Insecticide Target Research Articles

Enzymes in the juvenile hormone biosynthetic pathway can be potential targets for pest control.

Discovery of novel insecticides and targets has received global attention in recent years. Ten genes coding for enzymes involved in the juvenile hormone biosynthetic pathway of Manduca sexta were studied as potential insecticide targets. We determined the expression of genes encoding some critical enzymes in the JH biosynthetic pathway. Farnesol dehydrogenase (FOLD), Juvenile hormone acid O-methyltransferase (JHAMT) and Juvenile hormone epoxidase (CYP15C1) were selected as the candidate targets based on gene expression results. RNAi silencing and enzyme inhibitor tests were performed to validate whether these candidate genes could be the potential insecticide targets. The down-regulation of FOLD, JHAMT and CYP15C1 resulted in a 68%, 82% and 79% reduction in the rates of JH biosynthesis in vitro, respectively. In addition, RNA interference and inhibitor studies of these enzymes following oral administration demonstrated the potential application in pest management, with respect to high mortality and effects on growth. Based on our study, FOLD, JHAMT and CYP15C1 could be potential targets for pest control as a consequence of their important roles in insect development. © 2019 Society of Chemical Industry.

Crystal structure of diamondback moth ryanodine receptor Repeat34 domain reveals insect-specific phosphorylation sites

BackgroundRyanodine receptor (RyR), a calcium-release channel located in the sarcoplasmic reticulum membrane of muscles, is the target of insecticides used against a wide range of agricultural pests. Mammalian RyRs have been shown to be under the regulatory control of several kinases and phosphatases, but little is known about the regulation of insect RyRs by phosphorylation.ResultsHere we present the crystal structures of wild-type and phospho-mimetic RyR Repeat34 domain containing PKA phosphorylation sites from diamondback moth (DBM), a major lepidopteran pest of cruciferous vegetables. The structure has unique features, not seen in mammalian RyRs, including an additional α-helix near the phosphorylation loop. Using tandem mass spectrometry, we identify several PKA sites clustering in the phosphorylation loop and the newly identified α-helix. Bioinformatics analysis shows that this α-helix is only present in Lepidoptera, suggesting an insect-specific regulation. Interestingly, the specific phosphorylation pattern is temperature-dependent. The thermal stability of the DBM Repeat34 domain is significantly lower than that of the analogous domain in the three mammalian RyR isoforms, indicating a more dynamic domain structure that can be partially unfolded to facilitate the temperature-dependent phosphorylation. Docking the structure into the cryo-electron microscopy model of full-length RyR reveals that the interface between the Repeat34 and neighboring HD1 domain is more conserved than that of the phosphorylation loop region that might be involved in the interaction with SPRY3 domain. We also identify an insect-specific glycerol-binding pocket that could be potentially targeted by novel insecticides to fight the current resistance crisis.ConclusionsThe crystal structures of the DBM Repeat34 domain reveals insect-specific temperature-dependent phosphorylation sites that may regulate insect ryanodine receptor function. It also reveals insect-specific structural features and a potential ligand-binding site that could be targeted in an effort to develop green pesticides with high species-specificity.

Combining genetic crosses and pool targeted DNA-seq for untangling genomic variations associated with resistance to multiple insecticides in the mosquito Aedes aegypti.

In addition to combating vector‐borne diseases, studying the adaptation of mosquitoes to insecticides provides a remarkable example of evolution‐in‐action driving the selection of complex phenotypes. Actually, most resistant mosquito populations show multi‐resistance phenotypes as a consequence of the variety of insecticides employed and of the complexity of selected resistance mechanisms. Such complexity makes the identification of alleles conferring resistance to specific insecticides challenging and prevents the development of molecular assays to track them in the field. Here we showed that combining simple genetic crosses with pool targeted DNA‐seq can enhance the specificity of resistance allele's detection while maintaining experimental work and sequencing effort at reasonable levels. A multi‐resistant population of the mosquito Aedes aegypti was exposed to three distinct insecticides (deltamethrin, bendiocarb and fenitrothion), and survivors to each insecticide were crossed with a susceptible strain to generate three distinct lines. F2 individuals from each line were then segregated based on their survival to two insecticide doses. Hundreds of genes covering all detoxifying enzymes and insecticide targets together with more than 7,000 intergenic regions equally spread over mosquito genome were sequenced from pools of F0 and F2 individuals unexposed or surviving insecticide. Differential coverage analysis identified 39 detoxification enzymes showing an increased gene copy number in association with resistance. Combining an allele frequency filtering approach with a Bayesian F ST‐based genome scan identified multiple genomic regions showing strong selection signatures together with 50 nonsynonymous variations associated with resistance. This study provides a simple and cost‐effective approach to improve the specificity of resistance allele's detection in multi‐resistant populations while reducing false positives frequently arising when comparing populations showing divergent genetic backgrounds. The identification of novel DNA resistance markers opens new opportunities for improving the tracking of insecticide resistance in the field.

