Ti3C2 MXene/MoS2@AuNPs ternary nanocomposite for highly sensitive electrochemical detection of phoxim residues in fruits.

Phoxim, extensively utilized in agriculture as an organothiophosphate insecticide, has the potential to cause neurotoxicity and pose human health hazards. In this study, an electrochemical enzyme biosensor based on Ti3C2 MXene/MoS2@AuNPs/AChE was constructed for the sensitive detection of phoxim. The two-dimensional multilayer structure of Ti3C2 MXene provides a robust framework for MoS2, leading to an expansion of the specific surface area and effectively preventing re-stacking of Ti3C2 MXene. Additionally, the synergistic effect of self-reduced grown AuNPs with MoS2 further improves the electrical conductivity of the composites, while the robust framework provides a favorable microenvironment for immobilization of enzyme molecules. Ti3C2 MXene/MoS2@AuNPs electrochemical enzyme sensor showed a significant response to phoxim in the range of 1 × 10-13 M to 1 × 10-7 M with a detection limit of 5.29 × 10-15 M. Moreover, the sensor demonstrated excellent repeatability, reproducibility, and stability, thereby showing its promising potential for real sample detection.

Improving Sphenophorus levis Adult Mortality Through Solid Insecticide Applications and Increased Insecticide Dose.

The sugarcane weevil (Sphenophorus levis Vaurie, 1978) is currently considered the most important sugarcane pest in Brazil, causing significant yield losses. Application methods of insecticides for S. levis control have not been effective, mostly due to the insect's cryptic behavior below the soil surface which suppresses the correct placement of insecticide active ingredients on target. Two experiments were conducted using an innovative bioassay methodology that simulates sugarcane field conditions to effectively evaluate S. levis adult mortality and insecticide residues in the soil under different treatments. The first experiment aimed to assess the efficacy of two liquid- and solid-applied insecticides, while the second aimed to examine the effect of increasing the dose of lambda-cyhalothrin + thiamethoxam on S. levis adult control. The novel bioassays simulated liquid and solid insecticide applications on sugarcane by exposing S. levis adults to chemical residuals on rhizomes and in soil after insecticide application. In the first experiment, low S. levis adult control was detected (< 53% mortality) across all treatments, where both solid and liquid applications of lambda-cyhalothrin + thiamethoxam provided greater efficacy levels than imidacloprid and control treatments, respectively. Solid applications maintained higher insecticide concentrations in the soil for longer periods than liquid insecticide applications, providing maximum insect control levels 7 days after application. The second experiment revealed that solid applications at higher insecticide doses significantly improved control of S. levis adult (76.7% mortality) and resulted in greater insecticide concentrations in the soil compared to the recommended label rate (58.8% mortality).

No impact of cyantraniliprole on the hibernation success of bumble bees (Bombus terrestris audax) in a soil-mediated laboratory exposure study.

Increasing evidence shows that wild bees, including bumble bees, are in decline due to a range of stressors, including pesticides. Our knowledge of pesticide impacts has consequently grown to enable the design of increasingly realistic risk assessment methods. However, one area where knowledge gaps may still hinder our ability to assess the full range of bee-pesticide interactions is the field of exposure. Exposure has historically been linked to either direct contact with pesticides or the ingestion of contaminated pollen and nectar by bees. However, bumble bees, and other wild bees, may also be exposed to pesticides while using contaminated soil as an overwintering substrate. Yet knowledge of how soil-mediated exposure affects bumble bee health is lacking. Here we take one of the first steps towards addressing this knowledge gap by designing a method for testing the effects of soil-mediated pesticide exposure on bumble bee queen hibernation success. We measured hibernation survival, body weight change and abdominal fat content and found that none of these responses were affected by a field realistic soil exposure to the novel insecticide cyantraniliprole. Our study may help in developing a standardised method to test the effects of the soil-mediated pesticide exposure route in bumble bee queens.

Characterization of ATP-binding cassette transporters associated with emamectin benzoate tolerance: from the model insect Drosophila melanogaster to the agricultural pest Spodoptera frugiperda.

