limitations. First, the assumption of a steady state implies that the concentration gradient across each layer of the integument is linear, and that this could not occur instantaneously. Second, the analysis assumes that material diffuses from a pool on the surface at some external concentration, whereas, ideally, it is applied as a plane source of infinite concentration. If the steady-state assumption is rejected, the diffusion equation can be solved exactly, using Green's function, for diffusion of a topically applied dose across the integument modeled as a homogeneous plate, if the concentration at the internal face is assumed to remain zero. An analytical expression is obtained for concentration as a function of time and position in the integument, and from it are calculated flux into the insect and disappearance of material recoverable in a solvent wash, where material is assumed to be recoverable to an arbitrary depth. For small values of recovery depth, recovery falls precipitously following application, then progressively more slowly as the experiment proceeds. This phenomenon has been seen in much published data, and is explained adequately for the first time by the theory presented here. Predicted flux into the insect rises with a delay and then falls, but its fall is not exponential, as is often assumed. Peak flux is predicted to be directly related to the diffusion coeficient. The impact of these results on the design of penetration experiments is discussed.     

新烟碱类杀虫剂抗药性研究进展

新烟碱类杀虫剂是一类新开发的杀虫剂。研究表明，害虫野外种群对其敏感性差弄较大，现已有多种害虫对吡虫啉和啶虫脒产生了抗性。初步研究显示，马铃薯叶甲对吡虫啉抗性以不完全隐性的常染色体遗传；抗性似不稳定，交互抗性谱随虫种而变化，抗性形成可能与多功能氧化酶和酯酶有关。合理轮用和高剂量杀死策略是治理其抗性的有效措施。     

菊酯类农药对水田生物影响研究进展

拟除虫菊酯类农药是一个重要的杀虫剂类别,对有害昆虫具有良好的触杀作用.但是,在中国一直被禁止应用于水田害虫防治.本文综述了菊酯类农药对各种水田生物的急性和慢性作用机制、影响因子,提出拟除虫菊酯类农药能否在水田应用需要进行深入系统的研究论证.     

杀虫剂分子靶标烟碱型乙酰胆碱受体研究进展

昆虫烟碱型乙酰胆碱受体(nicotinic acetylcholine receptors,nAChRs)广泛分布于昆虫中枢神经系统,是杀虫剂作用的主要靶标。目前昆虫中该受体的天然亚基组成尚不完全明确。果蝇的任意α亚基与脊椎动物的一个β亚基共表达是目前最好的异源表达模型,但仍然急需新的研究工具,研究表明一些与受体相关的蛋白质影响着表达。胞内磷酸化的调节作用为今后受体药理学特性的研究提供了新方向。受体亚基上一些关键氨基酸在新烟碱杀虫剂对受体的选择作用中起重要作用。在对吡虫啉抗性的褐飞虱种群中找到了与抗性相关的突变位点,这为新烟碱类杀虫剂靶标不敏感性研究提供了直接证据。对昆虫烟碱型乙酰胆碱受体的分子多样性、功能表达、胞内调节机制、受体与杀虫剂的选择作用及其抗性分子机理等的研究进展进行了综述。     

昆虫对拟除虫菊酯类杀虫剂的代谢抗性机制研究进展

随着拟除虫菊酯类杀虫剂在卫生和农业害虫防治中的广泛应用,昆虫对此类杀虫剂产生抗性的报道越来越多。目前已明确昆虫对拟除虫菊酯类杀虫剂的抗性机制包括表皮穿透率下降、靶标抗性以及代谢抗性,其中代谢抗性机制较为普遍,而且其与昆虫对多种杀虫剂的交互抗性关系密切。目前,随着基因组、转录组以及蛋白质组学等新技术的发展及应用,昆虫对拟除虫菊酯类杀虫剂的代谢抗性机制研究也取得了很多新进展。昆虫体内细胞色素P450酶（P450s）、羧酸酯酶（CarE）及谷胱甘肽S-转移酶（GSTs）等重要解毒酶系的改变均与昆虫对拟除虫菊酯类杀虫剂的代谢抗性有关,其中这3类解毒酶的活性及相关基因表达量的变化是昆虫对此类杀虫剂产生代谢抗性的主要原因。明确昆虫对拟除虫菊酯类杀虫剂的代谢抗性机制,对合理使用此类杀虫剂及延缓抗药性的产生均具有重要意义。本文在总结拟除虫菊酯类杀虫剂代谢路径及相关生物酶研究概况的基础上,综述了近年来有关昆虫对此类杀虫剂代谢抗性机制研究的主要进展。     

