Current Research in Insect Science最新文献

The increasing global burden of mosquito-borne diseases require targeted, environmentally friendly, and sustainable approaches for effective vector control without endangering the non-target beneficial insect population. Biological interventions such as biopesticides, Wolbachia-mediated biological controls, or sterile insect techniques are used worldwide. Here we review Binary or BinAB toxin—the mosquito-larvicidal component of WHO-recognized Lysinibacillus sphaericus bacterium employed in mosquito control programs. Binary (BinAB) toxin is primarily responsible for the larvicidal effect of the bacterium. BinAB is a single-receptor-specific toxin and is effective against larvae of Culex and Anopheles, but not against Aedes aegypti. The receptor in Culex, the Cqm1 protein, has been extensively studied. It is a GPI-anchored amylomaltase and is located apically in the lipid rafts of the larval-midgut epithelium. The interaction of the toxin components with the receptor is crucial for the mosquito larvicidal activity of the BinAB toxin. Here we extend support for the pore formation model of BinAB toxin internalization and the role of toxin-glycan interactions in the endoplasmic reticulum in mediating larval death. BinAB is phylogenetically safe for humans, as Cqm1-like protein is not expected in the human proteome. This review aims to initiate targeted R&D efforts, such as applying fusion technologies (chimera of BinA, chemical modification of BinA), for efficient mosquito control interventions. In addition, the review also examines other areas such as bioremediation and cancer therapeutics, in which L. sphaericus is proving useful and showing potential for further development.

A unique aspect of metabolic detoxification in insects compared to other animals is the presence of xenobiotic phosphorylation, about which little is currently understood. Our previous work raised the hypothesis that members of the taxonomically restricted ecdysteroid kinase-like (EcKL) gene family encode the enzymes responsible for xenobiotic phosphorylation in the model insect Drosophila melanogaster (Diptera: Ephydroidea)—however, candidate detoxification genes identified in the EcKL family have yet to be functionally validated. Here, we test the hypothesis that EcKL genes in the rapidly evolving Dro5 clade are involved in the detoxification of plant and fungal toxins in D. melanogaster. The mining and reanalysis of existing data indicated multiple Dro5 genes are transcriptionally induced by the plant alkaloid caffeine and that adult caffeine susceptibility is associated with a novel naturally occurring indel in CG31370 (Dro5-8) in the Drosophila Genetic Reference Panel (DGRP). CRISPR-Cas9 mutagenesis of five Dro5 EcKLs substantially decreased developmental tolerance of caffeine, while individual overexpression of two of these genes—CG31300 (Dro5-1) and CG13659 (Dro5-7)—in detoxification-related tissues increased developmental tolerance. In addition, we found Dro5 loss-of-function animals also have decreased developmental tolerance of the fungal secondary metabolite kojic acid. Taken together, this work provides the first compelling functional evidence that EcKLs encode detoxification enzymes and suggests that EcKLs in the Dro5 clade are involved in the metabolism of multiple ecologically relevant toxins in D. melanogaster. We also propose a biochemical hypothesis for EcKL involvement in caffeine detoxification and highlight the many unknown aspects of caffeine metabolism in D. melanogaster and other insects.

True fruit flies (Tephritidae) are among the most destructive agricultural pests in the world, attacking a wide range of fruits and vegetables. The Mediterranean fruit fly Ceratitis capitata is a highly polyphagous species but, being widely established in the Mediterranean region, is not considered as a EU quarantine pest. Hence, it is important to discriminate Ceratitis capitata from non-EU tephritid species, present in imported fruit and vegetables, as non-EU species have a quarantine status. However, morphological identification of tephritid larvae, the most frequently intercepted stage in non-EU produce, is difficult and an easy-to-use molecular diagnostic tool would be helpful for rapid species identification. Therefore, a loop-mediated isothermal amplification (LAMP) method was developed for C. capitata and non-EU tephritids Ceratitis cosyra group1 and Ceratitis species from the FARQ complex, C. fasciventris, C. anonae, C. rosa and C. quilicii. LAMP assays were run with DNA from ILVO collected specimens and DNA samples collected during previous research surveys. LAMP primers were species-specific, with LAMP amplification occurring within 45 minutes for the targeted species. In addition, LAMP assays were successful for all C. capitata life stages or a limited amount of tissue. To conclude, the LAMP assays developed in this study were able to distinguish C. capitata from non-EU Tephritidae species and could be a useful tool for the rapid identification of C. capitata.

