Acetylcholinesterase inhibition by nootkatone and carvacrol in arthropods

The essential oils from many botanicals have been screened for insecticidal activity. Two constituents of the Alaskan yellow cedar tree, the monoterpenoid carvacrol and the sesquiterpenoid nootkatone, both are toxic against several arthropods. The mode of action through which nootkatone and carvacrol exert their insecticidal activity remains uncertain. It has been hypothesized that they may inhibit acetylcholinesterase enzyme activity. The degree of acetylcholinesterase inhibition of carvacrol and nootkatone was compared to that of carbaryl, a known acetylcholinesterase inhibitor, in the house fly (Musca domestica), yellow fever mosquito (Aedes aegypti), American dog tick (Dermacentor variabilis) and American cockroach (Periplaneta americana). The concentration of carbaryl, at which 50% of the acetylcholinesterase activity was inhibited (IC₅₀), was less than 2 μM in all four arthropod models. Carvacrol was observed to cause slight inhibition of the acetylcholinesterase enzyme in house flies, ticks and cockroaches, but it did not inhibit the mosquito acetylcholinesterase enzyme. Nootkatone did not inhibit the acetylcholinesterase enzyme in any of the four arthropod models tested. From this study, we conclude that the acetylcholinesterase inhibition is not likely the primary mode of action for insecticidal activity by nootkatone or carvacrol.     

Cross-Resistance to Imidacloprid in Strains of German Cockroach (Blattella germanica) and House Fly (Musca domestica)

The toxicity of a promising new insecticide, imidacloprid, was evaluated against several susceptible and resistant strains of German cockroach and house fly. Imidacloprid rapidly immobilized German cockroaches followed by a period of about 72 h during which some cockroaches recovered. After 72 h there was no further recovery. Imidacloprid-treated houseflies were immobilized more slowly than treated cockroaches, with the maximum effect observed after 72 h, and there was no recovery. Based upon 72-h LD₅₀ values imidacloprid was moderately toxic to German cockroaches (LD₅₀ values were 6–8 ng mg^-1) and had only low toxicity to house flies (LD₅₀ 140 ng mg^-1). Piperonyl butoxide (PBO) blocked the observed recovery in German cockroaches. PBO also greatly enhanced the 72-h LD₅₀ of imidacloprid from 43- to 59-fold in cockroaches and 86-fold in house flies. Two strains of German cockroach (Baygon-R and Pyr-R) showed >4-fold cross-resistance to imidacloprid. This cross-resistance could not be suppressed by PBO, suggesting that P450 monooxygenase-mediated detoxication is not responsible for this cross-resistance. Variation in the level of synergism observed with PBO (between strains) suggests the ‘basal’ level of monooxygenase-mediated detoxication of imidacloprid is quite variable between strains of German cockroach. The AVER and LPR strains of house fly showed significant cross-resistance to imidacloprid. PBO reduced the level of cross-resistance in AVER from >4·2-fold to 0·5-fold (i.e. the AVER strain LD₅₀ was half that of the susceptible strain when both were treated with PBO), but PBO did not suppress the cross-resistance in LPR. These data suggest monooxygenases are the mechanism responsible for cross-resistance to imidacloprid in AVER, but not in the LPR strain. © of SCI.     

Actions of formamidines,local anesthetics,octopamine and related compounds upon the electrical activity of neurohaemal organs of the stick insect (Carausius morosus) and sense organs of fly larvae (Musca domestica; Calliphora erythrocephala)

