神经递质释放与家蝇对拟除虫菊酯抗性关系研究

通过生物测定比较溴氰菊酯、氯菊酯和DDT对Dec-R,2C1-R,DDT-R和敏感(SP)家蝇Musca domestica vicina的毒力,表明三个抗性品系对溴氰菊酯、氯菊酯和DDT均有很高的抗性,抗性倍数分别达120 912,6 032和112.2倍,并对上述三种杀虫剂有明显的交互抗性和抗击倒效应。杂交试验表明Dec-R对溴氰菊酯的抗性是一个隐性基因,电生理试验表明抗性家蝇中枢神经系统(CNS)对药剂敏感度的降低是其产生抗性和交互抗性的重要机制。研究结果表明Dec-R和2CLR家蝇品系中存在有击倒抗性因子(Kdr)。当用1×10^-7mol/L溴氰菊酯对SP家蝇脑突触体在提高K+浓度去极化后,可加强3H-胆碱的释放,而在Dec-R品系中,溴氰菊酯浓度提高到1×10^-4m0l/L也未能加强³H-胆碱的释放,表明溴氰菊酯与神经递质的释放和钠通道亲和性的降低是抗性的主要机制。     

家蝇对拟除虫菊酯抗性机理研究进展

本文概述了家蝇对拟除虫菊酯的抗性机制 ,特别是结合我国室内培育的抗拟除虫菊酯家蝇品系 (Dec R和 2Cl R) ,探讨了Na+通道、神经递质释放、ATPase、蛋白质磷酸化等与家蝇Kdr抗性的关系 ,从多方面证明了神经敏感性降低是家蝇对拟除虫菊酯产生抗性的重要机制。     

拟除虫菊酯对家蝇Na-K-ATPase抑制作用的研究

通过对家蝇神经系统Na-K-ATPase性质的研究,表明Na-K-ATPase反应的适宜pH值为7.0～8.0,适温为35～40℃,K_m为0.22 mmol/L,V_max为555.56 nmol/(mg·min)。比较测定了家蝇敏感品系、Del-R、2Cl-R抗性品系的Na-K-ATPase活性及溴氰菊酯和氯菊酯对该酶的抑制作用。实验证明,敏感与抗性品系间Na-K-ATPase活力无显著的差异,但溴氰菊酯和氯菊酯对不同家蝇品系Na-K-ATPase的抑制作用有明显区别,两种拟除虫菊酯可抑制敏感家蝇品系Na-K-ATPase的活性,而对抗性品系无明显的抑制作用。     

拟除虫菊酯抗性家蝇的交互抗药性研究

拟除虫菊酯的广泛使用使家蝇普遍产生了抗药性,为有效的控制家蝇的危害,需要了解家蝇对轮换或新杀虫剂的交互抗性状况。作者用点滴法测定了两个实验室汰选的拟除虫菊酯抗性家蝇品系对几种杀虫剂的交互抗性,结果表明:二氯苯醚菊酯和溴氰菊酯之间存在较高程度的交互抗药性;拟除虫菊酯抗性较高的家蝇对作用机制不同的新农药(多杀菌素、氟虫腈)表现较低程度的交互抗性。     

棉铃虫对拟除虫菊酯抗性稳定性研究

研究棉铃虫Helicoverpa armigera (Hubner)对三种拟除虫菊醌（氰戊菊酯、溴氰菊酯、功夫菊酯)的抗性稳定性及敏感性恢复表明,即使棉铃虫对氰戊菊酯的抗性达到3166.3倍以上,抗性仍不稳定,经14代室内饲养后抗性下降为61.4倍;对一系列田间抗性种群的抗性稳定性研究后发现,棉铃虫对这三种拟除虫菊酯的抗性不稳定,在没有杀虫剂选择的情况下,开始几代抗性下降较快,当下降到一定水平(2～9倍)后,抗性比较稳定,很难完全恢复对拟除虫菊酯的敏感性。     