Mode of entry of oxime carbamates into insects

The outcome of our earlier work suggested that the transport of contact insecticides into insects does not involve the haemolymph, but that the chemical probably reaches the internal organs by migrating in and/or over integumental tissue. This applies to various insects having quite different integumental organisation, and to various groups of insecticides differing widely in physical and chemical characteristics. The present work has shown that carbamates (1-alkylthioacetaldoxime carbamates) with high water solubility are no exception. The permeability of the body wall of the housefly for the methyl homologue (methomyl) which is the most water-soluble of the series, although relatively high, is not high enough to account for the toxicity of the compound if it followed the haemolymph route. Methomyl when injected with water instead of an organic solvent was also found to be less toxic than the equivalent amount topically applied. Lateral migration in the integument could be demonstrated with all the carbamates tested and dissimilarities in rates were apparent. Methomyl migrated the most rapidly and the cyanomethylene homologue the most slowly. Differences in mobility may be partly responsible for inequality in toxic effect, but it is thought that metabolic detoxication is of over-riding importance.     

Insect resistance to insecticides

The past 40 years have seen insect resistance to insecticides develop from a scientific curiosity to an immense practical problem that threatens man's ability to control not only the insect pests of agriculture but also the insect vectors that transmit major human and animal diseases. The spread of genes for cross and multiple resistance among insect pests has rendered most of our present insecticides obsolescent and very few novel insecticides are under development as substitutes. The most feasible strategy to maintain adequate control of insect pests is integrated pest management or I P M, in which insecticide management is a useful component. However, much of our present planning for the future of insect control is carried out in ignorance of past failures. We must learn from the past if we are to retain the use of chemical insecticides as a viable component of IPM.     

14种农药对美洲斑潜蝇的防治效果

本研究对爱福丁、阿巴丁、绿菜宝、农地乐等１４种农药在大田中防治美洲斑潜蝇效果进行了综合评价。结果表明：爱福丁、阿巴丁、灭虫灵、农地乐、绿菜宝、双齐乳油、灭蝇蛾等农药对斑潜蝇的控制效果好;农哈哈在山东及华北地区、绿保素在西南地区、杀虫双在华南地区对斑潜蝇有较好的控制效果。     

Toxicity of insecticides to the sweetpotato whitefly (Hemiptera: Aleyrodidae) and its natural enemies

Efficient chemical control is achieved when insecticides are active against insect pests and safe to natural enemies. In this study, the toxicity of 17 insecticides to the sweetpotato whitefly, Bemisia tabaci (Gennadius), and the selectivity of seven insecticides to natural enemies of this insect pest were evaluated. To determine the insecticide toxicity, B. tabaci adults were exposed to abamectin, acephate, acetamiprid, cartap, imidacloprid, malathion, methamidophos, bifenthrin, cypermethrin, deltamethrin, esfenvalerate, fenitrothion, fenpropathrin, fenthion, phenthoate, permethrin and trichlorphon at 50 and 100% of the field rate (FR), and to water (untreated control). To determine the insecticide selectivity, adults of Encarsia sp., Acanthinus sp., Discodon sp. and Lasiochilus sp. were exposed to abamectin, acephate, acetamiprid, cartap, imidacloprid, malathion and methamidophos at 50 and 100% FR, and to water. Groups of each insect species were exposed to kale leaves preimmersed in each treatment under laboratory conditions. Mortality of exposed individuals was recorded 24 h after treatment. Cartap and imidacloprid at 50 and 100% FR and abamectin and acetamiprid at 100% FR showed insecticidal activity to B. tabaci adults. Abamectin at 50 and 100% FR was the least insecticidal compound to the natural enemies Acanthinus sp., Discodon sp. and Lasiochilus sp. The present results suggest that abamectin at 100% FR may decrease B. tabaci field populations but can still be harmless to predators. Implications of these results within an integrated pest management context are discussed.     