BACKGROUND: Multiple families of detoxification genes, including the increasingly recognized family of ATP-binding cassette (ABC) transporters, work together to influence the toxicity of synthetic insecticides and thus their resistance. Effective management of insecticide resistance requires identification of all toxicity-affecting members from each family of toxicity-related genes. RESULTS: Here, we used emamectin benzoate (EB), ABC transporters and Spodoptera frugiperda as a working case to test whether the strategy of 'from the model insect Drosophila melanogaster to agricultural pests' can identify all or most ABC transporter members related to EB tolerance in S. frugiperda. After confirming the involvement of ABC transporters in the toxicity of EB against fruit fly with the ABC inhibitor verapamil, four ABC transporter genes (DmCG3327, DmCG11147, DmCG4822, and DmCG7627) were found to be involved in EB tolerance using RNA interference-based family-wide functional screening. A combination of phylogenic analysis and a reciprocal TBLASTN search identified five S. frugiperda ABC transporter members as homologs (SfABCC4, SfABCG1, and SfABCG23) or one-way best hits (SfABCG4 and SfABCG20) of the four fly ABC genes. Real-time quantitative polymerase chain reaction (qPCR) analysis found that all five S. frugiperda ABC transporter genes were inducible by EB, and expressed in all the developmental stages and larval tissues, but with significant quantitative differences among stages and tissues. A cytotoxicity assay of ABC-overexpressing Sf9 cell lines showed that all the five S. frugiperda ABC transporter genes made Sf9 cells tolerant to EB. CONCLUSIONS: This study not only identifies nine ABC transporter genes related to EB tolerance from D. melanogaster (four genes) and S. frugiperda (five genes), but also demonstrates the utility and effectiveness of the 'model to pests' strategy to identify most toxicity-affecting members from a given family of toxicity-related genes. © 2024 Society of Chemical Industry.

Leveraging long-lasting insecticide-incorporated netting to improve fumigation efficacy against stored product insects.

BACKGROUND: Long-lasting insecticide-incorporated netting (LLIN) has successfully been used to impair mobility and prevent infestation of stored grain by stored product beetles. Understanding how to integrate LLIN with existing integrated pest management (IPM) tactics, such as phosphine fumigation, can further enhance IPM programs. RESULTS: We used three 110 metric tons (MT) capacity grain bins, and in each, 60 perforated buckets (e.g., miniature silos) were filled with 500 g of uninfested wheat. Miniature silos were protected by LLIN (0.3% α-cypermethrin, Carifend®, BASF), positive control (without insecticide), or negative control (no netting). Half of each treatment was randomly assigned to phosphine fumigation treatment, while the remainder were not fumigated. Monthly samples of 100 g of grain from four silos from each treatment in four blocks from three-grain bins were taken between June and October both in 2022 and 2023. We determined whether phosphine fumigation could be reduced with the use of LLIN over the season. Overall, we found that silos protected with LLIN showed insect dispersal and progeny production that was reduced by 83-99% and 89-99%, respectively, compared with insecticide-free netting and no-netting controls. Additionally, damage in silos was reduced by 37-99% compared with controls. Importantly, the total number of fumigations could be reduced by 68-91% by using LLIN compared with controls. CONCLUSION: Our study demonstrates that LLIN is consistently effective for existing pest management tactics such as phosphine fumigation in bulk storage structures. © 2024 Society of Chemical Industry. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.

Cryptic population structure and insecticide resistance in Anopheles gambiae from the southern Democratic Republic of Congo.