北京地区家蝇对拟除虫菊酯的抗药性与α-乙酸萘酯(α-NA)酯酶的关系

 通过1988年以来对北京地区家蝇(Musca domestica L.)抗药性的研究表明,北京朝阳、回龙观、宣武、景陵地区的家蝇种群与海口种群LD_(50)比较,对溴氰菊酯的抗性为5.4—17.2倍,对二氯苯醚菊酯的抗性为2460—3080倍。不同抗性水平的家蝇种群中,α-NA确酶活性大于0.5(A600值(0.01头·15分))的个体频率与其溴氰菊酯和二氯苯醚菊酯的抗性程度具有显著相关(相关系数分别为0.94和0.98)。抗性程度较高的宣武种群平均酯酶活性为280μmol/(mg蛋白质·分),是海口敏感种群的10.5倍。海口种群α-NA酯酶的米氏常数值(Km)为4.49mol/L,分别是朝阳、回龙观、宣武种群的4.7、4.2和2.3倍,说明抗性种群α-NA酯酶对底物的亲合力高于相对敏感的海口种群。     

拟除虫菊酯的结构与害虫抗药性的关系

以家蝇Musca domestica vicina L.为试虫,用汰选方法研究了家蝇对六种不同拟除虫菊酯的抗性发展.结果表明,家蝇对不同化学结构和不同光学异构体组分的拟除虫菊酯抗性差别很大.对溴氧菊酯、顺式氯氰菊酯抗性发展很快,氯氰菊酯次之,氰戊菊酯、氰戍菊酯A和氰戊菊酯A_σ抗性发展较慢.用抗溴氰菊酯家蝇品系和点滴法测定了十种拟除虫菊酯和七种有机磷杀虫剂的毒力,讨论了它们之间的交互抗性和结构与抗性的关系.溴氰菊酯(抗性比值24.00)、氯氰嫡酯(抗性比值20.11)、顺式氯氰菊酯(抗性比值38.10)和二氯苯醚菊酯(抗性比值11.04),结构相近的交互抗性比较严重.氰戊菊酯(抗性比值4.64)、氰戊菊酯A(抗性比值5.97)、戊菊酯(抗性比值4.49)和氟氰菊酯(抗性比值4.12)化学结构中醇部分与溴氰菊酯相同、酸部分不同,它们与溴氰菊酯交互抗性水平较低.联苯菊酯(抗性比值1.98)化学结构中酯和醇部分都与溴氰菊酯不同,其交互抗性水平较低,杀螟松等七种有机磷杀虫剂,除敌敌畏与溴氰菊酯有轻微交互坑性外,其它均无交互抗性.     

家蝇对DDT和拟除虫菊酯的重要抗性机制——中枢神经系统的不敏感性

本文以电生理技术研究了四个品系的家蝇Musca domestica vicina Macq.中枢神经系统(CNS)对DDT、二氯苯醚菊酯和澳氰菊酯的敏感性,结果表明:三种抗性家蝇,DDT高抗品系(DDT-R)、二氯苯醚菊酯高抗品系(2C1-R)和溴氰菊酯高抗品系(Dec-R)的中枢神经系统(CNS)对三种杀虫剂的敏感性与敏感家蝇相比均明显降低,而且,GNS的不敏感性随杀虫剂LD₅₀的升高有逐渐上升的趋势.我们认为,CNS不敏感性是家蝇对DDT相拟除虫菊酯产生抗性的一个重要机制,也是产生交互抗性的一个重要原因.     

家蝇对DDT和拟除虫菊酯的重要抗性机制——中枢神经系统的不敏感性

本文以电生理技术研究了四个品系的家蝇Musca domestica vicina Macq.中枢神经系统(CNS)对DDT、二氯苯醚菊酯和澳氰菊酯的敏感性,结果表明:三种抗性家蝇,DDT高抗品系(DDT-R)、二氯苯醚菊酯高抗品系(2C1-R)和溴氰菊酯高抗品系(Dec-R)的中枢神经系统(CNS)对三种杀虫剂的敏感性与敏感家蝇相比均明显降低,而且,GNS的不敏感性随杀虫剂LD₅₀的升高有逐渐上升的趋势.我们认为,CNS不敏感性是家蝇对DDT相拟除虫菊酯产生抗性的一个重要机制,也是产生交互抗性的一个重要原因.     

