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The Mossie-Buster: a hose-driven insecticide delivery tool for the control of container-breeding mosquitoes.

A novel insecticide delivery tool, the Mossie-Buster, was recently developed to control larval populations from urban breeding sites in Townsville, Australia. This functional and user-friendly control device directly targets the main breeding sites of Aedes aegypti (L.) with a focused delivery of insecticides. The Mossie-Buster comprises a commercially available device and attachments that mix an insecticide solution into the flow of water emitted from a hose. Attached to the device is a trigger for controlled insecticide release. Preliminary laboratory and field trials demonstrated the tool to be effective in eliminating all Ae. aegypti present in various typical breeding containers in different environmental conditions for a minimum of 2 wk in exposed areas to 3 months in an unexposed area.

Aedes↗

Monitoring insecticide resistance in house flies (Diptera: Muscidae) from New York dairies.

House flies were collected from dairies across New York state and the levels of resistance to seven insecticides were measured using standard laboratory assays with three to five diagnostic concentrations. The highest levels of resistance were found for tetrachlorvinphos, permethrin and cyfluthrin. Although levels of resistance to methomyl were somewhat lower, they were among the highest ever reported for field-collected house flies. Resistance to pyrethrins was limited primarily to the lowest diagnostic concentration. House flies were susceptible to fipronil at all dairies, suggesting that this material would be highly effective for fly control. The levels of resistance were similar at all the dairies, irrespective of their insecticide use, suggesting substantial movement of flies between facilities. Relative to resistance levels found at New York dairies in 1987, resistance levels had increased for permethrin, were unchanged for tetrachlorvinphos and had decreased for dimethoate. To identify a single diagnostic concentration that could be used in the laboratory assays to assess accurately resistance levels in future studies, we carried out a 'simulated' field bioassay using formulated materials. A diagnostic concentration for each insecticide is proposed on the basis of a comparison of our bioassays.

Adaptation, Physiological↗

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.

Animals↗

Temporal and spatial dynamics of insecticide resistance in Myzus persicae (Hemiptera: Aphididae).

Advances in understanding insecticide resistance in the peach-potato aphid, Myzus persicae (Sulzer), at the genotypic, biochemical and molecular levels have led to rapid and precise methods for the detection of several resistance mechanisms (elevated carboxylesterase, modified acetylcholinesterase or MACE, and knockdown resistance or kdr) in individual insects, and for monitoring their frequencies over space and time. This paper summarises the results of two long-term surveys of resistance dynamics in M persicae in England, based on samples collected directly from field and glasshouse crops or from four 12.2-m suction traps. The study showed marked fluctuations in resistance frequencies that probably reflect the counteracting forces of selection imposed by insecticides for aphids possessing more copies of esterase resistance genes, and selection against these forms when insecticide use is relaxed. There is growing evidence that several different resistance mechanisms in M persicae have associated fitness costs. In the case of esterase and MACE, these costs are apparently strong enough to effect a decline in resistance frequency over winter, and a more prolonged decline over successive cropping seasons when aphid numbers are insufficient to trigger intensive chemical applications. Changes in the overall frequency of resistance genotypes may also be influenced by the predominance of year-round parthenogenesis in M persicae in the UK, leading to non-random associations between mechanisms and selection operating on clonal lineages rather than individual genotypes.

Acetylcholinesterase↗

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.

Animals↗

Regional reversion of insecticide resistance in Helicoverpa armigera (Lepidoptera: Noctuidae) is associated with the use of Bt cotton in northern China.

A continuous programme for monitoring resistance of Helicoverpa armigera (Hubner) to commonly used insecticides was undertaken for assessing the impact of Bt cotton planting on the evolution of the pest resistance from 1994 to 2002 in China. The bioassay results showed that the resistance levels per year in field populations of H armigera to lambda-cyhalothrin, phoxim and endosulfan decreased, respectively, to 9-15-fold, 11-14-fold and 6-8-fold in 2001-2002 from 197-262-fold, 52-74-fold and 18-38-fold before the cultivation of Bt cotton in 1997. This significant increase in susceptibility to insecticides is expected to result in a reduction in insecticide application for H armigera control in Bt cotton. It is concluded that Bt cotton is playing an important role in the long-term management of H armigera by increasing the potential for natural and chemical control of the pest.