The pesticides, chlordimeform and amitraz, and their metabolites, demethylchlordimeform, N¹-(2,4-dimethylphenyl)-N-methylformamidine, and 2,4-dimethylformanilide, are effective at concentrations as low as 3 μM in raising the firing rate of endogenously active neurosecretory fibres in the isolated neurohaemal organs of Carausius morosus. Molecules such as bunamidine and cetrimide, with cationic detergent properties, produced sporadic bursting which did not elevate the overall firing rate to any great extent. Indeed, bunamidine could induce complete block of action potentials at concentrations as low as 30 μM. The local anesthetics, procaine, lidocaine, and benzocaine, do not induce block of activity at least up to a level of 1 mM. They have no effect at concentrations lower than 100 μM. Between 100 μM and 1 mM lidocaine and benzocaine produce a small increase in firing rate. Procaine produced a pronounced increase in the frequency of firing. The phenolic amines, octopamine, synephrine, and tyramine, markedly increased electrical activity. The catecholamines, dopamine, noradrenaline, and adrenaline, by contrast, only produced a weak excitation. Neither α- nor β-adrenergic blocking agents were effective in antagonizing the electrical activity induced by chlordimeform or phenolic amines until relatively high concentrations of about 1 mM were used. Chlordimeform was able to induce high-frequency bursts from sense organs associated with the epidermis and body-wall musculature in larvae of Musca domestica and Calliphora erythocephala. Octopamine did not induce any similar bursting activity in these sense organs. These results are discussed in relationship to current views on the mode of action of the N-aryl amidines. It is concluded that the excitatory effects of these compounds upon neurohaemal organs and sense organs are more likely to result from a direct action upon voltage sensitive channels of the nerve membranes, rather than by an effect mediated by interactions with octopamine receptors.     

Symptomatic and neurophysiological activities of new synthetic non-ester pyrethroids,ethofenprox, MTI-800, and related compounds

Ethofenprox (MTI-500), MTI-800, and related compounds, which have a m-phenoxybenzyl moiety but lack ester bonding, were compared with DDT-type compounds and pyrethroid insecticides for their lethal and convulsive activities toward American cockroaches. The most potent among them ranked between phenothrin and cyphenothrin. Neurophysiological effects were also examined by extra- and intracellular recording and voltage clamp techniques. Some derivatives induced repetitive discharges in the excised central nerve cord of the American cockroach. The after-potential recorded intracellularly from the crayfish giant axon was markedly increased by some compounds. Voltage clamp experiments with the crayfish giant axon showed that ethofenprox decreased the peak sodium current and induced a large residual current during a step depolarization. It also induced a large and prolonged tail current after a step repolarization of the membrane. The effects of the test compounds on the action potential and the sodium current were similar to those of DDT-type compounds and the pyrethroids such as allethrin and phenothrin. A shift of sodium channel population from normal to modified was thought to result in modifications of the sodium current which, in turn, cause the increase in after-potential and the induction of repetitive discharges.     

Mode of action of insecticidal dihydropyrazoles: Selective block of impulse generation in sensory nerves

When injected into cockroaches (Periplaneta americana), dihydropyrazoles (pyrazolines), e.g. RH 3421, methyl 1-(N-(α,α,α-trifluoro-p-tolyl)carbamoyl-3-(4-chlorophenyl)-4-methyl-2-pyrazolin-4-ylcarboxylate, caused incoordination, followed by prostration and violent tremors. After several hours, prostrate cockroaches became very still, appearing paralyzed, but for several days they retained the ability to produce violent tremors when physically disturbed. The paralysis was associated with block of spontaneous activity in sensory nerves of cockroaches, tobacco hornworm larvae (Manduca sexta) and housefly larvae (Musca domestica). Block of spontaneous central nervous system activity was also seen in some cases. Current-clamp experiments on the cell body of the crayfish slowly-adapting stretch receptor neuron revealed that spike initiation in response to injected current was blocked by dihydropyrazoles, with no change in resting potential or input resistance, suggesting that the sodium current was suppressed. Hyperpolarization of blocked stretch receptors could reverse block, and crayfish giant axons, which were not normally sensitive to block by dihydropyrazoles, were rendered sensitive when artificially depolarized by elevation of extracellular potassium ion concentration, leading to the conclusion that the block of sodium current was strongly voltage dependent. The proposed mode of action, while similar to that of local anesthetics, class I anticonvulsants and class I antiarrhythmics, is novel for an insecticide.     

