Pest Management Science

BACKGROUNDInsect pests present a global threat to crops, with plant resistance representing a key breeding goal. The cabbage stem flea beetle (Psylliodes chrysocephala; CSFB) is the most damaging pest of oilseed rape (Brassica napus; OSR) within Europe; however, CSFB resistance is yet to be found within B. napus. To address this, we examine CSFB larval development over time, explore antibiosis across a diverse Brassica panel, and test whether larvae can develop within model Brassica relatives (Brassica rapa and Arabidopsis thaliana). RESULTSCSFB larvae completed development from four weeks post-infestation, undergoing a 20-fold size increase, with larval recovery after two weeks allowing semi-high throughput resistance phenotyping. Applying this method to 98 Brassica genotypes (97 B. napus and a single Sinapis alba), we found weak evidence for genotype effects on larval survival, however phenotype validation with resistant and susceptible B. napus genotypes showed no differences in larval survival or adult emergence. Larval antibiosis was consistently observed in S. alba. Finally, we showed that B. rapa and A. thaliana represent suitable hosts for CSFB, with larvae increasing 8-10x in size after two weeks. CONCLUSIONCSFB larval antibiosis appears absent within B. napus, possibly due to bottlenecks experienced during domestication. However, larval antibiosis is present within S. alba, and future work should study the basis of this resistance. Further, CSFB larval screening in Brassica relatives presents an opportunity to explore CSFB resistance genetics, informing breeding progress for insect resistance in B. napus.

Neonicotinoid insecticides used in seed treatment present phytotoxic potential that may accelerate seed deterioration during storage; however, how this effect interacts with genotype remains poorly understood. We evaluated the physiological quality of five maize genotypes sourced from the Maize Breeding Programme of the Federal University of Lavras, comprising three inbred lines (L44, L91 and L64) and two half-sib hybrids (H44 and H91), treated with a neonicotinoid-based insecticide formulation (thiamethoxam and cyantraniliprole) and stored for up to nine months at 25{degrees}C. Physiological quality was assessed through germination on rolled paper + vermiculite, cold test, primary root length, a phytotoxicity index, projection pursuit multivariate analysis, and scanning electron microscopy of pericarp and aleurone layer. Insecticide treatment reduced germination and increased phytotoxicity indices, with inbred line L44 showing the most severe response, reaching phytotoxicity values up to 15.89 percentage points above hybrid H91 at six months of storage. Scanning electron microscopy revealed that L44 presented a thinner pericarp and pronounced aleurone layer disorganisation following treatment, whilst L91 remained structurally preserved. Tolerance to post-treatment storage is highly genotype-dependent, and pericarp thickness may represent a useful morphological marker for selecting tolerant genotypes in maize breeding programmes.

Ultra-low volume (ULV) insecticide spraying with deltamethrin as the active ingredient is widely used in mosquito control programs, yet its effectiveness against target mosquitoes and its ecological side effects remain poorly quantified under field conditions in Central Europe. Here, we experimentally evaluated the short-term impact of ground ULV spraying on both mosquito populations and non-target flying insects in Hungary using a paired before-after-control-impact (BACI) design. Mosquitoes were sampled with BG Sentinel traps, while non-target insects were collected using malaise traps. ULV treatment resulted in a significant reduction in mosquito abundance at treated sites, with an average decline of approximately 45%. Native and invasive mosquito species, including Aedes albopictus and Aedes koreicus, showed similar proportional decreases. However, treatment effectiveness varied substantially among sites and was influenced by initial mosquito abundance and wind conditions. In parallel, malaise trap samples revealed a marked decline in non-target flying insects, with reductions exceeding 40% across multiple taxonomic groups, particularly among small- and medium-sized insects, and also when considering pollinator taxa together. Our results indicate that while ULV spraying can temporarily reduce mosquito abundance, it also imposes considerable short-term impacts on non-target insect communities, highlighting trade-offs between vector control and insect conservation within mosquito management programs.

