Susceptible Insects Research Articles

Comparative transcriptional analysis between susceptible and resistant populations of Aedes (Stegomyia) aegypti (Linnaeus, 1762) after malathion exposure.

Aedes aegypti is an important vector of arboviruses, including dengue, chikungunya and Zika. The application of synthetic insecticides is a frequently used strategy to control this insect. Malathion is an organophosphate insecticide that was widely used in Brazil in the 1980s and 1990s to control the adult form of A. aegypti. In situations where resistance to currently used insecticides is detected, the use of malathion may be resumed as a control measure. Many studies have confirmed resistance to malathion, however, comparative studies of differential gene expression of the entire transcriptome of resistant and susceptible insects are scarce. Therefore, understanding the molecular basis of resistance to this insecticide in this species is extremely important. In this paper, we present the first transcriptomic description of susceptible and resistant strains of A. aegypti challenged with malathion. Guided transcriptome assembly resulted in 39,904 transcripts, where 2133 differentially expressed transcripts were detected, and three were validated by RT-qPCR. Enrichment analysis for these identified transcripts resulted in 13 significant pathways (padj < 0.05), 8 associated with down-regulated and 5 with up-regulated transcripts in treated resistant insects. It was possible to divide the transcripts according to the mechanism of action into three main groups: (i) genes involved in detoxification metabolic pathways; (ii) genes of proteins located in the membrane/extracellular region; and (iii) genes related to DNA integration/function. These results are important in advancing knowledge of genes related to resistance mechanisms in this insect, enabling the development of effective technologies and strategies for managing insecticide resistance.

Horizontal transmission of a rice bacterial pathogen Pantoea ananatis among plants by rice planthoppers.

Pantoea ananatis is a bacterium commonly found in various agronomic crops and agricultural pests. In this study, we present findings on a genome-reduced strain of P. ananatis, known as Lstr, which was initially isolated from Laodelphax striatellus (small brown rice planthopper, SBPH). We identified Lstr as a plant pathogen causing disease in rice using Koch's postulates. The pathogenicity of Lstr on rice is comparable to that of Xanthomonas oryzae pv. oryzae, the main causative agent of rice bacterial blight. Through a series of experiments involving live insects, molecular investigations, and microscopy, we find that Lstr can accumulate within SBPH. Subsequently, Lstr can be transmitted from SBPH to rice plants, resulting in leaf blight, and can also be transmitted to other SBPH individuals. Collectively, our results suggest that SBPH serves as a vector for P. ananatis Lstr in rice plants. P. ananatis may encounter susceptible insect populations and become endemic through horizontal transmission from these insects. This could also be valuable for predicting future occurrences of bacterial leaf blight in rice and other crops caused by P. ananatis.

Dietary potassium and cold acclimation additively increase cold tolerance in Drosophila melanogaster

In the cold, chill susceptible insects lose the ability to regulate ionic and osmotic gradients. This leads to hemolymph hyperkalemia that drives a debilitating loss of cell membrane polarization, triggering cell death pathways and causing organismal injury. Biotic and abiotic factors can modulate insect cold tolerance by impacting the ability to mitigate or prevent this cascade of events. In the present study, we test the combined and isolated effects of dietary manipulations and thermal acclimation on cold tolerance in fruit flies. Specifically, we acclimated adult Drosophila melanogaster to 15 or 25 °C and fed them either a K+-loaded diet or a control diet. We then tested the ability of these flies to recover from and survive a cold exposure, as well as their capacity to protect transmembrane K+ gradients, and intracellular Na+ concentration. As predicted, cold-exposed flies experienced hemolymph hyperkalemia and cold-acclimated flies had improved cold tolerance due to an improved maintenance of the hemolymph K+ concentration at low temperature. Feeding on a high-K+ diet improved cold tolerance additively, but paradoxically reduced the ability to maintain extracellular K+ concentrations. Cold-acclimation and K+-feeding additively increased the intracellular K+ concentration, aiding in maintenance of the transmembrane K+ gradient during cold exposure despite cold-induced hemolymph hyperkalemia. There was no effect of acclimation or diet on intracellular Na+ concentration. These findings suggest intracellular K+ loading and reduced muscle membrane K+ sensitivity as mechanisms through which cold-acclimated and K+-fed flies are able to tolerate hemolymph hyperkalemia.