Response of insecticide-resistant and susceptible houseflies (Musca domestica) to a commercial granular bait formulation containing methomyl

Female houseflies (Musca domestica L.) from a susceptible and a multi-insecticide-resistant strain were used to evaluate the relative toxicity of an insecticide bait formulation of the carbamate insecticide methomyl. Individual flies were allowed to feed on bait granules for an unrestricted period or for 5 s. Resistant flies took longer than susceptible flies to initiate a feeding response. When allowed to feed continuously, those from the resistant strain spent longer feeding than susceptible ones. The time taken to knock-down (KD), including feeding times, was significantly greater for resistant than susceptible flies (P less than 0.001), but once the proboscis was withdrawn from the granule there was no difference in KD times between the strains. All flies from both strains were knocked down, and only a very small number of resistant and susceptible flies recovered. The toxic effects of methomyl on flies which were restricted to a 5 s feed ranged from no observed effect to KD in less than 1 min. After feeding for 5 s, 81% of resistant and 98% of susceptible flies developed signs of methomyl poisoning. More resistant than susceptible flies recovered from KD, giving final mortalities of 46% and 88% respectively. With both feeding regimes, some flies of both strains which had apparently recovered from KD had lost their ability to fly. Observations have also shown that 8% of resistant flies may have been repelled by methomyl granules. The implication of these results on the survival of M. domestica in intensive animal units following exposure to methomyl bait is also discussed.     

有机磷对抗拟除虫菊酯家蝇的毒力

测定敏感、抗溴氰菊酯(Del-R)、抗氯菊酯(2Cl-R)的家蝇品系对有机磷杀虫剂敌敌畏、辛硫磷及马拉硫磷的LD₅₀,α-乙酸萘酯（α-NA）酯酶动力学,酯酶的活性和酯酶的抑制作用。Del-R和2Cl-R的家蝇品系对三种有机磷杀虫剂的抗性倍数为0.966～7.190倍,均为低抗水平。三个家蝇品系的羧酸酯酶活性水平与抑制中浓度存在正相关性,说明羧酸酯酶在抗拟除虫菊酯家蝇对有机磷杀虫剂的抗性中起一定的作用。     

Lipophorin in the larval and adult stages of Musca domestica

The major Musca domestica hemolymph lipoprotein, lipophorin, was purified from larval and from adult animals. The housefly lipophorin is composed of two apoproteins, apolipophorin I (M_r ∽ 253,000) and apolipophorin II (M_r ∽ 85,000). The lipophorin contains about 3.9% carbohydrates and reacts positively with concanavalin A. The density of larval lipophorin is equal to 1.152 g/ml and of adult lipophorin to 1.106 g/ml. The amount of lipophorin per animal increases during the larval stage, is constant during pupal stage, and suffers a great reduction at the pharate adult stage. The amount of lipophorin remains stable during the whole first gonotrophic cycle of the housefly. Lipophorin is not detected in the eggs of this insect.     

家蝇攻击素(Attacin)基因的克隆与表达

通过同源克隆策略并结合cDNA末端快速扩增(RACE)技术,克隆到了编码家蝇攻击素(Attacin)基因的全长cDNA序列(GenBank登陆号：AY460106),并对相应的氨基酸序列进行了序列比对,对系统关系等进行了生物信息学分析。家蝇Attacin的cDNA全长778bp,其中包括627bp的开放式读码框(ORF),44bp的5’末端非编码区(5’UTR)和107bp的3’末端非编码区(3’UTR),相应的蛋白产物含208个氨基酸,与其他双翅目昆虫的Attacin具有很高的相似性,约50％-70％。以邻接法(Neighbor-Joining,NJ)所构建的系统关系表明,家蝇的Attacin与其他双翅目昆虫的Attacin起源于共同的祖先。应用半定量RT-PCR的方法,研究家蝇幼虫在受到外源刺激后Attacin基因的表达,结果表明,在家蝇幼虫体内Attacin基因呈诱导型表达,表达水平随诱导时间和诱导源的不同而变化。     

Resistance to organophosphorus insecticides in the SKA strain of houseflies (Musca domestica)

The response of a susceptible strain and the diazinon-selected strain of houseflies, the SKA, to 20 organophosphorus insecticides was measured by topical application and injection.The SKA strain was very resistant to diazinon, ethyl chlorthion, chlorthion and parathion, moderately resistant to five other insecticides and only slightly resistant to the remainder. It was more resistant to the diethoxy than dimethoxy compounds and was only very slightly resistant to malathion. The compounds, especially the phosphates, were more toxic when injected than when applied topically. Additions or substitutions in the phenyl ring of the parathion molecule resulted in compounds less toxic than parathion, particularly to susceptible flies.