Transcriptome-based identification and characterization of genes responding to imidacloprid in Myzus persicae

Myzus persicae is a serious and widespread agricultural pest, against which, imidacloprid remains an effective control measure. However, recent reports indicate that this aphid has evolved and developed resistance to imidacloprid. This study aimed to elucidate the underlying mechanisms and genetic basis of this resistance by conducting comparative transcriptomics studies on both imidacloprid-resistant (IR) and imidacloprid-susceptible (IS) M. persicae. The comparative analysis identified 252 differentially expressed genes (DEGs) among the IR and IS M. persicae transcriptomes. These candidate genes included 160 and 92 genes that were down- and up-regulated, respectively, in the imidacloprid-resistant strain. Using functional classification in the GO and KEGG databases, 187 DEGs were assigned to 303 functional subcategories and 100 DEGs were classified into 45 pathway groups. Moreover, several genes were associated with known insecticide targets, cuticle, metabolic processes, and oxidative phosphorylation. Quantitative real-time PCR of 10 DEGs confirmed the trends observed in the RNA sequencing expression profiles. These findings provide a valuable basis for further investigation into the complicated mechanisms of imidacloprid resistance in M. persicae.

Suppression of Gene Juvenile Hormone Diol Kinase Delays Pupation in Heortia vitessoides Moore.

Juvenile hormone diol kinase (JHDK) is a critical enzyme involved in juvenile hormone degradation in insects. In this study, HvJHDK in the Heortia vitessoides Moore (Lepidoptera: Crambidae) transcriptional library was cloned. Stage-specific expression patterns of HvJHDK, HvJHEH, and HvJHE as well as juvenile hormone titers were determined. The three tested enzymes participated in juvenile hormone degradation. Moreover, juvenile hormone titers peaked after larval–larval molts, consistent with a role for juvenile hormone in inhibition of metamorphosis. HvJHDK was subsequently suppressed using RNA interference (RNAi) to reveal its functions. Different concentrations of dsJHDK elicited the optimal interference efficiency at different life stages of H. vitessoides. Suppression of HvJHDK decreased HvJHDK content and increased the juvenile hormone titer, thereby resulting in reduced triglyceride content, sharply declined survival rate, clearly lethal phenotypes, and extended larval growth. Moreover, suppression of HvJHDK upregulated HvJHEH and HvJHE expression levels, suggesting that there is feedback regulation in the juvenile hormone metabolic pathway. Taken together, our findings provide molecular references for the selection of novel insecticidal targets.

Biochemical and Molecular Analysis of Acetylcholine Esterase to Rule out Organophosphorus Detoxification Level in Field Populations of Culex Quiquefasciatus Say