Late fall synthetic acaricide application is effective at reducing host-seeking adult and nymphal Ixodes scapularis (Ixodida: Ixodidae) abundances the following spring.

Based on increases in reported cases of tick-borne illnesses, expanding ranges of native ticks, and repeated documentation of arrivals of nonnative tick species, there is a clear need for their effective management in the United States. Synthetic acaricides have proven efficacious in tick management, but real/perceived negative impacts to the environment and nontarget, beneficial insects must be addressed. We sought to determine whether late fall synthetic acaricide application, when most susceptible beneficial insects are presumably dormant or have migrated, could effectively manage host-seeking spring Ixodes scapularis Say abundances as compared to traditional spring application. We compared results of delivery of Demand CS (lambda-cyhalothrin) via truck-mounted high-pressure spray and powered backpack blower as well as delivery of granular Demand G to experimental control (water) in peridomestic habitats in fall 2021, spring 2022, and combined fall 2021/spring 2022. High-pressure fall delivery of Demand CS and backpack delivery of Demand G significantly reduced host-seeking adult I. scapularis abundances within-season and the following spring combined by 100% and 94%, respectively. No host-seeking nymphal I. scapularis were documented in spring after fall only, spring only, or fall and spring combined delivery of Demand CS via high-pressure or powered backpack blower. No adult I. scapularis were documented at any time posttreatment on locations that received high-pressure delivery of Demand CS. We conclude that high-pressure delivery of Demand CS in late fall successfully eliminated multiple stages of host-seeking I. scapularis through the following spring while likely limiting exposure of beneficial insects to synthetic pyrethroids.

CgSSR marker based trait associations for Sitotroga cerealella (Olivier) resistance in stored paddy rice

Rice is stored for a longer time and many primary and secondary insect pests attack grains, causing both quantitative and qualitative losses. Physicochemical characteristics of grain influence stored grain insect susceptibility and resistance. Identification of gene/Quantitative Trait Loci (QTLs) and introgression in the cultivars would provide the desired solution for varietal resistance against S. cerealella. A group of 80 rice varieties was used for genetic analysis for the identification of candidate genes responsible for Sitotroga cerealella resistance. Heat maps derived from values in the kinship matrix were used to group the varieties into three subpopulations. The largest subpopulation has 34 varieties, while the lowest had 19. The first three principal components (PCs) explained 16, 9, and 6 percent of total variation, respectively, as indicated by the Principal Component Analysis (PCA). The PCA results complemented the heat map results in defining population structure. Genome-wide association mapping can detect marker-trait associations/QTLs in a heterogeneous population of breeding lines, despite population structure and unequal family relatedness. As shown by the Genomic Association and Prediction Integrated Tool (GAPIT) heat map, there are three subpopulations in the association mapping panel. Two cgSSR markers, related to gene SDG725 and FLO2, were linked to the attributes and responsible for conferring resistance against S. cerealella infestation. This is the first report of association and that would help in the development of cultivar with S. cerealella resistance.

A chromosome-level genome assembly of the soybean pod borer: insights into larval transcriptional response to transgenic soybean expressing the pesticidal Cry1Ac protein

BackgroundGenetically modified (GM) crop plants with transgenic expression of Bacillus thuringiensis (Bt) pesticidal proteins are used to manage feeding damage by pest insects. The durability of this technology is threatened by the selection for resistance in pest populations. The molecular mechanism(s) involved in insect physiological response or evolution of resistance to Bt is not fully understood.ResultsTo investigate the response of a susceptible target insect to Bt, the soybean pod borer, Leguminivora glycinivorella (Lepidoptera: Tortricidae), was exposed to soybean, Glycine max, expressing Cry1Ac pesticidal protein or the non-transgenic parental cultivar. Assessment of larval changes in gene expression was facilitated by a third-generation sequenced and scaffolded chromosome-level assembly of the L. glycinivorella genome (657.4 Mb; 27 autosomes + Z chromosome), and subsequent structural annotation of 18,197 RefSeq gene models encoding 23,735 putative mRNA transcripts. Exposure of L. glycinivorella larvae to transgenic Cry1Ac G. max resulted in prediction of significant differential gene expression for 204 gene models (64 up- and 140 down-regulated) and differential splicing among isoforms for 10 genes compared to unexposed cohorts. Differentially expressed genes (DEGs) included putative peritrophic membrane constituents, orthologs of Bt receptor-encoding genes previously linked or associated with Bt resistance, and those involved in stress responses. Putative functional Gene Ontology (GO) annotations assigned to DEGs were significantly enriched for 36 categories at GO level 2, respectively. Most significantly enriched cellular component (CC), biological process (BP), and molecular function (MF) categories corresponded to vacuolar and microbody, transport and metabolic processes, and binding and reductase activities. The DEGs in enriched GO categories were biased for those that were down-regulated (≥ 0.783), with only MF categories GTPase and iron binding activities were bias for up-regulation genes.ConclusionsThis study provides insights into pathways and processes involved larval response to Bt intoxication, which may inform future unbiased investigations into mechanisms of resistance that show no evidence of alteration in midgut receptors.