Insecticide resistance in mosquitoes, largely mediated by cytochrome P450 monooxygenases (CYPs), compromises the efficacy of vector control tools. In this study, chemically-wise selected natural extracts and compounds were screened for their CYP inhibition potential. Among 37 tested plant extracts and fractions, a decarboxylated acidic fraction of industrial hemp (Cannabis sativa Linnaeus var. Futura 75) emerged as a promising hit, and phytochemical profiling identified cannabidiol (CBD) as its major component (IC = 18.37 M for CYP9K1). CBD was used as a scaffold to generate semisynthetic analogues; of which a piperazinyl analogue outperformed the natural scaffold demonstrating significantly greater potency (IC = 2.50 M for CYP9K1). Docking studies using homology-derived CYP9K1 models also supported a stronger binding affinity of the piperazinyl analogue relative to CBD. Toxicity assays using pyrethroid-resistant Anopheles gambiae Giles adults confirmed that neither CBD nor the piperazinyl analogue had intrinsic toxicity, yet the semisynthetic analogue significantly enhanced deltamethrin efficacy, showing a threefold synergistic effect. The safety profile of the cannabis compounds for non-target organisms was evaluated through human cell line cytotoxicity tests and bee toxicity assays, suggesting low non-target organism toxicity. Our study describes the identification of a plant-derived synergist lead with strong potential as an insecticide additive to combat metabolic resistance in malaria-transmitting mosquitoes.

Istocheta aldrichi (Diptera: Tachinidae) is a specialist parasitoid of the invasive Japanese beetle, Popillia japonica (Coleoptera: Scarabaeidae). Research and releases for biological control depend on field collecting parasitized hosts and rearing the parasitoid through diapause to obtain I. aldrichi adults. This study investigated how rearing practices before, during and after the seasonal overwintering period affected the proportion of I. aldrichi pupae that emerged as adults, the timing of parasitoid emergence, and their longevity. Increasing cold exposure duration during overwintering increased adult I. aldrichi emergence from puparia and reduced development time after transfer to warm conditions. Adult I. aldrichi emergence from overwintered puparia depended on interactions between overwintering environment (indoors vs. outdoors), spring thermal regime, and the timing of host collection in the previous season. Burying puparia in the soil in late summer/early fall resulted in higher subsequent adult I. aldrichi emergence. Manipulating spring temperatures in controlled environments allowed parasitoid emergence to be staggered over several weeks without reducing emergence success. Emergence under outdoor spring conditions was unreliable. Adult longevity was affected by temperature and diet: cooler conditions extended lifespan, honey-water increased longevity relative to pollen alone or honey-water and pollen together. These results provide a foundation to further improve I. aldrichi rearing techniques for use in experimental research and applied biological control of P. japonica.

Istocheta aldrichi (Diptera: Tachinidae), a specialist parasitoid of the invasive Japanese beetle, Popillia japonica (Coleoptera: Scarabaeidae), was released to eastern North America in the 1920s as part of a classical biological control program. Further releases are being considered in different regions of North America and Europe where P. japonica is establishing. Successful releases of the biocontrol agent depend on identifying efficient techniques for collecting parasitized hosts from the field and rearing the parasitoid through diapause to obtain I. aldrichi adults. In this study, we evaluated how the collection date, the collection method (hand-picking vs. regular traps vs. modified traps) and rearing conditions (food provision and substrate type) of parasitized hosts influence I. aldrichi pupariation and emergence. The proportion of parasitized beetles yielding I. aldrichi puparia decreased considerably as the season progressed. Rearing conditions immediately after collection influenced both puparium yield and quality: withholding food from parasitized P. japonica slightly increased puparium yield but reduced puparium weight, while the effect of food provision on subsequent overwintering survival depended on rearing substrate. Finally, simple modifications to commercial traps (larger, ventilated, containers with added food source and substrate) collected more beetles than regular traps and promoted successful development of the parasitoid to the puparium stage. Our results are used to suggest basic guidelines for collecting and rearing I. aldrichi in experimental research and applied biological control of P. japonica.

