Characterization of the hepatic mixed function oxidase system in endrin-resistant and -susceptible pine voles

Studies were conducted to assess the contribution of the hepatic microsomal mixed function oxidase system to a 7.2-fold difference in susceptibility to the lethal effects of endrin between endrin-resistant and -susceptible pine voles, Microtus pinetorum. Evaluations of microsomal enzyme systems were conducted for basal and endrin-treated pine voles of both strains. The microsomal activity of ICR white mice was investigated to provide a species comparison. Maximal microsomal mixed function oxidase activities were determined in in vitro incubations for the model substrates ethylmorphine, aniline, and benzo(a)pyrene. V_max values were estimated for the rate of disappearance of benzo(a)pyrene in in vitro incubations. No significant strain differences in basal microsomal enzyme activity were found for the model substrates investigated, although activity was invariably higher in the resistant strain. The concentration of cytochrome P-450 was significantly higher in the resistant vole though actually less than 20% different. The occurrence of significant strain differences in the levels of microsomal enzyme activity induced by endrin were rare. Significant endrin treatment effects on the levels of microsomal enzyme activity for the pine vole were observed but the degree and direction of change depended on the substrate used. A marked species difference in microsomal mixed function oxidase activity was noted between pine voles and white mice. This was particularly evident for endrin-treated animals. The microsomal activity of endrin-treated white mice was greatly induced relative to basal levels. The degree of induction depended on the substrate used. The small strain differences in microsomal enzyme activity for the systems investigated were judged to be insufficient to explain the strain difference in susceptibility to endrin.     

Acetylcholinesterase of Aphis citricola: Properties and significance in determining toxicity of systemic organophosphorus and carbamate compounds

In apterous adults of the spirea aphid, Aphis citricola van der Goot, the optimum conditions for determining acetylcholinesterase (AChE) activity consist of reaction mixture of 0.1 M phosphate buffer (pH 7.5), 10^?3M acetylthiocholine (ASCh), and enzyme extract equivalent to 80 ± 3 μg protein incubated for 15 min at 30°C. The K_m value for ASCh (6.7 × 10^?5M) was much lower than that of butyrylthiocholine (BuSCh) (1.25 × 10^?2M). The enzyme activity was almost completely inhibited by 10^?6M paraoxon or 10^?5M eserine and was 84% inhibited by 10^?5M BW284C51 (a specific AChE inhibitor). DTNB was found to inhibit the enzyme activity and was therefore added at the end of the reaction. AChE activity of A. citricola was inhibited in vitro and in vivo by dimethoxon > dimethoate, and aldicarb sulfoxide > aldicarb > aldicarb sulfone. The in vivo effect correlates well with the toxicity level of the various toxicants. A neurotoxicity index which combines both mortality and in vivo inhibition of the aphid AChE activity is suggested as a measure for determining the toxicity of organophosphorus and carbamate compounds toward aphids.     

Hydrolysis of paraoxon and malaoxon in three strains of Myzus persicae with different degrees of parathion resistance

Homogenates of three strains of Myzus persicae, A, R, and E, with an LD₅₀ for topically applied parathion of 9, 93, and 263 ng per aphid, showed an in vitro hydrolytic degradation of paraoxon of 2.3, 4.7, and 8.6 pmol/mg aphid/h, respectively. These values represent V_max; K_m was <10^?7M. The three strains showed a malaoxon degradation of 2.4, 11.9, and 18.8 pmol/mg/h at 10^?6M substrate concentration. V_max for R and E was 21 and 27 pmol, respectively and K_m 7 and 4 × 10^?7M. Activity in strain A was too low to estimate these entities. The breakdown product of paraoxon was mainly diethyl phosphoric acid, that of malaoxon mainly dimethyl phosphoric acid. No hydrolysis of the carboxylester groups of malaoxon was found. Hydrolysis of paraoxon and malaoxon was inhibited by isopropyl and n-propyl paraoxon and by the salioxon-analog K₂. The two latter compounds were shown to act as synergists with parathion when added in amounts that caused little mortality when given alone. The hydrolytic enzyme is soluble and retains its activity during incubations of several hours. It is likely that it is responsible for at least part of the resistance. Resistance was maintained without selection over a period of three years. There was no correlation between degree of resistance and carboxylesterase activity of the strains.     

