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。     

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.     

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。     

抗苄呋菊酯淡色库蚊对其它拟除虫菊酯的交互抗性

1978年秋末从上海闸北区采得淡色库蚊用苄呋菊酯选育获得高达960倍的抗性品系(Res品系),它对被测定的其它拟除虫菊酯具有不同程度的交互抗性,抗性倍数从7倍(天然除虫菊酯)至7,342倍(戊氰菊酯)。交互抗性大于90倍以上的是醇部位中带有3-苯氧基苄基的二氯苯醚菊酯及其顺、反异构体或醇部位中带有`α``-氰基-3-苯氧基苄基的溴氰菊酯和戊氰菊酯。具有高抗性的4种杀虫剂(包括苄呋菊酯)分别与增效剂增效醚(Pb)或磷酸三苯酯(TPP)以1:1的比例混用后的结果表明,抗性机制与多功能氧化酶和酯酶皆有密切关系,但氧化酶抑制剂增效醚(Pb)的增效效果明显地要强于酯酶抑制剂磷酸三苯酯(TPP)的增效效果。     

甜菜夜蛾对三种拟除虫菊酯类杀虫剂的抗性遗传力及风险评估

在室内抗性选育的基础上,应用数量遗传学中的域性状分析法,分别研究了甜菜夜蛾(Spodoptera exigua)对三氟氯氰菊酯、氰戊菊酯和顺式氯氰菊酯3种拟除虫菊酯类杀虫剂的抗性现实遗传力(h²),并对3种药剂在不同杀死率下的抗性发展速率进行了预测.结果表明,用三氟氯氰菊酯、氰戊菊酯、顺式氯氰菊酯分别连续汰选12、10和10代后,甜菜夜蛾对3种药剂的抗性分别提高了18.1、27.6和45.4倍,抗性现实遗传力分别为0.2567、0.3571和0.4239.假设遗传力为实验室汰选估算值的一半、药剂杀死率为50%～90%、预计抗性增长10倍时,三氟氯氰菊酯需要约9～20代,氰戊菊酯需要约6～14代,顺式氯氰菊酯需要约5～12代.甜菜夜蛾对以上3种拟除虫菊酯类杀虫剂均存在抗性风险.相比之下,顺式氯氰菊酯和氰戊菊酯的抗性风险大于三氟氯氰菊酯.     

离体鼠肝细胞及P-450酶系与拟除虫菊酯类杀虫剂的相互作用

经研究表明顺式氯氰菊酯、甲氰菊酯、氟氰菊酯、氰戊菊酯在正辛醇——水相中的分配系数(logP)分别为6.13,5.50,5.55和4.02。离体培养的大白鼠原代肝细胞对上述杀虫剂(除氟氰菊酯外)的吸收系数(q值)分别为322.6,281.1,201.9。上述杀虫剂在离体培养的原代肝细胞中降解很快,温育30分钟后降解速率分别为90.0％(顺式氯氰菊酯)、86.2％(甲氰菊酯)、69.5％(氰戊菊酯)。同时有明显的减少离体培养的鼠肝细胞中P-450含量的作用,其中以溴氰菊酯和顺式氯氰菊酯作用较明显。在溴氰菊酯处理组中,随杀虫剂浓度的升高,这种减少P-450含量的作用明显增强。检查温育反应90分种后的细胞悬浮反应液,对照组仍维持高的细胞存活率(74.4％和91.2％),而杀虫剂处理组,除甲氰菊酯处理组仍有42.4％的存活率外,其余各杀虫剂(溴氰菊酯各浓度组和顺式氯氰菊酯处理组)均未检出存活的细胞。结果表明,试验所用的几种拟除虫菊酯类杀虫剂具有一定的细胞毒性。它们在鼠肝细胞中的降解代谢可能主要不是由于多功能氧化酶起作用。另外,杀虫剂的亲脂性与q值及在肝细胞中的降解速率三者间有一定的正相关。     

