Insecticide toxicity, synergism and resistance in the German cockroach (Dictyoptera: Blattellidae)

The toxicity of synergism of and resistance to insecticides in four strains of German cockroach, Blattella germanica (L.), were investigated. Toxicity of nine insecticides by topical application to the susceptible strain varied greater than 2,000-fold, with deltamethrin (LD50 = 0.004 micrograms per cockroach) and malathion (LD50 = 8.4 micrograms per cockroach) being the most and least toxic, respectively. Resistance to pyrethrins (9.5-fold) in the Kenly strain was unaffected by the synergists piperonyl butoxide (PBO) or S,S,S-tributylphosphorotrithioate (DEF), suggesting that the metabolism is not involved in this case. Malathion resistance in the Rutgers strain was suppressible with PBO, implicating oxidative metabolism as a resistance mechanism. The Ectiban-R strain was resistant to all the pyrethroids tested, and cypermethrin resistance was not suppressible with PBO or DEF. These findings support results of previous studies that indicated this train has a kdr-like mechanism. Bendiocarb resistance in both the Kenly and Rutgers strains was partially suppressed by either PBO or DEF, suggesting that oxidative and hydrolytic metabolism are involved in the resistance. Trends between the effects of the synergists on the susceptible versus resistant strains are discussed.     

Pyrethroid resistance and synergism in a field strain of the German cockroach (Dictyoptera: Blattellidae).

A field-collected strain of the German cockroach, Blattella germanica (L.), was highly resistant to 10 pyrethroid insecticides (cyfluthrin, cyhalothrin, cypermethrin, fenvalerate, esfenvalerate, fluvalinate, permethrin, resmethrin, sumithrin, tralomethrin) based on topical applications and comparison with a known susceptible strain. Resistance ratios ranged from 29 to 337. In general, pyrethroid compounds with an alpha-cyano functional group were more toxic than those lacking this moiety, but resistance ratios were similar for both classes of compound. The metabolic inhibitors DEF and PBO were tested for synergism in conjunction with cypermethrin (alpha-cyano) and permethrin (non alpha-cyano). Application of synergists resulted in partial elimination of resistance, suggesting that the basis of resistance involves enhanced metabolism as well as target site insensitivity. These results suggest that pyrethroid insecticides may have a very short functional life in German cockroach control unless they are used judiciously.     

Effects of the synergists piperonyl butoxide and S,S,S-tributyl phosphorotrithioate on propoxur pharmacokinetics in Blattella germanica (Blattodea: Blattellidae).

Effects of the synergists piperonyl butoxide (PBO) and S,S,S-tributyl phosphorotrithioate (DEF) on propoxur pharmacokinetics were examined in the German cockroach, Blattella germanica (L.). Treatment of adult male German cockroaches with the cytochrome P450 monooxygenase inhibitor, PBO, or the esterase inhibitor, DEF, increased propoxur toxicity by 2- and 6.8-fold, respectively, implicating hydrolysis as a major detoxification route of propoxur in the German cockroach. However, significant hydrolytic metabolism could not be demonstrated conclusively in vitro resulting in a conflict between in situ bioassay data and in vitro metabolic studies. In vitro propoxur metabolism with NADPH-fortified microsomes produced at least nine metabolites. Formation of metabolites was NADPH-dependent; no quantifiable metabolism was detected with cytosolic fractions. However, microsomal fractions lacking an NADPH source did produce a low, but detectable, quantity of metabolites (1.6 pmol). PBO inhibited NADPH-dependent propoxur metabolism in a dose-dependent fashion, implicating cytochrome P450 monooxygenases as the enzyme system responsible for the metabolism. Interestingly, DEF also inhibited the NADPH-dependent metabolism of propoxur, albeit to a lower extent. Treatment with PBO or DEF also caused a significant reduction in the cuticular penetration rate of propoxur. The data demonstrate that unanticipated effects are possible with synergists and that caution must be exercised when interpreting synergist results.     