Pyrethroid synergists suppress esterase-mediated resistance in Indian strains of the cotton bollworm, Helicoverpa armigera (Hübner)

The role of esterase in pyrethroid resistance was studied in the final larval instar of different strains of the cotton bollworm, Helicoverpa armigera. The resistant strains viz., Nagpur strain and the Delhi strain were found to have elevated midgut esterase activity in comparison to the susceptible strain. Nagpur strain and Delhi strain have 2.24 and 1.73-fold higher esterase activity, respectively, than that of the susceptible strain. The Native PAGE displayed important differences in the midgut esterase isozyme pattern between the susceptible and the pyrethroid-resistant strains. Out of the 10 esterase isozyme observed, susceptible strain lacked three bands, E2, E6 and E10 that were found in the resistant strains. The potency of the synergists piperonyl butoxide (PBO) and dihydrodillapiole (DDA) as esterase inhibitor were also studied both in vitro and in vivo. The in vitro results clearly show that both PBO and DDA inhibited esterase activity in the two resistant strains, while there was almost no esterase inhibition in the homogenate of the susceptible strain. The in vivo inhibition studies (topical application of PBO and DDA followed by biochemical analysis) illustrated that PBO- and DDA-esterase binding is rather slow and non permanent process. Esterase inhibition did not occur immediately after the synergist treatment but at 4 and 8 h post treatment in case of PBO and DDA, respectively. Native PAGE revealed that the in vivo esterase inhibition caused by both PBO and DDA was due to the binding of the synergist with the E6 isozyme which was not present in the susceptible strain.     

The biochemical basis of malathion resistance in the sheep blowfly,Lucilia cuprina

The in vivo and in vitro metabolism of [¹⁴C]malathion was studied in susceptible (LS) and malathion resistant (RM) strains of the sheep blowfly, Lucilia cuprina (Wiedemann). No difference was found between strains in the penetration, excretion, storage, or inhibitory potency of the insecticide. However, RM degraded malathion to its α- and β-monocarboxylic acid metabolites more rapidly than LS, both in vivo and in vitro. This enhanced degradation of [¹⁴C]malathion occurred in vitro in both mitochondrial and microsomal fractions of resistant flies. Kinetic analysis revealed that these fractions degraded malathion by discrete mechanisms. The enzymes from the mitochondria of both strains had the same K_m, whereas the microsomal enzyme from the RM strain had a fivefold higher K_m than that from the LS strain. Studies of esterase activities and the effect of enzyme inhibitors showed that both the mitochondrial and microsomal resistance mechanisms were the result of enhanced carboxylesterase activity. It was concluded that increased carboxylesterase detoxification of malathion adequately explained the high level of malathion resistance in RM if rate-limiting factors such as cuticular penetration were taken into account.     

Bimodal effect of methylenedioxyphenyl compounds on detoxifying enzymes in the housefly

Thirteen methylenedioxyphenyl (MDP) compounds, including commercial insecticide synergists and juvenile hormone analogs, were compared in their effect on detoxifying enzymes in the housefly (Musca domestica). Flies were fed a diet containing 1% of the compounds for 3 days. Enzymes were then assayed in vitro for their activity using aldrin and DDT as substrates. Piperonyl butoxide (PB), sesamex, propyl isome, sulfoxide, safrole, isosafrole, 6,7-epoxy-3,7-diethyl-1-[3-4(methylenedioxy) phenoxy]-2-octene (MDP-JH I) and 6,7-epoxy-3-methyl-7-ethyl-1-[3,4-(methylenedioxy) phenoxy]-2-octene (MDP-JH II) all caused a bimodal effect, inhibiting microsomal epoxidase and inducing DDT-dehydrochlorinase in the resistant Isolan-B strain. Two of these, PB and MDP-JH I, gave similar results with the susceptible strain, stw;w⁵ and two resistant strains, Fc-B and Orlando-DDT. However, o-safrole, piperonylic acid, piperonal, 3,4-methylenedioxybenzyl acetate and methyl-(3,4-methylenedioxy) benzoate had little or no effect on the enzyme systems studied. The standard susceptible strain (WHO-SRS) responded to these compounds very differently. Among those tested, piperonyl butoxide, sesamex, safrole, and isosafrole were inducers of microsomal epoxidase, a 4-fold increase occurring after treatment with sesamex. Only MDP-JH II showed a marked inhibition of the epoxidase. These treatments did not effect DDT-dehydrochlorinase activity in this strain.The enhancement of DDT-dehydrochlorinase activity by the MDP compounds is associated with an increased rate of DDT dehydrochlorination in vivo. The stimulatory effect could be blocked by treatment with actinomycin D or cycloheximide.     

