Identification of a mutation in the para-sodium channel gene of the cattle tick Rhipicephalus (Boophilus) microplus associated with resistance to synthetic pyrethroid acaricides

Resistance against synthetic pyrethroid (SP) products for the control of cattle ticks in Australia was detected in the field in 1984, within a very short time of commercial introduction. We have identified a mutation in the domain II S4-5 linker of the para-sodium channel that is associated with resistance to SPs in the cattle tick Rhipicephalus (Boophilus) microplus from Australia. The cytosine to adenine mutation at position 190 in the R. microplus sequence AF134216, results in an amino acid substitution from leucine in the susceptible strain to isoleucine in the resistant strain. A similar mutation has been shown to confer SP resistance in the whitefly, Bemisia tabaci, but has not been described previously in ticks. A diagnostic quantitative PCR assay has been developed using allele-specific Taqman® minor groove-binding (MGB) probes. Using the assay to screen field and laboratory populations of ticks showed that homozygote allelic frequencies correlated highly with the survival percentage at the discriminating concentration of cypermethrin

Knockdown Resistance to DDT and Pyrethroids in the House Fly (Diptera: Muscidae): From Genetic Trait to Molecular Mechanism

Knockdown resistance to DDT and pyrethrins was the 1st insecticide resistance trait involving reduced target site sensitivity to be identified and isolated genetically. Almost 5 decades later, knockdown resistance continues to threaten the continued effectiveness of pyrethroid insecticides in the control of numerous agricultural pests and vectors of human disease. In this review we summarize progress in the characterization of knockdown resistance in the house fly, Musca domestica L., culminating in the identification of the specific sodium channel gene mutations that cause the reduced neuronal sensitivity to DDT and pyrethroids in knockdown-resistant insect

Otimização e validação do diagnóstico molecular da resistência a pesticidas piretroides em populações brasileiras da mosca-dos-chifres.

A fim de validar a aplicabilidade de utilização das sequências nucleotídicas diagnósticas identificadas para as populações brasileiras da mosca-dos-chifres, análises fenotípicas da resistência a piretroides foram realizadas para a triagem de espécimes resistentes ao pesticida, os quais posteriormente foram submetidos à análises moleculares a fim de caracterizar a presença das mutações tipo kdr e skdr nas populações brasileiras de H. irritans.bitstream/item/122197/1/CT-130-moscadoschifres.pd

Ion channels as insecticide targets.

Published onlineJournal ArticleIon channels remain the primary target of most of the small molecule insecticides. This review examines how the subunit composition of heterologously expressed receptors determines their insecticide-specific pharmacology and how the pharmacology of expressed receptors differs from those found in the insect nervous system. We find that the insecticide-specific pharmacology of some receptors, like that containing subunits of the Rdl encoded GABA receptor, can be reconstituted with very few of the naturally occurring subunits expressed. In contrast, workers have struggled even to express functional insect nicotinic acetylcholine receptors (nAChRs), and work has therefore often relied upon the expression of vertebrate receptor subunits in their place. We also examine the extent to which insecticide-resistance-associated mutations, such as those in the para encoded voltage-gated sodium channel, can reveal details of insecticide-binding sites and mode of action. In particular, we examine whether mutations are present in the insecticide-binding site and/or at sites that allosterically affect the drug preferred conformation of the receptor. We also discuss the ryanodine receptor as a target for the recently developed diamides. Finally, we examine the lethality of the genes encoding these receptor subunits and discuss how this might determine the degree of conservation of the resistance-associated mutations found

Gain of function mutants: Ion channels and G protein-coupled receptors

Many ion channels and receptors display striking phenotypes for gain-of-function mutations but milder phenotypes for null mutations. Gain of molecular function can have several mechanistic bases: selectivity changes, gating changes including constitutive activation and slowed inactivation, elimination of a subunit that enhances inactivation, decreased drug sensitivity, changes in regulation or trafficking of the channel, or induction of apoptosis. Decreased firing frequency can occur via increased function of K+ or Cl- channels. Channel mutants also cause gain-of-function syndromes at the cellular and circuit level; of these syndromes, the cardiac long-QT syndromes are explained in a more straightforward way than are the epilepsies. G protein-coupled receptors are also affected by activating mutations

Occurrence of target-site resistance to neonicotinoids in the aphid Myzus persicae in Tunisia, and its status on different host plants

This is the peer reviewed version of the following article: Kamel Charaabi, Sonia Boukhris-bouhachem, Mohamed Makni, and Ian Denholm, ‘Occurrence of target‐site resistance to neonicotinoids in the aphid Myzus persicae in Tunisia, and its status on different host plants’, Pest Management Science, Vol. 74(6): 1297-1301, June 2018, which has been published in final form athttps://doi.org/10.1002/ps.4833 Under embargo until 19 December 2018. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving.BACKGROUND: The R81T mutation conferring target-site resistance to neonicotinoid insecticides in Myzus persicae was first detected in France and has since spread across much of southern Europe. In response to recent claims of control failure with neonicotinoids in Tunisia, we have used a molecular assay to investigate the presence and distribution of this target-site mutation in samples collected from six locations and six crops attacked by M. persicae. RESULTS: The resistance allele containing R81T was present at substantial frequencies (32–55%) in aphids collected between 2014 and 2016 from northern Tunisia but was much rarer further south. It occurred in aphids collected from the aphid's primary host (peach) and four secondary crop hosts (potato, pepper, tomato and melon). Its absence in aphids from tobacco highlights complexities in the systematics of M. persicae that require further investigation. CONCLUSION: This first report of R81T from North Africa reflects a continuing expansion of its range around the Mediterranean Basin, although it remains unrecorded elsewhere in the world. Loss of efficacy of neonicotinoids presents a serious threat to the sustainability of aphid control.Peer reviewe

