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1.
Four major esterases in one susceptible (CSMA) and two resistant (Hirokawa, E1) house fly strains were separated by chromatofocusing. Of the four esterases, those with pI's of 5.1 and 5.3 accounted for 90% of the p-nitrophenyl butyrate hydrolyzing activity in the three house fly strains. They also accounted for 70% (Hirokawa, E1) and 40% (CSMA) of the paraoxon-hydrolyzing activity as well as 87% (Hirokawa), 39% (E1) and 66% (CSMA) of the malathion-hydrolyzing activity in microsomes as measured by esterase-antibody interaction. In the Hirokawa strain, the pI 5.1 esterase was the predominant esterase and was more active than that of the the CSMA strain. Different substrate specificities and a different Km toward acetylthiocholine, as well as different rates of malathion and paraoxon hydrolysis between the Hirokawa and CSMA strains, suggest a qualitative difference in the pI 5.1 esterase. For the pI 5.1 esterase from the E1 strain, a different substrate specificity, a different Km for p-nitrophenyl butyrate, a different sensitivity to inhibitors, and a different rate of paraoxon hydrolysis suggest that it is a modified esterase. This esterase is not a phosphorotriester hydrolase, nor does it lack nonspecific esterase activity. It is a modified esterase which has a different substrate specificity when compared to the esterases from the other strains. The molecular weight of the esterases studied was approximately 220,000, with pH optima of about 7.0.The ratio of malathion α-monoacid to β-monoacid formation was about 9.0 for the pI 5.1 and 5.3 esterases and 1.5 for the pI 4.8 and 5.6 esterases. The existence of a higher αβ ratio for the pI 5.1 and 5.3 esterases and their significant rate of malathion hydrolysis in the Hirokawa strain indicate that an increase in the αβ ratio in house flies reported was due to the increase in the pI 5.1 esterase in the resistant strain.  相似文献   

2.
The role of esterases in malathion resistance in Culex tarsalis has been investigated. When larvae of a resistant and a sensitive strain were placed in water containing [14C]malathion, malathion penetrated to give initially similar internal levels. With resistant mosquitoes, after 15 min the internal malathion concentration decreased to low levels while the monoacid degradation products accumulated in the larvae and were excreted into the surrounding water, whereas in susceptible larvae the internal malathion level stayed high and was lethal. It is suggested that the decrease in internal malathion and the resulting resistance were caused by an active malathion carboxylesterase in the resistant strain. A specific assay for malathion carboxylesterase with [14C]malathion showed 55 times more activity in resistant than in susceptible larvae, whereas when general esterase activity was assayed with α-naphthyl acetate only 1.7 times the activity was found. Analyses by starch gel electrophoresis showed a peak of malathion carboxylesterase, 60-fold higher from resistant than from susceptible larvae, in a gel zone which did not stain for general esterase activity. General esterases that did not hydrolyze malathion showed different electrophoretic patterns in the two populations, which are likely due to the nonisogenic character of the strains. These results show that use of a specific assay and the demonstration of degradation of malathion in vivo are essential for assessment of the contribution of esterase activity to the malathion-resistant phenotype in mosquito populations.  相似文献   

3.
Malathion resistance in Anopheles stephensi from Pakistan was synergized by triphenyl phosphate, primarily a carboxylesterase inhibitor. There was a slight degree of antagonism with piperonyl butoxide. The major metabolite of malathion in larvae of both the resistant and susceptible strains was malathion monocarboxylic acid. Resistant larvae produced about twice as much of this product as the susceptible larvae. This suggests that a qualitative or a quantitative change in a carboxylesterase enzyme may be the basis of malathion resistance in this strain. Analysis of general esterase levels to α- and β-naphthyl acetate showed that there was no quantitative change in the amount of carboxylesterase enzyme present in the resistant strain as compared to the susceptible.  相似文献   

4.
Six to seven esterases from mouse, rat, and rabbit liver microsomes were resolved by chromatofocusing in the pH range 7–4. Each esterase peak showed a different substrate specificity pattern with the substrates evaluated. Malathion and paraoxon hydrolysis always corresponded with p-nitrophenyl acetate and methylthiobutyrate hydrolysis, whereas the pattern of fenvalerate hydrolysis was more complicated. Phosphorotriester hydrolase activity was isolated, and was found to be more specific toward paraoxon than toward the other insecticides. Time-course studies of paraoxon hydrolysis indicated that the hydrolysis of paraoxon by carboxylesterase was an inhibitory reaction. This reaction and phosphorotriester hydrolase activity can serve as a detoxication reaction toward organophosphate insecticides.  相似文献   