B型烟粉虱田间种群对毒死蜱和敌敌畏抗性的生化机制

通过增效剂生物测定和生化分析,探讨了采自福建省的B型烟粉虱Bemisia tabaci 6个田间种群对毒死蜱和敌敌畏抗性的生化机制。结果表明：与敏感品系SUD-S相比,6个田间种群对毒死蜱和敌敌畏分别具有54.53～78.43倍和6.23～11.25倍的抗性。TPP、PBO和DEM对毒死蜱的增效比分别为3.61～24.94倍、1.14～1.76倍和1.04倍,对敌敌畏的增效比分别为1.67～2.64倍、1.33～1.65倍和1.09倍,表明羧酸酯酶的解毒代谢在烟粉虱对毒死蜱的抗性中起着重要作用。烟粉虱抗性种群乙酰胆碱酯酶的K_m值是敏感品系的1.83～4.0倍,V _max值是敏感品系的0.34～0.62倍; 敏感品系乙酰胆碱酯酶的活性在底物浓度大于1.0 mmol/L时受抑制,抗性种群乙酰胆碱酯酶的活性在底物浓度大于16 mmol/L时受抑制;抗性种群乙酰胆碱酯酶对敌敌畏和毒死蜱的敏感度分别比敏感品系低119.92～161.33倍和10.11～14.24倍,表明烟粉虱田间抗性种群乙酰胆碱酯酶可能已发生了变构,由变构引起的乙酰胆碱酯酶不敏感是烟粉虱田间种群对毒死蜱和敌敌畏产生抗性的重要原因。结果提示,乙酰胆碱酯酶不敏感性和羧酸酯酶的解毒代谢在烟粉虱对毒死蜱的抗性中均起着重要作用,而乙酰胆碱酯酶不敏感性在对敌敌畏的抗性中起重要的作用,多功能氧化酶和谷胱甘肽S转移酶在烟粉虱对毒死蜱和敌敌畏抗性中所起的作用不大。     

Biochemical characterization of hydrolytic and oxidative enzymes in insecticide resistant and susceptible strains of the German cockroach (Dictyoptera: Blattellidae).

We have identified resistance mechanisms in the German cockroach, Blattella germanica (L.), for propoxur and chlorpyrifos in strains of cockroaches that display multiresistance to several organophosphate and carbamate insecticides. The resistance mechanisms involve the combined effects of increased oxidative and hydrolytic metabolism and both strains are resistant to chlorpyrifos and propoxur. Experiments designed to test for similarity in metabolic enzymes suggest that, although the mechanisms involve similar processes, the enzymes responsible for insecticide detoxification are different in the two strains. Both resistant strains exhibited enhanced activity toward alpha-naphtholic esters relative to a standard susceptible strain; however, analysis of the progeny from resistant X susceptible crosses suggests that this general esterase activity is inherited differently than propoxur or chlorpyrifos resistance. Hybrids of the propoxur-resistant strain displayed the highest activity of all cockroaches tested, in contrast to hybrids of the chlorpyrifos-resistant strain, which were similar to the susceptible strain. Native gel electrophoresis of cytosolic preparations provided further evidence for differences in the pattern of hydrolytic enzymes and inheritance of resistance in the two strains. Analysis of components of the cytochrome P450-dependent monooxygenase system and activities toward model substrates indicate that the two resistance mechanisms also involve different oxidative processes. The propoxur-resistant strain displayed significantly higher levels of total cytochrome P450, but no other components were correlated with resistance. In contrast with the chlopyrifos-resistant strain, which was similar to the susceptible strain in all parameters measured, activity toward model substrates was higher in the propoxur-resistant strain than in any of the other strains and hybrids tested.(ABSTRACT TRUNCATED AT 250 WORDS)     

Insecticide resistance and cross-resistance in the house fly (Diptera: Muscidae)