The role of GABA and glutamate receptors in susceptibility and resistance to chloride channel blocker insecticides

Despite a point mutation in the pore-forming segment of the Rdl GABA receptor subunit that is widespread and persistent in insect populations and confers high levels of resistance to dieldrin and other polychlorocycloalkane (PCCA) insecticides, the phenylpyrazole insecticide fipronil, which binds at same site, has proven to be effective in controlling many insects, including dieldrin-resistant populations. Fipronil and its major sulfone metabolite are unique among chloride channel blocking insecticides in that they also potently block GluCls. We present here a patch clamp study of the action of fipronil sulfone on native GABA receptors and GluCl receptors from susceptible and dieldrin-resistant German cockroaches, to provide a better understanding of the effect of the Rdl mutation on the function and insecticide sensitivity of these two targets, and its role in resistance. Dieldrin blocked GABA currents with an IC₅₀ of 3 nM in wild-type cockroaches, and 383 nM in resistant insects, yielding a resistance ratio of 128. Fipronil sulfone blocked GABA currents with an IC₅₀ of 0.8 nM in susceptible insects and 12.1 nM, or 15-fold higher, in resistant insects. While both GluClD (desensitizing) and GluClN (non-desensitizing) receptors were found in German cockroach neurons, GluClN receptors were rare and could not be included in this study. GluClD receptors from resistant insects had reduced sensitivity to glutamate and a lower rate of desensitization than those from susceptible insects, but their sensitivity to block by fipronil sulfone was not significantly changed, with an IC₅₀ of 38.5 ± 2.4 nM (n = 8) in the susceptible strain and 40.3 ± 1.0 nM (n = 7) in the resistant strain. Fipronil sulfone also slowed the decay time course of GluClD currents. These results suggest that GluClD receptors contain the Rdl subunit, but their sensitivity to fipronil sulfone is not altered in resistant insects.     

The A302S mutation in Rdl that confers resistance to cyclodienes and limited cross-resistance to fipronil is undetectable in field populations of house flies from the USA

Fipronil is a relatively new insecticide with great potential for insect control, however widespread use of cyclodiene insecticides has selected for an A302S mutation in the Rdl (GABA gated chloride channel) allele. This mutation gives resistance to cyclodienes and limited cross-resistance to fipronil. Given the concern over the possible reduction in efficacy and/or lifetime that fipronil might be used for pest control (given the extensive use of cyclodienes in the past), it is important to know the frequency of the A302S Rdl mutation in field populations. To ascertain the relative frequency of the A302S Rdl mutation in house fly populations we used three experimental approaches. First, we attempted to select for fipronil resistance by initially treating 33,100 field collected flies and selecting 14 additional generations. We were unable to produce a highly resistant strain. Second, we directly sequenced field collected flies. Third, we tested field collected house flies with a diagnostic dose of dieldrin and then genotyped the survivors. Out of the 4750 flies tested, there were no Rdl resistance alleles detected. We conclude that the resistant Rdl allele is rare in house flies in the US due to decades without cyclodiene use and a fitness disadvantage (in the absence of cyclodienes) of the 302S Rdl allele. The limited cross-resistance provided by the cyclodiene resistant Rdl allele, combined with the very low frequency of this allele in field populations, suggests that fipronil could be a promising insecticide for house fly control.     

Dihydrooxadiazines: Octopaminergic system as a potential site of insecticidal action

Dihydrooxadiazines are structural analogs of octopamine and were compared with octopamine for their ability to compete with [³H]dihydroergocryptine ([³H]DHE) for binding sites on DHE-sensitive receptors, to stimulate adenylate cyclase activity in nervous system homogenates of Periplaneta americana L., and to modulate the action of the peptide proctolin on the oviducal muscles of Locusta migratoria L. [³H]DHE binding was inhibited by low concentrations (μM range) of octopamine, phentolamine, N-demethylchlordimeform (DCDM) and several dihydrooxadiazines. The tested dihydrooxadiazines acted as aminergic agonists in stimulating cyclic AMP production in cockroach nervous system homogenates and did not show additive effects with octopamine, whereas additivity was observed with 5-hydroxytryptamine. The relative potency of octopaminergic antagonists, including mianserin, cyproheptadine, phentolamine, and gramine, to block octopamine-mediated elevation of cyclic AMP production was similar to the rank-order potency of the same antagonists to inhibit dihydrooxadiazine-mediated elevation of cyclic AMP production. Octopamine, 2-(4-bromophenyl)-5,6-dihydro-4H-1,3,4-oxadiazine (4-Br-PDHO), and 8-Br-cyclic AMP caused increased phosphorylation of proteins that are phosphorylated by exogenously added cyclic AMP-dependent protein kinase. These results indicate that the dihydrooxadiazine-induced rise in cyclic AMP levels in homogenates of the cockroach nervous system results directly in activation of an endogenous cyclic AMP-dependent protein kinase. 4-Br-PDHO behaved similarly to octopamine in modulating the action of proctolin-induced contractions in locust oviducal muscles. These observations suggest that dihydrooxadiazines act as octopamine agonists and have an octopaminergic action in modulating the action of proctolin. Thus, it is proposed that dihydrooxadiazines exert at least part of their insecticidal and miticidal actions through interaction with the octopaminergic system.     

