Search

Insects' host preferences are regulated by multiple factors whose interactions are only partly understood. Here we make use of an in-depth, untargeted metabolomic approach combining molecular networking (MN) and supervised Analysis of variance Multiblock Orthogonal Partial Least Squares (AMOPLS) to untangle egg-laying preferences of Drosophila suzukii , an invasive, highly polyphagous and destructive fruit pest originating from Southeast Asia. Based on behavioural experiments in the laboratory as well as field observation, we selected eight genetically related Vitis vinifera cultivars (e.g., Ancellotta, Galotta, Gamaret, Gamay, Gamay précoce, Garanoir, Mara and Reichensteiner) exhibiting significant differences in their susceptibility toward D. suzukii . The two most and the two least attractive red cultivars were chosen for further metabolomic analyses of their grape skins. The combination of MN and statistical AMOPLS findings with semi-quantitative detection information enabled us to identify flavonoids as interesting markers for differences in the attractiveness of the four studied grape cultivars towards D. suzukii . Overall, dihydroflavonols were accumulated in unattractive grape cultivars, while attractive grape cultivars were richer in flavonols. Crucially, both dihydroflavonols and flavonols were abundant metabolites in the semi-quantitative analysis of the extracted molecules from the grape skin. We discuss how these two flavonoid classes might influence the egg-laying behaviour of D. suzukii females and how they could serve as potential markers for D. suzukii infestations in grapes that can be potentially extended to other fruits. We believe that our novel, integrated analytical approach could also be applied to the study of other biological relationships characterised by multiple evolving parameters., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Marcellin-Gros, Hévin, Chevalley, Boccard, Hofstetter, Gindro, Wolfender and Kehrli.)

Background: Spinosyns are a group of naturally occurring and semi-synthetic insecticides with widespread utility in agriculture, including organic production systems. One example is spinetoram (Delegate), which is the only registered insecticide in New York State (for control of Drosophila melanogaster in vineyards) to which vinegar flies have not yet evolved high levels of resistance. However, low levels of resistance have been found in vineyard populations of D. melanogaster, and a highly resistant strain was obtained after only five selections (in the laboratory). We identified the nAChR α6 mutation (G275A) responsible for the resistance and developed a rapid, high-throughput assay for resistance., Results: Surveys of collections made in 2023 show low levels of the resistance allele in four populations. A correlation was observed between vineyard use of spinetoram and frequency of the resistance allele, but not between county-wide use of spinosyns and frequency of the resistance allele., Conclusions: One of the sites we monitored was previously surveyed in 2019 and the frequency of the resistance allele detected in 2023 had increased. Implications of these findings to resistance management of D. melanogaster are discussed. © 2024 Society of Chemical Industry., (© 2024 Society of Chemical Industry.)