Utilization of Diverse Molecules as Receptors by Cry Toxin and the Promiscuous Nature of Receptor-Binding Sites Which Accounts for the Diversity.

By 2013, it had been shown that the genes cadherin-like receptor (Cad) and ATP-binding cassette transporter subfamily C2 (ABCC2) were responsible for insect resistance to several Cry1A toxins, acting as susceptibility-determining receptors, and many review articles have been published. Therefore, this review focuses on information about receptors and receptor-binding sites that have been revealed since 2014. Since 2014, studies have revealed that the receptors involved in determining susceptibility vary depending on the Cry toxin subfamily, and that binding affinity between Cry toxins and receptors plays a crucial role. Consequently, models have demonstrated that ABCC2, ABCC3, and Cad interact with Cry1Aa; ABCC2 and Cad with Cry1Ab and Cry1Ac; ABCC2 and ABCC3 with Cry1Fa; ABCB1 with Cry1Ba, Cry1Ia, Cry9Da, and Cry3Aa; and ABCA2 with Cry2Aa and Cry2Ba, primarily in the silkworm, Bombyx mori. Furthermore, since 2017, it has been suggested that the binding sites of BmCad and BmABCC2 on Cry1Aa toxin overlap in the loop region of domain II, indicating that Cry toxins use various molecules as receptors due to their ability to bind promiscuously in this region. Additionally, since 2017, several ABC transporters have been identified as low-efficiency receptors that poorly induce cell swelling in heterologously expressing cultured cells. In 2024, research suggested that multiple molecules from the ABC transporter subfamily, including ABCC1, ABCC2, ABCC3, ABCC4, ABCC10, and ABCC11, act as low-efficiency receptors for a single Cry toxin in the midgut of silkworm larvae. This observation led to the hypothesis that the presence of such low-efficiency receptors contributes to the evolution of Cry toxins towards the generation of highly functional receptors that determine the susceptibility of individual insects. Moreover, this evolutionary process is considered to offer valuable insights for the engineering of Cry toxins to overcome resistance and develop countermeasures against resistance.

Wolbachia infection negatively impacts Drosophila simulans heat tolerance in a strain- and trait-specific manner.

The susceptibility of insects to rising temperatures has largely been measured by their ability to survive thermal extremes. However, the capacity for maternally inherited endosymbionts to influence insect heat tolerance has been overlooked. Further, while some studies have addressed the impact of heat on traits like fertility, which can decline at temperatures below lethal thermal limits, none have considered the impact of endosymbionts. Here, we assess the impact of three Wolbachia strains (wRi, wAu and wNo) on the survival and fertility of Drosophila simulans exposed to heat stress during development or as adults. The effect of Wolbachia infection on heat tolerance was generally small and trait/strain specific. Only the wNo infection significantly reduced the survival of adult males after a heat shock. When exposed to fluctuating heat stress during development, the wRi and wAu strains reduced egg-to-adult survival but only the wNo infection reduced male fertility. Wolbachia densities of all three strains decreased under developmental heat stress, but reductions occurred at temperatures above those that reduced host fertility. These findings emphasize the necessity to account for endosymbionts and their effect on both survival and fertility when investigating insect responses to heat stress.

A winning formula: sustainable control of three stored-product insects through paired combinations of entomopathogenic fungus, diatomaceous earth, and lambda-cyhalothrin.