A house fly strain, ALHF, was collected from a poultry farm in Alabama after a control failure with permethrin, and further selected in the laboratory with permethrin for five generations. The level of resistance to permethrin in ALHF was increased rapidly from an initial 260-fold to 1,800-fold after selection. Incomplete suppression of permethrin resistance by piperonyl butoxide (PBO) and S,S,S,-tributylphosphorotrithioate (DEF) reveals that P450 monooxygenase- and hydrolase-mediated detoxication, and one or more additional mechanisms are involved in resistance to permethrin. The ALHF strain showed a great ability to develop resistance or cross-resistance to different insecticides within and outside the pyrethroid group including some relatively new insecticides. Resistance to beta-cypermethrin, cypermethrin, deltamethrin, and propoxur (2,400-4,200-, 10,000-, and > 290-fold, respectively, compared with a susceptible strain, aabys) in ALHF house flies was partially or mostly suppressed by PBO and DEF, indicating that P450 monooxygenases and hydrolases are involved in resistance to these insecticides. Partial reduction in resistance with PBO and DEF implies that multiresistance mechanisms are responsible for resistance. Fifteen- and more than fourfold resistance and cross-resistance to chlorpyrifos and imidacloprid, respectively, were not effected by PBO or DEF, indicating that P450 monooxygenases and hydrolases are not involved in resistance to these two insecticides. Forty-nine-fold cross-resistance to fipronil was mostly suppressed by PBO and DEF, revealing that monooxygenases are a major mechanism of cross-resistance to fipronil. Multiresistance mechanisms in the ALHF house fly strain, however, do not confer cross-resistance to spinosad, a novel insecticide derived from the bacterium Saccharopolyspora spinosa. Thus, we propose that spinosad be used as a potential insecticide against house fly pests, especially resistant flies.     

Biochemical characterization and purification of esterases from three strains of German cockroach,Blattella germanica (Dictyoptera: Blattellidae)

Three strains of German cockroach, Blattella germanica (L.) showed varying levels of resistance to chlorpyrifos, methyl parathion, propoxur, bendiocarb, and cypermethrin. The general esterase activity was at least twofold higher than susceptible strain. The subcellular distribution studies revealed that the majority of the esterase activity is present in the 100,000g cytosolic fraction. Only a small portion of the activity was membrane bound. Using non-denaturing gel electrophoresis, ten isozymes were identified in German cockroaches. These isozymes were isolated individually from the gels and analyzed for differences in activity. The isozymes E5, E6, and E7 of resistant strains had significantly higher specific activities when compared with the susceptible strain. The purification process using various column chromatography and preparative gel electrophoresis resulted in 9–11% of total esterase recovery. About double the amount of E6 was recovered from the resistant strains when compared with the susceptible strain. Kinetic analyses of E6 did not indicate differences in K_m and V_max values between the resistant and susceptible strains. Also, inhibition of esterase activity by paraoxon, chlorpyrifos, and propoxur did not suggest any structural differences in esterase E6 between strains. The results suggest that the increased production of E6 esterase contributes to insecticide resistance in German cockroaches. The role of E6 may be sequestration of toxic molecules rather than hydrolysis. © 1996 Wiley-Liss, Inc.     

Effects of hydroprene exposure on the physiology and insecticide susceptibility of German cockroaches (Orthoptera: Blattellidae).

We determined nonlethal physiological effects of hydroprene on the German cockroach, Blattella germanica L., and the effects of hydroprene exposure on German cockroach susceptibility to traditional insecticides. Live and dry weights of German cockroach adults exposed as nymphs to the juvenile hormone analog hydroprene increased significantly (6-35%) in both sexes. The change in weight was affected by hydroprene dose and time after adult emergence. Hydroprene treatment did not affect total body lipids and carbohydrates consistently. However, total body urates of males and females increased significantly. The susceptible and multiresistant German cockroach strains were significantly more susceptible to propoxur at the lower hydroprene application rate (0.697 mg per tub) than the untreated cockroaches. Hydroprene application also significantly lowered the LD50 for chlorpyrifos of the multiresistant strain at the lower hydroprene application rate. However, hydroprene failed to affect chlorpyrifos susceptibility of the susceptible strain.     