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。     

我国白纹伊蚊对三种拟除虫菊酯类杀虫剂抗性检测诊断剂量的建立

【目的】建立中国白纹伊蚊Aedes albopictus成蚊对溴氰菊酯、氯菊酯和高效氯氟氰菊酯杀虫剂抗性检测的诊断剂量。【方法】应用溴氰菊酯、氯菊酯和高效氯氟氰菊酯原药制作不同浓度的药膜滤纸,接触筒法测定白纹伊蚊实验室敏感品系成蚊对3种拟除虫菊酯类杀虫剂的敏感性,记录1 h的击倒数和24 h的死亡数。应用Excel 2007和SPSS20.0进行数据统计处理,并制作杀虫剂的毒力回归线,计算各自的LC_(50)和LC_(99)值。以2倍LC_(99)值作为区分抗性和敏感种群的诊断剂量,制作药膜滤纸,接触筒法生物测定海口市白纹伊蚊现场种群成蚊对杀虫剂的抗药性。【结果】溴氰菊酯、高效氯氟氰菊酯和氯菊酯杀虫剂对白纹伊蚊实验室敏感品系成蚊的LC_(50)值分别为0.00619%,0.01403%和0.05009%,LC_(99)值分别为0.05175%,0.11859%,和0.53165%,相对应的诊断剂量分别为0.1035%,0.2372%和1.0633%。应用上述溴氰菊酯、氯菊酯和高效氯氟氰菊酯诊断剂量测定的海口市白纹伊蚊现场种群的死亡率分别为22.58%,36.29%和40.83%,表明该种群对这3种菊酯类杀虫剂均已产生了抗性。【结论】本研究建立的白纹伊蚊对3种拟除虫菊酯类杀虫剂的诊断剂量可作为该蚊成蚊抗药性监测的参考。     

不同杀虫剂选育对家蝇抗药性水平及kdr基因频率的影响

采用杀虫剂（溴氰菊酯和甲基嘧啶磷）筛选及不接触药物自然衰退的方法,研究了家蝇Musca domestica氯氟氰菊酯高抗品系(Cyh-R)对杀虫剂的抗性变化,探讨蝇类抗药性治理的方法。用点滴法测定氯氟氰菊酯对不同家蝇品系的毒力,比较抗药性的变化,结合特异性等位基因PCR扩增（PASA）技术检测了不同家蝇品系的kdr基因频率,探讨kdr基因频率与抗性水平之间的关系。结果表明:甲基嘧啶磷筛选后,氯氟氰菊酯对第2~8代Cyh-R品系的LD₅₀呈递减趋势,从F₀的2.8434 μg/蝇降为F₈的0.4404 μg/蝇,但第8~18代Cyh-R品系的LD₅₀呈逐代递增趋势;溴氰菊酯筛选后,氯氟氰菊酯对Cyh-R品系第2~16代的LD₅₀呈上升趋势,从F₀的2.8434 μg/蝇升至F₁₆的24.3249 μg/蝇;表明了施用有机磷杀虫剂可降低其对氯氟氰菊酯的抗药性,而施用拟除虫菊酯药剂则有助于其对氯氟氰菊酯抗药性的增长;不使用任何杀虫剂也能降低其对氯氟氰菊酯的抗药性,但下降速率低于甲基嘧啶磷。PASA技术检测表明Cyh-R品系的kdr抗性基因频率为88.8%,不经过任何药剂筛选其kdr抗性基因频率下降程度最大,达到69.7%;甲基嘧啶磷筛选后其结果降为78.8%,而经溴氰菊酯筛选后kdr抗性基因频率则明显升高,达到98.9%。通过对kdr抗性基因频率和抗性水平进行相关和回归分析表明kdr抗性基因频率与家蝇对氯氟氰菊酯的LD₅₀呈对数关系,即LD₅₀值高的品系其kdr抗性基因频率相应的也较高。建议在家蝇防治中考虑轮换用药。     

Resistance and synergistic effects of insecticides in Bactrocera dorsalis (Diptera: Tephritidae) in Taiwan