Vinegar flies are vectors of pathogens causing fruit rots of grapes, so control of these insects is important for preventing vineyard yield loss. Recent outbreaks of sour rots may be linked to greater challenges controlling vinegar flies, so we investigated the insecticide susceptibility of populations collected from commercial vineyards across Michigan. We first determined the discriminating concentration for phosmet, malathion, methomyl, and zeta-cypermethrin using a laboratory susceptible (Canton-S) strain of D. melanogaster females. The discriminating concentrations were determined as 252.08, 2.58, 0.96, and 1.68 ppm of the four insecticides, respectively. These concentrations were first tested in 2020 against populations from the two major counties for grape production. In 2021, we expanded monitoring to twenty-three populations collected from vineyards across six counties. All populations had significantly lower sensitivity to all four insecticides compared with Canton-S strain, with up to 98.8% lower mortality for phosmet. The LC50, LC90, and LC99 values of the four insecticides for the two populations tested in 2020 were 7-1,157-fold higher than the Canton-S strain. For the twenty-three populations collected in 2021, mortality ranged from 56.3 to 100% when the flies were screened using a 10x concentration of the discriminating concentration of the insecticides, whereas it ranged from 82.4 to 100% when the flies were screened using a 20x concentration. Our results suggest variable levels of resistance to insecticides from multiple chemical classes in D. melanogaster populations in Michigan vineyards, highlighting the need to implement integrated sour rot management approaches that are less dependent on insecticides for control of this species., (© The Author(s) 2022. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Background: Over the past 14 years, the invasive vinegar fly, spotted-wing drosophila (Drosophila suzukii), has become one of the most damaging fruit pests in the United States. With regional economic losses estimated as high as $500 million for moderate infestations, D. suzukii control represents an often-untenable cost to growers. Management relies heavily on chemical control, which may be applied up to nine times in one season. The widespread use of chemical controls has led to concerns about insecticide resistance, and resistant field populations have already been documented in California and Michigan., Results: We cultured sub-populations of three different Minnesota field populations of D. suzukii in the laboratory and exposed them to increasing concentrations of two commonly-used insecticides, zeta-cypermethrin (pyrethroid) and spinetoram (spinosyn). Over the exposure period, the sub-populations experienced an 8- to 45-fold increase in insecticide concentration. We saw significant increases in the median lethal concentration (LC 50 ) values of one sub-population exposed to zeta-cypermethrin and one exposed to spinetoram. Across the spinetoram exposures, we also observed significant reductions in the top mortality values for three different sub-populations., Conclusion: Our results suggest that field populations of D. suzukii can develop resistance to zeta-cypermethrin and spinetoram in short periods of time under laboratory selection but that resistance to spinosyns occurs more readily than to pyrethroids. These results support other studies that have documented spinosyn resistance in field populations and in laboratory selections. Resistance evolution to spinosyns is a particularly important issue, as they represent one of few organic insecticide options for D. suzukii. © 2022 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry., (© 2022 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.)

Over the last decade, numerous companies have marketed aqueous ozone sprayers for insect and disease management, but little to no data has been published on their efficacy. Thus, we evaluated the potential of both gaseous and aqueous ozone as a potential preharvest insecticide against the adult life stage of the invasive fruit pest, spotted-wing drosophila, Drosophila suzukii (Matsumura). Gaseous ozone was applied at two dosages, 14,600 and 30,100 ppmv, for varying durations and the respective concentration-time (CT) exposure responses were modeled for sex-specific mortalities recorded at 0, 24, 48, and 72 h following treatment. We found that gaseous ozone primarily caused mortality immediately following exposure, with slight increases 72 h following ozone treatments. The female and male lethal concentration-time (LCT) 50 estimates were significantly different at 0, 24, 48, and 72 h after 30,100 ppmv treatments, where males observed an increased mortality response. However, the LCT 99 estimates confidence intervals (95%) of adult female and male D. suzukii were similar at 0, 24, 48, and 72 h after 14,600 or 30,100 ppmv ozone treatments. In contrast, ozone dissolved in distilled water at 18.52 ppm (mg/L) did not provide any mortality after total immersion of subjects for 30 s. While gaseous ozone may have some utility as a fumigant for D. suzukii in closed vessels where concentrations could be maintained, we did not identify any insecticidal potential for ozone dissolved in aqueous solution when simulating a preharvest treatment under optimal laboratory conditions., (© The Author(s) 2022. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Many insect species exhibit basal social behaviors such as aggregation, which play important roles in their feeding and mating ecologies. However, the evolutionary, genetic, and physiological mechanisms that regulate insect aggregation remain unknown for most species. Here, we used natural populations of Drosophila melanogaster to identify the genetic architecture that drives larval aggregation feeding behavior. By using quantitative and reverse genetic approaches, we have identified a complex neurogenetic network that plays a role in regulating the decision of larvae to feed in either solitude or as a group. Results from single gene, RNAi-knockdown experiments show that several of the identified genes represent key nodes in the genetic network that determines the level of aggregation while feeding. Furthermore, we show that a single non-coding variant in the gene CG14205 , a putative acyltransferase, is associated with both decreased mRNA expression and increased aggregate formation, which suggests that it has a specific role in inhibiting aggregation behavior. Our results identify, for the first time, the genetic components which interact to regulate naturally occurring levels of aggregation in D. melanogaster larvae.

