Metabolic mechanisms of indoxacarb resistance in field populations of Choristoneura rosaceana (Harris) (Lepidoptera: Tortricidae).

Synergism and metabolic studies were conducted to identify the resistance mechanism against indoxacarb in two Choristoneura rosaceana (Harris) field populations compared to a susceptible population. The synergism study was carried out using diet incorporation bioassay for indoxacarb and the three synergists PBO, DEM, and DEF. The metabolic study consists of indoxacarb in vitro reaction with fifth instar larvae 12,000 g midgut supernatant or with pre-inhibited (in vivo by the esterases inhibitor DEF) fifth instar larvae 12,000 g midgut supernatant at different incubation times. In both susceptible and cherry populations, only DEF significantly synergized indoxacarb with a synergism ratio (SR) of 6.5 and 22.6-fold respectively indicating an involvement of esterases in the both populations. In the apple population, all synergists PBO, DEM, and DEF significantly synergized indoxacarb with SR of 9.6, 7.7, and 285.6-fold respectively indicating a complex resistance case with the possible involvement of all three metabolic resistance mechanisms with the central role of esterase enzymes. In vitro, the indoxacarb (DPX-JW062) was very rapidly metabolized within 5 min into small molecules in the lower portion of the metabolic pathway when it reacted with the midgut supernatant of each population. None of the metabolites in the upper portion of the metabolic pathway were detected at any incubation time including the potent sodium channel blocker DCJW metabolite. The two field populations showed significantly higher rates of metabolism of DPX-JW062 compared to the susceptible population at five min of incubation and that may explain the presence of indoxacarb resistance. In the second part of the in vitro study, the bio-transformation of DPX-JW062 was remarkably decreased when it reacted with the pre-inhibited (by DEF) midgut supernatant of each population. Additionally, the degradation of metabolites in the upper portion of the metabolic pathway remarkably decreased, which resulted in accumulation of DCJW and MP819 metabolites. The accumulation of DCJW metabolite under the pre-inhibited midgut supernatants treatment provided a persuasive explanation of the synergistic impact of esterase inhibitor DEF on indoxacarb in C. rosaceana.

Defining terms for proactive management of resistance to Bt crops and pesticides.

Evolution of pest resistance to pesticides is an urgent global problem with resistance recorded in at least 954 species of pests, including 546 arthropods, 218 weeds, and 190 plant pathogens. To facilitate understanding and management of resistance, we provide definitions of 50 key terms related to resistance. We confirm the broad, long-standing definition of resistance, which is a genetically based decrease in susceptibility to a pesticide, and the definition of "field-evolved resistance," which is a genetically based decrease in susceptibility to a pesticide in a population caused by exposure to the pesticide in the field. The impact of field-evolved resistance on pest control can vary from none to severe. We define "practical resistance" as field-evolved resistance that reduces pesticide efficacy and has practical consequences for pest control. Recognizing that resistance is not "all or none" and that intermediate levels of resistance can have a continuum of effects on pest control, we describe five categories of field-evolved resistance and use them to classify 13 cases of field-evolved resistance to five Bacillus thuringiensis (Bt) toxins in transgenic corn and cotton based on monitoring data from five continents for nine major pest species. We urge researchers to publish and analyze their resistance monitoring data in conjunction with data on management practices to accelerate progress in determining which actions will be most useful in response to specific data on the magnitude, distribution, and impact of resistance.

Resistance of codling moth, Cydia pomonella (L.) (Lepidoptera: Tortricidae), larvae in Michigan to insecticides with different modes of action and the impact on field residual activity.

BACKGROUND: The codling moth is one of the principal pests of apple in the world. Resistance monitoring is crucial to the effective management of resistance in codling moth. Three populations of codling moth in neonate larvae were evaluated for resistance to seven insecticides via diet bioassays, and compared with a susceptible population. In addition, apple plots were treated with labeled field rate doses of four insecticides. Treated fruit were exposed to neonate larvae of two populations from commercial orchards. RESULTS: Two populations of codling moth expressed two- and fivefold resistance to azinphos-methyl, seven- and eightfold resistance to phosmet, six- and tenfold resistance to lambda-cyhalothrin, 14- and 16-fold resistance to methoxyfenozide and sixfold resistance to indoxacarb, but no resistance to acetamiprid and spinosad. The impact of the resistance to azinphos-methyl, measured as fruit damage, increased as the insecticide residues aged in the field. In contrast, fruit damage in methoxyfenozide- and lambda-cyhalothrin-treated fruit was observed earlier for resistant codling moth. No differences in efficacy were found for acetamiprid. CONCLUSIONS: Broad-spectrum insecticide resistance was detected for codling moth. Resistance to azinphos-methyl, lambda-cyhalothrin and methoxyfenozide was associated with reduced residual activity in the field. Broad-spectrum resistance presents serious problems for management of the codling moth in Michigan.