Oriental fruit flies, Bactrocera dorsalis (Hendel), were treated with 10 insecticides, including six organophosphates (naled, trichlorfon, fenitrothion, fenthion, formothion, and malathion), one carbamate (methomyl), and three pyrethroids (cyfluthrin, cypermethrin, and fenvalerate), by a topical application assay under laboratory conditions. Subparental lines of each generation treated with the same insecticide were selected for 30 generations and were designated as x-r lines (x, insecticide; r, resistant). The parent colony was maintained as the susceptible colony. The line treated with naled exhibited the lowest increase in resistance (4.7-fold), whereas the line treated with formothion exhibited the highest increase in resistance (up to 594-fold) compared with the susceptible colony. Synergism bioassays also were carried out. Based on this, S,S,S-tributyl phosphorotrithioate displayed a synergistic effect for naled, trichlorfon, and malathion resistance, whereas piperonyl butoxide displayed a synergistic effect for pyrethroid resistance. All 10 resistant lines also exhibited some cross-resistance to other insecticides, not only to the same chemical class of insecticides but also to other classes. However, none of the organophosphate-resistant or the methomyl-resistant lines exhibited cross-resistance to two of the pyrethroids (cypermethrin and fenvalerate). Overall, the laboratory resistance and cross-resistance data developed here should provide useful tools and information for designing an insecticide management strategy for controlling this fruit fly in the field.     

Correlated insecticide cross-resistance in azinphosmethyl resistant codling moth (Lepidoptera: Tortricidae)

Resistance to several classes of insecticides was correlated with azinphosmethyl resistance in codling moth, Cydia pomonella (L.), in California. In tests of laboratory and field populations, cross-resistance was positively correlated with azinphosmethyl and two organophosphates (diazinon, phosmet), a carbamate (carbaryl), a chlorinated hydrocarbon (DDT), and two pyrethroids (esfenvalerate and fenpropathrin). Additionally, negatively correlated cross-resistance was identified between azinphosmethyl and two other organophosphates, chlorpyrifos and methyl parathion. Patterns of resistance observed in laboratory colonies were confirmed with field bioassays. In bioassays of field populations, azinphosmethyl resistance was observed to increase from 1991 to 1993, although levels of resistance remained < 13-fold. Because orchards with azinphosmethyl resistance have had difficulties with suppression of codling moth, and cross-resistance was found for all tested classes of insecticides, strategies for managing resistance will need to be developed so as to protect current and future control tactics. The two insecticides with negatively correlated cross-resistance are discussed as potential tools for resistance management.     

Evidence of field-evolved resistance to organophosphates and pyrethroids in Chrysoperla carnea (Neuroptera: Chrysopidae)

The toxicity of some of the most commonly used insecticides in the organophosphate and pyrethroid classes were investigated against different Chrysoperla carnea (Stephens) (Neuroptera: Chrysopidae) populations collected over three consecutive years (2005-2007). The populations were tested using leaf dip bioassays for residual effects and topical applications to measure the response of larvae that would come into direct contact with field application of insecticides. In leaf dip assays, the LC50 (micrograms per milliliter; 120 h) values for chlorpyrifos and profenofos were in the range of 59.3-1,023 and 180.02-1,118 respectively. The LC50 values for lambda-cyhalthrin, alphamethrin, and deltamethrin were 359.08-2,677, 112.9-923.5, and 47.81-407.03, respectively. The toxicity for the above insecticides in topical application was similar to toxicity in leaf dip assays. The susceptibility of a laboratory population, which was locally developed and designated as (Lab-PK), to deltamethrin was comparable with another susceptible laboratory population. Resistance ratios for five field populations were generally low to medium for deltamethrin, but high to very high for chlorpyrifos, profenofos, lambda-cyhalthrin and alphamethrin compared with the Lab-PK population. Our data also suggested that the five field populations had multiple resistance to two classes of insecticides. The populations showed resistance to two organophosphates tested and to lambda-cyhalthrin and alphamethrin; however, resistance to deltamethrin was only found at two locations. This pattern indicates occurrence of two divergent patterns of resistance within pyrethroids. The resistance to the insecticides was stable across 3 yr, suggesting field selection for general fitness had also taken place in various populations of C. carnea. The broad spectrum of resistance and stability of resistance to insecticides in C. carnea in the current study suggested that it could be a prime candidate for mass releases and compatible with most spray programs.     