Susceptibility of adult cat fleas (Siphonaptera: Pulicidae) to insecticides and status of insecticide resistance mutations at the Rdl and knockdown resistance loci

This is an Open Access article. © 2015 The Author(s). Published by Springer Berlin Heidelberg.The susceptibility of 12 field-collected isolates and 4 laboratory strains of cat fleas, Ctenocephalides felis was determined by topical application of some of the insecticides used as on-animal therapies to control them. In the tested field-collected flea isolates the LD50 values for fipronil and imidacloprid ranged from 0.09 to 0.35 ng/flea and 0.02 to 0.19 ng/flea, respectively, and were consistent with baseline figures published previously. The extent of variation in response to four pyrethroid insecticides differed between compounds with the LD50 values for deltamethrin ranging from 2.3 to 28.2 ng/flea, etofenprox ranging from 26.7 to 86.7 ng/flea, permethrin ranging from 17.5 to 85.6 ng/flea, and d-phenothrin ranging from 14.5 to 130 ng/flea. A comparison with earlier data for permethrin and deltamethrin implied a level of pyrethroid resistance in all isolates and strains. LD50 values for tetrachlorvinphos ranged from 20.0 to 420.0 ng/flea. The rdl mutation (conferring target-site resistance to cyclodiene insecticides) was present in most field-collected and laboratory strains, but had no discernible effect on responses to fipronil, which acts on the same receptor protein as cyclodienes. The kdr and skdr mutations conferring target-site resistance to pyrethroids but segregated in opposition to one another, precluding the formation of genotypes homozygous for both mutations.Peer reviewedFinal Published versio

Voltage-gated sodium channels as targets for pyrethroid insecticides

The pyrethroid insecticides are a very successful group of compounds that have been used extensively for the control of arthropod pests of agricultural crops and vectors of animal and human disease. Unfortunately, this has led to the development of resistance to the compounds in many species. The mode of action of pyrethroids is known to be via interactions with the voltage-gated sodium channel. Understanding how binding to the channel is affected by amino acid substitutions that give rise to resistance has helped to elucidate the mode of action of the compounds and the molecular basis of their selectivity for insects vs mammals and between insects and other arthropods. Modelling of the channel/pyrethroid interactions, coupled with the ability to express mutant channels in oocytes and study function, has led to knowledge of both how the channels function and potentially how to design novel insecticides with greater species selectivity

The use of botanical synergists to increase the efficacy of natural pyrethrins

Insecticide resistance is an important aspect of pest control on both crop pests and vectors of animal diseases. Resistance can be caused by a number of mechanisms, one of which is enhanced detoxification of the insecticide by metabolic enzymes. Synergists may be used in insecticide formulations to inhibit metabolic defences in the insect, allowing the insecticide to reach its target site and kill the insect, thus enhancing the effect of the insecticide. This PhD project investigated the use of the synergist piperonyl butoxide (PBO) in combination with natural pyrethrins (tank mix) and as a pre-treatment prior to application of pyrethrins, as methods of enhancing the efficacy of the insecticide. The insects studied were Myzus persicae, Bemisia tabaci and Musca domestica. Results showed that the combination treatment (tank mix) was at least as good as, and sometimes better than, the pre-treatment. This is unlike the situation for synthetic pyrethroids where pre-treatments have been shown to be more effective than tank mixes. It is proposed that for natural pyrethrins, PBO aids the penetration of the pyrethrins into the insect, and this enhancement effect is greater than full inhibition of the metabolic enzymes. In some cases, the tank mix enabled less insecticide to be used to achieve 50% mortality in resistant insects, compared to a susceptible population treated with pyrethrins alone. A novel laboratory assay was developed to enable the screening of botanical extracts for their ability to inhibit esterase enzymes. This was used to test a range of compounds and those showing esterase inhibition were also screened for their ability to inhibit cytochrome P450 activity. The competency of some of these compounds as synergists was also tested in vivo with some showing potential activity both in vitro and in vivo

Novel mutations in the voltage-gated sodium channel of pyrethroid-resistant Varroa destructor populations from the Southeastern USA

The parasitic mite Varroa destructor has a significant worldwide impact on bee colony health. In the absence of control measures, parasitized colonies invariably collapse within 3 years. The synthetic pyrethroids tau-fluvalinate and flumethrin have proven very effective at managing this mite within apiaries, but intensive control programs based mainly on one active ingredient have led to many reports of pyrethroid resistance. In Europe, a modification of leucine to valine at position 925 (L925V) of the V. destructor voltage-gated sodium channel was correlated with resistance, the mutation being found at high frequency exclusively in hives with a recent history of pyrethroid treatment. Here, we identify two novel mutations, L925M and L925I, in tau-fluvalinate resistant V. destructor collected at seven sites across Florida and Georgia in the Southeastern region of the USA. Using a multiplexed TaqMan® allelic discrimination assay, these mutations were found to be present in 98% of the mites surviving tau-fluvalinate treatment. The mutations were also found in 45% of the non-treated mites, suggesting a high potential for resistance evolution if selection pressure is applied. The results from a more extensive monitoring programme, using the Taqman® assay described here, would clearly help beekeepers with their decision making as to when to include or exclude pyrethroid control products and thereby facilitate more effective mite management programmes