Glufosinate-ammonium (GA) has been widely used in Midwestern fields, and in recent years a growing number of failures to control waterhemp [Amaranthus tuberculatus (Moq.) Sauer] have raised concerns about the potential evolution of resistance. The goal of this study was to investigate four independent cases of suspected resistance to GA in A. tuberculatus from Illinois using greenhouse, field, and transcriptomics studies. Greenhouse dose-response experiments revealed resistance ratios ranging from 2.2- to 3.4-fold based on survival and from 1.3- to 2.8-fold based on dry biomass relative to a susceptible population. A subsequent field study where one of the populations originated confirmed that twenty percent of treated plants survived the labeled GA field-recommended rate. Screening for other herbicide sites of action revealed that most populations showed reduced sensitivity to atrazine, glyphosate, and imazethapyr, surviving up to three times the field-recommended rates, and to a lesser extent, lactofen and fomesafen. Transcriptomic analysis of plants surviving GA revealed no resistance-associated mutations or differential transcript abundance in the plastidic and cytosolic isoforms of glutamine synthetase. Among the four suspected resistant populations, there were 182 genes differentially expressed relative to two susceptible populations. Different sets of genes were differentially expressed among the populations studied, with only one gene (upregulated relative to two susceptible populations) shared among all four. Many of the differentially expressed genes, including cytochrome P450s, glutathione S-transferases, glycosyltransferases, transporters, and transcriptional regulators, are commonly associated with metabolic resistance. Gene ontology enrichment analyses indicated significant overrepresentation of stress response, defense regulation, and secondary metabolism categories across the populations. Together, these findings provide evidence for the evolution of GA resistance in populations of A. tuberculatus in Illinois. While more in-depth studies are needed to fully characterize the underlying mechanisms, the consistent differential expression of metabolism-related genes and no indication of target-site mechanisms points to a potential metabolic basis for resistance.

RNA interference (RNAi) shows great potential to protect crops against fungal diseases, yet reported protection efficiencies vary greatly, and our understanding of the factors responsible for this variance remains limited. In this meta-analysis, we evaluated 89 studies that compare the efficiency of host-induced gene silencing (HIGS) and spray-induced gene silencing (SIGS) in controlling fungal diseases, focusing on biotrophic, hemibiotrophic, and necrotrophic fungi, the use of formulations, and the dsRNA design as explanatory factors for differences between reported efficiency values. Our results indicate that SIGS is slightly more effective, particularly in biotrophs. Surprisingly, SIGS studies using formulations did not outperform those applying naked dsRNA. We also assessed parameters of RNA design. Differences in dsRNA length and the number of constructs, and number of targets showed no consistent significant effect on resistance in either HIGS or SIGS. Interestingly, however, HIGS studies reported significantly higher efficiency when targeting genes closer to the 3 end and SIGS when targeting genes closer to the 5 end. We discuss potential reasons for the reported patterns, such as variability in dsRNA uptake mechanisms, intercellular trafficking and Dicer processing, and conclude that more research is needed to understand the biological mechanisms determining RNAi efficiency for fungal control.

Developing grapevine cultivars with genetic resistance to pathogens is a key strategy to reduce fungicide use and enhance sustainability. The French INRAE-ResDur program aims to pyramid several resistance loci against Plasmopara viticola (Rpv), the causal agent of downy mildew, while integrating factors against Erysiphe necator (Ren/Run) which causes powdery mildew. We evaluated in field the performance of grapevine genotypes carrying single or pyramided Rpv loci during the exceptionally severe downy mildew epidemic of 2024. Disease severity was quantified as the proportion of leaf foliage exhibiting symptoms. Susceptible controls averaged 66.6 % symptomatic leaves, Rpv1/3.1 combination remained below 16.1 %. whereas the Rpv1/Rpv3.1/Rpv10 pyramid showed only 4.9 % symptomatic leaves. The single loci provided partial protection, but the effect varied with genetic background. Pyramiding improved resistance effectiveness and stability, indicating synergistic interactions among loci. These findings demonstrate that pyramiding Rpv loci is an effective strategy for durable downy mildew resistance and should be the preferred strategy in grapevine breeding programs and genetic resistance deployment strategies.