棉铃虫对氰戊菊酯抗性和敏感品系的选育

用氰戊菊酯对来自阳谷的棉铃虫(YG)Heliothis armigera(Hubncr) 进行抗性晶系的筛选。在15代期间经过9代的室内选育,获得抗性品系(Fcn-R),抗性倍数高达2“3倍,筛选后F₁₅代LD₅₀值(24.1412μg/头)比筛选前F1代lD₅₀值(0.2020μg/头)提高了119.5倍。对来自偃师的棉铃虫(YS)进行了连续两代单对筛选,得到敏感品系(Fen-S),敏感晶系的LD50值为0.0116μg/头,接近1983年东台敏感晶系的LD₅₀值(0.0098μg/头)Fcn-R抗性晶系筛选前后分别测定了七种杀虫剂的剂量-死亡回归线,发现Fen-R抗性品系对溴氰菊酯[LD₅₀(Fen-R)/LD₅₀(YG)=5.2X] 和氯氰菊酯(2.5X)具有一定程度的交互抗性;而对功夫菊酯(0.66X),氯菊酯(0.89x)、灭多威(0.74X)及久效磷(1.5x)没有交互抗性。氰戊菊酯加Pb的增效试验结果表明棉铃虫对氰戊菊酯的抗性主要是由于多功能氧化酶的代谢作用。毒理学资料还暗示抗性为多因子(基因)的。     

铃虫对氰戊菊酯抗性和敏感品系的选育

用氰戊菊酯对来自阳谷的棉铃虫（ＹＧ）Ｈｅｌｉｏｔｈｉｓａｒｍｉｇｅｒａ（Ｈｕｂｎｅｒ）进行抗性品系的筛选。在１５代期间经过９代的室内选育,获得抗性品系（Ｆｅｎ－Ｒ）,抗性倍数高达２４６３倍,筛选后Ｆ１,代ＬＤ５０值（２４．１４１２ｇ／头）比筛选前Ｆ１代ＬＤ５０值（０．２０２０　ｇ／头）提高了１１９．５倍。对来自偃师的棉铃虫（ＹＳ）进行了连续两代单对筛选,得到敏感品系（Ｆｅｎ－Ｓ）,敏感品系的ＬＤ５０值为０．０１１６ｇ／头,接近１９８３年东台敏感品系的ＬＤ５０值（０．００９８ｇ／头）。Ｆｅｎ－Ｒ抗性品系筛选前后分别测定了七种杀虫剂的剂量－死亡回归线,发现Ｆｅｎ－Ｒ抗性品系对溴氰菊酯［ＬＤ５０（Ｆｅｎ－Ｒ）ＬＤ５０（ＹＧ）＝５．２Ｘ］和氯氰菊酯（２．５Ｘ）具有一定程度的交互抗性;而对功夫菊酯（０．６６Ｘ）,氯菊酯（０．８７Ｘ）、灭多威（０．７４Ｘ）及久效磷（１．５Ｘ）没有交互抗性。氰戊菊酯加Ｐｂ的增效试验结果表明棉铃虫对氰戊菊酯的抗性主要是由于多功能氧化酶的代谢作用。毒理学资料还暗示抗性为多因子（基因）的。     

Development of resistance to spinosad in oriental fruit fly (Diptera: Tephritidae) in laboratory selection and cross-resistance