The Democratic Republic of Congo (DRC) suffers from one of the highest malaria burdens worldwide, but information on its Anopheles vector populations is relatively limited. Preventative malaria control in DRC is reliant on pyrethroid-treated nets, raising concerns over the potential impacts of insecticide resistance. We sampled Anopheles gambiae from three geographically distinct populations (Kimpese, Kapolowe and Mikalayi) in southern DRC, collecting from three sub-sites per population and characterising mosquito collections from each for resistance to pyrethroids using WHO tube bioassays. Resistance to each of three different pyrethroids was generally high in An. gambiae with < 92% mortality in all tests, but varied between collections, with mosquitoes from Kimpese being the most resistant. Whole genome sequencing of 165 An. gambiae revealed evidence for genetic differentiation between Kimpese and Kapolowe/Mikalayi, but not between the latter two sample sites despite separation of approximately 800 km. Surprisingly, there was evidence of population structure at a small spatial scale between collection subsites in Kimpese, despite separation of just tens of kilometres. Intra-population (H12) and inter-population (FST) genome scans identified multiple peaks corresponding to genes associated with insecticide resistance such as the voltage gated sodium channel (Vgsc) target site on chromosome 2L, a Cyp6 cytochrome P450 cluster on chromosome arm 2R, and the Cyp9k1 P450 gene on chromosome X. In addition, in the Kimpese subsites, the P450 redox partner gene Cpr showed evidence for contemporary selection (H12) and population differentiation (FST) meriting further exploration as a potential resistance associated marker.

Whole genome sequence analysis of population structure and insecticide resistance markers in Anopheles melas from the Bijagós Archipelago, Guinea-Bissau.

BACKGROUND: Anopheles melas is an understudied malaria vector with a potential role in malaria transmission on the Bijagós Archipelago of Guinea-Bissau. This study presents the first whole-genome sequencing and population genetic analysis for this species from the Bijagós. To our knowledge, this also represents the largest population genetic analysis using WGS data from non-pooled An. melas mosquitoes. METHODS: WGS was conducted for 30 individual An. melas collected during the peak malaria transmission season in 2019 from six different islands on the Bijagós Archipelago. Bioinformatics tools were used to investigate the population structure and prevalence of insecticide resistance markers in this mosquito population. RESULTS: Insecticide resistance mutations associated with pyrethroid resistance in Anopheles gambiae s.s. from the Bijagós were absent in the An. melas population, and no signatures of selective sweeps were identified in insecticide resistance-associated genes. Analysis of structural variants identified a large duplication encompassing the cytochrome-P450 gene cyp9k1. Phylogenetic analysis using publicly available mitochondrial genomes indicated that An. melas from the Bijagós split into two phylogenetic groups because of differentiation on the mitochondrial genome attributed to the cytochrome C oxidase subunits COX I and COX II and the NADH dehydrogenase subunits 1, 4, 4L and 5. CONCLUSIONS: This study identified an absence of insecticide-resistant SNPs common to An. gambiae in the An. melas population, but did identify structural variation over insecticide resistance-associated genes. Furthermore, this study presents novel insights into the population structure of this malaria vector using WGS analysis. Additional studies are required to further understand the role of this vector in malaria transmission.

Emamectin Poisoning in Thailand: Clinical Characteristics and Outcomes.

Emamectin benzoate (Emamectin) is a broad-spectrum insecticide. Current data regarding emamectin poisoning in humans are very limited. We performed a 10-year retrospective cross-sectional study (2011-2020) using data from the Ramathibodi Poison Center database to examine the clinical characteristics and outcomes in patients exposed to emamectin. Eighty-eight patients were included. Most of the patients were male (72.7%) and exposure was frequently oral (86.4%) and intentional (67.0%).Their mean age was 42.8 years. The clinical presentations included gastrointestinal tract symptoms (62.5%), neurological symptoms (27.3%) including seizures, respiratory symptoms (6.8%), and local effects (12.5%). At presentation, the majority of patients exhibited normal consciousness and vital signs. Eleven patients showed no obvious clinical effects. Initially, 15 patients had metabolic acidosis and 11 had hypokalemia. Overall, 46 and 52 patients were administered gastric lavage and activated charcoal, respectively. Most patients (78.4%) were hospitalized, with a median hospital stay of 40 h, and generally received supportive treatment. Eight patients were intubated for ventilator support and one received inotropic drugs. Most patients (90.9%) showed no or minor outcomes; however, two patients died. The presence of Glasgow Coma Scale (GCS) <15 differed significantly (p < 0.001) between patients with no or minor outcomes (n = 80) and those with moderate or fatal outcomes (n = 8). In conclusion, emamectin poisoning mainly caused no or minor clinical effects. A low GCS at presentation was associated with worse outcomes. Therefore, patients who present with low GCS should be closely observed, monitored, and properly managed during hospitalization.

