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Bactrocera dorsalis is one of the major invasive pests worldwide. The acetic acid-enriched sweet bait trapping is an important method for monitoring and controlling this fly. Several studies showed that acetic acid is perceived by ionotropic receptors (IRs). Thus, we annotated 65 IR genes in the B. dorsalis genome. We also investigated the IRs involved in acetic acid perception in this fly by behavioral, electrophysiological, and molecular methods. As the results indicated, the antennae are the main olfactory organs to sense acetic acid. Among the antennal IRs showed acetic acid-induced expression profiles, IR8a was proven to perceive acetic acid by CRISPR/Cas9-mediated mutagenesis. Additionally, calcium imaging showed that IR64a and IR75a are potential acetic acid receptors respectively co-expressed with IR76b and IR8a. This study represents the first comprehensive characterization of IRs in B. dorsalis at the whole-genome level, revealing the significant role of IRs in acetic acid perception.

Biotic and abiotic factors influence how insects respond to stimuli. This can make it challenging to interpret captures in traps used to monitor pest abundance in management programmes. To address this, the lure response of three pest fruit flies (Diptera: Tephritidae) was evaluated in a semi-field setting with respect to several physiological and environmental factors. Using standardised methods with known fly numbers in field cages, the response to Biolure (food-based lure) was evaluated for Ceratitis capitata, Ceratitis cosyra and Bactrocera dorsalis. Response to the male lures was tested: E.G.O PheroLure for C. capitata and C. cosyra, Trimedlure for C. capitata, and methyl eugenol for B. dorsalis. The physiological variables evaluated were fly age, sex, weight, and total body nutritional composition. The environmental effects of temperature, relative humidity and light intensity were also assessed. Protein-deprived adults responded more strongly to Biolure. The response to Biolure was not sex-specific. Fly age influenced the response of all species to all tested lures. However, this effect was species and lure specific. Temperature was the most influential environmental factor, with response generally increasing with temperature. Lower thresholds for lure response, despite the proximity of responsive flies, range from 12.21 to 22.95 °C depending on the species and lure tested. These results indicate that trapping systems and management activity thresholds must take physiological and environmental variation into account to increase their accuracy., (© 2024. The Author(s).)

Plants perceive and orchestrate defense responses when herbivorous insects are ovipositing. Fruits, as a crucial reproductive organ in plants, have rarely been researched on the responses to insect eggs. Here, we found that oviposition by the specialist insect Bactrocera minax in navel oranges activated the lignin synthesis pathway and cell division, causing mechanical pressure that crushed the eggs. Transcriptome and metabolome analyses revealed an enrichment of oviposition-induced genes and metabolites within the lignin synthesis pathway, which was confirmed by histochemical staining. Furthermore, hydrogen peroxide (H 2 O 2 ) accumulation was observed at the oviposition sites. Plant defense-related hormones jasmonic acid (JA) and salicylic acid (SA) exhibited rapid induction after oviposition, while indole-3-acetic acid (IAA) activation occurred in the later stages of oviposition. Additionally, secondary metabolites induced by prior egg deposition were found to influence larval performance. Our studies provide molecular evidence that host fruits have evolved defense mechanisms against insect eggs and pave the way for future development of insect-resistant citrus varieties.