Animals↗

GABA receptor minigene rescues insecticide resistance phenotypes in Drosophila.

A single point mutation within the GABA receptor gene Resistance to dieldrin (Rdl) confers a high level of resistance to cyclodiene insecticides in a wide range of insects. Previous studies have shown partial rescue of the susceptible phenotype via germline transformation of a 36 kb cosmid coding (or all four alternative Rdl splice forms. Here, we describe the construction of two Rdl promoter/cDNA minigenes, each coding for one of the splice forms alone. Single splice forms rescued both the insecticide susceptible and resistant phenotypes associated with the locus as effectively as the complete cosmid. The minigenes also rescue the lethality associated with homozygous re-arrangements disrupting the Rdl gene, and the level of rescue observed is not increased by the addition of more than one splice form. This demonstrates that only a single Rdl splice form is necessary both to confer insecticide sensitivity and also to rescue lethality. Methods by which phenotype rescue could be enhanced and the potential advantages of using Rdl as a selectable marker are discussed.

Animals↗

Cloning of the Drosophila cyclodiene insecticide resistance gene: a novel GABAA receptor subtype?

1. gamma-aminobutyric acid (GABA) is a major inhibitory neurotransmitter in both vertebrates and invertebrates. GABAA receptors are composed of a number of different subunits that assemble to form a chloride ionophore. 2. Several subunit types, alpha, beta, gamma, delta and rho have been cloned from vertebrates, but until recently these receptors have remained uncloned from invertebrates. 3. GABAA receptors form the proposed site of action of cyclodiene insecticides. Therefore a Drosophila mutant (Rdl), resistant to cyclodienes and the GABAA receptor ligand picrotoxin (PTX), was used to clone the gene responsible for resistance as a putative invertebrate GABAA receptor. 4. Analysis of the predicted amino acid sequence and gene structure shows that Rdl codes for a receptor subunit similar to vertebrate GABAA receptors, but sufficiently different that it may represent a novel class of GABAA receptor subtype. 5. Cyclodiene insecticide resistance accounts for over 60% of reported cases of insecticide resistance and is also found in vertebrates. 6. Therefore elucidating the molecular basis of cyclodiene resistance is not only important to our understanding of pesticide action on the GABAA receptor but also in examining the conservation of the resistance mechanism between vertebrates and invertebrates.

Animals↗

Toxicology, mode of action and target site-mediated resistance to insecticides acting on chloride channels.

1. The discovery that the avermectins and cyclodienes affected the chloride channels of excitable membranes generated intense research interest among academic and industrial scientists. 2. The results of biochemical and neurophysiological studies indicate that the gamma-aminobutyric acid (GABA)-gated chloride channel is an important, if not the primary site of action for these compounds. 3. The action of insecticides on the functional properties of the GABA receptor differs by structural class. The cyclodienes block the chloride ion channel and the avermectins activate it. 4. Blockage of the GABA-gated chloride channel by cyclodienes reduces neuronal inhibition, which leads to hyper-excitation of the central nervous system, convulsions, and death. For avermectins, activation of the channel suppresses neuronal activity, resulting in ataxia, paralysis and death. Although actions on the GABA-gated chloride channel can explain many of the effects of these compounds, there is evidence supporting the participation of other ligand- and voltage-gated chloride channels in the overall intoxication process. This consideration is especially true for the avermectins. 5. Several structural series of experimental insecticides have been synthesized which possess a blocking action on the GABA-gated chloride channel similar to that of the cyclodienes. 6. Resistance to cyclodienes usually occurs through an altered target site, and extends to all experimental compounds that block chloride channels. However, the resistance does not afford protection against the avermectins. 7. The continued search for new insecticides directed against chloride channels may lead to compounds with less environmental impact and greater selectivity than that of the cyclodienes. Given the pre-selection for resistance by the cyclodienes, new compounds with a similar mode of action must be used judiciously in order to suppress or delay the re-emergence of widespread resistance.