Background & Objectives: Acetylcholinesterase, responsible for neurotransmitter degradation at the cholinergic nerve synapse, is the target of both organophosphate and carbamate insecticides. The current study envisage to assess acetylcholine esterase levels and mutation in the acetylcholine esterase gene1 of Cx. quinquefasciatus of Kozhikode, Cochin, Malappuram, Thrissur, and Palakkad town areas of Kerala, India. Methods: The samples were collected from regularly insecticide spraying area and scarcely spraying area of the five locations in the year 2014, 15 and 16. The assays were done according to the protocol provided in techniques to detect insecticide resistance mechanism; Field and laboratory Manual. Site specific mutation in acetylcholine esterase gene 1 was identified by RFLP using Alu1 enzyme. Results: The % remaining activity in propoxur inhibited fraction in the regularly and scarcely insecticide treating field populations of Cx.quinquefasciatus were higher than 30% and lesser than 30% respectively. The value greater than 30% indicates the chance to develop resistance in field populations. The RFLP using Alu1 on the ace1 also showed the presence of heterozygous genotype in all the five field populations except in Ernakulam where the Cx.quinquefasciatus population possessed homozygous resistant genotype, which is an indication to the excessive and extensive usage of Temephos as a larvicide. Interpretation & Conclusion: The evidence of development of resistance to synthetic insecticides in mosquitoes observed in the present study points to the need of employing new or alternate insecticides which would be easily degradable and have less harmful effect on other organisms.

Identification of the ryanodine receptor mutation I4743M and its contribution to diamide insecticide resistance in Spodoptera exigua (Lepidoptera: Noctuidae).

Insect ryanodine receptors (RyRs) are the targets of diamide insecticides. Two point mutations G4946E and I4790M (numbering according to Plutella xylostella, PxRyR) in the transmembrane domain of the insect RyRs associated with diamide resistance have so far been identified in three lepidopteran pests, P. xylostella, Tuta absoluta and Chilo suppressalis. In this study, we identified one of the known RyR target site resistance mutations (I4790M) in a field-collected population of Spodoptera exigua. The field-collected WF population of S. exigua exhibited 154 fold resistance to chlorantraniliprole when compared with the susceptible WH-S strain. Sequencing the transmembrane domains of S. exigua RyR (SeRyR) revealed that the resistant WF strain was homozygous for the I4743M mutation (corresponding to I4790M in PxRyR), whereas the G4900E allele (corresponding to G4946E of PxRyR) was not detected. The 4743M allele was introgressed into the susceptible WH-S strain by crossing WF with WH-S, followed by three rounds of backcrossing with WH-S. The introgressed strain 4743M was homozygous for the mutant 4743M allele and shared about 94% of its genetic background with that of the recipient WH-S strain. Compared with WH-S, the near-isogenic 4743M strain showed moderate levels of resistance to chlorantraniliprole (21 fold), cyantraniliprole (25 fold) and flubendiamide (22 fold), suggesting that the I4743M mutation confers medium levels of resistance to all three diamides. Genetic analysis showed diamide resistance in the 4743M strain was inherited as an autosomal and recessive trait. Results from this study have direct implications for the design of appropriate resistance monitoring and management practices to sustainably control S. exigua.

Assessing Impacts of Insecticides on Different Embryonic Stages of the Nontarget Aquatic Insect Cheumatopsyche brevilineata (Trichoptera: Hydropsychidae).

Although the egg of aquatic insects is one of the main life stages that can be exposed to contaminants in water, little is known about the detailed impacts of contaminants on eggs of aquatic insects. The present study aimed to clarify the vulnerable embryonic stages of the caddisfly Cheumatopsyche brevilineata exposed to 2 insecticides, etofenprox and diflubenzuron, which are severely harmful to aquatic insects, and to assess the effects of exposure duration on toxicity of etofenprox to the embryonic stage. Eggs obtained from laboratory culture of the insect were exposed to etofenprox for different periods (2, 4, 6, or 8 d) and at different embryonic stages. In experiments with diflubenzuron, eggs were exposed for 2 d at different embryonic stages. These insecticides did not kill the embryos during exposure, but they inhibited hatching post exposure. Diflubenzuron also induced morphological abnormalities of hatchlings and reduced their survival. The overall median lethal concentration (LC50overall ) values varied significantly from 0.0560 to 5.19 μg/L for etofenprox among exposure durations and among embryonic stages, and from 0.442 to 2.89 μg/L for diflubenzuron between embryonic stages. The toxicity of etofenprox to the embryo was more dependent on the embryonic stage at the time of exposure than on the exposure duration. The vulnerable embryonic stage differed between the insecticides. Etofenprox more strongly inhibited hatching of embryos at later stages, whereas sensitivity to diflubenzuron was higher in the following order: stages E3 to E7 > stages E1, E2, and E8. The different responses of C. brevilineata embryos may be attributable to differences in insecticide mode of action and in functional development of insecticide target sites during embryogenesis. Environ Toxicol Chem 2019;38:1434-1445. © 2019 SETAC.