Genetic and biochemical mechanisms limiting fipronil toxicity in the LPR strain of house fly,Musca domestica

Fipronil is a new insecticide which exerts its toxic action by interacting with the insect GABA-gated chloride channel. Previous studies have shown that cyclodiene-resistant insects have low to moderate levels of cross-resistance to fipronil, while other resistant strains are usually susceptible. In contrast, we recently found a strain (LPR) of house fly (Musca domestica L) with 15-fold cross-resistance to fipronil that was not associated with cyclodiene resistance. Fipronil cross-resistance in LPR was inherited as an intermediately dominant, autosomal, multigenic trait. [¹⁴C]Fipronil was observed to penetrate into LPR flies more slowly than into susceptible flies. S,S,S-tributylphosphorotrithioate and diethyl maleate pretreatment did not reduce the level of fipronil cross-resistance, while piperonyl butoxide resulted in a slight decrease. These results indicate that decreased penetration and monooxygenase-mediated detoxification may be mechanisms contributing to fipronil cross-resistance in the LPR strain. © 1999 Society of Chemical Industry     

Insecticidal activity and microsomal metabolism of biooxidizable lindane analogs

The insecticidal activity of lindane analogs, in which some chlorine atoms were replaced by other groups susceptible to microsomal oxidative metabolism, was determined against mosquitos, house flies, and German cockroaches. When tested with a synergist, piperonyl butoxide, one of the methylthio analogs was as active as lindane, whereas several others were also highly active. By examining the ratio of synergized and unsynergized LD₅₀ values (synergistic ratio value), the highly insecticidal methylthio, methoxy, and methyl analogs appear to undergo metabolic detoxication effectively in house flies. By means of in vitro metabolism experiments using microsomal fraction from house fly abdomen, the methoxy, ethoxy, and methylthio analogs were shown to be metabolized rapidly at similar rates. The synergized insecticidal activities of these compounds against various insect species relate linearly with each other, suggesting that the oxidative degradation is inhibited by the synergist to a similar extent and that the transport process to the site of action is not a limiting factor in determining the relative insecticidal activity.     

Silica aerogel and boric acid against cockroaches: External and internal action

American cockroaches, Periplaneta americana (L.), and German cockroaches, Blattella germanica (L.), ingested large quantities of powder (silica aerogel and boric acid) in the process of preening their appendages. Their crops were greatly distended by the powder, but the denticles of the armarium of the proventriculus permitted the movement of only very small quantities of silica aerogel into the remainder of the alimentary tract. Boric acid likewise passed the armarium in very small quantities and apparently only in solution.There was no significant difference in the rate of mortality of both species of cockroaches confined with deposits of silica aerogel SG-68, whether their mouthparts were sealed with paraffin wax or left unsealed. Ingested powder did not increase the rate of water loss. Cockroaches with unsealed mouthparts died more rapidly on deposits of boric acid than those with sealed mouthparts, as might be expected of a toxic compound. However, cockroaches with sealed mouthparts were strikingly affected by boric acid, indicating cuticular penetration by that compound. Penetration within the integument, in the gross body contents, in the alimentary tract, and in the feces was confirmed by a boric acid flame test.     