Many ectotherms species grow faster but attain a smaller body size when reared under warmer conditions, a phenomenon known as the Temperature-Size Rule (TSR). This rule appears to be stronger in aquatic ectotherms than in terrestrial ectotherms. The difference could be related to difficulties for oxygen uptake in water, whereas on land, adaptive responses in body size may relate to seasonal time constraints. To assess the role of seasonal time constraints in temperature size response of terrestrial ectotherms, we reared the small copper Lycaena phlaeas at three temperatures (18 ˚C, 23˚C and 28˚C) and two photoperiods (16L: 8D and 12L: 12D). We examined whether differences in body size across treatments was related to (1) differences in growth and development, (2) differences in breakpoints during growth trajectories, or (3) differences in ommatidia size (as a proxy for cell size). We found a weak inverse relationship between developmental temperature and the body size of adult butterflies; adult size decreased by approximately 1% for every degree warmer. Under warmer temperatures, caterpillars developed more quickly and had higher growth rates but reached a smaller body size. Under a short photoperiod, both growth and development were slower, especially at the two lower temperatures, but the body size resulting from slow growth over a longer developmental period did not vary with photoperiod. Breakpoints in growth trajectories occurred when larvae reached ∼40% of their maximum mass and these breakpoints were strongly correlated with the size of the resulting adults, suggesting that adult size is predetermined at an early stage. Temperature did not appear to cause reductions in body size through reductions in cell size. Butterflies were largely able to buffer their body size by modulating larval growth and development in tandem. They appear to use photoperiod as a cue to gauge the availability of time (with 12L: 12D indicating less time available) while temperature speeds up growth and development and as such governs the amount of time they need to complete a developmental cycle. Temperature and photoperiod thus induce changes in voltinism to fit a discrete number of generations into a growing season.

Invasive species must often survive combinations of environmental conditions that differ considerably from their native range; however, for a given species it is unclear whether improved tolerance is the result of phenotypic plasticity or genetic adaptation (or both). Agrilus planipennis (Coleoptera: Buprestidae; the emerald ash borer) is an invasive pest of Fraxinus trees in North America and Europe. Previous studies in SW Ontario, Canada, showed that A. planipennis is freeze avoidant, preventing internal ice formation by accumulating Molar concentrations of glycerol in its hemolymph and depressing its supercooling point (SCP, the temperature at which it freezes). The cold tolerance of these SW Ontario animals was used to predict potential distribution, revealing that some Canadian cities should be too cold to allow populations to persist. However, a small population of A. planipennis has persisted in Winnipeg, Manitoba, Canada, through several severe ‘polar vortex’ events. In 2018/19, we collected A. planipennis larvae and prepupae from Winnipeg, MB and Southern Ontario, and found that individuals from Winnipeg were extremely cold tolerant – with SCPs as low as -52°C in prepupae (compared to -32°C in SW Ontario), and observed survival of unfrozen individuals exposed to -50°C for one hour. This cold tolerance was accompanied by higher hemolymph osmolality and glycerol concentration than in the SW Ontario individuals. To distinguish between phenotypic plasticity and local adaptation, in 2020/21 we overwintered Winnipeg-sourced individuals either outdoors in SW Ontario or in a simulated Winnipeg winter. Simulated Winnipeg winter individuals had cold tolerance similar to those overwintered in Winnipeg, while SW Ontario overwintered individuals had cold tolerance similar to those collected previously in the region. The simulated winter individuals had higher hemolymph glycerol concentrations than SW Ontario overwintered animals, at least in part due to greater dehydration. Thus, A. planipennis are cold-tolerant enough to survive some of the harshest winters where their host trees can grow, and most likely attain this cold tolerance via phenotypic plasticity. These findings raise the importance of delineating sensitivity of conclusions to unexpected phenotypic plasticity when predicting potential distributions of new invasives or responses to climate change.

Spodoptera frugiperda (fall armyworm) is a highly destructive invasive pest that feeds on numerous crops including maize and rice. It has developed sophisticated mechanisms to detoxify xenobiotics such as secondary plant metabolites as well as manmade insecticides. The aim of the study was to explore the detoxification response to plant secondary metabolites and insecticides employing a S. frugiperda Sf9 cell model exposed to indole 3-carbinol (I3C) and methoprene. The cell Inhibitory Concentration 50 (IC₅₀) for these molecules was determined and IC₁₀, IC₂₀ and IC₃₀ doses were used to monitor the induction profiles of detoxification genes. Cytochrome P450 monooxygenases (P450s) of the CYP9A subfamily were the most inducible genes of the seven examined. Our results also showed the induction of the transcription factor Cap‘n'collar isoform C (CncC). Transient transformation of Sf9 cells overexpressing CncC and its partner muscle aponeurosis fibromatosis (Maf) induces overexpression of CYP4M14, CYP4M15, CYP321A9 and GSTE1 while CYP9As were not induced. Next, we determined the capacity of recombinantly expressed CYP9A30, CYP9A31 and CYP9A32 to interact with methoprene and I3C. Fluorescence-based biochemical assays revealed an interaction of methoprene with functionally expressed CYP9A30, CYP9A31 and CYP9A32 whereas almost no interaction was detected for I3C, suggesting the ability of CYP9As to metabolize methoprene. Our results showed that Sf9 cells could be a useful model to decipher detoxification pathways of S. frugiperda.