5.
Resistance to malathion in Anopheles stephensi from Pakistan was measured at intervals during the first week of adult life. LT50 values for homozygous resistant females decreased four-fold during the first 7 days of adulthood. A decrease in resistance with age also occurred in heterozygotes; the LT50 values of males and females fell sevenfold during the first 5 days of adulthood. The sensitivity to malathion of a susceptible strain increased with age. A biochemical basis for the declining resistance levels was investigated. Resistant and susceptible adults were homogenized at intervals during the first week of adulthood and soluble extracts were incubated with [14C]malathion. The rate of malathion metabolism to mono- and dicarboxylic acids was faster in resistant than in susceptible mosquitoes. The rate of malathion metabolism decreased with age in both strains. A decrease in carboxylesterase activity with age in resistant and susceptible mosquitoes is thus responsible for the increasing sensitivity to malathion. Implications for the monitoring of resistance in the field by diagnostic dosages and for the future use of malathion in mosquito control are discussed.  相似文献   

6.
Several forms of carboxylesterase were observed in a malathion-resistant small brown planthopper, Laodelphax striatellus Fallén, by isoelectrofocusing. To study the mechanisms of increased esterase activity, esterases were purified and their biochemical characteristics were investigated in five active esterase isozymes of two resistant strains. These esterases have polymorphic characteristics and their molecular weights ranged from 66 to 70 kDa, due to variations in glycosylation. The pI values of these esterases ranged from 5.3 to 4.7. These esterases were immunologically related and NH2-terminal amino acids were identical in all isozymes regardless of pI or molecular weight. No differences have been found in kinetic parameters (Km and Vmax) to α-naphthylacetate and specific activity toward α-naphthylacetate and malathion as a substrate in all isozymes. Resistant individuals showed high ali- and malathion carboxylesterase activities and these enhancements were caused by quantitative differences of carboxylesterases with several different pI.  相似文献   

7.
Enzyme preparations from Drosophila melanogaster flies degraded [14C]malathion to α- and β-malathion monoacids and, hence, were considered to contain malathion carboxylesterase (ME) activity. Although ME- activity was stable during preincubation in the absence of malathion, it decreased dramatically during the course of the reaction, and could not be completely recovered by Sephadex G-25 chromatography. Furthermore, the protein fraction after chromatography still contained 14C, suggesting that the enzyme had become inhibited by a bound, 14C-labeled derivative. Extracts from a resistant (malathion-selected), an intermediate control, and the susceptible Canton S strains of D. melanogaster differed in the lability of ME activity during the reaction. This difference was partly attributed to the production of small amounts of malaoxon (2–8%) by the extracts from the more resistant strains. No consistent strain differences were found when the rate of malathion degradation was measured during the first minute of reaction, either with or without a microsomal oxidase inhibitor (metyrapone) present. These results, together with the cross-resistance of the malathion-selected strain to other insecticides and the lack of a synergistic effect of two carboxylesterase inhibitors (triphenyl phosphate and S,S,S-tributylphosphorotrithioate) suggested that malathion carboxylesterase does not contribute significantly to the observed differences in malathion resistance between strains.  相似文献   

8.
It had been reported that a Japanese multiple-resistant strain of house fly, Hirokawa, had a high malathion-carboxylesterase activity as well as a normal level of esterase activity to α-naphthylacetate (NA). This is different from the situation in several other malathion-resistant strains, where high malathion-carboxylesterase activity goes together with a low level of activity to α-NA. This had been explained by the so-called “mutant ali-esterase theory,” which assumed that the opposite changes in activity to malathion and α-NA were the result of one and the same change in an ali-esterase. In the Hirokawa strain the esterase degrading malathion seems to be responsible for about 64% of the activity to α-NA. This was concluded since the two activities were equally sensitive to denaturation and to two organophosphorus inhibitors. Moreover activity of malathion was inhibited by α-NA, and that of α-NA by malathion. Most of the latter activity was inhibited competitively. Inhibition of activity to malathion was lower, however, than to be expected on the basis of competitive mutual inhibition. This case of resistance to malathion therefore seems to involve a different kind of “mutant ali-esterase” than in other strains. Increased hydrolysis of the insecticide seems to be achieved without loss of activity to α-NA, although Km is different. The strain further showed an unusually high β-NA hydrolysis and malaoxon-carboxylesterase activity (about 3- and 200-fold, respectively, that of another malathion-resistant strain G).  相似文献   