Cyclodiene resistance at the insect GABA receptor/chloride channel complex confers broad cross resistance to convulsants and experimental phenylpyrazole insecticides

This study investigated the pharmacological profile of cyclodiene resistance in Drosophila melanogaster and the mode of action of a phenylpyrazole insecticide, JKU 0422. Toxicological studies were performed with a sucrose bait assay containing the synergist piperonyl butoxide. The Maryland strain of D. melanogaster was resistant to dieldrin, lindane, picrotoxinin, TBPS, p-CN-TBOB, and JKU 0422. In contrast, this strain was susceptible to cypermethrin and the avermectins MK-243, abamectin, and abamectin 8,9-oxide. Neurophysiological studies showed that both TBPS and JKU 0422 reversed the inhibitory action of GABA in central nerve preparations from susceptible D. melanogaster. However, the response to these compounds was attenuated in nerve preparations from the resistant Maryland strain, which indicated that the resistance was expressed at the level of the nerve. Topical toxicity bioassays with JKU 0422 on susceptible (CSMA) and cyclodiene-resistant (LPP) strains of German cockroach revealed a resistance ratio of 553-fold for this compound. These studies demonstrate that cyclodiene resistance in D. melanogaster confers broad cross resistance toward compounds thought to block the GABA-gated chloride channel in a manner similar to the cyclodienes. Moreover, the cross resistance extends to JKU 0422, and resistance to this compound is also present in a strain of cyclodiene-resistant German cockroach. These toxicological results, along with the neurophysiological studies, confirm that JKU 0422 has a mode of action that is similar to the cyclodienes and TBPS. These findings suggest that the introduction and use of new chloride channel antagonists as insecticides should be managed carefully in order to prevent the rapid development of resistance in the field. © 1994 Wiley-Liss, Inc.     

The Effect of Insecticide Synergists on the Response of Scabies Mites to Pyrethroid Acaricides

Background

Permethrin is the active component of topical creams widely used to treat human scabies. Recent evidence has demonstrated that scabies mites are becoming increasingly tolerant to topical permethrin and oral ivermectin. An effective approach to manage pesticide resistance is the addition of synergists to counteract metabolic resistance. Synergists are also useful for laboratory investigation of resistance mechanisms through their ability to inhibit specific metabolic pathways.

Methodology/Principal Findings

To determine the role of metabolic degradation as a mechanism for acaricide resistance in scabies mites, PBO (piperonyl butoxide), DEF (S,S,S-tributyl phosphorotrithioate) and DEM (diethyl maleate) were first tested for synergistic activity with permethrin in a bioassay of mite killing. Then, to investigate the relative role of specific metabolic pathways inhibited by these synergists, enzyme assays were developed to measure esterase, glutathione S-transferase (GST) and cytochrome P450 monooxygenase (cytochrome P450) activity in mite extracts. A statistically significant difference in median survival time of permethrin-resistant Sarcoptes scabiei variety canis was noted when any of the three synergists were used in combination with permethrin compared to median survival time of mites exposed to permethrin alone (p<0.0001). Incubation of mite homogenates with DEF showed inhibition of esterase activity (37%); inhibition of GST activity (73%) with DEM and inhibition of cytochrome P450 monooxygenase activity (81%) with PBO. A 7-fold increase in esterase activity, a 4-fold increase in GST activity and a 2-fold increase in cytochrome P450 monooxygenase activity were observed in resistant mites compared to sensitive mites.

Conclusions

These findings indicate the potential utility of synergists in reversing resistance to pyrethroid-based acaricides and suggest a significant role of metabolic mechanisms in mediating pyrethroid resistance in scabies mites.     