Hydrolysis of permethrin by pyrethroid esterases from resistant and susceptible strains of Amblyseius fallacis (Acari: Phytoseiidae)

Pyrethroid-hydrolyzing activity in whole body homogenates of three insecticide-resistant and one susceptible strain of the predator mite, Amblyseius fallacis Garman has been examined in vitro. The highest esterase activity was found in the synthetic pyrethroid-resistant GH-1 strain body homogenate. All three pyrethroid-resistant strains had esterases that hydrolyzed trans-permethrin two times faster than cis-permethrin but isomer specificity was not observed in the susceptible strain. The pyrethroid esterases of the GH-1 strain were very sensitive to inhibition by dichlorovos, S,S,S-tributhylphosphorotrithioate, and 3-octylthio-1,1,1-trifluoro-2-propanone. Carbaryl and tetraethylpyrophosphate exhibited moderate inhibition in the GH-1 strain. Eserine sulfate and piperonyl butoxide only inhibited permethrin hydrolysis at higher concentrations. Fifteen esterase bands were resolved from body homogenates by gradient gel electrophoresis in the GH-1 strain, and were identified as carboxylesterases. The major pyrethroid-hydrolyzing activity was located in E5–E12 bands from GH-1 and composite susceptible strain esterases. Six esterase bands exhibiting low pyrethroid-hydroloyzing activity were also obtained from the two spotted spider mite, Tetranychus urticae (Koch).     

In vitro inhibition of the carbonic anhydrase from saanen goat (Capra hircus) with pesticides

This study was conducted to determine the in vitro effects of four commonly used pesticides (Nuarimol™, Fenarimol™, Parathion-methyl™ and 2,4-D™) on erythrocyte carbonic anhydrase (CA) activity from Saanen goats (SG). The enzyme was 262.57-fold purified by affinity chromatography and the purity was confirmed by SDS-PAGE. Inhibitory effect of the pesticides on the purified enzyme was determined using the CO₂-hydratase activity method. IC₅₀ values of the pesticides that caused inhibition were determined by means of activity percentage diagrams. The concentrations of Nuarimol™, Fenarimol™, and 2,4-D™ that inhibited 50% of the enzymatic activity were 0.352, 0.924 and 2.04 mM, respectively. Conversely, the enzyme activity was increased by parathion-methyl.     

Triazophos resistance mechanisms in the rice stem borer (Chilo suppressalis Walker)

A field population of the rice stem borer (Chilo suppressalis Walker) with 203.3-fold resistance to triazophos was collected. After 8-generation of continuous selection with triazophos in laboratory, resistance increased to 787.2-fold, and at the same time, the resistance to isocarbophos and methamidophos was also enhanced by 1.9- and 1.4-fold, respectively, implying some cross-resistance between triazophos and these two organophosphate insecticides. Resistance to abamectin was slightly enhanced by triazophos selection, and fipronil and methomyl decreased. Synergism experiments in vivo with TPP, PBO, and DEM were performed to gain a potential indication of roles of detoxicating enzymes in triazophos resistance. The synergism results revealed that TPP (SR, 1.92) and PBO (SR 1.63) had significant synergistic effects on triazophos in resistant rice borers. While DEM (SR 0.83) showed no effects. Assays of enzyme activity in vitro demonstrated that the resistant strain had higher activity of esterase and microsomal O-demethylase than the susceptible strain (1.20- and 1.30-fold, respectively). For glutathione S-transferase activity, no difference was found between the resistant and the susceptible strain when DCNB was used as substrate. However, 1.28-fold higher activity was observed in the resistant strain when CDNB was used. These results showed that esterase and microsomal-O-demethylase play some roles in the resistance. Some iso-enzyme of glutathione S-transferase may involve in the resistance to other insecticides, for this resistant strain was selected from a field population with multiple resistance background. Acetylcholinesterase as the triazophos target was also compared. The results revealed significant differences between the resistant and susceptible strain. The V_max and K_m of the enzyme in resistant strain was only 32 and 65% that in the susceptible strain, respectively. Inhibition tests in vitro showed that I₅₀ of triazophos on AChE of the resistant strain was 2.52-fold higher. Therefore, insensitive AChE may also involved in triazophos resistance mechanism of rice stem borer.     