Animals↗

An amino acid substitution attributable to insecticide-insensitivity of acetylcholinesterase in a Japanese encephalitis vector mosquito, Culex tritaeniorhynchus.

A cDNA sequence encoding a Drosophila Ace-paralogous acetylcholinesterase (AChE) precursor of 701 amino acid residues was identified as the second AChE gene (Ace2) transcript from Culex tritaeniorhynchus. The Ace2 gene is tightly linked to organophosphorus insecticide (OP)-insensitivity of AChE on chromosome 2. The cDNA sequences were compared between an insecticide-susceptible strain and the resistant strain, TYM, that exhibits a 870-fold decrease in fenitroxon-sensitivity of AChE. Two amino acid substitutions were present in TYM mosquitoes. One is F455W whose homologous position in Torped AChE (Phe331) is located in the vicinity of the catalytic His in the acyl pocket of the active site gorge. The other substitution is located to a C-terminal Ile697 position that apparently seems to be excluded from the mature protein and is irrelevant to catalytic activity. The F455W replacement in the Ace2 gene is solely responsible for the insecticide-insensitivity of AChE in TYM mosquitoes.

Acetylcholinesterase↗

Collembola and macroarthropod community responses to carbamate, organophosphate and synthetic pyrethroid insecticides: direct and indirect effects.

Non-target effects on terrestrial arthropod communities of the broad-spectrum insecticides chlorpyrifos and cypermethrin and the selective insecticide pirimicarb were investigated in winter wheat fields in summer. Effects of chlorpyrifos on arthropod abundance and taxonomic richness were consistently negative whereas effects of cypermethrin were negative for predatory arthropods but positive for soil surface Collembola. Pirimicarb effects were marginal, primarily on aphids and their antagonists, with no effect on the Collembola community. Collembola-predator ratios were significantly higher following cypermethrin treatment, suggesting that cypermethrin-induced increases in collembolan abundance represent a classical resurgence. Observations in other studies suggest Collembola resurgences may be typical after synthetic pyrethroid applications. Collembola responses to insecticides differed among species, both in terms of effect magnitude and persistence, suggesting that coarse taxonomic monitoring would not adequately detect pesticide risks. These findings have implications for pesticide risk assessments and for the selection of indicator species.

Agriculture↗

Mutations in the acetylcholinesterase gene of Bactrocera dorsalis associated with resistance to organophosphorus insecticides.

Mutations in the gene encoding the enzyme acetylcholinesterase (AChE) of the oriental fruit fly, Bactrocera dorsalis, associated with resistance to an organophosphorus insecticide have been characterized. Three point mutations producing nonsynonymous changes in the predicted amino acid sequence of the product of the B. dorsalis ace gene in resistant vs. susceptible flies have been identified. One of these changes is unique to B. dorsalis while the other two occur at sites that are identical to mutations previously described for another Bactrocera species. Although the precise role of the third mutation is not clearly established, the independent origin of two identical alterations in these two species strongly supports the idea proposed previously that molecular changes associated with insecticide resistance in key genes and enzymes such as AChE are largely constrained to a limited number of sites. The results obtained here also suggest that the widespread use of organophosphorus insecticides will likely lead to a predictable acquisition of resistance in wild populations of B. dorsalis as well as other pest species. For surveys of B. dorsalis populations that may develop resistance, diagnostic tests using PCR-RFLP based methods for detecting the presence of all three mutations in individual flies are described.

Acetylcholinesterase↗

AaIT: from neurotoxin to insecticide.