Insect camera traps are a rapidly developing technology, enabling automated monitoring of insects. However, little has been reported on improving the attractants used for daytime flying insects on such cameras. This study compares the attractiveness of novel, 3D printed, artificial flowers with traditional methods of attracting insects (e.g. pan traps and solid coloured paper squares). We hypothesised that artificial flowers would attract a higher abundance and diversity of insects compared to traditional attractants by more accurately mimicking flowers. Additionally, we examined colour preference and average landing duration on the attractants. Artificial flowers, dry pan traps and paper squares, painted in yellow, white, or blue ultraviolet fluorescent paint, were filmed simultaneously to observe wild insect behavioural responses (landings and approaches). The results indicate overall preference for artificial flowers over the two traditional attractants when considering all insect groups together, and overall colour preferences for blue and yellow. When analysing insect groups separately, hoverflies preferred landing on artificial flowers over the other attractants. Bumblebees preferred approaching artificial flowers, and small insects preferred landing and approaching artificial flowers over the other attractants. Other flies preferred landing on pan traps and paper over artificial flowers. Hoverflies, small insects, wasps, and solitary bees responded more to yellow than the other colours, while bumblebees responded more to blue. Comparisons of landing durations revealed that hoverflies spent longer on the artificial flowers than paper. Other flies spent longer on the pan traps and paper. These results show that artificial flowers could offer an efficient attractant for insect camera traps as they attracted a higher abundance of key pollinating insects (hoverflies and bumblebees), and do not have worse attraction rates for the other insect groups (excluding other flies).

BACKGROUNDThe two-spotted spider mite, Tetranychus urticae Koch, is a major agricultural pest with a rapid propensity for developing acaricide resistance. Bifenazate targets mitochondrial cytochrome b (CYTB). While the G126S mutation is frequently associated with resistance, its independent role remains unclear as it often occurs with other substitutions. This study explores the molecular basis of bifenazate resistance in a Russian laboratory strain derived from a St. Petersburg greenhouse population. RESULTSDisruptive selection with increasing bifenazate concentrations generated resistant and susceptible isofemale lines. AlphaFold2 structural modeling of CYTB indicated that G126S causes a steric clash, leading to conformational destabilization, whereas other reported mutations primarily affect the ligand-binding pocket. Oxford Nanopore sequencing revealed a very low initial frequency of the G126S allele (<1%; 226/35,895 reads) in the unselected population. After one year of stepwise selection (0.00005-0.031% a.i.), the mutant allele frequency surged to 90% (7,272/8,056 reads). No other known resistance-associated mutations were found in the analyzed cytb fragment. CONCLUSIONWe report the first identification of the G126S mutation in a Russian T. urticae population and demonstrate its rapid fixation under bifenazate selection. Within this genetic background, G126S alone appears sufficient to confer high-level resistance, emphasizing the population-specific nature of resistance evolution and the critical need for local monitoring.

There is a growing concern that chronic, low-level exposure to organophosphate insecticides is a threat to human health. These synthetic chemicals are used in crop and livestock production all over the world, and the general population are exposed to them through consuming the residues that remain in food. Evidence is emerging that fermentation with lactic acid bacteria may be an effective way of reducing organophosphate insecticide residues. However, while several studies have investigated this topic, outcome measures have varied, and there has been no research to date which has consolidated this data to better understand the half-lives of organophosphate insecticides in fermented foods and the factors affecting degradation. The aim of this review was to synthesise the evidence on organophosphate insecticide degradation during lactic acid fermentation, and analyse organophosphate insecticide half-lives, in order to determine the effectiveness of lactic acid fermentation in reducing organophosphate insecticide residues in food. Furthermore, the study aimed to explore the factors that impact the rate of degradation. CAB Abstracts, Food Science and Technology Abstracts, Scopus and Web of Science were searched for eligible laboratory-based studies, which were published after 2000. The literature search and screening process resulted in the inclusion of 14 eligible studies. Studies were screened for Risk of Bias (ROB) using the RoBDMAT tool. Collated results showed that organophosphate insecticides degraded over time, and this was irrespective of fermentation. However, out of the 249 experiments that involved a controlled fermentation, 232 demonstrated that fermentation with lactic acid bacteria could speed up the degradation of organophosphate insecticides in food, beyond the rate of inherent degradation in the food matrix, leading to shorter half-lives. The half-lives of organophosphate insecticides in apple juice, milk and wheat ranged from 9.5 hours to 21 days in fermented foods and ranged from 21.4 hours to 36.5 days in non-fermented foods. Single species of lactic acid bacteria that demonstrated strong potential for organophosphate insecticide degradation were Lpb.plantarum subsp.plantarum, Lab.delbrueckii subsp.bulgaricus and Lvb. brevis, where the median percentage change in organophosphate insecticide half-life during fermentation was -42.3%, -25.0% and -22.9%, respectively. Organophosphate insecticide degradation during natural fermentation was less clear because of fewer studies and less consistent results. Whilst the collated data shows that fermentation with lactic acid bacteria is an effective method to reduce organophosphate insecticide residues in food, reflected in shorter half-lives, the small number of studies and variability among studies does limit the conclusions that can be drawn, and further research is needed to strengthen these findings. The results of our analysis may help to inform more reliable organophosphate exposure assessments for the population as well as provide novel insights for both consumers and food manufacturers, expanding the market potential for fermented foods.