Toxicity and metabolism of carbaryl in the European corn borer

Larvae from two strains of the European corn borer, Ostrinia nubilalis (Hübner), were compared for differences in their tolerance and metabolism of carbaryl (1-naphthyl N-methylcarbamate). The Geneva strain was about twice as susceptible to carbaryl, but both Valley and Geneva borers converted carbaryl to oxidative metabolites at similar rates in vivo and in vitro. Maximum carbaryl-metabolizing activity was present in last-instar larvae, particularly in the fat body and gut tissues. However, the specific activity of gut homogenates was highest in the Geneva strain and the specific activity of fat body was highest in the Valley strain. Other differences in the mixed-function oxidase systems of gut and fat body were also found. The major metabolite in vivo and in vitro was hydroxymethyl carbaryl.     

Comparative metabolism of atrazine and EPTC in proso millet (Panicum miliaceum L.) and corn

The purpose of this study was to examine the differential activities of proso millet (Panicum miliaceum L.) and corn (Zea mays L.) with respect to atrazine [2-chloro-4-(ethylamino)-6-(isopropylamino)-S-triazine] and EPTC (S-ethyldipropyl thiocarbamate) metabolism. GSH-S-transferase was isolated from proso millet shoots and roots. When assayed spectrophotometrically using CDNB (1-chloro 2,4-dinitrobenzene) as a substrate, the shoot enzyme had only 10% of the activity of corn shoot enzyme while the root enzyme had 33% the activity of corn root enzyme. However, when proso millet shoot GSH-S-transferase was assayed in vitro using ¹⁴C-ring-labeled atrazine, it degraded the atrazine to water-soluble products at the same rate as the corn shoot enzyme. Incubation of excised proso millet and corn roots with [¹⁴C]EPTC indicated that uptake of EPTC was similar in both plants. However, proso millet metabolized the EPTC to water-soluble products at only half the rate of corn. Glutathione levels of proso millet roots were 35.9 μg GSH/g fresh wt, compared with 65.4 μg GSH/g fresh wt for corn. However, a 2.5-day pretreatment with R-25788 (N,N-diallyl-2-2-dichloroacetamide) elevated proso millet GSH levels to 62.7 μg GSH/g fresh wt. R-25788 did not elevate the activity of proso millet GSH-S-transferase, in contrast to its effects on corn. We conclude that differences in response to atrazine and EPTC in proso millet and corn are a result of their differential metabolism.     

Studies on the metabolism of vamidothion and its thioanalog in insecticide-resistant and susceptible house fly strains

The in vivo and in vitro metabolism of vamidothion [O,O-dimethyl S-[2-(1-methylcarbamoyl)-ethylthio] ethylphosphorothiolate] as well as the in vitro metabolism of thiovamidothion [O,O-dimethyl S-[2-(1-methylcarbamoyl)ethylthio] ethylphosphorodithioate] was investigated in insecticide-resistant and susceptible house fly strains. Vamidothion was converted in vivo to the sulfoxide, the principle metabolite, and subsequently to the sulfone at a slower rate. Vamidothion and vamidothion sulfoxide were hydrolyzed at the PS and SC bond. The resulting primary alcohol metabolite was further oxidized to a carboxylic acid followed by decarboxylation. No metabolism of vamidothion or thiovamidothion occurred in vitro without the addition of NADPH. The addition of NADPH resulted in rapid conversion of vamidothion to the sulfoxide, and thiovamidothion was oxidatively metabolized to six metabolic products. No qualitative differences were found between resistant and susceptible strains, but there were signficant quantitative differences. The metabolism was highest in the Rutgers strain followed by Cornell-R, Hirokawa, and then CSMA strain. The route of vamidothion and thiovamidothion metabolism was via the cytochrome P-450-dependent monooxygenase system, and none of the resistant strains showed glutathione S-transferase activity toward vamidothion or thiovamidothion. No further oxidation of vamidothion sulfoxide to the sulfone was observed and also no hydrolysis products were formed, in vitro.     