Female insects commonly have more than one mate during a breeding period (‘polyandry’), storing and using sperm from multiple males. In addition to its evolutionary significance, insect polyandry has practical implications for pest management that relies on the sterile insect technique (SIT). The Queensland fruit fly, Bactrocera tryoni (Froggatt), is a major horticultural pest in Australia, and outbreaks are managed by SIT in some regions. The present study provides the first evidence for polyandry in female B. tryoni from field populations from New South Wales (NSW) and Queensland (QLD) through multi-locus genotyping (ten microsatellite markers in four fluorescent multiplexes) of the stored sperm in ovipositing females. Polyandry level was significantly higher in the NSW collection (80.0 %) than the QLD collection (26.1 %), suggesting substantial regional and/or temporal variation. These findings have important implications for the use of SIT to suppress B. tryoni populations and to eradicate outbreaks.

The western corn rootworm (WCR), Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae), is an economically important pest of corn (maize) in North America and Europe. Current management practices for WCR involve transgenic expression of insecticidal proteins to minimize larval feeding damage to corn roots. The evolution of resistant WCR populations to transgenic corn expressing insecticidal proteins (e.g. Cry3Bb1, Gpp34Ab1/Tpp35Ab1) necessitates efforts to discover and deploy new modes of action for WCR control. Here, we tested the hypothesis that the addition of short peptides selected for binding to the WCR gut would restore insecticidal activity of Cry3Bb1 to resistant insects. Phage display technology coupled with deep sequencing was used to identify peptides selected for binding to WCR brush border membrane vesicles and to recombinant putative receptors aminopeptidase and cadherin. The binding and specificity of selected peptides was confirmed by ELISA and pull-down assays, and candidate gut surface binding partners were identified. Although production of 284 novel Cry3Bb1 variants with these peptides did not restore activity against resistant WCR in artificial diet bioassays, 112 variants were active against susceptible insects. These results provided insights for the mechanism of Cry3Bb1 activity and toward engineering a new mode-of-action via receptor re-targeting in the context of protein structure and function.

Edible insects are currently promoted worldwide as an alternative animal protein source, but they are mostly still harvested from the wild where they are predisposed to contamination with agrochemicals. This study analysed six species of edible insects (Ruspolia differens, Rhynchophorus phoenicis, Schistocerca gregaria, Oryctes sp, Pachnoda ephippiata and Acanthoplus sp) collected from different habitats and/or reared in the laboratory in Kenya and Uganda for safety from agrochemical contaminants using liquid chromatography tandem mass spectrometry. The residue levels were statistically compared with the Codex Alimentarius Commission maximum residue limits (MRLs). Residues of only nine agrochemicals were detected in the insects out of 374 chemicals which were screened. The detected agrochemicals include two insecticides (aminocarb and pymetrozine), three herbicides (atraton, methabenzthiazuron and metazachlor) and four fungicides (carboxin, fenpropimorph, fludioxonil and metalaxyl). Ruspolia differens and adult Oryctes sp were free from detectable levels of any agrochemical. Whereas the pesticides residue levels in most insect samples were within maximum residue limits, some of them notably P. ephippiata from black soldier fly larval frass, R. phoenicis from oil palm and P. ephippiata from plant compost contained 2-, 8- and 49-fold higher levels of atraton, methabenzthiazuron and metazachlor, respectively, than MRLs. These findings demonstrate that edible insects may accumulate harmful residues of agrochemicals from the environment where they breed or forage, rendering them unsafe for human consumption or feeding animals. The mechanisms for possible bioaccumulation of these agrochemicals in the insects remains to be investigated. Development of methods for farming edible insects under regulated indoor conditions to ensure their safety as sources of food or feed is recommended.

Numerous plant-based repellents are widely used for personal protection against host-seeking mosquitoes. Vitiveria zizanioides (L.) Nash essential oil and its constituents have demonstrated various mosquito repellent activities. In this study, three chemical actions of vetiver oil and five constituents (terpinen-4-ol, α-terpineol, valencene, vetiverol and vetivone) were characterized against Aedes aegypti, Aedes albopictus and Culex quinquefasciatus by using the high-throughput screening assay system (HITSS). Significant contact escape responses in Ae. aegypti and Ae. albopictus to all test compounds at concentrations between 2.5 and 5% were observed. Spatial repellency responses were also observed in some tested mosquito populations depending upon concentrations. The most significant toxic response on mosquitoes was found at the highest concentration, except for vetivone which had no toxic effect on Ae. aegypti and Ae. albopictus. Results on phototoxic and genotoxic hazard revealed that vetiver oil and their constituents showed no phototoxic potential or any significant genotoxic response. In conclusion, vetiver oil and two constituents, valencene and vetiverol, are potentials as active ingredients for mosquito repellency and present no toxicity.