9.
Electrophysiological responses of labellar hairs of resistant and susceptible strains of the house fly were recorded at times following treatment of the hairs with DDT. Under the influence of DDT, the receptors of a hair discharged groups of impulses in high-frequency trains instead of the usual regular volley. The effect was observed to gradually spread to nontreated hairs. Among four strains chosen for gradation from high resistance to high susceptibility, in general the relative effectiveness of DDT corresponded to overall resistance as indicated by LD50 data. The highly resistant strain showed essentially no effects, and the other strains showed effects with some degree of recovery. Results for the highly susceptible strain were anomalous in not differing significantly from those of the moderately resistant strai. Unexpectedly small effects in the highly susceptible strain point to a strain characteristic not necessarily correlated with LD50 data.  相似文献   

10.
Levels of carboxylesterase activity in F1 clones of Myzus persicae, obtained by crossing sexuales from a resistant, high esterase clone with those from a susceptible, low esterase clone, fell into two distinct groups intermediate between the levels of carboxylesterase in the parent clones. When sexuales of F1 clones of the lower of these two intermediate levels of carboxylesterase activity were crossed, segregation ratios in the F2 generation indicated that this lower intermediate activity level (about 0.4 μmol mg?1 h?1). which is about twice the level in susceptible clones, is due to mutation at a single regulatory locus. The results obtained with backcrosses, between sexuales of an F1 clone having the higher intermediate level of carboxylesterase activity and a parent susceptible, low esterase clone, suggest that a second locus may be involved in the expression of higher levels of esterase activity.  相似文献   

11.
Resistance in a dual malathion- and permethrin-resistant head louse strain (BR-HL) was studied. BR-HL was 3.6- and 3.7-fold more resistant to malathion and permethrin, respectively, compared to insecticide-susceptible EC-HL. S,S,S-Tributylphosphorotrithioate synergized malathion toxicity by 2.1-fold but not permethrin toxicity in BR-HL. Piperonyl butoxide did not synergize malathion or permethrin toxicity. Malathion carboxylesterase (MCE) activity was 13.3-fold and general esterase activity was 3.9-fold higher in BR-HL versus EC-HL. There were no significant differences in phosphotriesterase, glutathione S-transferase, and acetylcholinesterase activities between strains. There was no differential sensitivity in acetylcholinesterase inhibition by malaoxon. Esterases from BR-HL had higher affinities and hydrolysis efficiencies versus EC-HL using various naphthyl-substituted esters. Protein content of BR-HL females and males was 1.6- and 1.3-fold higher, respectively, versus EC-HL adults. Electrophoresis revealed two esterases with increased intensity and a unique esterase associated with BR-HL. Thus, increased MCE activity and over-expressed esterases appear to be involved in malathion resistance in the head louse.  相似文献   

12.
Of six juvenile hormone analogs of the alkyl 3,7,11-trimethyl-2,4-dodecadienate type, only the isopropyl ester was strongly morphogenic in the house fly, Musca domestica L. In vitro assays revealed that house fly microsomes contain B-esterases as well as oxidases which metabolize such analogs. However, these esterases did not hydrolyze the isopropyl ester, ZR-515. Enzymes prepared from larvae, pupae, and adults were all active and there was evidence that in the late larval stage the esterase activity was cyclic, showing a minimum in the early third instar and a maximum a few hours later. When microsomes from two susceptible and two resistant house fly strains were compared for metabolic activity against the juvenile hormone analogs, those from the resistant strains were 1.3 to 20 × higher in oxidase activity but there was no difference in esterase activity. The oxidative metabolism of two analogs ZR-515 and 512 was greatly enhanced when the flies were induced with phenobarbital but there was no enhancement in metabolism of three of the remaining analogs and only a slight enhancement of a fourth. It is concluded that the insecticidal action of ZR-515 is largely due to its stability in the presence of the house fly esterases.  相似文献   