in vitro metabolism of azinphosmethyl in susceptible and resistant houseflies

The in vitro metabolism of [¹⁴C-methoxy] or [³²P]azinphosmethyl by subcellular fractions of abdomens from a resistant and a susceptible strain of houseflies was studied. The degradative activity in both strains was associated with the microsomal and soluble fractions and required NADPH and glutathione, respectively. The resistant strain possessed higher activity for both the mixed-function oxidases and the glutathione transferase than the susceptible strain, and both systems appear to be important in the resistance mechanism. The mixed-function oxidases were involved in the oxidative desulfuration as well as the dearylation of azinphosmethyl. A glutathione transferase located in the soluble fraction catalyzed the formation of desmethyl azinphosmethyl and methyl glutathione. This enzyme also demethylated azinphosmethyl oxygen analog. Although the soluble fraction exhibited both glutathione S-alkyltransferase and S-aryltransferase activity against noninsecticidal substrates, no evidence of the transfer of the benzazimide moiety from azinphosmethyl to glutathione was obtained. Sephadex G-100 chromatography of the soluble enzymes revealed a common eluting fraction responsible for both types of transferase activity.     

Relationship among expression, amplification, and methylation of FE4 esterase genes in Italian populations of Myzus persicae (Sulzer) (Homoptera: Aphididae)

The wide use of insecticides containing an esteric group selected resistant Myzus persicae populations characterised by the overproduction of one of two closely related carboxylesterases (E4 and FE4). In this paper, we present data collected from Italian population indicating that all the 22 populations analysed possess amplified FE4 gene only. The estimation of FE4 copy number, carried out by densitometric scanning of dot and Southern blots, puts in evidence that the different populations possess a gene copy number ranging from 6 to 104. Statistical analysis shows the existence of a high positive correlation between gene copy number and total esterase activity. In aphid strain with low FE4 copy number, these genes are almost totally methylated. On the contrary, aphid strains with high FE4 gene number evidenced highly variable methylation levels and absence of correlation between the number of genes and their methylation state. The same result has been observed when comparing FE4 methylation levels and esterase activity.     

Insecticide resistance in cotton aphid Aphis gossypii (Glov.) in the Sudan Gezira

The resistance to insecticides of three Sudanese strains of A. gossypii (Glov.) collected from cotton fields in the Sudan Gezira Scheme over three seasons (1988, 1989, 1990) and that of two French strains was studied in the laboratory. When compared with a known susceptible strain, the aphids were found to be resistant-to the eight insecticides tested. Evolution of resistance in Sudanese strains during the three crop seasons was observed. Assay of aphid homogcnate for carboxytesterase activity towards the substrates α-naphthyl acetate and β-naphthyl acetate showed that there was no enhancement of this class of enzyme and thus it was not a cause of resistance in this species. A study of interaction between the acetylcholinesterase (AchE) and pirimicarb established the kinetics of the inhibition process. I₅₀ values were found to be much higher for the Sudanese strains than for the susceptible strain. First-order inhibition kinetics revealed that resistance towards pirimicarb in Sudanese-strains was caused by a modified AchE which had a reduced affinity (higher K_a value) and poor carbamylation ability (lower K₂ value) for pirimicarb. The resistance mechanisms for the other insecticides remain to be studied.     

Interaction Between Entomopathogenic Fungi and Some Aphid Species in Thailand

Seven aphid species were screened with twelve entomopathogenic fungi for the selection of the most effective fungus. Beauveria bassiana CKB-048 caused the highest mortalities for all aphid species tested in their nymphal stage. The cowpea aphid (Aphis craccivora Koch) was chosen for further studies in its adult stage using B. bassiana CKB-048 as a biocontrol agent, and carbosulfan was used for a comparison in greenhouse testing. The mode of action of the fungal killing was studied by scanning electron microscopy. The endophytic colonization of the cowpea plant by B. bassiana CKB-048 was studied and confirmed using both stereo microscopy and transmission electron microscopy. The chitinase enzyme activity of cowpea plants after they were sprayed with the fungus or carbosulfan was found to fluctuate throughout five days. The lethal concentrations of B. bassiana CKB-048 suspensions that killed 50 % (LC₅₀) of A. craccivora was 6.69 × 10⁷ conidia/ml for the nymphs and 8.25 × 10⁷ conidia/ml for the adults. The time for lethality (LT₅₀) with a concentration of 2 × 10⁸ conidia/ml was 3.25 days for nymphs and 4 days for adults.     