The western honey bee (Apis mellifera) is an essential pollinator facing unprecedented threats from pesticide exposure. While pesticide resistance evolution is well documented in agricultural pests, our understanding of genetic variation in honey bee detoxification systems remains limited. This represents a missed opportunity, as harnessing naturally occurring detoxification diversity could provide new avenues for pollinator protection. Cytochrome P450 monooxygenases (CYPs), which are central to xenobiotic metabolism, offer a promising starting point. Here, we present the first comprehensive analysis of CYP genetic diversity in A. mellifera. We analysed the CYPome of 1,467 individuals representing 18 A. mellifera subspecies from 25 countries and identified 5,756 single-nucleotide polymorphisms (SNPs) in 46 CYP genes. Imputed McDonald-Kreitman testing revealed that 56% of non-synonymous CYP substitutions were driven by positive selection. Of the 1,302 haplotypes identified, 84% resided in CYP3, concentrated in the CYP9 and CYP6AS subfamilies implicated in xenobiotic detoxification. Population-level analysis of nucleotide diversity, Tajimas D selection signatures, FST-based differentiation, and McDonald-Kreitman testing pointed to CYP3 clan genes as the primary locus of adaptive variation. This work provides the first step toward building a comprehensive pharmacogenomic resource for honey bees, enabling the prediction of population-specific pesticide vulnerabilities and leveraging naturally occurring detoxification variants to enhance pollinator resilience - a critical step toward sustainable pollinator management.

Life-history traits play an important role in insect population dynamics and ecological processes. The azuki bean beetle Callosobruchus chinensis is a common pest of stored legumes and is also widely used as a model species in ecological and evolutionary research. In this study, we tested whether machine learning models could be used to estimate several traits of C. chinensis, including elytral length, development time and adult lifespan. Experimental data were obtained from laboratory populations. The dataset included biological and environmental variables such as strain, treatment condition, developmental day, sex, temperature, and CO2. Six different machine learning models were tested, including linear regression, random forest, support vector machine (SVM), neural network, gradient boosting and AdaBoost. Model performance was evaluated using cross-validation. The coefficient of determination (R2) and root mean square error (RMSE) were used to measure prediction accuracy. Prediction accuracy differed among traits. Elytral length showed relatively higher predictability than the other traits, while development time was difficult to estimate in most models. Lifespan was easier to predict than the other traits, and the neural network produced one of the highest prediction accuracies among the tested models. Feature importance analysis also showed that factors such as sex and treatment condition contributed to variation in several traits. Machine learning models therefore helped reveal relationships among biological variables and life-history traits in C. chinensis. Combining ecological experiments with machine learning analysis may help improve our understanding of insect traits and may support future studies in insect ecology and pest management.

It is commonplace in East Africa for 100% of cassava fields to be infected with Cassava mosaic disease (CMD) and/or Cassava brown streak disease (CBSD), resulting in annual losses of more than US$1.25 billion and reduced food and economic security for farming households. The vector of both diseases is the African cassava species of the whitefly Bemisia tabaci. Since the late 1990s, there has been an unprecedented increase in whitefly populations, to the extent that they are referred to as "super-abundant". Research efforts since the late 1990s has focused mainly on developing plant resistance to the viral pathogens and paid scant attention to understanding the root causes of disease epidemics or the control of whitefly infestation. Here, we aimed at developing long-term whitefly-control solutions using an in-planta RNA interference (RNAi) approach. First, transcriptome analysis identified candidate genes that play key roles in whitefly biology: osmoregulation, sugar metabolism and transport, symbiosis with endosymbiotic bacteria and detoxification of phytotoxins. Then, fifteen RNAi inverted repeat constructs were produced, designed to target the candidate genes and 140 independent transgenic lines were generated in cassava variety NASE 13. Whole plant bioassays showed insecticidal activity of transgenic plants, reaching 58% lethality for adults within 7 days and 75-90% lethality of nymphs after 25 days, compared to control plants. Target genes were confirmed to be downregulated by up to 2.5-fold in adult whiteflies and nymphs. We used population dynamics modelling to predict the potential of the RNAi technology to control whiteflies under field conditions in East Africa.