AaIT is a single chain neurotoxic polypeptide derived from the venom of the Buthid scorpion Androctonus australis Hector, composed of 70 amino acids cross-linked by four disulfide bridges. Its strict selectivity for insects has been documented by toxicity, electrophysiological and ligand receptor binding assays. These last have shown that various insect neuronal membranes possess a single class of non-interacting AaIT binding sites of high affinity (K(D) = 1-3(n)M) and low capacity (0.5-2.0 pmol/mg prot.). The fast excitatory paralysis induced by AaIT is a result of a presynaptic effect, namely the induction of a repetitive firing in the terminal branches of the insect's motor nerves resulting in a massive and uncoordinated stimulation of the respective skeletal muscles. The neuronal repetitive activity is attributed to an exclusive and specific perturbation of sodium conductance as a consequence of toxin binding to external loops of the insect voltage-dependent sodium channel and modification of its gating mechanism. From a strictly agrotechnical point of view AaIT involvement in plant protection has taken the following two complementary forms: firstly, as a factor for the genetic engineering of insect infective baculoviruses resulting in potent and selective bio-insecticides. The efficacy of the AaIT-expressing, recombinant baculovirus is attributed mainly to its ability to continuously provide and translocate the gene of the expressed toxin to the insect central nervous system; secondly, based on the pharmacological flexibility of the voltage-gated sodium channel, as a device for insecticide resistance management. Channel mutations conferring resistance to a given class of insecticidal agents (such as the KDR phenomenon) may greatly increase susceptibility to the AaIT expressing bioinsecticides. Thus the AaIT is a pharmacological tool for the study of insect neuronal excitability and chemical ecology and the development of new approaches to insect control.

Amino Acid Sequence↗

Horn fly (Diptera: Muscidae) resistance to organophosphate insecticides.

Insecticidal ear tags impregnated with organophosphate (OP) insecticides were used each year from 1989 to 1998 at Rosepine, LA. Weekly fly counts were conducted to evaluate control efficacy of the treatments, and bioassays were conducted at least twice per year to measure fly susceptibility to OP and pyrethroid insecticides. Between 1989 and 1992, the efficacy of 20% diazinon-impregnated ear tags was reduced from >20 to just 1 week of control. A high risk of control failure was observed when a resistance frequency of approximately 5% was measured in pre-season bioassays. Resistance to diazinon, fenthion, ethion, pirimiphos-methyl, and tetrachlorvinphos was observed. Esterase activity toward alpha-naphthyl acetate was significantly higher in flies collected at Rosepine in 1997 than in flies from a laboratory colony and from a susceptible field population.

Administration, Topical↗

Increased transcription of CYP6D1 causes cytochrome P450-mediated insecticide resistance in house fly.

Insecticide resistance is a major problem that continues to plague efforts to control pests of animals and crops. An important mechanism by which insects become resistant to insecticides is via increased detoxification mediated by the cytochrome P450 microsomal monooxygenases (monooxygenases). One of the fundamental gaps in our knowledge about this resistance mechanism is an understanding of how insects express high levels of the specific cytochrome P450(s) responsible for resistance. One such P450, CYP6D1, causes resistance to pyrethroid in the house fly and is expressed at 9-fold higher levels (mRNA and protein) in the Learn Pyrethroid Resistant (LPR) strain (compared to susceptible strains). The relative stability of CYP6D1 mRNA in resistant and susceptible strains was measured following inhibition of transcription with actinomycin D. The same time course of decrease in CYP6D1 mRNA abundance was detected in both strains indicating that the high level of expression of CYP6D1 in LPR is not due to increased stability of the mRNA. The comparative rates of transcription of CYP6D1 were measured using an in vitro run-on transcription assay. The relative amount of CYP6D1 transcript produced in this assay was 10-fold greater in the LPR strain compared to the susceptible strain. This demonstrates that increased transcription of CYP6D1 is an underlying cause of monooxygenase-mediated insecticide resistance. The increased rate of transcription of CYP6D1 in the resistant strain (LPR) is controlled by two factors: one on autosome 1 and another on autosome 2.

Animals↗

Increased fecundity of malathion-specific resistant beetles in absence of insecticide pressure.

Despite that resistance frequency is assumed to decline when selective pressure is relaxed, the stability of resistance frequency has been observed in some insects in the absence of insecticide. In the red flour beetle, Tribolium castaneum, the first case of malathion-resistance was reported in the early 1960s. The malathion-specific resistant phenotype has now almost completely replaced the susceptible one in red flour beetle populations. In the present study, several life-history traits that could influence the fitness of the insects were compared between insecticide-susceptible and malathion-specific resistant populations of the red flour beetle. On average, egg fertility and egg-to-adult development time did not differ between susceptible and resistant populations. However, the fecundity of resistant females was greater than that of susceptible ones. Generally, differences in development time between insecticide resistant and susceptible populations are considered as having more effect on fitness than do differences in fecundity. However, the observed increased female fecundity may participate, in combination with the previously observed increased male reproductive success, to the development and the stability of malathion-specific resistance in T. castaneum.