In this study, we assessed the potential for the development of resistance to the insecticide spinosad in a laboratory colony of the oriental fruit fly, Bactrocera dorsalis (Hendel) (Diptera: Tephritidae). Resistance was selected by using topical applications of spinosad. After eight generations of selection, the LD50 of the selected line was 408 times greater compared with that of the untreated parental colony. This spinosad-resistant line did not exhibit cross-resistance to 10 other insecticides tested, including six organophosphates (naled, trichlorfon, fenitrothion. fenthion, formothion, and malathion) one carbamate (methomyl), and three pyrethroids (cyfluthrin, cypermethrin, and fenvalerate). However, using lines previously selected for resistance to these same insecticides, two of the 10 lines tested (naled- and malathion-resistant) did show some cross-resistance to spinosad. Also, oriental fruit flies from different field collections where naled and malathion have been used for control purposes displayed some resistance to spinosad. In addition, the effects of direct ingestion of spinosad through dietary supplementation also were tested. Overall, the laboratory resistance and cross-resistance data developed in this study provide new information that will be useful for managing the development of resistance when spinosad is used to control B. dorsalis in the field.     

Evolution of insecticide resistance in non-target black flies (Diptera: Simuliidae) from Argentina

Black flies, a non-target species of the insecticides used in fruit production, represent a severe medical and veterinary problem. Large increases in the level of resistance to the pyrethroids fenvalerate (more than 355-fold) and deltamethrin (162-fold) and a small increase in resistance to the organophosphate azinphos methyl (2-fold) were observed between 1996-2008 in black fly larvae under insecticide pressure. Eventually, no change or a slight variation in insecticide resistance was followed by a subsequent increase in resistance. The evolution of pesticide resistance in a field population is a complex and stepwise process that is influenced by several factors, the most significant of which is the insecticide selection pressure, such as the dose and frequency of application. The variation in insecticide susceptibility within a black fly population in the productive area may be related to changes in fruit-pest control. The frequency of individuals with esterase activities higher than the maximum value determined in the susceptible population increased consistently over the sampling period. However, the insecticide resistance was not attributed to glutathione S-transferase activity. In conclusion, esterase activity in black flies from the productive area is one mechanism underlying the high levels of resistance to pyrethroids, which have been recently used infrequently. These enzymes may be reselected by currently used pesticides and enhance the resistance to these insecticides.     

Metabolic mechanisms only partially explain resistance to pyrethroids in Australian broiler house populations of lesser mealworm (Coleoptera: Tenebrionidae)

The susceptibility of six Australian broiler house populations and an insecticide susceptible population of lesser mealworm, Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae), to cyfluthrin, beta-cyfluthrin, gamma-cyhalothrin, and deltamethrin was investigated. One broiler house population had equivalent susceptibility to the susceptible to beta-cyfluthrin and beta-cyhalothrin, with higher susceptibility to cyfluthrin and deltamethrin. The remaining five populations demonstrated strong resistance to cyfluthrin (19-37-fold), the insecticide used most widely for management of A. diaperinus in Australia. Each cyfluthrin-resistant population demonstrated reduced susceptibility to beta-cyfluthrin (resistance ratios were 8-17-fold), deltamethrin (2.5-8-fold), and gamma-cyhalothrin (6-12-fold) compared with the laboratory population, but cross-resistance patterns varied considerably between populations. Adding piperonyl butoxide (PBO) had no effect on the susceptibility of the susceptible population to any of the insecticides, but it increased the susceptibility of each of the five cyfluthrin-resistant populations: to cyfluthrin (synergism ratio range, 1.9-5.0-fold), beta-cyfluthrin (1.6-4.1-fold), and y-cyhalothrin (1.7-2.0-fold). PBO had a more variable effect on susceptibility to deltamethrin, with three of the cyfluthrin-resistant populations being more susceptible to deltamethrin in the presence of PBO, but susceptibility of the remaining two populations was unaffected by adding PBO (synergism ratio range, 0.9-2.5-fold). Overall, the addition of PBO to the four pyrethroids had variable effects on their susceptibility. This variability indicated the presence of other resistance mechanisms in beetle populations apart from metabolic resistance. In addition, the relative importance of metabolic resistance in each beetle population varied widely between pyrethroids. Thus, it cannot be assumed that PBO will reliably synergize pyrethroids against cyfluthrin-resistant lesser mealworm populations when using it to mitigate insecticide resistance.     

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。