This research aimed to assess the effectiveness of Metarhizium robertsii, diatomaceous earth (Protect-It), and lambda-cyhalothrin, for the long-term protection of stored wheat against three destructive grain insect pests, Rhyzopertha dominica, Tribolium castaneum, and Trogoderma granarium. Different treatments were applied, both alone and in paired combinations in laboratory and persistence trials. Single treatments exhibited significantly lower mortality rates in comparison to the paired treatments for all tested insect species. Among the single treatments, lambda-cyhalothrin (Lamb) resulted in significantly higher mortality rates in laboratory trials, followed by diatomaceous earth (DE) and M. robertsii (Mr), with insignificant differences between Mr and DE. Evidently, DE exhibited the highest persistence after 120days of storage for all insect species and initial exposures, although variations in mortality rates among treatments were mostly insignificant. Overall, the most effective treatment in terms of mortality in laboratory, and persistence trials, and progeny production was DE + Lamb, followed by Mr + Lamb, and Mr + DE for all tested insect species. In general, the most susceptible insect species was R. dominica, followed by T. castaneum and T. granarium. This research highlights the effectiveness of M. robertsii, DE, and lambda-cyhalothrin in providing prolonged protection of stored wheat against all the examined grain insect species.

Effect of CO2 Concentrations on Entomopathogen Fitness and Insect-Pathogen Interactions

Numerous insect species and their associated microbial pathogens are exposed to elevated CO2 concentrations in both artificial and natural environments. However, the impacts of elevated CO2 on the fitness of these pathogens and the susceptibility of insects to pathogen infections are not well understood. The yellow mealworm, Tenebrio molitor, is commonly produced for food and feed purposes in mass-rearing systems, which increases risk of pathogen infections. Additionally, entomopathogens are used to control T. molitor, which is also a pest of stored grains. It is therefore important to understand how elevated CO2 may affect both the pathogen directly and impact on host-pathogen interactions. We demonstrate that elevated CO2 concentrations reduced the viability and persistence of the spores of the bacterial pathogen Bacillus thuringiensis. In contrast, conidia of the fungal pathogen Metarhizium brunneum germinated faster under elevated CO2. Pre-exposure of the two pathogens to elevated CO2 prior to host infection did not affect the survival probability of T. molitor larvae. However, larvae reared at elevated CO2 concentrations were less susceptible to both pathogens compared to larvae reared at ambient CO2 concentrations. Our findings indicate that whilst elevated CO2 concentrations may be beneficial in reducing host susceptibility in mass-rearing systems, they may potentially reduce the efficacy of the tested entomopathogens when used as biological control agents of T. molitor larvae. We conclude that CO2 concentrations should be carefully selected and monitored as an additional environmental factor in laboratory experiments investigating insect-pathogen interactions.

Two Point Mutations in CYP4CE1 Promoter Contributed to the Differential Regulation of CYP4CE1 Expression by FoxO between Susceptible and Nitenpyram-Resistant Nilaparvata lugens.

Four P450s were reported to be important for imidacloprid resistance in Nilaparvata lugens, a major insect pest on rice, which was confirmed in this study in an imidacloprid-resistant strain (ImiR). Here we found that only two (CYP4CE1 and CYP6ER1) from these four P450 genes were overexpressed in a nitenpyram-resistant strain (NitR) when compared to a susceptible strain (SUS). CYP4CE1 RNAi reduced nitenpyram and imidacloprid resistance in NitR and ImiR strains, with a greater reduction in nitenpyram resistance. The transcription factor FoxO mediated nitenpyram resistance in NitR and ImiR strains, but it was not differentially expressed among strains. The potential reason for the differential regulation of FoxO on CYP4CE1 expression was mainly from sequence differences in the CYP4CE1 promoter between susceptible and resistant insects. In six FoxO response elements predicted in the CYP4CE1 promoter, the single-nucleotide polymorphisms were frequently detected in over 50% of NitR and ImiR individuals. The luciferase reporter assays showed that two mutations, -650T/G and -2205T/A in two response elements at the positions of -648 and -2200 bp, mainly contributed to the enhanced regulation on CYP4CE1 expression by FoxO in resistant insects. The frequency was over 69% for both -650T/G and -2205T/A detected in NitR and ImiR individuals but less than 20% in SUS insects. In conclusion, CYP4CE1 overexpression importantly contributed to nitenpyram resistance in N. lugens, and two mutations in the CYP4CE1 promoter of resistant insects led to an enhanced regulation on CYP4CE1 expression by FoxO.