Host Tropism and Structural Biology of ABC Toxin Complexes.

ABC toxin complexes are a class of protein toxin translocases comprised of a multimeric assembly of protein subunits. Each subunit displays a unique composition, contributing to the formation of a syringe-like nano-machine with natural cargo carrying, targeting, and translocation capabilities. Many of these toxins are insecticidal, drawing increasing interest in agriculture for use as biological pesticides. The A subunit (TcA) is the largest subunit of the complex and contains domains associated with membrane permeation and targeting. The B and C subunits, TcB and TcC, respectively, package into a cocoon-like structure that contains a toxic peptide and are coupled to TcA to form a continuous channel upon final assembly. In this review, we outline the current understanding and gaps in the knowledge pertaining to ABC toxins, highlighting seven published structures of TcAs and how these structures have led to a better understanding of the mechanism of host tropism and toxin translocation. We also highlight similarities and differences between homologues that contribute to variations in host specificity and conformational change. Lastly, we review the biotechnological potential of ABC toxins as both pesticides and cargo-carrying shuttles that enable the transport of peptides into cells.

Knocking Out of UDP-Glycosyltransferase Gene UGT2B10 via CRISPR/Cas9 in Helicoverpa armigera Reveals Its Function in Detoxification of Insecticides.

The role of insect UDP-glycosyltransferases (UGTs) in the detoxification of insecticides has rarely been reported. A UGT gene UGT2B10 was previously found overexpressed in a fenvalerate-resistant strain of Helicoverpa armigera. Herein, UGT2B10 was cloned, and its involvement in insecticide detoxification was investigated. UGT2B10 was highly expressed in the larvae, mainly in the fat body and midgut. Treatment with UGT inhibitors 5-nitrouracil and sulfinpyrazone significantly enhanced the fenvalerate toxicity. Knocking down UGT2B10 by RNAi significantly increased the larvae mortality by 17.89%. UGT2B10 was further knocked out by CRISPR/Cas9, and a homozygous strain (HD-dUGT2B10) with a C-base deletion at exon 2 was obtained. The sensitivity of HD-dUGT2B10 to fenvalerate, deltamethrin, cyantraniliprole, acetamiprid, and lufenuron increased significantly, with sensitivity index increased 2.523-, 2.544-, 2.250-, 2.473-, and 3.556-fold, respectively. These results suggested that UGT2B10 was involved in the detoxification of H. armigera to insecticides mentioned above, shedding light upon further understanding of the detoxification mechanisms of insecticides by insect UGTs.

Assessment of sub-lethal effects of Celcron on Java barb through erythrocyte morphology and acetylcholinesterase activity: Implications for environmental health in aquatic ecosystems.

Industrialization and the extensive use of chemicals have raised significant concerns about their environmental impacts, particularly on aquatic ecosystems. This study evaluated the sub-lethal effects of Celcron (Cec), an organophosphate insecticide, on the Java barb (Barbonymus gonionotus) through erythrocyte morphology and acetylcholinesterase (AChE) activity, aiming to refine biomarkers for environmental health assessments. We hypothesized that sub-lethal Cec exposure would induce significant erythrocyte abnormalities and decrease AChE activity in Java barb, with variable recovery rates between gill and kidney tissues. To test this, we exposed the juvenile Java barbs to two sub-lethal Cec concentrations - 0.01 ppm (10 % of the LC50) and 0.05 ppm (50 % of the LC50) -for 60 days. After the exposure period, the fish were placed in pesticide-free water to allow for recovery. Results indicated a significant decline in AChE activity in both liver and kidney tissues, with activity levels showing gradual recovery over time. Erythrocyte abnormalities, including nuclear and cellular changes, were significantly elevated in response to Cec exposure. The frequency of nuclear abnormalities such as micronuclei and binucleation increased in a concentration- and duration-dependent manner, with the gill blood exhibiting higher sensitivity and slower recovery compared to kidney blood. Cellular abnormalities such as twin, teardrop and spindle-shaped cells were also more prevalent in Cec-treated fish. Recovery from these abnormalities was observed but varied between gill and kidney blood, with gill blood showing higher sensitivity and slower recovery compared to kidney blood. This study underscores the utility of AChE activity and erythrocyte abnormalities as biomarkers for assessing pesticide impacts on aquatic organisms. The findings highlight the sensitivity of fish erythrocytes to environmental contaminants and emphasize the need for continued research to better understand the long-term effects of pesticide exposure on aquatic health and ecosystem stability.