Animals↗

Structure and organization of amplicons containing the E4 esterase genes responsible for insecticide resistance in the aphid Myzus persicae (Sulzer).

Insecticide resistance in the aphid Myzus persicae results primarily from the amplification of genes encoding the insecticide-detoxifying esterase, E4. Here we report the analysis of flanking DNA co-amplified with the E4 gene. The 5' end of this gene has an untranslated leader sequence interspersed by two introns, and the promoter region lacks TATA and CAAT boxes. The DNA breakpoint involved in the generation of the amplification is just upstream (approx. 250 bp) of the putative E4 transcription start site; thus the E4 gene is very close to the 5' end of the approx. 24 kb amplicon. PCR primers specific to the 'novel joint' generated during the amplification have been used to show that a wide range of aphid clones have the same amplicons, arranged as a series of head-to-tail direct repeats. Long-distance mapping has revealed the structure of these repeats. This has important implications for understanding both the generation of the amplified genes and the origin and spread of insecticide resistance in M. persicae.

Animals↗

Insecticide resistance spectra and resistance mechanisms in populations of Japanese encephalitis vector mosquitoes, Culex tritaeniorhynchus and Cx. gelidus, in Sri Lanka.

Culex tritaeniorhynchus Giles and Cx. gelidus Theobald (Diptera: Culicidae), both vectors of Japanese encephalitis, were collected in 1984 and 1998 from two disease endemic localities in Sri Lanka: Anaradhapura and Kandy. Using wild-caught adult mosquitoes from light traps, log dosage-probit mortality curves for insecticide bioassays were obtained for three insecticides: malathion (organophosphate), propoxur (carbamate) and permethrin (pyrethroid). LD50 values showed that, in 1998, Cx. tritaeniorhynchus was -100-fold more resistant to malathion and 10-fold more resistant to propoxur than was Cx. gelidus. This difference was attributed to Cx. tritaeniorhynchus breeding mostly in irrigated rice paddy fields, where it would have been exposed to pesticide selection pressure, whereas Cx. gelidus breeds in other types of aquatic habitats less prone to pesticide applications. Resistance in Cx. tritaeniorhynchus increased between 1984 and 1998, whereas Cx. gelidus remained predominantly susceptible. Propoxur inhibition of acetylcholinesterase (AChE) activity (the target site of organophosphates and carbamates) indicated that in 1998, frequencies of insensitive AChE-based resistance were 9% in Cx. gelidus and 2-23% in Cx. tritaeniorhynchus, whereas in 1984 this resistance mechanism was detected only in 2% of the latter species from Anaradhapura. The AChE inhibition coefficient (ki) with propoxur was 1.86+/-0.24 x 10(5) M(-)1 min(-1) for Cx. tritaeniorhynchus from Anaradhapura in 1998. Both species were tested for activity levels of detoxifying glutathione S-transferases (GSTs) and malathion-specific as well as general carboxylesterases. High activities of GSTs and carboxylesterases were detected in Cx. tritaeniorhynchus but not Cx. gelidus. Malathion-specific carboxylesterase was absent from both species. Native polyacrylamide gel electrophoresis resolved two elevated general carboxylesterases, CtrEstbeta1 and CtrEstalpha1, from Cx. tritaeniorhynchus and none from Cx. gelidus. CtrEstbeta1 was the most intensely staining band. Gel inhibition experiments showed that both elevated esterases were inhibited by organophosphates and carbamates but not by pyrethroids. The major elevated esterase CtrEstbeta1 was partially purified (15-fold) by sequential Q-Sepharose and phenyl Sepharose column chromatography. The bimolecular rate constant (ka) and the deacylation rate constant (k3) for the malaoxon/enzyme interaction were 9.9+/-1.1 x 10(3) M(-1) min(-1) and 3.5+/-0.05 x 10(-4) M(-1) min(-1), respectively, demonstrating that the role of this enzyme in organophosphorus insecticide resistance is sequestration.

Animals↗