Soil microbes from conservation agriculture systems reduce growth of Bt-resistant western corn rootworm larvae

Sustainable agricultural management practices aimed at improving soil health can alter the soil microbiome, which can influence plant health and defenses against insects. The western corn rootworm (WCR), Diabrotica virgifera virgifera LeConte, is a major belowground pest of corn. Pest management relies heavily on the planting of transgenic crops expressing Bacillus thuringiensis (Bt) toxins. In this study, we ask how corn-WCR interactions via the soil microbiome are affected by a conservation management system (extended corn-soybean-wheat rotation with cover crops under no-till) compared with a conventional management system (corn-soybean rotation under mulch tillage and no cover crops) when combined with transgenic Bt corn. To do this, we applied soil microbes from the conservation and conventional management systems to two corn lines, one producing Bt and one non-Bt. We then reared Bt-resistant and Bt-susceptible WCR on inoculated seedlings to examine plant and insect changes in fitness. We found that Bt was effective against susceptible larvae in both soil treatments. Bt-resistant larvae were ~ 20% smaller when reared in the presence of soil microbes from the conservation management system. Thus, control of Bt-resistant WCR may be improved in a conservation system without sacrificing Bt effectiveness in susceptible insects. Comparing the microbial communities using 16S rRNA sequencing revealed that management practices influenced the microbiomes associated with the soil and the plant rhizosphere, but not WCR. Our findings suggest value for growers in utilizing conservation management practices, such as no-till and cover crops, in agricultural systems through bottom-up changes to plant–insect interactions via the soil microbiome.

Extended stage duration and diminished fecundity in deltamethrin-resistant Triatoma infestans (Klug, 1834) of the Argentinean Chaco.

Pyrethroid-resistance is an emergent trait in populations of various insect species. For Triatoma infestans (Klug, 1834) (Heteroptera: Reduviidae), the major vector of Chagas disease in the southern part of South America, hotspot areas of pyrethroid-resistance have recently been found in the Chaco Province of Argentina. Resistant condition can reduce fitness of individuals in the absence of insecticide exposure, that is, fitness costs. We evaluated the existence of developmental and/or reproductive costs in T. infestans collected from two areas of pyrethroid-resistance in Chaco Province, Argentina. Three toxicological groups were defined from field-collected insects: susceptible (survival <20%), moderately resistant (survival between 20% and 80%) and highly resistant (survival >80%). Cohorts of the three toxicological groups were followed-up to study life cycle and reproductive parameters. Additionally, we parameterized matrix population growth models. First and IV nymphal stages of the resistant groups exhibited a longer stage duration than susceptible ones. The reproductive days and hatching success showed significant lower values revealing reproductive costs for the resistant groups. Matrix analysis showed lower population growth rates for the resistant groups. Our results support developmental and reproductive costs for pyrethroid-resistant individuals. This trait could be interpreted as lower population recovery ability for pyrethroid-resistant individuals compared to susceptible insects after alternative vector control actions.

Effects of Mixed Baculovirus Infections in Biological Control: A Comprehensive Historical and Technical Analysis.

Baculoviruses are insect-specific DNA viruses that have been exploited as bioinsecticides for the control of agricultural and forest pests around the world. Mixed infections with two different baculoviruses have been found in nature, infecting the same host. They have been studied to understand the biology of virus interactions, their effects on susceptible insects, and their insecticidal implications. In this work, we summarize and analyze the in vivo baculovirus co-infections reported in the literature, mainly focusing on pest biocontrol applications. We discuss the most common terms used to describe the effects of mixed infections, such as synergism, neutralism, and antagonism, and how to determine them based on host mortality. Frequently, baculovirus co-infections found in nature are caused by a combination of a nucleopolyhedrovirus and a granulovirus. Studies performed with mixed infections indicated that viral dose, larval stage, or the presence of synergistic factors in baculovirus occlusion bodies are important for the type of virus interaction. We also enumerate and discuss technical aspects to take into account in studies on mixed infections, such as statistical procedures, quantification of viral inocula, the selection of instars, and molecular methodologies for an appropriate analysis of baculovirus interaction. Several experimental infections using two different baculoviruses demonstrated increased viral mortality or a synergistic effect on the target larvae compared to single infections. This can be exploited to improve the baculovirus-killing properties of commercial formulations. In this work, we offer a current overview of baculovirus interactions in vivo and discuss their potential applications in pest control strategies.

Insecticidal toxicity of ω-Atypitoxin-Cs1a and its inhibitory effects on insect voltage-gated calcium channels.