Recent innovations in crop protection research.

As the world's population continues to grow and demand for food increases, the agricultural industry faces the challenge of producing higher yields while ensuring the safety and quality of harvests, operators, and consumers. The emergence of resistance, pest shifts, and stricter regulatory requirements also urgently calls for further advances in crop protection and the discovery of new innovative products for sustainable crop protection. This study reviews recent highlights in innovation as presented at the 15th IUPAC International Congress of Crop Protection Chemistry held in New Delhi, in 2023. The following new products are discussed: the insecticides Indazapyroxamet, Dimpropyridaz and Fenmezoditiaz, the fungicides Mefentrifluconazole and Pyridachlomethyl, the nematicide Cyclobutrifluram, the herbicides Rimisoxafen, Dimesulfazet, and Epyrifenacil as well as the abiotic stress management product Anisiflupurin. In addition, the latest innovative research areas and discovery highlights in all areas of crop protection will be presented, including insecticidal alkyl sulfones and 1,3,4-trisubstituted pyrazoles, fungicidal picolinamides, herbicidal ketoenols, and trifluoromethylpyrazoles, as well as the latest advances in crop enhancement and green pest control research. © 2024 The Author(s). Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Monitoring and Detection of Insecticide Resistance in Spodoptera frugiperda (Lepidoptera: Noctuidae): Evidence for Field-Evolved Resistance in Egypt.

Spodoptera frugiperda (J.E. Smith) (Noctuidae: Lepidoptera) is a notable insect pest that invades major cereal crops, causing significant damage and loss. Resistances of 2nd instar larvae of two Egyptian field populations of S. frugiperda, collected from the Fayoum and Giza governments, were measured against eight insecticides, including traditional insecticides (profenofos and cypermethrin), bio-insecticides (emamectin benzoate, spinosad, and Bacillus thuringiensis), and insect growth regulators (IGRs) (lufenuron, diflubenzuron, and methoxyfenozide). In addition, the synergistic effects of three synergists (Piperonyl butoxide (PBO), diethyl maleate (DEM), and triphenyl phosphate (TPP) were assessed, and the activities of detoxification enzymes (acetylcholine esterase (AChE), cytochrome P-450 (CYP-450), carboxylesterase (CarE), and glutathione-s-transferase (GST) were also determined. Resistance surveillance revealed that the Fayoum field population showed moderate resistance to cypermethrin (RR = 5.75-fold), followed by spinosad (RR = 2.62-fold), and lufenuron (2.01-fold). On the other hand, the Giza population exhibited significant resistance to cypermethrin only (RR = 3.65-fold). Our results revealed that emamectin benzoate was the most effective insecticide, with an LC50 value of 0.003 mg/L for the Fayoum population and 0.001 mg/L for the Giza population, compared to the susceptible strain (0.005 mg/L). Among the biological insecticides, Bacillus thuringiensis was the least toxic insecticide of all the tested strains. Synergism assays indicated that DEM and TPP had the most synergistic effect on spinosad (SR = 8.00-fold for both), followed by PBO (SR = 5.71-fold) for the Fayoum population, compared with spinosad alone. The assay of detoxification enzymes showed that GST activity significantly (p < 0.05) increased in the two field strains compared to the susceptible strain. However, no significant changes were observed among the tested strains in CYP-450, CarE, or AChE. The findings of this study provide substantial insights into tracking and managing the development of insecticide resistance in S. frugiperda in Egypt.

Effect of the Localized Insecticides Spray Technique to Control Halyomorpha halys in Actinidia chinensis Orchards.