Can Indirect Herbicide Exposure Modify the Response of the Colorado Potato Beetle to an Organophosphate Insecticide?

Organisms live in complex multivariate environments. In agroecosystems, this complexity is often human-induced as pest individuals can be exposed to many xenobiotics simultaneously. Predicting the effects of multiple stressors can be problematic, as two or more stressors can have interactive effects. Our objective was to investigate whether indirect glyphosate-based herbicide (GBH) exposure of the host plant has interactive effects in combination with an insecticide (azinphos-methyl) on an invasive pest Colorado potato beetle (Leptinotarsa decemlineata Say). We tested the effects of GBH and insecticide on the survival, insecticide target genes expression (acetylcholinesterase genes) and oxidative status biomarkers (glutathione S-transferase [GST], glucose-6-phosphate dehydrogenase [G6PDH], glutathione reductase homolog [GR], glutathione peroxidase homolog [GPx], total glutathione [totGSH], glutathione reduced-oxidized [GSH: GSSG], catalase [CAT], superoxide dismutase [SOD], lipid hydroperoxides). We found that exposure to indirect GBH has no single or interactive effects in combination with the insecticide on larval survival. However, prior exposure to GBH inhibits Ldace1 gene expression by 0.55-fold, which is the target site for the organophosphate and carbamate insecticides. This difference disappears when individuals are exposed to both GBH and insecticide, suggesting an antagonistic effect. On the other hand, oxidative status biomarker scores (PCAs of GPx, GR, and CAT) were decreased when exposed to both stressors, indicating a synergistic effect. Overall, we found that indirect GBH exposure can have both antagonistic and synergistic effects in combination with an insecticide, which should be considered when aiming for an ecologically relevant risk assessment of multiple human-induced stressors.

Investigasi resistensi Anopheles sp. terhadap insektisida piretroid dan kemungkinan terjadinya mutasi gen voltage gated sodium channel (VGSC)

Penggunaan insektisida golongan piretroid secara luas dan terus menerus dalam pencegahan penularan malaria dapat menimbulkan mutasi gen voltage gate sodium channel (VGSC) pada vektor nyamuk yang dikenal dengan mutasi knockdown resistance (kdr). Mutasi gen ini membuat sensitifitas ikatan protein VGSC dengan insektisida menurun sehingga menyebabkan resistensi. Timbulnya resistensi insektisida piretroid pada nyamuk vektor dapat menjadi penghambat keberhasilan pemutusan penularan penyakit malaria sehingga deteksi adanya mutasi mutlak diperlukan. Tujuan penelitian ini untuk mengetahui status resistensi insektisida piretroid dan mengidentifikasi mutasi pada gen VGSC kodon 1014 penanda resistensi nyamuk Anopheles sp. yang menjadi vektor malaria di Provinsi Sumatera Selatan. Sampel diambil di tiga Kabupaten, yaitu Muara Enim, Ogan Komering Ulu (OKU), dan Lahat. Uji awal kerentanan terhadap insektisida piretroid (permetrin 0,75%) dilakukan sesuai standar WHO tahun 2016 pada spesies dominan, yaitu Anopheles vagus Dönitz. Selanjutnya, identifikasi mutasi gen VGSC pada kodon 1014 dilakukan pada An. vagus dan spesies Anopheles barbirostris van der Wulp yang tertangkap, dengan teknik seminested-PCR dan dilanjutkan dengan analisis sekuensing. Hasil uji kerentanan Anopheles sp. di Kabupaten Muara Enim menunjukkan resisten terhadap piretroid, sedangkan di Kabupaten Lahat dan OKU masih rentan terhadap insektisida piretroid. Sementara, analisis sekuensing PCR menunjukkan tidak terjadi perubahan basa alel kdr pada target site insektisida gen VGSC sehingga dapat disimpulkan secara molekuler belum ditemukan adanya mutasi pada gen VGSC kdr allel penanda resistensi insektisida piretroid.

Biophysical study on the interaction of cartap hydrochloride and hemoglobin: Heme degradation and functional changes of protein.