Excessive use of chemical insecticides raises concerns about insecticide resistance, urging the development of novel insecticides. Peptide neurotoxins from spider venom are an incredibly rich source of ion channel modulators with potent insecticidal activity. A neurotoxin U1-Atypitoxin-Cs1a from the spider Calommata signata was annotated previously. It was of interest to investigate its insecticidal activity and potential molecular targets. Cs1a was heterologously expressed, purified and pharmacologically characterized here. The recombinant neurotoxin inhibited high-voltage-activated calcium channel currents with an median inhibitory concentration (IC50 ) value of 0.182 ± 0.026 μm on cockroach DUM neurons and thus was designated as ω-Atypitoxin-Cs1a. The recombinant Cs1a was toxic to three insect pests of agricultural importance, Nilaparvata lugens, Spodoptera frugiperda and Plutella xylostella with median lethal concentration (LD50 ) values of 0.121, 0.172 and 0.356 nmol g-1 , respectively, at 24 h postinjection. Cs1a was equivalently toxic to both insecticide-susceptible and -resistant insects. Cs1a exhibited low toxicity to Danio rerio with an LD50 of 2.316 nmol g-1 . Our results suggest that ω-Atypitoxin-Cs1a is a potent CaV channel inhibitor and an attractive candidate reagent for pest control and resistance management. © 2023 Society of Chemical Industry.

The Response of Susceptible and Pyrethroid-Resistant Blattella germanica (Dyctioptera: Blattellidae) to Shelter-Associated Cues.

In this work, it was studied the role of faeces in the location and permanence in a shelter in susceptible and pyrethroid-resistant individuals of Blattella germanica (Linnaeus 1767). Additionally, the effect of different concentrations of palmitic acid on the modulation of these behaviours was tested. A shelter constituted by a square cardboard structure was offered to susceptible as well as to resistant specimens. The shelter bases were treated with faecal extracts obtained from susceptible or resistant cockroaches, or with solutions of palmitic acid. The behaviour of susceptible as well as resistant specimens was analysed using infrared videography software. Susceptible's faecal extract attracted both specimens since the time spent by cockroaches to locate the treated shelters was lower, whereas the faecal extract from resistant insects did not elicit any effect on both strains. Faecal extracts showed an arrestant effect on both strains, suggested by the time spent inside the shelter that was significantly higher in their presence. On the other hand, treatment with palmitic acid produced an attractant or a repellent effect depending on the concentration and strain. The tested lower concentration was attractant to susceptible insects, but did not produce any effect on resistant ones. In addition, the higher concentrations did not produce any effect on susceptible individuals, but resulted repellent for resistant ones. Palmitic acid did not produce an arrestant effect on the strains as there was not an increase in time spent inside the shelter in the presence of this substance. An increase in the number of visits to the shelter and to the periphery was also observed in shelters treated with the faecal extract and with the lower concentration of palmitic acid. These results show that compounds of the susceptible faeces were attractant to cockroaches of both strains, while faecal extracts from resistant insects were not. Moreover, a dual effect of palmitic acid was observed, being attractant at low concentrations and repellent as concentration increased. Additionally, a difference in the concentration threshold at which the effect of this substance changes was observed between strains.

Resistance to deltamethrin in Triatoma infestans (Hemiptera: Reduviidae): Does it influence the phenotype of antennae, wings, and heads?

In vector control terms, insecticide resistance is the development of the capacity, of an insect population, to tolerate doses of an insecticide that are lethal to most individuals in a typical population of the same species. The genetic changes that determine resistance may have adaptive costs in the resistant phenotype or, conversely, may result in an adaptive advantage when compared to susceptible insects in the environment without insecticides. Triatoma infestans is one of the main vectors of Trypanosoma cruzi in the southern cone of South America.High insecticide resistance in T. infestans was detected in Argentina in Salta and Chaco provinces. The objective of this study was to determine the possible morphometric changes in wings, heads, and the antennal phenotype of deltamethrin-resistant T. infestans (RR) males and females compared to susceptible insects (SS), evaluating its implication in adaptive processes such as olfactory capacity, dispersion, and probability of colonizing new habitats, among others. Nine type I landmarks were marked on wings, 5 type II landmarks on heads, and 10 antennal sensilla were counted on 106 adults of both sexes (resistant and susceptible from first and second laboratory generations). Morphological divergence was observed between the two groups (RR and SS). The RR insects showed smaller sizes of wings and heads and shape compatible with lower dispersal potential and different active dispersal behaviors. Antennae also revealed sensory simplification in RR and divergence between RR and SS, although more marked in females. This study characterizes for the first time T. infestans RR and SS through wings, heads, and antennae. The results suggest a lower dispersive potential in resistant insects and the differences described lay the foundations for the identification of a resistance biomarker in triatomines.