13.
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14.
Malathion resistance of a field-collected population of Rhizopertha dominica (Coleoptera: Bostrichidae) from Mexico was evaluated and the resistance mechanisms were characterized both in vivo and in vitro. The Mexican population showed a resistance level of 50-fold at LC50 as compared with that of a susceptible laboratory population. Malathion bioassays with the synergists triphenyl phosphate, piperonyl butoxide and diethyl maleate suggested that esterases were likely to contribute to the resistance whereas cytochrome P450 monooxygenases and glutathione S-transferases were not. In-vitro assays of esterases indicated that the general esterase activity was 1·3-fold higher in the Mexican population than in the susceptible population. However, the phosphotriesterase activity in the resistant population was 3·7-fold higher than in the susceptible population. Significantly higher phosphotriesterase activity in the resistant population was further indicated by 3·4-fold increase of Vmax in enzyme kinetics and higher frequency of individuals with high phosphotriesterase activity in this population. All these findings suggested that phosphotriesterases play a role in malathion resistance in the Mexican population of lesser grain borer. © 1998 SCI  相似文献   

15.
The metabolic fate of six 3H-ring-substituted ethoxychlor analogs with altered aliphatic moieties and [14C]p,p′-DDT was investigated in susceptible and DDT-resistant strains of the house fly Musca domestica Linnaeus. The chloroalkane analogs, dichloroethane, chloropropane, and dichloropropane were primarily metabolized to the corresponding dehydrochlorinated products. This pathway was relatively more prominent in the resistant strain than in the susceptible strain. Biotransformation and detoxication of the isobutane, nitropropane, and neopentane derivatives was through microsomal oxidation (O-deethylation) of aryl ethoxy degradophores, and oxidation of the aliphatic moieties to produce the corresponding benzophenones, with no substantial differences between the resistant and susceptible strain. There was a strong correlation between the Taft (σ1) values for the altered aliphatic moieties of chloroalkane analogs and their rate of dehydrochlorination in both the strains. These results suggest the importance of altered aliphatic moieties in developing resistance-proof DDT derivatives.  相似文献   

16.
The hydrolysis of malation by rabbit liver oligomeric and monomeric carboxylesterases (CE's) (EC 3.1.1.1) results in the formation of a mixture of α- and β-monoacids. A new chromatographic procedure was utilized to investigate the formation of α- and β-monoacids. The oligomeric carboxylesterase (oCE) produced an αβ ratio of monoacids of 4.55, and the monomeric carboxylesterase (mCE) produced an αβ ratio of monoacids of 2.33. The ratios of α- and β-monoacids were independent of the initial concentration of malathion and remained constant over the time course of the reaction. Kinetic studies demonstrated that the Km values were the same for the corresponding reactions which produced either α-monoacid or β-monoacid with the same enzyme. Since both carboxylesterases are electrophoretically pure, the kinetic data strongly supports the theory that the reactions which produced α- and β-monoacids are catalyzed by the same active site. Comparison of the kcat and Km values governing the hydrolysis of malathion by the two esterases, together with their relative abundance in liver, indicated that the oCE would be responsible for about 80 to 98% of the hydrolytic detoxication of malathion by rabbit liver.  相似文献   

17.
We have examined the chromosomal basis for reproductive and developmental defects that are associated with malathion resistance in a laboratory-selected population of Drosophila melanogaster. Strains homozygous for second or third chromosomes from this population were more resistant to malathion and had greater mixed-function oxidase activity, decreased fertility, and lower egg production when compared with first chromosome or susceptible strains. Some of the strains carrying resistant third chromosomes were developmentally delayed and required a significantly longer time to pupate. Delayed pupation was not associated with increased in vitro degradation of ecdysone by larvae having increased mixed-function oxidase activity, nor could it be reversed by feeding larvae ecdysone. Differences in mixed function oxidase activity among strains homozygous for second or third chromosomes were strongly correlated with malathion resistance but not with fitness. Although both second and third chromosome strains had high mixed-function oxidase activity, only fly extracts from the third chromosome strains oxidatively degraded [3H]juvenile hormone in vitro to a significant extent. A deficit of vitellogenic oocytes and increased egg laying by females in response to topically applied juvenile hormone-I supported the hypothesis that juvenile hormone titer was lower than normal in these strains. The results indicate that different polygenic systems control malathion resistance and associated fitness defects in this selected population of D. melanogaster. Although these systems are partly independent, they overlap due to pleiotropic effects of third chromosomal genes controlling mixed-function oxidase activity on female reproduction.  相似文献   