Kiwifruit (Actinidia chinensis) cultivation is expanding worldwide, with China, New Zealand, and Italy being the major producing countries. Halyomorpha halys, the brown marmorated stink bug, is raising serious concerns to kiwifruit cultivation both in China and Italy. This study aimed at improving the chemical control efficacy against this pest by comparing two insecticide spray techniques (a conventional ray atomizer and a trumpet-modified atomizer adapted for localized spray application) in kiwifruit. In fact, kiwifruit is often grown with a 'pergola' training system, which may reduce the effectiveness of insecticide penetration into the canopy. Experiments were performed in naturally infested orchards of both Actinidia chinensis var. chinensis 'Jintao' and A. chinensis var. deliciosa 'Hayward'. Furthermore, mesh cages containing H. halys adults were deployed within orchards to assess the insects' mortality at 1, 3, 7, and 10 days after an insecticide application with two spray techniques during two consecutive seasons. In the cultivar 'Jintao', the two systems performed similarly, while in the cultivar 'Hayward', an overall significantly higher insect mortality was recorded with the trumpet atomizer (94-100%) compared to the conventional atomizer (59-78%). Crop damage was also evaluated on both cultivars, simulating the grower insecticide applications with the two spray techniques. At harvest, no difference emerged between the spray techniques, which provided a significantly better protection compared to the untreated control (12-17% compared to 33-47% of injured fruits). Further investigations in this direction are needed also considering the restriction of insecticidal active substances ongoing in the European Union and the need to maximize the efficacy of the available tools.

Anatomy of a pest control failure: introgression of cytochrome P450 337B3 alleles from invasive old-world bollworm into native corn earworm (Lepidoptera: Noctuidae).

The establishment of invasive species populations can threaten the ecological balance in naïve habitats and impact agricultural production practices. Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) (old-world bollworm, OWBW) and Helicoverpa zea (corn earworm, CEW) were geographically separated prior to the 2013 report of OWBW invasion into South America. Introgression of OWBW-specific cytochrome P450 337B3 (CYP337B3) gene into CEW was repeatedly detected across South America and the Caribbean. Two hybrids were documented from Texas in 2019. In this study, screening insects collected in Olathe, CO, USA, where a failure of pyrethroids to control CEW damage to conventional sweetcorn in 2023 detected 28.6% of insects with the OWBW-specific CYP337B3 marker. Nucleotide sequencing of the CYP337B3 gene identified 73.1% and 26.9% of insects carried CYP337B3v2 and CYP337B3v6 alleles, respectively, and 0.15 overall frequency of CYP337B3 alleles. Based on prior data for distinct phylogeographic origins of CYP337B3v2 and v6 alleles, our results indicate Olathe samples were derived from 2 different introductions: An uncertain source of the v6 allele that was initially reported in West Africa and possibly South American or Caribbean origin of the globally distributed v2 allele. One of the 1618 individuals screened also carried a ribosomal RNA internal transcribed spacer 1 derived from OWBW. Local selection pressures at the Olathe location imposed by repeated pyrethroid exposures are likely attributed to the prevalence of CYP337B3, where control practices hasten the accumulation of phenotypic resistance by adaptive introgression. Pyrethroid and other resistance factors carried by invasive OWBW may continue to impact CEW management tactics across the Americas.

Identification and biophysical characterization of potential phytochemical inhibitors of carboxyl/choline esterase from Helicoverpa armigera for advancing integrated pest management strategies.