The European honey bee (Apis mellifera L.) is a key agricultural pollinator frequently exposed to pesticide residues, yet the molecular basis of its chemical adaptation, particularly glutathione S-transferases (GSTs) involved in xenobiotic detoxification, remain incompletely understood. In this study, AmGSTO1 was structurally and functionally characterized to evaluate its role in agrochemical interaction and protection against oxidative stress. The crystal structure of AmGSTO1 in complex with glutathione revealed its 3D architecture and key active-site residues were identified by structural analysis and site-directed mutagenesis. Fluorescence binding assays demonstrated measurable affinity for multiple agrochemicals, including TCP, fenoprop, 2,4-D, tetramethrin, nicotine, and 3-phenoxybenzaldehyde. However, HPLC analysis showed no detectable substrate depletion, suggesting ligand binding to AmGSTO1 without catalytic turnover. AmGSTO1 exhibited antioxidant activity toward cumene hydroperoxide, hydrogen peroxide, and paraquat, as well as dehydroascorbate reductase activity. These findings indicate that AmGSTO1 may contribute to agrochemical tolerance through ligand sequestration and redox protection mechanisms.

Australia is the largest producer of Pyrethrum (Tanacetum cinerariifolium) globally. Amongst the constraints on production are the fungal pathogens Didymella tanaceti and Stagonosporopsis tanaceti, which pose a significant threat to the industry, causing substantial yield losses. While the infection biology of S. tanaceti is well characterised, knowledge of D. tanaceti and its potential interaction with S. tanaceti on plants remains limited, hindering disease management. We developed fluorescently labelled strains of both pathogens via Agrobacterium tumefaciens-mediated transformation (ATMT). Binary vectors carrying the mNeonGreen or tdTomato fluorescent protein genes were introduced into D. tanaceti and S. tanaceti, respectively, and expression of the fluorescent proteins was confirmed by microscopy. Genome sequencing revealed single-copy T-DNA insertions in all transformants, with minor genomic rearrangements at insertion sites. Detached leaf assays demonstrated that transformed strains retained pathogenicity, producing disease symptoms indistinguishable from those of the wild type. These fluorescently labelled variants enabled detailed visualisation of D. tanaceti infection biology and its interactions with S. tanaceti, including co-infection dynamics. Co-infection assays using fluorescent strains further facilitated simultaneous visualisation and differentiation of both pathogens within host tissues. Importantly, these tools also allowed the first description of the early stages of infection by D. tanaceti in pyrethrum leaves. This study represents the first successful transformation of D. tanaceti and S. tanaceti, providing valuable resources to investigate their infection processes.

O_LIAlthough ecological research has long focused on the effects of temperature on population growth, arthropod pests are exposed to a wide variety of environmental factors that affect their performance, such as chemical pesticides targeted against them. Moreover, these environmental factors likely do not act in isolation. Identifying the extent to which abiotic factors interact to affect pest population dynamics can strengthen current and future pest management programs. C_LIO_LIHere, we investigated the extent to which temephos, a common pesticide applied to aquatic environments for mosquito control, influences the thermal performance of juvenile survival and development rate, as well as the intrinsic population growth rate, of the invasive mosquito pest, Aedes aegypti. We implemented a response surface experimental design to measure these traits across seven temperatures and five temephos concentrations and fit temperature- and insecticide-dependent performance curves to assess impacts on the overall performance and the thermal optimum, minimum, and maximum. C_LIO_LITemephos exposure profoundly altered the thermal performance of juvenile survival by reducing survival across all temperatures, shrinking the thermal breadth, and shifting the thermal optimum to warmer temperatures. Through this, temephos also altered the thermal performance of population growth primarily by reducing its thermal breadth. C_LIO_LISynthesis and applications: Our findings demonstrate that interactions between temperature and insecticide exposure can fundamentally reshape pest population dynamics, rather than acting as independent stressors. By quantifying this interaction, we showed that temphos is most effective below the pests thermal optimum, suggesting that larvicides may yield the greatest population suppression in cooler regions or during cooler periods of the year. Incorporating such temperature-dependent efficacy into pest management strategies could improve the timing and spatial targeting of control efforts. More broadly, these results highlight the need to integrate anthropogenic stressors with climatic drivers when predicting pest risk and optimizing management under ongoing environmental change. C_LI