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Zusammenfassung Die Reaktion eines anfälligen und des auf Diazinonresistenz ausgelesenen Stammes SKA der Stubenfliege auf Organophosphor-Insektizide wurde nach topischer Applikation und Injektion gemessen.Der SKA-Stamm war sehr widerstandsfähig gegen Diazinon, Äthylchlorthion, Chlorthion und Parathion, mäßig resistent gegen fünf andere Insektizide und nur schwach resistent gegen die übrigen. Er war gegen die Diäthoxy-Verbindungen widerstandsfähiger als gegen Dimethoxy-Verbindungen und gegen Malathion nur sehr schwach resistent. Die Verbindungen, insbesondere die Phosphate, wirkten injiziert giftiger als bei äußerer Applikation. Additionen und Substitutionen im Phenyl-Ring des Parathion-Moloküls ergaben Verbindungen, die besonders gegen anfällige Fliegen weniger toxisch wirken.

     

家蝇抗菌肽的研究与应用

近年来,对家蝇Muscadomestica体内外抗菌活性物质的研究受到人们广泛的关注,其中研究较多的是抗菌肽。家蝇抗菌肽对细菌、真菌、肿瘤癌细胞和病毒等具有生物活性,可通过针刺、带菌针刺、超声波、放射性射线、高频电磁场和生理盐水等方法诱导增量产生,能耐极端的温度和pH值的溶液,在高浓度盐溶液中也很稳定。文章还对家蝇抗菌肽的结构特点、作用机理与分子生物学研究做了概述,同时对家蝇抗菌肽的应用前景进行了讨论。     

Laboratory evaluation of cyromazine against insecticide-resistant field strains of Musca domestica

A screening programme was undertaken to examine the possibility of cross resistance occurring between cyromazine and conventional insecticides. The responses of nine strains of Musca domestica to treatment with cyromazine, trichlorphon, methomyl and pyrethrins with piperonyl butoxide were measured. No tolerance to cyromazine was found, neither was there a direct correlation between larval responses to cyromazine and adult responses to other insecticides. Cyromazine is a potent larvicide against M. domestica and the results of these tests show that it has good potential for the control of houseflies with high levels of resistance to other insecticides.     

Superoxide dismutase in the housefly,Musca domestica (L.)

Four superoxide dismutase (SOD) (E.C. 1.15.1.1) isozymes were present in whole tissue homogenates of Musca domestica when examined by polyacrylamide gel electrophoresis. One of the isozymes contained manganese, and the other three contained copper and zinc. All were observed in each of the body tagma (head, abdomen, and thorax) and at each developmental stage (egg to adult). The copper- and zinc-containing isozymes purified from newly emerged, adult M. domestica had a relative molecular weight of 34,800 as determined by gel filtration chromatography but consisted of two equal-size subunits of 16,000 as measured by sodium dodecylsulfate polyacrylamide gel electrophoresis. An isoelectric point between 4.8 and 5.1 was measured. Approximately 2 mol each of copper and zinc were present per dimer. The three copper, zinc isozymes were identified as charge variants. The amino acid composition of the enzyme was similar to that of copper, zinc-containing superoxide dismutases from other sources. Purified housefly copper, zinc superoxide dismutase was neither deactivated nor able to protect lactic dehydrogenase against deactivation in the presence of light and rose bengal, a known generator of singlet oxygen. The role of SOD in the phototoxic reaction involving rose bengal is discussed.     

Tracking and chasing in houseflies (Musca)

The flight trajectories of free flying female and male houseflies have been analyzed in 3 dimensions. Both female and male flies track other flies. The turning velocity α (around the vertical axis) is linearly dependent upon the horizontal angle ψ_F (that is the angle between the trajectory of the tracking fly and the target) for small values of ψ_F in females and for the whole range of ψ_F in males. The 3-dimensional velocity υ _xyz of the chasing fly is linearly dependent upon the distance between leading and chasing fly in males but not in females. Male chasing thus appears to be more efficient than female tracking. It is shown that earlier assumptions on visual control of flight in female flies derived from experiments on fixed flying flies are justified.