Ian Denholm

Analogous pleiotropic effects of insecticide resistance genotypes in peach–potato aphids and houseflies

We show that single-point mutations conferring target-site resistance (kdr) to pyrethroids and DDT in aphids and houseflies, and gene amplification conferring metabolic resistance (carboxylesterase) to organophosphates and carbamates in aphids, can have deleterious pleiotropic effects on fitness. Behavioural studies on peach–potato aphids showed that a reduced response to alarm pheromone was associated with both gene amplification and the kdr target-site mutation. In this species, gene amplification was also associated with a decreased propensity to move from senescing leaves to fresh leaves at low temperature. Housefly genotypes possessing the identical kdr mutation were also shown to exhibit behavioural differences in comparison with susceptible insects. In this species, resistant individuals showed no positional preference along a temperature gradient while susceptible genotypes exhibited a strong preference for warmer temperatures.

Comparative survival of insecticide-susceptible and resistant peach-potato aphids, Myzus persicae (Sulzer) (Hemiptera: Aphididae), in low temperature field trials

The survival of Myzus persicae (Sulzer) clones, representing the full range of the recognized levels of insecticide resistance, was measured after 3–4 week exposure to the winter climate in the field. Ten separate trials were carried out using known numbers of first/second instar nymphs on unsprayed oilseed rape and groundsel. These predominantly showed a negative association between resistance level and the proportion of aphids recovered alive after exposure. The strength of the relationship correlated closely with three meteorological variables: the length of time that aphids spent below 2°C, mean rainfall and mean windspeed. We conclude that selection for resistance to insecticides is subject to counteracting selection by cold, wet and windy conditions in the UK winter. Revertant clones, that had spontaneously lost extreme resistance to insecticides despite retaining resistance genes, survived no better than aphids that had kept their high resistance, indicating that overproduction of esterase was not the reason for poor survival.

Reduced alarm response by peach–potato aphids, Myzus persicae (Hemiptera: Aphididae), with knock-down resistance to insecticides ( kdr )may impose a fitness cost through increased vulnerability to natural enemies

Termites were sampled using randomized soil pits in 64 cropping plots, each 25 x 25 m, forming an experimental agrisilvicultural system in both a 6- and an 18-year-old Terminalia ivorensis plantation, in which canopy cover, crop, cropping system and land preparation were the principal treatment variables. The treatments were established in April 1995 and sampling was carried out in November 1995, February 1996 and July 1996. A total of 82 termite species were found, of which 67 were soil-feeders. Overall termite abundance and the abundance of soil-feeders increased between November 1995 and July 1996, reaching a mean of nearly 6000 m -2 . Pooling termite data from these sampling dates, in the old plantation, the high canopy cover treatment (192 stems ha -1 ) had a greater abundance of termites, compared with the low canopy cover treatment (64 stems ha -1 ) and this effect was independent of crop type (plantain or cocoyam), cropping system (single stands or mixed crops) and land preparation (mulch retained or burned, plantain only). The young tree plantation (same tree densities as in the old plantation) showed no significant difference in termite abundance between high and low canopy (levels of tree foliage) densities, though the high canopy sheltered a greater number of termites. Analysis of covariance showed that crop yield (both plantain and cocoyam) was not directly linked to the abundance of all termite populations, but that the cocoyam yield was positively correlated with the abundance of soil-feeding termites (the majority in the assemblage) in the young plantation. This may be due to the beneficial conditioning of soil resulting from the foraging and construction activities of soil-feeders.

A microtitre plate assay for characterizing insensitive acetylcholinesterase genotypes of insecticide-resistant insects

A rapid technique is described for characterizing and monitoring, in single insects, the insensitivity of acetylcholinesterase (AChE) to organophosphorus and carbamate insecticides. Ninety-six insects are homogenized simultaneously in a microtitre plate and portions (e.g. 0·05 for Musca domestica L.) assayed colorimetrically with acetylthiocholine in the presence and absence of diagnostic concentrations of insecticide. Reactions are monitored by a kinetic microplate reader linked to a microcomputer that determines mean AChE activities automatically by linear regressions of absorbance-time data. Mean inhibited activity is then expressed as a percentage of uninhibited activity. Several inhibitors can be tested against the same insect to yield an ‘insensitivity profile‘ of individuals and strains. In tests on M. domestica adults of known AChE genotype, the assay clearly distinguished not only between a sensitive and two slightly (3-15-fold) insensitive AChE variants but between all six genotypic combinations of these three alleles.

The role of B-type esterases in conferring insecticide resistance in the tobacco whitefly, Bemisia tabaci (Genn)

Separation of non-specific esterases on electrophoretic gels has played a key role in distinguishing between races or biotypes of the tobacco whitefly, Bemisia tabaci. One intensively staining esterase in particular (termed E0.14) has assumed significance as a diagnostic of B-type whiteflies (aka Bemisia argentifolii), despite any knowledge of its biological function. In this study, a whitefly strain (B-Null) homozygous for a null allele at the E0.14 locus that had been isolated from a B-type population was used to demonstrate a significant role for E0.14 in resistance of B-type populations to pyrethroids but not to organophosphates (OPs). Bioassays with pyrethroids, following pre-treatment with sub-lethal doses of the OP profenofos (to inhibit esterase activity), coupled with metabolism studies with radiolabelled permethrin, supported the conclusion that pyrethroid resistance in a range of B-type strains expressing E0.14 was primarily due to increased ester hydrolysis. In the same strains, OP resistance appeared to be predominantly conferred by a modification to the target-site enzyme acetylcholinesterase. © 2000 Society of Chemical Industry