In the realm of disease vectors and agricultural pest management, insecticides play a crucial role in preserving global health and ensuring food security. The pervasive use, particularly of organophosphates (OPs), has given rise to a substantial challenge in the form of insecticide resistance. Carboxylesterases emerge as key contributors to OP resistance, owing to their ability to sequester or hydrolyze these chemicals. Consequently, carboxylesterase enzymes become attractive targets for the development of novel insecticides. Inhibiting these enzymes holds the potential to restore the efficacy of OPs against which resistance has developed. This study aimed to screen the FooDB library to identify potent inhibitory compounds targeting carboxylesterase, Ha006a from the agricultural pest Helicoverpa armigera. The ultimate objective is to develop effective interventions for pest control. The compounds with the highest scores underwent evaluation through docking studies and pharmacophore analysis. Among them, four phytochemicals-donepezil, protopine, 3',4',5,7-tetramethoxyflavone, and piperine-demonstrated favorable binding affinity. The Ha006a-ligand complexes were subsequently validated through molecular dynamics simulations. Biochemical analysis, encompassing determination of IC50 values, complemented by analysis of thermostability through Differential Scanning Calorimetry and interaction kinetics through Isothermal Titration Calorimetry was conducted. This study comprehensively characterizes Ha006a-ligand complexes through bioinformatics, biochemical, and biophysical methods. This investigation highlights 3',4',5,7-tetramethoxyflavone as the most effective inhibitor, suggesting its potential for synergistic testing with OPs. Consequently, these inhibitors offer a promising solution to OP resistance and address environmental concerns associated with excessive insecticide usage, enabling a significant reduction in their overuse.

Whole-genome sequencing of major malaria vectors reveals the evolution of new insecticide resistance variants in a longitudinal study in Burkina Faso.

BACKGROUND: Intensive deployment of insecticide based malaria vector control tools resulted in the rapid evolution of phenotypes resistant to these chemicals. Understanding this process at the genomic level is important for the deployment of successful vector control interventions. Therefore, longitudinal sampling followed by whole genome sequencing (WGS) is necessary to understand how these evolutionary processes evolve over time. This study investigated the change in genetic structure and the evolution of the insecticide resistance variants in natural populations of Anopheles gambiae over time and space from 2012 to 2017 in Burkina Faso. METHODS: New genomic data have been generated from An. gambiae mosquitoes collected from three villages in the western part of Burkina Faso between 2012 and 2017. The samples were whole-genome sequenced and the data used in the An. gambiae 1000 genomes (Ag1000G) project as part of the Vector Observatory. Genomic data were analysed using the analysis pipeline previously designed by the Ag1000G project. RESULTS: The results showed similar and consistent nucleotide diversity and negative Tajima's D between An. gambiae sensu stricto (s.s.) and Anopheles coluzzii. Principal component analysis (PCA) and the fixation index (FST) showed a clear genetic structure in the An. gambiae sensu lato (s.l.) species. Genome-wide FST and H12 scans identified genomic regions under divergent selection that may have implications in the adaptation to ecological changes. Novel voltage-gated sodium channel pyrethroid resistance target-site alleles (V402L, I1527T) were identified at increasing frequencies alongside the established alleles (Vgsc-L995F, Vgsc-L995S and N1570Y) within the An. gambiae s.l. POPULATIONS: Organophosphate metabolic resistance markers were also identified, at increasing frequencies, within the An. gambiae s.s. populations from 2012 to 2017, including the SNP Ace1-G280S and its associated duplication. Variants simultaneously identified in the same vector populations raise concerns about the long-term efficacy of new generation bed nets and the recently organophosphate pirimiphos-methyl indoor residual spraying in Burkina Faso. CONCLUSION: These findings highlighted the benefit of genomic surveillance of malaria vectors for the detection of new insecticide resistance variants, the monitoring of the existing resistance variants, and also to get insights into the evolutionary processes driving insecticide resistance.

Phytochemical Investigations, Antioxidant and Insecticidal Properties of Essential Oil and Extracts from the Aerial Parts of Pelargonium graveolens from Morocco.

The essential oil and the aqueous and ethanolic extracts obtained from the aerial parts of Pelargonium graveolens cultivated in Morocco were studied for their antioxidant and insecticidal activity against rice weevils (Sitophylus oryzae). The total phenolic content of the extracts was determined by a spectrophotometric method and the phenolic compounds were extensively characterized by HPLC-PDA/ESI-MS. To evaluate antioxidant potential, three in vitro assays were used. In the DPPH test, the ethanolic extract was the most active, followed by the aqueous extract and the essential oil. In the reducing power assay, excellent activity was highlighted for both extracts, while in the Fe2+ chelating activity assay, weak activity was observed for both the essential oil and the ethanolic extract and no activity for the aqueous extract. Concerning insecticide activity, the toxicity of the essential oil and the extracts was tested against rice weevils; the lethal concentrations LC50 and LC99 were determined, as well as the lethal time required for the death of 50% (LT50) and 99% (LT99) of the weevils. The essential oil had the highest activity; 100% mortality of S. oryzae was observed around 5, 9, and 8 days for the essential oil and the aqueous and ethanolic extracts, respectively.