线粒体复合体Ⅰ呼吸抑制剂的研究

线粒体复合体Ⅰ呼吸抑制剂不仅在医药上有着重要的研究价值,在农药方面也有着特殊的意义。这类药剂的作用机制比较特殊,害虫不易产生抗性,是一类非常有前途的杀虫药剂。从植物,微生物等生物体中发现了许多线粒体复合体Ⅰ呼吸抑制剂,如鱼藤酮、粉蝶霉素A、辣椒碱,番荔枝内酯、myxalamid等,它们可以作为农药的先导化合物进行药物合成。根据不同的作用方式,线粒体复合体Ⅰ呼吸抑制剂可分3种类型,分别以粉蝶霉素A、鱼藤酮、辣椒碱为代表。     

Biophysical aspects of nervous activity in relation to studies on the mode of action of insecticides

The spread to the nervous system of topically applied neurotoxic insecticides is discussed. During spread different insecticides may distribute themselves between fluid and solid phases within the insect's tissues in greatly differing ratios. These differences are reflected in their concentrations in the haemolymph, which is probably the medium mainly responsible for distributing insecticides within the insect, and also the main source from which insecticides enter the nervous system. Evidence for spread via more restricted pathways, such as tracheae or the nerves themselves, is conflicting. The mode of action of neurotoxic insecticides is briefly reviewed, including the role of secondarily released neuroactive stress substances, and the extent to which resistance is expressed at the site of action is also discussed. Finally, the nervous system is examined for new sites of insecticidal action which, if attacked, could give greater specificity.     

Scorpion neurotoxins: structure/function relationships and application in agriculture

Continued use of non‐specific chemical insecticides poses potential risks to the environment and to human health resulting from non‐target toxicity and increased insect resistance to these agents. Scorpions produce anti‐insect selective polypeptide toxins that bind to and modulate voltage‐sensitive ion channels in excitable tissues, thus offering alternative, environmentally safer means for insect pest control. Despite this potential, little is known about their structural elements dictating anti‐insect preference, which may be useful for the design of selective insecticides. We used a bacterial system for expression and genetic dissection of two pharmacologically distinct scorpion toxins: alpha and excitatory. By exploiting a multi‐disciplinary approach consisting of mutagenesis, protein chemistry, electrophysiology, binding and toxicity assays, and structural studies, we elucidated the bioactive surface of two anti‐insect toxins, LqhαIT and Bj‐xtrIT. In both polypeptides the bioactive surface is composed of residues surrounding the C‐terminal region. In addition, a direct, immediate approach in using the toxin genes was demonstrated by engineering baculoviruses with cDNAs encoding LqhIT2 (depressant toxin), and LqhIT1 (excitatory toxin) resulting in viral vectors with significantly improved insecticidal efficacy. © 2000 Society of Chemical Industry     

Prospects for the development of a genetically engineered baculovirus insecticide

Baculoviruses are insect-specific pathogens which have been employed for the natural control of insect pests for many years. They present an alternative to the use of chemical agents for crop protection. Their restricted host range enables particular pest species to be targeted, and this prevents any loss of beneficial insects from a treated area. However, their large-scale development as insect control agents has been limited because they take longer to kill the host following infection than rapidly-acting chemical insecticides. The aim of genetic modification of baculoviruses is to increase their speed of action, while preserving their beneficial characteristics, so creating a fast-acting, environmentally friendly insecticide. The prospects for achieving this are discussed in this paper, with consideration of scientific progress and the influence of economic and legislative factors.