Cartap hydrochloride is a mildly perilous insecticide known as "Padan" which is used largely in agricultural farms to control weevil and caterpillars. The over use of cartap causes harmful effects on human health. Since the blood may acts as a target and carrier for insecticides, the effect of these compounds on blood in mammalian toxicology is very important. Hemoglobin is a tetramer protein that play critical role in oxygen transport. The aim of this study was to analyze and compare the function and structural changes of hemoglobin in the presence of different concentrations of cartap by employing different spectroscopic techniques. The obtained results show that cartap has a high hemolytic effect which is increased with cartap concentration and reduces the thermal midpoint of hemoglobin. Fluorescence measurements reveal heme degradation at different concentrations of cartap. In consequence of theoretical and experimental results, cartap has an undesirable effect on hemoglobin structure and function.

Functional Coupling of K+-Cl- Cotransporter (KCC) to GABA-Gated Cl- Channels in the Central Nervous System of Drosophila melanogaster Leads to Altered Drug Sensitivities.

GABAergic signaling is the cornerstone for fast synaptic inhibition of neural signaling in arthropods and mammals and is the molecular target for insecticides and pharmaceuticals, respectively. The K+-Cl- cotransporter (KCC) is the primary mechanism by which mature neurons maintain low intracellular Cl- concentration, yet the fundamental physiology, comparative physiology, and toxicological relevance of insect KCC is understudied. Considering this, we employed electrophysiological, genetic, and pharmacological methods to characterize the physiological underpinnings of KCC function to the Drosophila CNS. Our data show that genetic ablation or pharmacological inhibition of KCC results in an increased spike discharge frequency and significantly ( P < 0.05) reduces the CNS sensitivity to γ-aminobutyric acid (GABA). Further, simultaneous inhibition of KCC and ligand-gated chloride channel (LGCC) complex results in a significant ( P < 0.001) increase in CNS spontaneous activity over baseline firing rates that supports functional coupling of KCC to LGCC function. Interestingly, 75% reduction in KCC mRNA did not alter basal neurotransmission levels indicating that only a fraction of the KCC population is required to maintain the Cl- ionic gradient when at rest, but prolonged synaptic activity increases the threshold for GABA-mediated inhibition and reduces nerve sensitivity to GABA. These data expand current knowledge regarding the physiological role of KCC in a model insect and provides the necessary foundation to develop KCC as a novel biochemical target of insecticides, as well as complements existing research to provide a holistic understanding of the plasticity in mammalian health and disease.

The evolution of multiple-insecticide resistance in UK populations of tomato leafminer, Tuta absoluta.

The tomato leafminer, Tuta absoluta, is an economically important pest of tomatoes in Europe, Africa, Asia and South America. In the UK this species is controlled using an integrated pest management (IPM) programme which incorporates the insecticides spinosad and chlorantraniliprole. In response to UK grower concerns of loss of efficacy of these compounds at certain sites, insecticide bioassays were performed on five populations collected from four commercial glasshouses and potential mechanisms of resistance investigated. We observed high levels of resistance to spinosad in four of the strains, and in two of these tolerance to chlorantraniliprole. Selection of one of these strains with chlorantraniliprole rapidly resulted in a line exhibiting potent resistance to this compound. Sequencing of messenger RNA encoding the nicotinic acetylcholine receptor (nAChR) α6 subunit, target of spinosad, revealed Taα6 transcripts in the spinosad-resistant strains that lack exon 4 and encode a highly truncated protein, or contain a triplet deletion in the predicted first transmembrane domain resulting in the loss of a highly conserved amino acid. Sequencing of the ryanodine receptor gene, encoding the target of diamide insecticides, of the chlorantraniliprole-selected line revealed an amino acid substitution (G4903V) that has been previously linked to diamide resistance in populations of T. absoluta in the Mediterranean and South America. Taken together our results reveal emerging resistance in UK populations of T. absoluta to two of the most important insecticides used as part of IPM, with significant implications for the control of this species in the UK. © 2019 Society of Chemical Industry.