Can infection with Trypanosoma cruzi modify the toxicological response of Triatoma infestans susceptible and resistant to deltamethrin?

Chemical control plays a central role in interrupting the vector transmission of Chagas disease. In recent years, high levels of resistance to pyrethroids have been detected in the main vector Triatoma infestans, which were associated with less effectiveness in chemical control campaigns in different regions of Argentina and Bolivia. The presence of the parasite within its vector can modify a wide range of insect physiological processes, including toxicological susceptibility and the expression of resistance to insecticides. This study examined for the first time the possible effects of Trypanosoma cruzi infection on susceptibility and resistance to deltamethrin in T. infestans. Using WHO protocol resistance monitoring assays, we exposed resistant and susceptible strains of T. infestans, uninfected and infected with T. cruzi to different concentrations of deltamethrin in fourth-instar nymphs at days 10–20 post-emergence and monitored survival at 24, 48, and 72 h. Our findings suggest that the infection affected the toxicological susceptibility of the susceptible strain, showing higher mortality than uninfected susceptible insects when exposed to both deltamethrin and acetone. On the other hand, the infection did not affect the toxicological susceptibility of the resistant strain, infected and uninfected showed similar toxic responses and the resistance ratios was not modified. This is the first report of the effect of T. cruzi on the toxicological susceptibility of T. infestans and triatomines in general and, to our knowledge, one of the few on the effect of a parasite on the insecticide susceptibility of its insect vector.

Eggsplorer: a rapid plant–insect resistance determination tool using an automated whitefly egg quantification algorithm

BackgroundA well-known method for evaluating plant resistance to insects is by measuring insect reproduction or oviposition. Whiteflies are vectors of economically important viral diseases and are, therefore, widely studied. In a common experiment, whiteflies are placed on plants using clip-on-cages, where they can lay hundreds of eggs on susceptible plants in a few days. When quantifying whitefly eggs, most researchers perform manual eye measurements using a stereomicroscope. Compared to other insect eggs, whitefly eggs are many and very tiny, usually 0.2 mm in length and 0.08 mm in width; therefore, this process takes a lot of time and effort with and without prior expert knowledge. Plant insect resistance experiments require multiple replicates from different plant accessions; therefore, an automated and rapid method for quantifying insect eggs can save time and human resources.ResultsIn this work, a novel automated tool for fast quantification of whitefly eggs is presented to accelerate the determination of plant insect resistance and susceptibility. Leaf images with whitefly eggs were collected from a commercial microscope and a custom-built imaging system. A deep learning-based object detection model was trained using the collected images. The model was incorporated into an automated whitefly egg quantification algorithm, deployed in a web-based application called Eggsplorer. Upon evaluation on a testing dataset, the algorithm was able to achieve a counting accuracy as high as 0.94, r2 of 0.99, and a counting error of ± 3 eggs relative to the actual number of eggs counted by eye. The automatically collected counting results were used to determine the resistance and susceptibility of several plant accessions and were found to yield significantly comparable results as when using the manually collected counts for analysis.ConclusionThis is the first work that presents a comprehensive step-by-step method for fast determination of plant insect resistance and susceptibility with the assistance of an automated quantification tool.

Refuges of conventional host plants counter dominant resistance of cotton bollworm to transgenic Bt cotton

Transgenic crops have revolutionized insect pest control, but evolution of resistance by pests threatens their continued success. The primary strategy for combating pest resistance to crops producing insecticidal proteins from Bacillus thuringiensis (Bt) uses refuges of non-Bt host plants to allow survival of susceptible insects. The prevailing paradigm is that refuges delay resistance that is rare and recessively inherited. However, we discovered refuges countered resistance to Bt cotton that was neither rare nor recessive. In a 15-year field study of the cotton bollworm, the frequency of a mutation conferring dominant resistance to Bt cotton surged 100-fold from 2006 to 2016 yet did not rise from 2016 to 2020. Computer simulations indicate the increased refuge percentage from 2016 to 2020 is sufficient to explain the observed halt in the evolution of resistance. The results also demonstrate the efficacy of a Bt crop can be sustained by non-Bt refuges of other crops.