Unusual features of the toxic action of dimethoate on Periplaneta americana and their causes

Dimethoate and its oxygen analog dimethoxon take more than six times longer than diazoxon (whose speed of action is more typical of organophosphorus insecticides in general) to cause irreversible symptoms of poisoning in cockroaches, although all are equally toxic to the insects if enough time is allowed. Dimethoxon was 200 times less active than diazoxon, both against cockroach cholinesterase and in electrophysiological tests against the central nervous systems of cockroaches. Other tests established that neither restricted penetration into the nervous system nor detoxication within it contributed to the weak neurotoxicity of dimethoxon. This weak neurotoxicity is compensated for by relatively high stability to detoxication, enabling dimethoxon to persist long enough within insects to achieve high overall toxicity and explaining why its overal toxicity is similar to that of diazoxon, which is more rapidly detoxified by the insects.     

The action of formamidines on octopamine receptors in the locust

The actions of a range of formamidines have been investigated biochemically and physiologically on octopamine receptors in the locust Schistocerca gregaria. All the formamidines tested [chlordimeform (CDM), demethylchlordimeform (DCDM), amitraz, and UK 16353] mimicked the action of octopamine by (1) increasing the amplitude and relaxation rate of slow motorneurone tension in the extensor-tibiae muscle of the hind leg and (2) changing the levels of cyclic AMP in this muscle. UK 16353 was most effective in changing these parameters, followed by DCDM then amitraz and CDM. The formamidine-induced increase in cyclic AMP levels was reduced or completely blocked by phentolamine, an antagonist of insect octopamine receptors. The time course for the increase in cyclic AMP was followed for 30 min by incubating muscles in 10^?5M DCDM. The increase was potentiated by the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine. In the presence of this compound, the response peaked within 5 min, before declining to a lower plateau after 12 min. The response to 10^?5M CDM was lower and the maximum increase occurred after 7 min, then rapidly declined. Both formamidines increased cyclic AMP in a dose-dependent way with a threshold of between 5 × 10^?7 and 10^?8M. The results are discused in terms of the relationship between the biochemical and physiological actions of the formamidines in this preparation.     

Acute toxicity of chlorantraniliprole to non‐target crayfish (Procambarus clarkii) associated with rice–crayfish cropping systems

BACKGROUND: Chlorantraniliprole, a novel anthranilic diamide insecticide, was recently introduced into the United States where rice–crayfish crop rotations are practiced to control rice water weevil (Lissorhoptrus oryzophilus Kuschel) infestations. Chlorantraniliprole has high margins of mammalian safety and excellent insecticidal efficacy, but its toxicity to non‐target crayfish is uncertain. In this study, the acute toxicity of chlorantraniliprole to the red swamp crayfish Procambarus clarkii Girard was determined using aquatic and feeding assays. RESULTS: The aquatic 96 h median lethal toxicity (LC₅₀) data indicate that technical‐grade chlorantraniliprole is highly toxic (US EPA category) to crayfish with an LC₅₀ of 951 µg L^?1 (95% CL = 741–1118 µg L^?1). A no observed effect concentration (NOEC) of 480 µg L^?1 was recorded. Neither the 36 day chronic feeding study, where crayfish fed on chlorantraniliprole‐treated rice seed in aquaria, nor the 144 h acute feeding test, where crayfish fed on rice seeds treated with chlorantraniliprole, produced mortality or abnormal behavior. CONCLUSION: Chlorantraniliprole is three orders of magnitude less acutely toxic to P. clarkii than lambda‐cyhalothrin and etofenprox, two pyrethroid insecticides also used in rice, and is less likely to cause acute crayfish toxicity in rice pond ecosystems. Based on acute toxicity data, the use of chlorantraniliprole should be more compatible with rice–crayfish crop rotations than pyrethroids. Copyright © 2010 Society of Chemical Industry     

Quantitative structure-activity studies of substituted benzyl chrysanthemates: 9. Calcium uptake inhibition in crayfish nerve cord and lobster axon homogenates in vitro by sunthetic pyrethroids