Neural actions of imidacloprid and their involvement in resistance in the Colorado potato beetle, Leptinotarsa decemlineata (Say).

BACKGROUND: The development of resistance to imidacloprid in eastern US populations of the Colorado potato beetle (CPB), Leptinotarsa decemlineata (Say), threatens this critical use for neonicotinoid insecticides. Previous pharmacokinetic studies with resistant adult CPBs provided no explanation for the high resistance level (over 200-fold) to topically applied imidacloprid. The authors assessed the neural activity of imidacloprid by recording spontaneous activity from a motor nerve leaving the isolated central nervous system to compare the sensitivity of the latter to imidacloprid between susceptible and resistant CPBs. RESULTS: On the isolated central nervous system, imidacloprid was initially neuroexcitatory, and neuroinhibitory at higher concentrations. The neuroexcitatory action of imidacloprid was blocked by coapplication of a specific nAChR antagonist, methyllycaconitine, indicating that it is a result of action on nAChRs. The sensitivity to the neuroexcitatory and inhibitory activities of imidacloprid varied independently among individuals in each population. The sensitivity of the central nervous system of resistant CPBs to excitation by imidacloprid did not differ from that of susceptible insects, but the sensitivity to inhibition by imidacloprid was reduced 52- to 58-fold, indicating a possible change in the sensitivity of at least one subgroup of nAChRs. CONCLUSION: This study provides evidence that reduced nerve sensitivity to the blocking action of imidacloprid is associated with imidacloprid resistance in the CPB.

Agonist actions of neonicotinoids on nicotinic acetylcholine receptors expressed by cockroach neurons.

The agonist actions of seven commercial neonicotinoid insecticides and nicotine were studied on nicotinic acetylcholine receptors (nAChRs) expressed by neurons isolated from the three thoracic ganglia of the American cockroach, Periplaneta americana. Single electrode voltage clamp recording was used to measure agonist-induced inward currents. Acetylcholine, nicotine and all neonicotinoids tested, except thiamethoxam, caused inward currents which were blocked reversibly by methyllycaconitine, a nAChR antagonist. Based on maximum inward currents, neonicotinoids could be divided into two subgroups: (1) those with a heterocyclic ring in their electronegative pharmacophore moiety (i.e. nicotine, imidacloprid and thiacloprid) were relatively weak partial agonists causing only 20-25% of the maximum ACh current and (2) open chain compounds (i.e. acetamiprid, dinotefuran, nitenpyram, and clothiandin) which were much more effective agonists producing 60-100% of the maximum ACh current. These compounds also elicited different symptoms of poisoning in American cockroaches with excitatory responses evident for the low efficacy agonists but depressive and paralytic responses predominating for the most efficacious agonists. No correlation was observed between agonist affinity and efficacy on these nAChRs. Thiamethoxam, even at 100 microM, failed to cause an inward current and showed no competitive interaction with other neonicotinoids on nAChRs, indicating that it is not a direct-acting agonist or antagonist. Despite the probable presence of multiple subtypes of nAChRs on cockroach neurons, competition studies between neonicotinoids did not reveal evidence that separate binding sites exist for the tested compounds. The size of inward currents induced by co-application of neonicotinoid pairs at equal concentration (100 microM) were predominantly determined by the one with higher binding affinity as indicated by EC(50) values, rather than by the one with higher binding efficacy as indicated by maximal current (I(max)). Agonist efficacy, but not affinity, was positively correlated with insecticidal activity. These findings indicate that: (1) agonist affinity and efficacy vary independently with neonicotinoid structure; (2) high agonist efficacy is dependent on the presence of an acyclic electronegative pharmacophore group; (3) agonist efficacy is a significant factor in the insecticidal activity of neonicotinoids to cockroaches; (4) lower efficacy compounds cause excitatory symptoms (Type A), while high efficacy compounds cause depressive/paralytic symptoms (Type B).

Resistance and cross-resistance to neonicotinoid insecticides and spinosad in the Colorado potato beetle, Leptinotarsa decemlineata (Say) (Coleoptera: Chrysomelidae).