Carboxylamidases inSpodoptera exigua: properties and distribution in the larval body

Resistance to many insecticides demonstrated by the beet armyworm,Spodoptera exigua (Hübner), can be caused by the action of carboxylamidases. A colorimetric method, based on the hydrolysis of 4-nitroacetanilide to 4-nitroaniline by carboxylamidases, was used for evaluating biochemical properties of these detoxifying enzymes in beet armyworm. The optimum pH and temperature were 7.5 and 38°C, respectively. K_m (Michaelis constant) and V_max (maximal velocity) at 28°C were 2.3 X 10^-4 M and 2.06 nmol min^-1 mg protein^-1, respectively. The enzyme activity was evaluated in several body parts and located mostly (66.2%) in the midgut. The soluble fraction (supernatant of 105,000g) contained the highest enzyme activity relative to the total (69.3%), and exhibited the highest specific activity. Carboxylamidase activity was totally inhibitedin vitro at a concentration of 10^-6 M methomyl. The analysis of thein vitro inhibition kinetics indicated the ability of methomyl and diflubenzuron to inhibit carboxylamidases noncompetitively. Over 95% inhibitionin vivo was obtained when the larvae were fed with castor bean leaves dipped in 250 mgl ^-1 of methomyl. Thein vivo enzyme activity could be reduced to half with a pretreatment of 15 mgl ^-1 diflubenzuron.     

Insensitivity of acetylcholinesterase as a factor in resistance of houseflies to the organophosphate Rabon

Acetylcholinesterase from an organophosphate-resistant strain of housefly exhibited a decrease in sensitivity to Rabon in vitro, to the extent of 206-fold for the soluble enzyme and 38-fold for the particulate. These effects upon the bimolecular reaction constant, k_i, were due to a 573-fold decrease in affinity of the inhibitor for the soluble and 103-fold for the particulate enzyme, coupled with a small increase in reactivity.     

Resistance selection and biochemical mechanism of resistance to two Acaricides in Tetranychus cinnabarinus (Boiduval)

A Tetranychus cinnabarinus strain was collected from Chongqing, China. After 42 generations of selection with abamectin and 20 generations of selection with fenpropathrin in the laboratory, this T. cinnabarinus strain developed 8.7- and 28.7-fold resistance, respectively. Resistance to abamectin in AbR (abamectin resistant strain) and to fenpropathrin in FeR (fenpropathrin resistant strain) was partially suppressed by piperonyl butoxide (PBO), diethyl maleate (DEM) and triphenyl phosphate (TPP), inhibitors of mixed function oxidase (MFO), glutathione S-transferases (GST), and hydrolases, respectively, suggesting that these three enzyme families are important in conferring abamectin and fenpropathrin resistance in T. cinnabarinus. The major resistant mechanism to abamectin was the increasing activities of carboxylesterases (CarE), glutathione-S-transferase (GST) and mixed function oxidase (MFO), and the activity in resistant strain developed 2.7-, 3.4- and 1.4-fold contrasted to that in susceptible strain, respectively. The activity of glutathione-S-transferase (GST) in the FeR strain developed 2.8-fold when compared with the susceptible strain, which meant the resistance to fenpropathrin was related with the activity increase of glutathione-S-transferase (GST) in T. cinnabarinus. The result of the kinetic mensuration of carboxylesterases (CarE) showed that the structure of CarE in the AbR has been changed.     

Metabolic mechanisms involved in the resistance of field populations of Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) to spinosad

Tuta absoluta (Meyrick) resistance to insecticides has become a significant problem in many tomato production areas in South America. New insecticides are now available for the management of this pest (i.e. spinosad), however there is scarce information about their efficacy on field populations.With the aim of determining the susceptibility of T. absoluta to spinosad we evaluated the response of second instar larvae, from five field populations (Azapa 1, Azapa 2, Lluta, Colín and Valdivia) and a laboratory reference strain (S), to a diagnostic concentration of the insecticide. We also determined the activity of the detoxifying enzymes mixed-function oxidases (MFO), glutathione-S-transferases (GST) and esterases (EST) in the same larval stage. Larval mortality in field populations was significantly lower in Azapa 1 (50.0%), Azapa 2 (44.9%), Lluta (39.9%) and Colín (53.5%) when compared to the laboratory strain (91.7%). MFO activities in field populations were between 1.8 and 4.6 times higher than those observed in the S strain, while for EST, the ratio varied from 1.7 to 14.7. The lowest ratios were observed for the GST (0.5-2.7), however, significant differences were detected for the three enzyme systems. We conclude that the evaluated mechanisms would be involved in spinosad resistance of populations of T. absoluta, presenting an increased MFO activity in all populations.