Inheritance and synergism of resistance to imidacloprid in the Colorado potato beetle (Coleoptera: Chrysomelidae)

Colorado potato beetle, Leptinotarsa decemlineata (Say), adults and larvae collected from Long Island, NY, were 100.8 and 13.2 times more resistant to imidacloprid, respectively, compared with a susceptible strain. This high level of resistance appeared in only the third field season of imidacloprid use. Analysis of probit lines from F1 reciprocal crosses indicated that resistance to imidacloprid in adults was inherited autosomaly as an incompletely recessive factor. The degree of dominance of the resistance was -0.23 and -0.10, respectively, 3 and 7 d after treatment (incompletely recessive). The chi2 analysis of response ratio statistics from F1 x susceptible back crosses compared with a monogenic model suggested that more than one locus is responsible for resistance to imidacloprid. Synergism studies with piperonyl butoxide suggested that mixed-function oxidase mediated detoxification is responsible for the resistance to imidacloprid in adults. Synergism studies with S,S,S-tributyl phosphorotrithioate (DEF) indicated that esterase mediated detoxification may be an additional resistance factor. Mixed-function oxidase mediated detoxification is probably also one of the mechanisms of resistance to imidacloprid in larvae. Because the synergists used did not completely eliminate resistance in the resistant strain, there may be additional mechanisms involved. Refugia and crop rotation decrease the frequency of homozygous resistant genotypes and may be effective resistance management strategies, because of the recessive nature of the resistance.     

Effect of synergists on the oral and topical toxicity of azamethiphos to organophosphate-resistant houseflies (Diptera: Muscidae).

Dermal and oral toxicities of azamethiphos were determined in two organophosphate-resistant and one susceptible strain of houseflies, Musca domestica L. The 594vb strain was 1,967-fold more resistant to azamethiphos when compared with the susceptible Chemical Specialties Manufacturers Association (CSMA) strain by dermal application. When the compound was administered orally to the 594vb strain, we observed only a 15-fold resistance. In contrast, the Yachiyo strain, which show 1,500-fold resistance to diazinon and which has known multiple mechanisms for organophosphate resistance, showed only 6-fold resistance to azamethiphos by topical application and 4-fold resistance by oral administration. Azamethiphos administered dermally and orally was equally toxic to the CSMA and Yachiyo strains. However, when azamethiphos was administered to the 594vb strain, the insecticide was 71 times more toxic orally than by the dermal route. This result indicated involvement of a cuticular penetration factor in the resistance mechanism. The effect on azamethiphos toxicity of piperonyl butoxide (PB), an inhibitor of the monooxygenases, and tributylphosphorotrithioate (DEF), an esterase inhibitor, was investigated in the three strains. Pretreatment of the flies with PB, DEF, or PB+DEF before topical application of azamethiphos resulted in a significant decrease in LD50s in all the strains. The degree of synergism, however, varied depending upon the strains and the synergists. Similar results were obtained when azamethiphos was administered orally following pretreatment of the flies with PB+DEF. We attribute the high level of azamethiphos resistance in the 594vb strain partly to increased degradation by oxidative and hydrolytic enzymes. The hydrolytic enzymes are more important, but other factors including reduced cuticular penetration and insensitive acetylcholinesterase may be involved.(ABSTRACT TRUNCATED AT 250 WORDS)     

甜菜夜蛾抗氯氟氰菊酯品系相对适合度、抗性生化机理及抗性遗传方式

比较了甜菜夜蛾Spodoptera exigua 抗氯氟氰菊酯品系和敏感品系的繁殖和生长发育特征。结果表明：抗性品系幼虫发育历期延长、蛹重减轻、化蛹率和产卵量降低,抗性品系的适合度为0.61,抗性品系在繁殖和生长发育上存在明显的生存劣势。用两品系3龄幼虫分别测定胡椒基丁醚（PBO）、增效磷SV₁）、脱叶磷（DEF）和顺丁烯二酸二乙酯（DEM）对氯氟氰菊酯的增效作用,抗性品系增效倍数与敏感品系增效倍数之比分别为14.1、14.8、2.3和2.3倍,胡椒基丁醚和增效磷对氯氟氰菊酯增效作用最明显,表明多功能氧化酶参与了甜菜夜蛾对氯氟氰菊酯的抗性。抗性品系3龄幼虫酯酶和谷胱甘肽S-转移酶的活性分别为敏感品系的1.05倍和0.91倍, 抗性品系5龄幼虫多功能氧化酶O-脱甲基活性为敏感品系的1.05倍,两品系间3种酶的活性差异不显著,表明甜菜夜蛾对氯氟氰菊酯的抗性与酯酶、谷胱甘肽S-转移酶及多功能氧化酶O-脱甲基酶活性无关。用剂量对数死亡机率值回归线分析法研究甜菜夜蛾对氯氟氰菊酯的抗性遗传规律,表明甜菜夜蛾对氯氟氰菊酯的抗性为常染色体遗传、多基因控制;正、反交后代的显性度分别为0.61和0.43,抗性遗传为不完全显性。     