Twenty-five synthetic pyrethroids and related chemicals were assessed for their effects on the uptake of Ca²⁺ (as ⁴⁵Ca²⁺) by crude homogenates prepared from crayfish (Procambarus clarkii) nerve cords and 20 were assessed on the uptake by homogenates prepared from lobster (Panulirus japonicus) axons. A parabolic relationship was demonstrated between inhibition of Ca²⁺ uptake for this series of chemicals and lipophilicity (log P) in both species when tested at 5 μM. Optimal log P for maximal inhibition was located at about 6.6 and 84(±6)% inhibition was obtained with resmethrin. Compounds of higher or lower log P were either weaker inhibitors, not inhibitors or occasionally resulted in slightly increased levels of Ca²⁺ uptake. No direct correlations between the potential for the pyrethroids to inhibit Ca²⁺ uptake and the potential for these agents to increase the frequency of spontaneous discharges in crayfish nerve cords, to induce repetitive firing in American cockroaches, or the lethality to cockroaches or to any other neurophysiological or toxicity parameter could be established. It was concluded that although some synthetic pyrethroids are moderately potent inhibitors of Ca²⁺ uptake into nerve cord and axonic preparations (i.e., I₅₀ for trans-resmethrin equals 1 μM) this inhibition alone does not relate to the neurophysiological changes in isolated nerve preparations or to the toxicity of these agents in insects.     

Comparison of symptomatic and neurophysiological activities of enantiomers of the insecticide 3-phenoxybenzyl 1-(4-ethoxyphenyl)-2,2-dichlorocyclopropane-1-carboxylate

The insecticidal activities of optical isomers of 3-phenoxybenzyl 1-(4-ethoxyphenyl)-2,2-dichlorocyclopropanecarboxylate and related compounds were measured with American cockroaches and their knockdown activities were evaluated with house flies. The activities of the S(?)-isomer of the dichlorocyclopropanecarboxylate were higher than those of the R(+)-isomer. The effects of the compounds on the inward membrane currents induced by a stepdepolarizing pulse in crayfish axonal membranes were examined under voltage clamp conditions by the sucrose gap method. The compounds induced a tail current upon step repolarization of the membrane. The tail current decayed to zero in each record, but developed with time after the start of application of the compound until a steady level was reached. The rate of decay of the tail current observed in axonal membranes treated with the S(?)-isomer was slower than with the R(+)-isomer. The rates of development of the tail current induced by the two isomers were not very different.     

DDT and pyrethroids: Receptor binding in relation to knockdown resistance (kdr) in the house fly

The nature of target site or knockdown resistance (kdr) to DDT and pyrethroids was studied by investigating specific binding of [14_C] DDT and [14_C] cis-permethrin to the previously established membrane receptors from the heads of susceptible (sbo) and resistant (kdr) strains of the house fly, Musca domestica L. In vivo studies showed the heads from sbo flies bound two to three times more DDT than those from kdr flies at all doses tested. Reduced binding was also observed in kdr flies in in vitro [14_C] DDT binding assays. Scatchard analysis indicated that kdr flies have the same affinity but fewer receptors per milligram protein in the CNS than sbo flies. Assays with [14_C] cis-permethrin also showed binding was much reduced in kdr flies in comparison with sbo flies. Based on these results, the nature of the target site insensitivity of kdr flies may relate to their having a reduced number of receptors for the insecticides.     

Insecticidal activity of plant-derived extracts against different economically important pest insects

With the aim of selecting potential botanical insecticides, seven plant extracts (Daphne mucronata (Family: Thymelaeaceae), Tagetes minuta (Asteraceae), Calotropis procera (Apocynaceae), Boenninghausenia albiflora (Rutaceae), Eucalyptus sideroxylon (Myrtaceae), Cinnamomum camphora (Lauraceae) and Isodon rugosus (Lamiaceae)) were screened for their toxic effects against four important agricultural pest insects, each representing a separate insect order; pea aphids of Acyrthosiphon pisum (Hemiptera), fruit flies of Drosophila melanogaster (Diptera), red flour beetles of Tribolium castaneum (Coleoptera), and armyworms of Spodoptera exigua (Lepidoptera). Aphids were the most susceptible insect with 100% mortality observed after 24 h for all the plant extracts tested. Further bioassays with lower concentrations of the plant extracts against aphids, revealed the extracts from I. rugosus (LC₅₀ 36 ppm and LC₉₀ 102 ppm) and D. mucronata (LC₅₀ 126 ppm and LC₉₀ 198 ppm) to be the most toxic to aphids. These most active plant extracts were further fractionated into different solvent fractions on polarity basis and their insecticidal activity evaluated. While all the fractions showed considerable mortality in aphids, the most active was the butanol fraction from I. rugosus with an LC₅₀ of 18 ppm and LC₉₀ of 48 ppm. Considering that high mortality was observed in aphids within 24 h of exposure to a very low concentration of the butanol fraction from I. rugosus, we believe this could be exploited and further developed as a potential plant-based insecticide against sucking insect pests, such as aphids.     

