Esterase inhibition by synergists in the western flower thrips Frankliniella occidentalis.

BACKGROUND: Western flower thrips (WFT), Frankliniella occidentalis (Pergande), is among the most important crop pests in the south-eastern region of Spain. Its increasing resistance to insecticides constitutes a serious problem, and understanding the mechanisms involved is therefore of great interest. Use of synergists to inhibit the enzymes involved in insecticide detoxification is widely used to determine their responsibility for insecticide resistance. However, they do not always act as intended or expected, and caution must be exercised when interpreting synergist results. RESULTS: Laboratory-selected strains of WFT were used to analyse the effects of the synergists piperonyl butoxide (PBO), S,S,S-tributyl phosphorotrithioate (DEF) and methiocarb on total esterase activity. Significant differences were found, indicating esterase activity inhibition by DEF, a lower effect for methiocarb and a small inhibition of the activity by PBO. Esterase isoenzyme inhibition by these compounds showed a similar result; this assay revealed an extreme sensitivity of Triplet A (resistance-associated esterases) to DEF. In an in vivo assay carried out with these compounds at different incubation times, only DEF caused posterior in vitro esterase activity inhibition, with a maximum effect 1 h after treatment. CONCLUSION: In this work, only DEF shows true synergistic inhibition of WFT esterases.

Esterase isoenzymes and insecticide resistance in Frankliniella occidentalis populations from the south-east region of Spain.

BACKGROUND: Frankliniella occidentalis (Pergande) is among the most important crop pests in the south-east region of Spain; its increasing resistance to insecticides constitutes a serious problem, and understanding the mechanisms involved is therefore of great interest. To this end, F. occidentalis populations, collected from the field at different locations in south-east Spain, were studied in terms of total esterase activity and esterase isoenzyme pattern. RESULTS: Individual thrips extracts were analysed by native polyacrylamide gel electrophoresis (PAGE) and stained for esterase activity with the model substrate alpha-naphthyl acetate. Significant correlations were found between resistance to the insecticides acrinathrin and methiocarb and the presence of a group of three intensely stained bands, named Triplet A. For each individual thrips extract, total esterase activity towards the substrates alpha-naphthyl acetate and alpha-naphthyl butyrate was also measured in a microplate reader. Insects possessing Triplet A showed a significantly higher alpha-naphthyl acetate specific activity and alpha-naphthyl acetate/alpha-naphthyl butyrate activity ratio. This observation allowed a reliable classification of susceptible or resistant insects either by PAGE analysis or by total esterase activity determination. CONCLUSION: The PAGE and microplate assays described can be used as a monitoring technique for detecting acrinathrin- and methiocarb-resistant individuals among F. occidentalis field populations.

Relationship between esterase activity and acrinathrin and methiocarb resistance in field populations of western flower thrips, Frankliniella occidentalis.

The western flower thrips, Frankliniella occidentalis (Pergande), is a serious pest in the south-east of Spain owing to its direct feeding on crops, transmission of the tomato spotted wilt virus and its very high level of resistance to insecticides. Mechanisms of resistance were examined using field populations of F. occidentalis with different susceptibilities to acrinathrin, methiocarb (selective insecticides), endosulfan, metamidophos and deltamethrin (broad-spectrum insecticides). Esterase activity towards alpha-naphthyl acetate and p-nitrophenyl acetate in resistant strains was significantly higher than in the reference strain (MLFOM) for both model substrates. This higher activity was significantly correlated with acrinathrin and methiocarb resistance.

Vitellogenin polypeptide levels in one susceptible and one cadmium-resistant strain of Oncopeltus fasciatus (Heteroptera: Lygaeidae), and its role in cadmium resistance.

The heavy metal Cd has recently been shown to have endocrine disrupting activity in different vertebrate species. In the insect Oncopeltus fasciatus, Cd exposure delays ovarian maturation and inhibits vitellogenesis, probably through a reduction in vitellogenin (Vg) polypeptide synthesis. An O. fasciatus strain was selected in the laboratory for Cd resistance with gradually increasing Cd concentrations for 50 generations. Cd-selected (OCdR) insects were 2.7-2.3 times more resistant to chronic exposure to Cd than susceptible (OS) insects. OCdR females showed increased (up to 90%) Vg polypeptide levels in hemolymph. Vg polypeptide synthesis was equally decreased in OCdR and OS females after acute Cd exposure, indicating that resistance in the OCdR strain was not due to a reduced sensitivity of the target of Cd. Vg polypeptide concentrations and ovarian maturation in acutely exposed OCdR females, however, were equal to those of control OS females. OCdR females chronically exposed to Cd showed a 30% decrease in Vg polypeptide concentrations, relative to control OS females. It is concluded that increased Vg polypeptide levels play an important role in resistance to the toxic effects of Cd on reproduction in the OCdR strain, and that this novel Cd resistance mechanism has the potential to evolve in vertebrate species in which Cd inhibits Vg synthesis by disruption of the endocrine system.

Vitellogenesis inhibition in Oncopeltus fasciatus females (Heteroptera: Lygaeidae) exposed to cadmium.

Newly moulted females of the insect Oncopeltus fasciatus were exposed to cadmium (Cd) dissolved in the drinking water (50-400 mg l(-1) Cd) for 5 days. Cd exposure delayed ovarian maturation and inhibited egg production. Exposure to Cd, moreover, decreased hemolymph levels of the two major vitellogenin polypeptides of O. fasciatus, VG1 and VG2, in a concentration-dependent way, probably by a reduction in their synthesis. The ovarian levels of VG1 and VG2 were also decreased in Cd-exposed females. It was next investigated whether Cd effects might be a consequence of the endocrine disruption of vitellogenin synthesis, which is controlled by juvenile hormone (JH). JH replacement therapy did not restore VG1 or VG2 levels in Cd-exposed females, but did so in starved females. Our results do not therefore support a disturbance of JH production or a reduction in feeding as the cause of the reduced vitellogenin polypeptide levels, but rather point to the site of action of JH, the JH receptor, as the target of Cd effects.

Evaluation of metabolic detoxifying enzyme activities and insecticide resistance in Frankliniella occidentalis.

The western flower thrips Frankliniella occidentalis (Pergande) is a very significant pest of a number of different agricultural crops in the south-east of Spain. The importance of thrips as a pest is not due mainly to the direct damage inflicted on the plant, but to the loss in commercial value which occurs as a consequence of the development of dark spots caused by the tomato spotted wilt virus (TSWV) which they transmit. The economic threshold is therefore almost zero, which enhances the problems of resistance management. The present work is part of a global project that attempts to evaluate the status of insecticide resistance in field populations of thrips obtained from several agricultural crops. We have studied, in either individual or pooled insects, some enzyme systems classically related to detoxification of insecticides: esterase and glutathione-S-transferase (GST). The activity of these enzymes from laboratory populations selected with various classes of insecticides has also been measured using several appropriate substrates. An increase in GST mean activity was found in two field-collected strains. Differences in frequency distributions of esterase and GST activities were found for both field-collected strains and for a laboratory strain selected with acrinathrin. These activities were compared with those of a wild-type reference strain.