New approaches to mitigate the reduction of crop yields by plant parasitic nematodes are needed in the face of increasing concerns of the impact of nematicides on precious ecosystems. One approach is to target receptors in the parasitic nematode that are vital for their survival that less widely expressed in non-target organisms. Nematodes express a phylogenetically restricted 5-HT-gated chloride channel, MOD-1, activation of which causes paralysis in Caenorhabditis elegans. We show that MOD-1 is expressed in the motor nervous system of the plant parasitic nematode Globodera pallida and its functional characterisation is validated by 5-HT activation when reconstituted in Xenopus laevis oocytes. To evaluate MOD-1 as a nematicide target we utilised a previously described platform called PhaGeM4 for PharmacoGenetic targeting of M4 neurone in which MOD-1 is expressed in transgenic C. elegans and nematode development in the face of MOD-1 chemical modulation is tracked. We screened Pathogen Box, a chemical library of 400 diverse drug-like molecules, using PhaGeM4. This identified 10 putative hits for C. elegans MOD-1. These hits were pursued through a sequential, iterative pipeline encompassing mod-1 dependent C. elegans motility and G.pallida motility assays in combination with pharmacological interrogation of G. pallida mod-1 in PhaGeM4. This approach highlights 3 compounds with a mod-1 dependent action (quipazine, our benchmark compound; MMV687251, a vancomycin-like compound; MMV688774, an antifungal with common name posaconazole) and one compound that acts through an undetermined target (MMV002816, also known as the antifilarial drug, diethylcarbamazine). Each of these compounds had a significant inhibitory effect on G. pallida J2 root invasion. Overall, this lends confidence that the PhaGeM4 screening platform can delivery new chemical leads for crop protection and highlights four new chemistries of interest. More generally, this approach could be applied to other ligand-gated ion channels of interest as targets.

Insecticide resistance is threatening malaria control. While the evolution and spread of resistance has been linked to scale-up in the distribution of public health insecticides, the role of environmental pollutants such as the polyaromatic hydrocarbons (PAHs) from industrial and agricultural use remains largely uncharacterized. The PAHs are potent ligands of the aryl hydrocarbon receptor (Ahr) transcription factors involved in the regulation of xenobiotic metabolizing enzymes, and potentially involved in insecticide resistance. Here, using field insecticide-resistant (Auyo) An. coluzzii and a laboratory-susceptible colony (Ngousso), we conducted a multi-generational selection experiment using naphthalene, fluorene and a mixture of both PAHs. After ten generations, the changes in susceptibility to insecticides were monitored using WHO bioassays and whole-transcriptome analysis (RNASeq) was conducted. Compared with the non-selected colony lines, PAH exposures significantly reduced pyrethroid and DDT resistance in the field population, suggesting fitness cost associated with established resistance. In contrast, Ngousso showed a significant increase in DDT resistance (p = 0.01) at the tenth generation. A significant increase in permethrin resistance was also observed at the seventh generation (p = 0.03). Several candidate genes from the major detoxification classes were overexpressed in the selected lines (including GSTe2, CYP6Z1, and CYP6P4); the most consistent were CYP6M4 and CYP4C27, as well as those from the Ahr pathway. Heterologous expression of CYP6M4 revealed its ability to metabolise pyrethroids, including permethrin, deltamethrin, and -cypermethrin, as well as PAHs (naphthalene and fluorene). These findings establish the role of environmental pollutants as additional drivers of metabolic insecticide resistance in An, coluzzii.