Comparative acute toxicity of neonicotinoid and pyrethroid insecticides to non‐target crayfish (Procambarus clarkii) associated with rice–crayfish crop rotations

BACKGROUND: Most insecticides used to control rice water weevil (Lissorhoptrus oryzophilus Kuscel) infestations are pyrethroids. However, pyrethroids are highly toxic to non‐target crayfish associated with rice–crayfish crop rotations. One solution to the near‐exclusive reliance on pyrethroids in a rice–crayfish pest management program is to incorporate neonicotinoid insecticides, which are insect specific and effective against weevils but not extremely toxic to crayfish. This study aimed to take the first step to assess neonicotinoids as alternatives to pyrethroids in rice–crayfish crop rotations by measuring the acute toxicities of three candidate neonicotinoid insecticides, clothianidin, dinotefuran and thiamethoxam, to juvenile Procambarus clarkii (Girard) crayfish and comparing them with the acute toxicities of two currently used pyrethroid insecticides, lambda‐cyhalothrin and etofenprox. RESULTS: Neonicotinoid insecticides are at least 2–3 orders of magnitude less acutely toxic (96 h LC₅₀) than pyrethroids to juvenile Procambarid crayfish: lambda‐cyhalothrin (0.16 µg AI L^?1) = etofenprox (0.29 µg AI L^?1) ? clothianidin (59 µg AI L^?1) > thiamethoxam (967 µg AI L^?1) > dinotefuran (2032 µg AI L^?1). CONCLUSION: Neonicotinoid insecticides appear to be much less hazardous alternatives to pyrethroids in rice–crayfish crop rotations. Further field‐level neonicotinoid acute and chronic toxicity testing with crayfish is needed. Copyright © 2009 Society of Chemical Industry     

Antifeedant effect,biological efficacy and high affinity binding of imidacloprid to acetylcholine receptors in Myzus persicae and Myzus nicotianae

It is known from laboratory studies that tobacco-associated forms of Myzus persicae (Sulzer) and the closely related tobacco aphid Myzus nicotianae (Blackman) are often somewhat less susceptible to imidacloprid than non-tobacco strains of M. persicae. Choice tests (floating leaf technique) showed that tobacco aphids were also less susceptible to the antifeedant potential of imidacloprid in contact bioassays. Synergists like piperonyl butoxide or DEF did not enhance the susceptibility of tobacco-associated morphs of Myzus ssp. to imidacloprid, thus providing evidence that neither oxidative detoxication nor hydrolytic metabolization took place. However, in an attempt to study the influence of endosymbiotic bacteria on the efficacy of imidacloprid, we allowed small populations of tobacco aphids to feed on diets containing the antibiotic chlortetracycline prior to imidacloprid treatment. While the effectiveness of imidacloprid, i.e. lower LC₅₀ values, could be improved in all strains, including the susceptible reference strain, there was no change in overall tolerance factors. In order to investigate any possible alteration of the target site, the affinity of imidacloprid and nicotine to nicotinic acetylcholine receptors in whole-aphid homogenates was measured. All strains (and clones) showed the same high-affinity binding sites and no detectable difference. Studies using the FAO dip method revealed that the lower susceptibility of M. nicotianae is not restricted to chloronicotinyls like imidacloprid or acetamiprid, because other insecticides with different modes of action such as pymetrozine and fipronil were also affected in laboratory studies. It is considered that the observed tolerance to chloronicotinyls in certain strains of Myzus ssp. is a natural variation in response, probably not coupled with any known mechanism of resistance in this species complex. © 1998 SCI