The invasive insect pest Drosophila suzukii causes extensive damage to soft-skinned fruit crops as they ripen. Current control methods involve the application of chemical pesticides, but this approach is ineffective and environmentally hazardous. To investigate the potential of bacterial pathogens carried by D. suzukii as biocontrol agents, we characterized bacteria associated with D. suzukii larvae in two parts of Hesse, Germany, by collecting infested fruits and culturing individual bacteria from moribund specimens for taxonomic classification by 16S rDNA sequencing. Among the bacteria we detected, some had a detrimental effect on the host whereas others were neutral or beneficial. When the detrimental and beneficial bacteria were presented simultaneously, we observed complex tripartite interactions that modulated the insect's innate immune response. Our study provides insight into the complex relationships within the microbiome and pathobiome of D. suzukii and may lead to the isolation of bacteria that can be used as biological control agents., (Copyright © 2020. Published by Elsevier Inc.)

The biochemical and molecular mechanisms underlying insect cold acclimation prior to cold stress are relatively well explored, but the mechanisms linked to recovery and repair after cold stress have received much less attention. Here we focus on recovery from cold stress in the larvae of the vinegar fly (Drosophila melanogaster) that were exposed to two physiologically distinct cold stress situations: supercooling (S, survival > 95%) and freezing (F, survival < 10%), both at -5 °C. We analysed the metabolic and transcriptomic responses to cold stress via GC-MS/LC-MS and whole-genome microarrays, respectively. Both stresses (S and F) caused metabolic perturbations which were transient in supercooled larvae but deeper and irreversible in frozen larvae. Differential gene expression analysis revealed a clear disparity in responses to supercooling and freezing (less than 10% of DE genes overlapped between S and F larvae). Using GO term enrichment analysis and KEGG pathway mapping, we identified the stimulation of immune response pathways as a strong candidate mechanism for coping with supercooling. Supercooling caused complex transcriptional activation of innate immunity potential: from Lysozyme-mediated degradation of bacterial cell walls, recognition of pathogen signals, through phagocytosis and lysosomal degradation, Toll and Imd signaling, to upregulation of genes coding for different antimicrobial peptides. The transcriptomic response to freezing was instead dominated by degradation of macromolecules and death-related processes such as autophagy and apoptosis. Of the 45 upregulated DE genes overlapping in responses to supercooling and freezing, 26 were broadly ascribable to defense and repair functions., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The ability to detect and identify substances based on the volatile compounds (odors) they emit is relied upon heavily for numerous investigative purposes. Animals have an innate olfactory sensitivity and selectivity that out-performs current instrumentation. This has led to immense interest in their employment as chemical sensors for a range of applications, including forensic science, both as whole organisms and as sensing elements in biosensors. Using electrophysiological and calcium imaging assays, this research examined the response of Drosophila melanogaster olfactory receptors (ORs) to odor compounds significant in forensic science and assessed their potential utility as volatile compound sensors. This investigation illustrated the different sensitivities, selectivities, and sensing features of individual ORs and demonstrated that their employment for detection purposes is feasible. While further research expanding on this study will be required to demonstrate the performance characteristics that an OR-based detection system will ultimately possess, this research provides an encouraging first step towards the goal of utilizing isolated biological ORs as volatile compound sensors in forensic science.