西花蓟马抗辛硫磷种群的抗性机制及交互抗性

为明确西花蓟马对辛硫磷的抗性风险,研究了西花蓟马抗辛硫磷种群对其他杀虫剂的交互抗性及其对辛硫磷的抗性机制.交互抗性测定结果表明,西花蓟马抗辛硫磷种群对辛硫磷与毒死蜱、高效氯氟氰菊酯和灭多威存在中等水平的交互抗性,对溴虫腈、吡虫啉、甲维盐和多杀菌素存在低水平交互抗性,对啶虫脒和阿维菌素不存在交互抗性.酶抑制剂与辛硫磷的增效剂测定结果表明,胡椒基丁醚(PBO)、三丁基三硫磷酸酯(DEF)和磷酸三苯酯(TPP)对西花蓟马抗辛硫磷种群(XK)、田间种群(BJ)和敏感种群(S)均起到了显著的增效作用(P＜0.05),马来酸二乙酯对西花蓟马抗辛硫磷种群和敏感种群增效作用均不显著,但对田间种群增效作用显著(P＜0.05).生化测定发现:除田间种群西花蓟马乙酰胆碱酯酶活性提高不显著外,西花蓟马抗辛硫磷种群和田间种群的细胞色素P450含量(2.79和1.48倍)、细胞色素b5含量(2.88和1.88倍)及O-脱甲基酶活性(2.60和1.68倍)、羧酸酯酶活性(2.02和1.61倍)和乙酰胆碱酯酶活性(3.10倍)均显著高于敏感种群(P＜0.05);谷胱甘肽-S-转移酶酶活性也有一定程度提高(1.11和1.20倍),但不显著(P＞0.05).表明其体内解毒代谢酶和靶标酶活性提高是西花蓟马对辛硫磷产生抗性的重要原因.     

Cytochrome P450 MA expression in insecticide-resistant German cockroaches (Dictyoptera: Blattellidae).

Cytochrome P450 monooxygenases are detoxification enzymes commonly involved in insecticide resistance by insects. Recently, an overexpressed form of this enzyme, P450 MA, was purified from an insecticide-resistant strain of German cockroach, Blattella germanica (L.), and polyclonal antisera (anti-P450 MA) was produced. To test hypotheses that the overexpressed condition of P450 MA has evolved in > 1 geographic location and that P450 MA might be involved in insecticide resistance to specific insecticides, investigations were conducted using 4 insecticide-resistant and 1 susceptible German cockroach strains. In western blots that used anti-P450 MA antiserum as a probe, substantial differences in expression of P450 MA were observed. Strains showing the highest P450 MA expression had both the highest tolerance to the organophosphate insecticide chlorpyrifos and cytochrome P450-mediated demethylation activity. Results support the hypothesis that cytochrome P450 MA is potentially overexpressed in insecticide-resistant populations on a global scale.     

Strain differences in the response of German cockroaches (Dictyoptera: Blattellidae) to emulsifiable concentrates.