The Colorado potato beetle, Leptinotarsa decemlineata (Say), has developed resistance to many insecticides used for its control, recently including imidacloprid, a neonicotinoid compound. Other neonicotinoids are now being deployed to control this pest. A key point in the strategies of resistance management is the monitoring of resistance and cross-resistance. In the summer of 2003, imidacloprid-resistant adult Colorado potato beetles collected from Long Island, New York, USA were bioassayed using topical applications of imidacloprid and nine other neonicotinoids. Compared to a standard susceptible strain, the Long Island beetles showed 309-fold resistance to imidacloprid, and lower levels of cross-resistance to all other neonicotinoids, despite these never having been used in the field, i.e., 59-fold to dinotefuran, 33-fold to clothianidin, 29-fold to acetamiprid, 28-fold to N-methylimidacloprid, 25-fold to thiacloprid, 15-fold to thiamethoxam, 10-fold to nitenpyram, but less than 2-fold to nicotine. In injection bioassays, high resistance to imidacloprid was also found (116-fold). Piperonyl butoxide partially suppressed resistance to imidacloprid, but the resistance level was still over 100-fold, indicating that other mechanisms were primarily responsible for resistance. Low levels of resistance (8- to 10-fold) were found to the nicotinic activator, spinosad, in an imidacloprid-resistant strain collected from the same field in 2004. The cross-resistance seen with all the neonicotinoids tested suggests that the rotation of imidacloprid with other neonicotinoids may not be an effective long-term resistance management strategy. Rotation with spinosad also carries some risk, but it is unlikely that spinosad resistance in this case is mechanistically related to that for the neonicotinoids.

Synergism of insecticides provides evidence of metabolic mechanisms of resistance in the obliquebanded leafroller Choristoneura rosaceana (Lepidoptera: Tortricidae).

The interactions between six insecticides (indoxacarb, cypermethrin, chlorpyrifos, azinphosmethyl, tebufenozide and chlorfenapyr) and three potential synergists, (piperonyl butoxide (PBO), S,S,S-tributyl phosphorotrithioate (DEF) and diethyl maleate (DEM)) were studied by dietary exposure in a multi-resistant and a susceptible strain of the obliquebanded leafroller, Choristoneura rosaceana (Harris). The synergists did not produce appreciable synergism with most of the insecticides in the susceptible strain. Except for tebufenozide, PBO synergized all the insecticides to varying degrees in the resistant strain. A very high level of synergism by PBO was found with indoxacarb, which reduced the resistance level from 705- to 20-fold when PBO was administered alone and to around 10-fold when used in combination with DEF. DEF also synergized indoxacarb, cypermethrin, chlorpyrifos, azinphosmethyl and tebufenozide in the resistant strain. DEM produced synergism of indoxacarb, chlorpyrifos, azinphos-methyl and chlorfenapyr in the resistant strain. DEM was highly synergistic to cypermethrin, and to some extent to tebufenozide in both the susceptible and resistant strains equally, implying that detoxification by glutathione S-transferases was not a mechanism of resistance for these insecticides. The high level of synergism seen with DEM in the case of cypermethrin may be due to an increase in oxidative stress resulting from the removal of the antioxidant, glutathione. These studies indicate that enhanced detoxification, often mediated by cytochrome P-450 monooxygenases, but with probable esterase and glutathione S-transferase contributions in some cases, is the major mechanism imparting resistance to different insecticides in C. rosaceana.

Broad-spectrum insecticide resistance in obliquebanded leafroller Choristoneura rosaceana (Lepidoptera: Tortricidae) from Michigan.

Nineteen insecticides, belonging to nine chemical classes, were bioassayed by dietary exposure against two strains of obliquebanded leafroller, Choristoneura rosaceana, collected from Michigan apple orchards. Berrien is a putatively organophosphate-resistant strain from a commercial orchard with a history of insecticide use, and Kalamazoo a susceptible strain from an isolated and unsprayed orchard. The Berrien strain was moderately resistant (about 25-fold) to organophosphates such as azinphos-methyl and chlorpyrifos. Very low resistance (< 10-fold) was also observed to pyrethroids such as cypermethrin, zeta-cypermethrin, bifenthrin, deltamethrin and esfenvalerate, to the ecdysone agonists tebufenozide and methoxyfenozide, and to the chlorinated pyrrole chlorfenapyr. Endosulfan and carbamates such as thiodicarb, methomyl and carbaryl had low intrinsic toxicities against both strains, with little difference in sensitivity between them. There was no resistance to spinosad. Emamectin benzoate was found to be the most toxic insecticide against C rosaceana, with slightly higher lethal doses required for the Berrien strain. Unexpectedly, Berrien exhibited a very high level of resistance (> 700-fold) to indoxacarb, which has never been used in Michigan to control this insect pest. The active metabolite of indoxacarb, DCJW, was considerably more toxic than the parent compound, but the resistance against DCJW was comparable to that seen with indoxacarb. This indicates that a failure to activate indoxacarb was not the mechanism of resistance in Berrien. The low level of resistance to several chemistries recorded in Michigan C rosaceana can be managed at this stage by adopting a rotation of chemistries having different modes of action.