Nanoencapsulated deltamethrin as synergistic agent potentiates insecticide effect of indoxacarb through an unusual neuronal calcium-dependent mechanism

The use of neurotoxic chemical insecticides has led to consequences against the environment, insect resistances and side-effects on non-target organisms. In this context, we developed a novel strategy to optimize insecticide efficacy while reducing doses. It is based on nanoencapsulation of a pyrethroid insecticide, deltamethrin, used as synergistic agent, combined with a non-encapsulated oxadiazine (indoxacarb). In this case, the synergistic agent is used to increase insecticide efficacy by activation of calcium-dependant intracellular signaling pathways involved in the regulation of the membrane target of insecticides. In contrast to permethrin (pyrethroid type I), we report that deltamethrin (pyrethroid type II) produces an increase in intracellular calcium concentration in insect neurons through the reverse Na/Ca exchanger. The resulting intracellular calcium rise rendered voltage-gated sodium channels more sensitive to lower concentration of the indoxacarb metabolite DCJW. Based on these findings, in vivo studies were performed on the cockroach Periplaneta americana and mortality rates were measured at 24 h, 48 h and 72 h after treatments. Comparative studies of the toxicity between indoxacarb alone and indoxacarb combined with deltamethrin or nanoencapsulated deltamethrin (LNC-deltamethrin), indicated that LNC-deltamethrin potentiated the effect of indoxacarb. We also demonstrated that nanoencapsulation protected deltamethrin from esterase-induced enzymatic degradation and led to optimize indoxacarb efficacy while reducing doses. Moreover, our results clearly showed the benefit of using LNC-deltamethrin rather than piperonyl butoxide and deltamethrin in combination commonly used in formulation. This innovative strategy offers promise for increasing insecticide efficacy while reducing both doses and side effects on non-target organisms.

Chemical inhibition of Kir channels reduces salivary secretions and phloem feeding of the cotton aphid, Aphis gossypii (Glover).

The unique feeding biology of aphids suggests novel insecticide targets are likely to exist outside of the nervous system. We therefore aimed to directly test the hypothesis that pharmacological inhibition of inward rectifier potassium (Kir) channels would result in salivary gland failure and reduced sap ingestion by the cotton aphid, Aphis gossypii. The Kir inhibitors VU041 and VU590 reduced the length of the salivary sheath in a concentration dependent manner, indicating that the secretory activity of the salivary gland is reduced by Kir inhibition. Next, we employed the electrical penetration graph (EPG) technique to measure the impact Kir inhibition has to aphid sap feeding and feeding biology. Data show that foliar application of VU041 eliminated the E1 and E2 phases (phloem feeding) in all aphids studied. Contact exposure to VU041 after foliar applications was found to be toxic to A. gossypii at 72 and 96 h post-infestation, indicating mortality is likely a result of starvation and not acute toxicity. Furthermore, VU041 exposure significantly altered the feeding behavior of aphids, which is toxicologically relevant for plant-virus interactions. These data suggest Kir channels are critical for proper function of aphid salivary glands and the reduced plant feeding justifies future work in developing salivary gland Kir channels as novel mechanism aphicides. Furthermore, products like VU041 would add to a very minor arsenal of compounds that simultaneously reduce vector abundance and alter feeding behavior. © 2019 Society of Chemical Industry.

Spatiotemporally different expression of alternatively spliced GABA receptor subunit transcripts in the housefly Musca domestica.

Insect γ-aminobutyric acid (GABA) receptors are important as major inhibitory neurotransmitter receptors and targets for insecticides. The housefly GABA receptor subunit gene MdRdl is alternatively spliced at exons 3 (a or b) and 6 (c or d) to yield the variants of ac, ad, bc, and bd combinations. In the present study, the expression of the MdRdl transcript in the body parts and in the developmental stages of the housefly Musca domestica was examined by quantitative polymerase chain reaction using specific primers that amplify the combinations of alternative exons. The results indicated that the transcripts of MdRdl, including four combinations, were highly expressed in the adult stage. MdRdlbd was the most abundant in the adult head. The expression pattern did not change in the adult stage over 7 days after eclosion. The expression level of the MdRdl bd transcript in the female head was similar to that of the male head. In contrast, MdRdl bc was the predominant transcript in the pupal head and the adult leg. Because the homomeric Rdl bc GABA receptor has a high affinity for GABA, our results provide grounds for designing agonist or competitive-antagonist insecticides that target the orthosteric site of the GABA receptor containing this Rdl variant.