The sense of smell influences many behaviors, yet how odors are represented in the brain remains unclear. A major challenge to studying olfaction is the lack of methods allowing activation of specific types of olfactory neurons in an ethologically relevant setting. To address this, we developed a genetic method in Drosophila called olfactogenetics in which a narrowly tuned odorant receptor, Or56a, is ectopically expressed in different olfactory neuron types. Stimulation with geosmin (the only known Or56a ligand) in an Or56a mutant background leads to specific activation of only target olfactory neuron types. We used this approach to identify olfactory sensory neurons (OSNs) that directly guide oviposition decisions. We identify 5 OSN-types (Or71a, Or47b, Or49a, Or67b, and Or7a) that, when activated alone, suppress oviposition. Projection neurons partnering with these OSNs share a region of innervation in the lateral horn, suggesting that oviposition site selection might be encoded in this brain region., (Copyright © 2018 The Author(s). Published by Elsevier Inc. All rights reserved.)

Despite the comprehensive knowledge on odor coding, our understanding of the relationship between sensory input and behavioral output in Drosophila remains weak. Here, we measure the behavioral responses generated by larval and adult flies in response to 34 fruit odors and find that larval preference for fruit odors differs from that of adult flies. Next, we provide a functional analysis of the full repertoire of the peripheral olfactory system using the same comprehensive stimulus spectrum. We find that 90% and 53% of larval and adult olfactory receptors tested here, respectively, are involved in evaluating these fruit odors. Finally, we find that the total amount of olfactory neuronal activity correlates strongly positively with behavioral output in larvae and correlates weakly negatively in adult flies. Our results suggest that larval and adult flies have evolved different mechanisms for detection and computation of fruit odors, mechanisms likely mirroring the different lifestyles of both developmental stages., (Copyright © 2018 The Author(s). Published by Elsevier Inc. All rights reserved.)

Members of the natural resistance-associated macrophage protein (NRAMP) family are evolutionarily conserved metal ion transporters that play an essential role in regulating intracellular divalent cation homeostasis in both prokaryotes and eukaryotes. Malvolio (Mvl), the sole NRAMP family member in insects, plays a role in food choice behaviors in Drosophila and other species. However, the specific physiological and cellular processes that require the action of Mvl for appropriate feeding decisions remain elusive. Here, we show that normal food choice requires Mvl function specifically in the dopaminergic system, and can be rescued by supplementing food with manganese. Collectively, our data indicate that the action of the Mvl transporter affects food choice behavior via the regulation of dopaminergic innervation of the mushroom bodies, a principle brain region associated with decision-making in insects. Our studies suggest that the homeostatic regulation of the intraneuronal levels of divalent cations plays an important role in the development and function of the dopaminergic system and associated behaviors., (© 2017 John Wiley & Sons Ltd and International Behavioural and Neural Genetics Society.)

Conspicuously colored dendrobatid frogs sequester alkaloid defenses from dietary arthropods, resulting in considerable alkaloid variation among populations; however, little is known about how variation is perceived as a defense against predators. Previous studies have found variable alkaloids in the dendrobatid Oophaga pumilio to be associated with differences in toxicity to laboratory mice, suggesting variable defenses are important. Arthropods are natural predators that use chemoreception to detect prey, including frogs, and may therefore perceive variation in alkaloid profiles as differences in palatability. The goal of the present study is to determine how arthropods respond to variable alkaloid defenses in O. pumilio. Frog alkaloids were sampled from individual O. pumilio from ten geographic locations throughout the Bocas del Toro region of Panama and the Caribbean coast of Costa Rica. Alkaloid extracts were used in feeding bioassays with the vinegar fly Drosophila melanogaster and the ant Ectatomma ruidum. Both species of arthropods fed significantly less on frog alkaloid extracts when compared to controls, and differences in alkaloid palatability were observed among frog populations, as well as between sexes and life stages within a population. Differences in alkaloid quantity, richness, and type were the main predictors of arthropod palatability. Our findings also represent the first direct evidence of a palatability spectrum in a vertebrate that sequesters chemical defenses from dietary sources. Further, the presence of a palatability spectrum suggests that variable alkaloid defenses in O. pumilio are ecologically relevant and play an important role in natural predator-prey interactions, particularly with respect to arthropod predators.