The response of a susceptible and two pyrethroid-resistant field strains of the German cockroach, Blattella germanica, to four emulsifiable concentrates (ECs)--Tempo (cyfluthrin), Demon (cypermethrin), Dursban (chlorpyrifos) LO, and XRM 5184 (chlorpyrifos)--was investigated. Application rates were 0.4, 0.1, 0.5, and 0.5%, respectively. Susceptible cockroaches avoided dried formulations of the pyrethroids Tempo and Demon, but resistant cockroaches did not; avoidance was greater with Demon than with Tempo. Filter papers freshly impregnated (wet) with Demon and Tempo flushed susceptible cockroaches (Demon greater than Tempo). Resistant cockroaches were flushed only by Demon, but less so than susceptible strain cockroaches. No mortality occurred in experiments with the pyrethroids. Dried formulations of Dursban LO and XRM 5184 had little, if any, repellency to nymphs of either the susceptible or resistant strains. Initially (1-3 h), most cockroaches settled on treated papers. Subsequent movement off treated papers was a step in the process leading to knockdown. At 24 h, mortality of susceptible cockroaches was 100% in the experiment with Dursban LO and 95% in the experiment with XRM 5184. Highest mortality in a resistant strain (40%) was in the contact experiment with Dursban LO. In contact repellency and flushing experiments, mortality was higher with Dursban LO than with XRM 5184. Neither chlorpyrifos formulation flushed cockroaches effectively. Mortality in flushing experiments was less than in the contact repellency experiments.     

Effects of spirodiclofen on reproduction in a susceptible and resistant strain of <Emphasis Type="Italic">Tetranychus urticae</Emphasis> (Acari: Tetranychidae)

In this study the reproductive capacity of a laboratory-selected spirodiclofen resistant strain was investigated after treatment with spirodiclofen. Firstly, females were exposed to different concentrations of spirodiclofen (200 and 1,000 mg/l) during 6, 12 or 24 h. In contrast to the susceptible parental strain, the fecundity and fertility of resistant mites was not affected by treatment with these concentrations after any time of exposure tested. Secondly, pre-treatment of the resistant females with the synergists PBO or DEF could increase the inhibitory effect of spirodiclofen on reproduction, demonstrating the possible involvement of monooxygenases and esterases in metabolic detoxification of the acaricide. Because spirodiclofen interferes with lipid biosynthesis, total lipid content was measured in female adults. There were no significant differences between treated and non-treated female adults, both in the susceptible and resistant strain. However, the total lipid content in the resistant females was significantly higher than in susceptible females. Our data shows that the detection of spirodiclofen resistance should not be limited to mortality bioassays with eggs or larvae, but should be combined with inhibitory studies on female fertility and fecundity.     

Complete maternal inheritance of bifenazate resistance in Tetranychus urticae Koch (Acari: Tetranychidae) and its implications in mode of action considerations

Bifenazate is a selective hydrazine carbazate acaricide launched in 1999 and reported to be neurotoxic, since preliminary studies on the mode of action suggested that bifenazate may act on GABA-gated chloride channels. However, this information has not yet been supported by mechanistic studies. Therefore bifenazate is still considered as a neuronal inhibitor, but with unknown mode of action. Here we report an alternative hypothesis on the mode of action of bifenazate, i.e. its possible interference with a non-neuronal target site. An acaricide susceptible strain of Tetranychus urticae Koch (Acari: Tetranychidae), LS-VL, was artificially selected for bifenazate resistance, and after 36 generations an extremely high resistance ratio (RR) of >164,000 was obtained. This bifenazate-resistant strain (BR-VL) lacks cross-resistance to many different chemical classes and modes of action of other acaricides. In order to check for metabolic resistance mechanisms, synergists known to inhibit well-known detoxification routes were used together with in vitro enzymatic assays. No synergism or highly increased detoxification activity was observed in the resistant strain. However, the organophosphorous esterase inhibitor S,S,S-tributylphosphorotrithioate (DEF) applied to the susceptible strain could completely antagonise the acaricidal efficacy of bifenazate, suggesting that bifenazate is a pro-acaricide, not active by itself, that needs in vivo activation by esterases. Reciprocal crosses of diploid females and haploid males of strains LS-VL (susceptible) and BR-VL (bifenazate resistant) revealed that bifenazate resistance was inherited completely maternally, i.e. resistance is fully dominant when susceptible males were crossed with resistant females, and fully recessive when resistant males were crossed with susceptible females. Such an inheritance pattern has to our knowledge never been observed before in the case of insecticide/acaricide resistance. This observation may suggest a target-site for bifenazate encoded by the mitochondrial genome. Further evidence supporting such a hypothesis was obtained when measuring the ATP-level in spider mites treated with bifenazate. The ATP content in bifenazate treated mites declined progressively between 0 and 4h after treatment, similarly to mites treated with the complex I inhibitor fenpyroximate, an acaricide known to interfere with mitochondrial function. The obtained results suggest a target-site other than GABA-gated chloride channels, most likely encoded by and located in the mitochondria.     