Intensification of insecticide resistance in UK field populations of the peach-potato aphid, Myzus persicae (Hemiptera: Aphididae) in 1996

The well-established carboxylesterase-based resistance to insecticides in Myzus persicae Sulzer has recently been accentuated by the appearance of aphids with a modified acetylcholinesterase (MACE) insensitive to pirimicarb and the novel aphicide, triazamate. This target site resistance mechanism was found in M. persicae from crops in the UK for the first time in 1996, together with especially large proportions of aphids with R 2 and R 3 levels of carboxylesterases, a combination that was associated with serious insecticide failures. This paper describes the incidence of both mechanisms and discusses the implications for future recommendations for aphid control in the UK.

Use of biochemical and DNA diagnostics for characterising multiple mechanisms of insecticide resistance in the peach-potato aphid, Myzus persicae (Sulzer).

The peach-potato aphid Myzus persicae (Sulzer) can resist a range of insecticides by over-producing detoxifying esterase and having mutant-insensitive forms of the target proteins, acetylcholinesterase (AChE), and the sodium channel. Using a combination of bioassays, biochemical and DNA diagnostics, it is now possible to diagnose all three mechanisms in individual aphids, and thereby establish their spatial distributions and temporal dynamics. A survey of 58 samples of wide geographic origin showed that all 46 resistant clones had amplified esterase genes (E4 or FE4) conferring broad-spectrum resistance to pyrethroids, organophosphates and carbamates. These occurred in combination with insensitive AChE (11 clones), conferring resistance to pirimicarb and triazamate, and/or mutant sodium channel genes (25 clones), conferring knockdown (kdr) resistance to pyrethroids and DDT. Amplified esterase genes were in linkage disequilibrium with both insensitive AChE and the kdr mutation, reflecting tight physical linkage, heavy selection favouring aphids with multiple mechanisms, and/or the prominence of parthenogenesis in many M. persicae populations. An ability to monitor individual mechanisms with contrasting cross-resistance profiles has important implications for the development of resistance management recommendations.

Insecticide resistance in the currant-lettuce aphid, Nasonovia ribisnigri (Hemiptera: Aphididae) in the UK

Bioassay data for a reference strain of Nasonovia ribisnigri (Mosely), exhibiting similar responses to proven susceptible strains of Myzus persicae (Sulzer) and Aphis gossypii Glover, were used to assess insecticide resistance in two suspected resistant strains and nine field strains of N. ribisnigri originating from lettuce in the UK. Results showed widespread but varied levels of resistance to pirimicarb, lower and also varied resistance to pyrethroids and organophosphates, and no significant differences in response to imidacloprid. In some strains, resistance was associated with an intensely-staining esterase band disclosed by polyacrylamide gel electrophoresis (PAGE). However, no direct relationship between esterases and resistance has yet been established. There was no biochemical evidence of an altered acetylcholinesterase contributing to pirimicarb resistance in these strains.

Factors affecting resistance to insecticides in house-flies, Musca domestica L. (Diptera: Muscidae). I. Long-term control with bioresmethrin of flies with strong pyrethroid-resistance potential

The assumption that by minimising contact with pyrethroids, satisfactory control of Musca domestica L. could be retained without eliciting resistance was tested on a pig farm in southern England where permethrin had failed through rapid development of resistance. Flies were satisfactorily controlled in enclosed buildings for 12 months by space spraying bioresmethrin (2 mg a.i./m 3 ) at approximately fortnightly intervals when numbers reached an arbitrary nuisance level. Throughout this period, bioassays revealed no increase in tolerance of pyrethroids, although selection experiments in the laboratory confirmed the strong pyrethroid-resistance potential of the fly population. Thus effective control can be retained in spite of strong resistance potential when non-persistent insecticides are used intermittently. The results are discussed in the light of a published theoretical study of the influence of pesticide persistence on the evolution of resistance.

The response of pyriproxyfen-resistant and susceptible Bemisia tabaci Genn (Homoptera: Aleyrodidae) to pyriproxyfen and fenoxycarb alone and in combination with piperonyl butoxide

Pyriproxyfen was effective against susceptible Bemisia tabaci eggs at a LC 50 of 0.003 mg litre -1 and against nymphs at 0.02 mg litre -1 . In comparison, eggs of a laboratory selected, pyriproxyfen-resistant B tabaci strain, originating in an Israeli greenhouse, exhibited 6500-fold resistance and nymphs exhibited 1100-fold resistance. Eggs and nymphs of a strain from an Israeli sunflower field exhibited 450 and 210-fold resistance in comparison to the susceptible standard. Fenoxycarb was generally less effective than pyriproxyfen against B tabaci eggs and nymphs but was unaffected by pyriproxyfen resistance. Piperonyl butoxide (PB) was antagonistic to pyriproxyfen, and this increased with increasing pyriproxyfen resistance. PB had no effect on the toxicity of fenoxycarb. Collectively, these data imply that the modes of action of pyriproxyfen and fenoxycarb are distinct, despite the structural similarities of these molecules. Possible reasons for the antagonism of PB against pyriproxyfen are discussed.