18.
The mechanisms of resistance and cross resistance to the juvenoids methoprene and R-20458 in the house fly, Musca domestica, were examined. Radiolabeled methoprene was found to be metabolized faster in resistant and cross-resistant house fly larvae than in susceptible larvae, and methoprene and R-20458 penetrated more slowly into larvae of the resistant strain. In vivo and in vitro metabolism of methoprene was largely by oxidative pathways followed by conjugation in all strains examined, and little or no ester change of methoprene was noted in vitro. In vitro oxidative metabolism of methoprene, R-20458, juvenile hormone I, and several model substrates was higher in resistant and cross-resistant larvae than in susceptible larvae. Juvenoid functionalities susceptible to metabolic attack by resistant strains are indicated.  相似文献   

19.
Fipronil resistance mechanisms were studied between the laboratory susceptible strain and the selective field population of rice stem borer, Chilo suppressalis Walker in the laboratory. The borer population was collected from Wenzhou county, Zhejiang province. After five generations of selection, fipronil resistance ratio was 45.3-fold compared to the susceptible strain. Synergism experiments showed that the synergistic ratios of PBO, TPP and DEF on fipronil in susceptible and resistant strains of C. suppressalis were 7.55-, 1.93- and 2.91-fold, respectively, and DEM showed no obvious synergistic action on fipronil. Activities of carboxylesterase and microsomal-O-demethylase in the resistant strain were 1.89- and 1.36-fold higher that in susceptible strain, and no significant difference of glutathione-S-transferase activity was found between the resistant and susceptible strains. The Km and Vmax experiments also demonstrated that fipronil resistance of C. suppressalis was closely relative to the enhanced activities of carboxylesterase and microsomal-O-demethylase. Moreover, cross-resistance between fipronil and other conventional insecticides and the multiple resistant properties of the original Wenzhou’s population were also discussed.  相似文献   

20.
Populations of the housefly Musca domestica isolated from farms in different German districts with strong resistance problems were compared to laboratory strains with varying resistance spectra. Resistance against pyrethroids, organophosphates and carbamates was tested using impregnated filter papers, and by topical application using a susceptible housefly strain (origin WHO) for comparison. The multi-resistant fly strains tested had a strong resistance against these insecticide groups, ranging from 37- to >10000-fold for organophosphates and 150- to >6600-fold for pyrethroids. The constituent enantiomer pairs of the α-cyano-pyrethroid cyfluthrin were tested, as was beta-cyfluthrin. With respect to multi-resistant fly strains, the isomers II and IV had the best activity, with LD50 values of 0·012 and 0·014 μg per fly, respectively. In addition, different groups of insect growth regulators (juvenile hormone analogues, chitin synthesis inhibitors and one triazine derivative) were tested in a special larvicidal test. The chitin synthesis inhibitors were quite effective against multi-resistant M. domestica strains except for one strain with strong resistance against chitin synthesis inhibitors, developed after extensive treatments with benzoylphenylureas for several years. The fly strains tested were not resistant against cyromazine. Additionally, the insecticides were combined with the synergists piperonyl butoxide, tributylphosphorotrithioate (DEF) and Cibacron blue and tested against the fly strain with the strongest resistance spectrum (‘Grimm’) in comparison to the susceptible strain (‘WHO-N’). Piperonyl butoxide had the greatest effect on the efficacy of cyfluthrin followed by Cibacron blue and DEF. In a parallel investigation with susceptible and resistant house fly strains, different enzyme activities related with resistance mechanisms were tested, e.g. glutathione S-transferase (3·5-fold) and mixed-function oxidase (2·3-fold). Implications of these results for management of insecticide resistance in M. domestica are discussed.  相似文献   

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