Role of Enterococcus mundtii in gut of the tomato leaf miner (Tuta absoluta) to detoxification of Chlorantraniliprole.

Chlorantraniliprole (CAP) is applied worldwide for the control of caterpillars (Lepidoptera). However, with the overuse of CAP, the resistance problem in pest control is becoming increasingly serious. Recent studies have indicated a central role of the gut symbiont in insect pest resistance to pesticides and these may apply to the tomato leaf miner Tuta absoluta, is one of the most destructive insects worldwide. Here, we successfully isolated seven strains of tolerant CAP bacterium from the CAP-resistant T. absoluta gut, of which Enterococcus mundtii E14 showed the highest CAP tolerance, with a minimum inhibitory concentration (MIC) of 1.6 g/L and CAP degradation rate of 42.4%. Through transcriptomics and metabolism analysis, we studied the detoxification process of CAP by the E. mundtii E14, and found that CAP can be degraded by E. mundtii E14 into non-toxic compounds, such as 3,4-dihydroxy-2-(5-hydroxy-3,7-dimethylocta-2,6-dien-1-yl) benzoic acid and 2-pyridylacetic acid. Additionally, 2-pyridylacetic acid was detected both intracellular and extracellular in E. mundtii E14 treated with CAP. Meanwhile, we identified 52 up-regulated genes, including those associated with CAP degradation, such as RS11670 and RS19130. Transcriptome results annotated using KEGG indicated significant enrichment in up-regulated genes related to the glyoxylate cycle, nitrogen metabolism, and biosynthesis of secondary metabolites. Additionally, we observed that reinfection with E. mundtii E14 may effectively enhance resistance of T. absoluta to CAP. The LC50 values of the antibiotic treatment population of T. absoluta reinfection with E. mundtii E14 is 0.6122 mg/L, which was 18.27 folds higher than before reinfection. These findings offer new insights into T. absoluta resistance to CAP and contribute to a better understanding of the relationship between insecticide resistance and gut symbionts of T. absoluta, which may play a pivotal role in pest management.

Plant versus pollinator protection: balancing pest management against floral contamination for insecticide use in Midwestern US cucurbits.

Controlling crop pests while conserving pollinators is challenging, particularly when prophylactically applying broad-spectrum, systemic insecticides such as neonicotinoids. Systemic insecticides are often used in conventional agriculture in commercial settings, but the conditions that optimally balance pest management and pollination are poorly understood. We investigated how insecticide application strategies control pests and expose pollinators to insecticides with an observational study of cucurbit crops in the Midwestern United States. To define the window of protection and potential pollinator exposure resulting from alternative insecticide application strategies, we surveyed 62 farms cultivating cucumber, watermelon, or pumpkin across 2 yr. We evaluated insecticide regimes, abundance of striped and spotted cucumber beetles (Acalymma vittatum [Fabricius] and Diabrotica undecimpunctata Mannerheim), and insecticide residues in leaves, pollen, and nectar. We found that growers used neonicotinoids (thiamethoxam and imidacloprid) at planting in all cucumber and pumpkin and approximately half of watermelon farms. In cucumber, foliar thiamethoxam levels were orders of magnitude higher than the other crops, excluding nearly all beetles from fields. In watermelon and pumpkin, neonicotinoids applied at planting resulted in 4-8 wk of protection before beetle populations increased. Floral insecticide concentrations correlated strongly with foliar concentrations across all crops, resulting in high potential exposure to pollinators in cucumber and low-moderate exposure in pumpkin and watermelon. Thus, the highest-input insecticide regimes maintained cucumber beetles far below economic thresholds while also exposing pollinators to the highest pollen and nectar insecticide concentrations. In cucurbits, reducing pesticide inputs will likely better balance crop protection and pollination, reduce costs, and improve yields.