Insecticide Resistance in Drosophila melanogaster (Diptera: Drosophilidae) is Associated with Field Control Failure of Sour Rot Disease in a New York Vineyard.

Sour rot is a complex disease of grapes caused by an interaction of yeast, acetic acid bacteria, and Drosophila spp. Application of insecticides (most commonly zeta-cypermethrin) targeting Drosophila has previously provided substantial control of sour rot in wine grapes of New York vineyards. In harvest season of 2018, a control failure of sour rot and high populations of Drosophila, mostly Drosophila melanogaster, were observed in a vineyard in the Finger Lakes region, NY, despite repeated applications of zeta-cypermethrin (Mustang Maxx). To determine if resistance was responsible for the control failure, we quantified the toxicity of zeta-cypermethrin and the four other insecticides registered for Drosophila control in NY vineyards. Diagnostic concentrations (susceptible strain LC95, 4 × LC95, and 16 × LC95) were used to evaluate percentage survival of the field flies relative to the susceptible Canton-S strain. Resistance to zeta-cypermethrin, acetamiprid, and malathion, but not to spinosad and spinetoram, was observed in the field-collected flies. This study provides evidence that insecticide resistance of Drosophila is associated with control failure of sour rot in some vineyards, and directly influencing grape production. The implications of these results to insecticide resistance monitoring and management are discussed.

Toxicity of the insecticides spinosad and indoxacarb to the non-target aquatic midge Chironomus riparius

Spinosad and indoxacarb are two relatively new insecticides mainly used in agriculture to control insect pests. However, at their current application rates, non-target aquatic insect species may also be impacted. In this study, larvae of the non-biting midge Chironomus riparius were exposed in the laboratory to both insecticides and their effects evaluated at the organismal level, using standard ecotoxicological tests, and at the biochemical level, by monitoring specific oxidative stress, neuronal, and energy metabolism biomarkers. Chronic exposure to both insecticides compromised growth and emergence of C. riparius. Short-term exposures revealed alterations at biochemical level that might be related to the toxicological targets of both insecticides. Growth and development time were the most sensitive endpoints at individual level for both pesticides, while at the biochemical level, the electron transport system activity was the most sensitive biomarker for spinosad exposure, suggesting an increase in energy demands associated with the activation of defense mechanisms. Glutathione-S-transferase was the most sensitive biomarker for indoxacarb exposure, underlining the role of this enzyme in the detoxification of indoxacarb. Additionally, changes in lactate dehydrogenase and glutathione peroxidase activities were observed for both insecticides, and evidences of oxidative damage were found for spinosad. This study contributes to the growing knowledge on sublethal effects of novel insecticides on non-target aquatic invertebrates and strengthens the usefulness of biochemical biomarkers to support the interpretation of their potentially deleterious effects on aquatic insects near agricultural fields.

Molecular cloning, characterization and functional analysis of GluCl from the oriental armyworm, Mythimna separata Walker

Glutamate-gated chloride channels (GluCls) mediate inhibitory synaptic transmission in invertebrate nervous systems, and only one GluCl gene has been found in insects. Therefore, insect GluCls are one of the major targets of insecticides including avermectins. In the present study, a 1347 bp full-length cDNA encoding a 449-amino acid protein (named MsGluCl, GenBank ID: MK336885) was cloned from the oriental armyworm, Mythimna separata, and characterized two alternative splicing variants of MsGluCl. The protein shares 76.9–98.6% identity with other insect GluCl isoforms. Spatial and temporal expression analysis revealed that MsGluCl was highly expressed in the 3rd instar and adult head. Dietary ingestion of dsMsGluCl significantly reduced the mRNA level of MsGluCl and decreased abamectin mortality. Thus, our results reveal that MsGluCl could be the molecular target of abamectin and provide the basis for further understanding the resistance mechanism to abamectin in arthropods.

Bioinformatics of New Selective Insecticide Target in Insect Species

Bioinformatics of New Selective Insecticide Target in Insect Species