Monitoring pyrethroid resistance in field collected Blattella germanica Linn. (Dictyoptera: Blattellidae) in Indonesia

The German cockroach, Blattella germanica , is a major and the most common pest in public areas in Indonesia. Although intensive control measures have been carried out to control the populations of this pest, results have been far from successful, which is believed to be because of its resistance to insecticides. A standard World Health Organization (WHO) glass jar test was carried out to determine the resistance level of this insect to pyrethroid insecticides, the most commonly used insecticides for cockroach control in Indonesia. A susceptible S1 strain collected from Tembagapura Papua was compared with four strains collected from Bandung, West Java: strain S2, from a local restaurant; strain S3, from the Bandung train station; and strains S4 and S5, from two different hotels. All strains showed low resistance to the pyrethroid, except the S5 strain, which had a Resistance Ratio (RR)₅₀ of 95 for permethrin. The addition of piperonyl butoxide (PBO) suggests that the detoxifying enzyme mixed function oxidases (MFO) played an important role in the development of resistance to permethrin in the S5 strain, suggested by the high Synergist Ratio (SR) of 70.4. However, the low level of resistance to cypermethrin was not affected by PBO, suggesting that other mechanisms of pyrethroid resistance are involved. Our study is the first report of German cockroach resistance to permethrin in Indonesia, and the findings can be used in formulating potential strategies for cockroach resistance management.     

棉蚜啶虫脒抗性种群交互抗性和增效剂增效作用的研究

【目的】明确棉蚜Aphis gossypii Glover啶虫脒抗性品系与其它杀虫剂的交互抗性现状以及增效剂的增效作用,为延缓和治理棉蚜对啶虫脒的抗性提供依据。【方法】采用单头反选育和群体汰选的方式,获得了棉蚜啶虫脒敏感和抗性品系;采用叶片药膜法测定了13种杀虫剂对啶虫脒的交互抗性以及增效剂对啶虫脒的增效作用。【结果】经过室内棉蚜敏感和抗性品系的筛选,获得了相对抗性倍数为82.33倍的棉蚜啶虫脒抗性品系。棉蚜啶虫脒抗性品系的交互抗性谱的研究表明,交互抗性倍数小于5的药剂为:吡蚜酮,甲基阿维菌素;交互抗性倍数在5~10倍的药剂为:噻虫嗪,联苯菊酯,毒死蜱,马拉硫磷,丙溴磷,辛硫磷;交互抗性倍数在10~15倍的药剂为:硫丹,阿维菌素,高效氯氰菊酯,三唑磷,氧化乐果;交互抗性倍数大于1 5倍的药剂为:吡虫啉。增效剂实验表明,TPP和PBO在啶虫脒敏感品系中增效作用不明显,但在抗性品系中增效作用显著。在啶虫脒抗性品系中的增效比为1.77、1.61,在啶虫脒敏感品系中的增效比为1.02、1.03。DEM在啶虫脒抗性、敏感品系中的增效作用均不明显,增效比为1.04、1.02。TPP和PBO对啶虫脒有很好的增效作用。以室内棉蚜敏感品系(LC_(50)为0.180 mg/L)为基础,对新疆各主要棉区的棉蚜种群进行了啶虫脒药剂的抗性调查,结果表明新疆各主要棉区棉蚜对啶虫脒的相对抗性倍数为6.1~22.0倍。【结论】由此说明新疆主要棉区棉蚜对啶虫脒具有一定的抗性风险,生产中可以利用无交互抗性的吡蚜酮和甲基阿维菌素来治理抗性棉蚜种群。