Mating and the transfer of seminal fluid components including male accessory glands (MAGs) proteins can affect oviposition behavior in insects. After oviposition, some species of fruit flies deposit a host-marking pheromone (HMP) on the fruit that discourages oviposition by other females of the same or different species or genus and reduces competition between larvae. However, we know very little about how mating, receiving seminal fluid, or male condition can affect female host marking behavior. Here, we tested how the physiological state of females (mated or unmated), the receipt of seminal fluid, and the condition of the male (wild or sterile) affect oviposition and host-marking behavior (HMB) in Anastrepha ludens (Diptera: Tephritidae). We also determined the efficiency of the host-marking pheromone from mated or unmated females in deterring oviposition. In a further examination of how seminal fluid may be affecting HMB we assessed if there were differences in the size of wild or sterile MAGs and the protein quantity transferred during mating. Our results indicate that receiving seminal fluid increased egg laying and increased time invested in host-marking (HM). Unmated females laid fewer eggs than mated females but invested the same amount of time in depositing host-marking pheromone, which had similar effectiveness in deterring oviposition as that of mated females. Females that mated with sterile males laid the same number of eggs as females that mated with wild males but spent less time depositing host-marking pheromone, which suggests that females detect the condition of the male and invest less in marking hosts. Finally, sterile males had larger accessory glands and transferred more MAGs proteins during mating compared to wild males. Seminal proteins could be manipulating HM behavior and female investment into their current reproductive effort. We are only beginning to understand how male condition and seminal fluid can affect female physiology and maternal investment in HMP., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Background: An innovative version of the sterile insect technique (SIT) for pest control, called boosted SIT, relies on the use of sterile males coated with a biocide to control a target wild pest population of the same species. The objective of the present study was to assess the relevance of such technology to control the fruit fly Bactrocera dorsalis and fruit losses in mango orchards using. An agent-based simulation model named BOOSTIT was used to explore the reduction of fruit losses thank to sterile male fruit flies control and economic benefits according to different strategies of sterile male release. The simulation considered a landscape of 30.25 ha made up of four mango orchards., Results: The SIT and the boosted SIT reduced fruit losses when releases were made before the mango fruiting period. According to model simulations, releases should be performed at least seven times at 2-week intervals and with a sterile/wild male ratio of at least 10:1. Considering the benefit/cost ratio (BCR), few releases should be done with a late start date. The BCR showed economic gains from the two control methods, the number of saved fruits and BCR being higher for SIT., Conclusion: Our simulations showed that SIT would have better results than the boosted SIT to contribute to an effective control of Bactrocera dorsalis at the scale of a small landscape. We highlight the need for laboratory studies of other types of pathogen to find a suitable one with higher incubation time and lower cost. © 2024 The Author(s). Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry., (© 2024 The Author(s). Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.)

Background: The olive fruit fly (OFF), Bactrocera oleae (Rossi), is the main insect pest of olive trees worldwide. Legislation limits to the use of some synthetic larvicidal insecticides is leading to the development of new control options for preventive control of adult flies. In the present study, the biological activity of four short-chain aliphatic aldehydes, namely hexanal, (E)-2-hexenal, heptanal and (E)-2-heptenal, previously reported as repellents to the OFF adults was investigated., Results: Electroantennography (EAG) recordings showed that antennae of OFF males and females are able to perceive the test compounds in a wide range of doses. In field trapping experiments, reservoir-type polypropylene (PP) membrane dispensers loaded with individual compounds did not elicit a significant attraction of OFF males and females. On the contrary, a significant reduction of male catches was noticed when sex pheromone dispensers and PP membrane dispensers, loaded with one of the test compounds, were applied on the same white sticky traps ≈20 cm apart. Likewise, male and female catches in yellow sticky traps baited with ammonium carbonate (AC) dispensers as food attractant were significantly reduced by the presence of PP membrane dispensers of individual aliphatic aldehydes on the same traps. In small plots control trials, solid formulations of the four aldehydes into a bentonite clay support induced a significant reduction of the OFF active infestation mainly when C 6 and C 7 aldehyde-activated bentonites were used., Conclusion: Short-chain aliphatic aldehydes showed inhibitory effects on sex pheromone and food attractant-mediated attraction of OFF. Results of field trials suggest potential of short-chain aliphatic aldehydes to develop new semiochemical-based OFF control options. © 2024 Society of Chemical Industry., (© 2024 Society of Chemical Industry.)

Effective pest population monitoring is crucial in precision agriculture, which integrates various technologies and data analysis techniques for enhanced decision-making. This study introduces a novel approach for monitoring lures in traps targeting the Mediterranean fruit fly, utilizing air quality sensors to detect total volatile organic compounds (TVOC) and equivalent carbon dioxide (eCO 2 ). Our results indicate that air quality sensors, specifically the SGP30 and ENS160 models, can reliably detect the presence of lures, reducing the need for frequent physical trap inspections and associated maintenance costs. The ENS160 sensor demonstrated superior performance, with stable detection capabilities at a predefined distance from the lure, suggesting its potential for integration into smart trap designs. This is the first study to apply TVOC and eCO 2 sensors in this context, paving the way for more efficient and cost-effective pest monitoring solutions in smart agriculture environments.

Background: The melon fly, Zeugodacus cucurbitae (Coquillett), is an invasive Tephritidae pest with robust fertility. The male accessory glands (MAGs) form a vital organ that ensures insect reproductive efficiency. Most of the secreted proteins by MAGs exhibit a male bias expression. Takeout, one of these proteins, is abundantly present in the MAGs of many insects., Results: In this study, we identified 32 takeout genes in Z. cucurbitae. The phylogenetic analysis and multiple sequence alignment results showed that Zctakeout1 is the most related homolog to the MAGs-specific takeout in Tephritidae. The real-time quantitative PCR results showed that Zctakeout1 was exclusively expressed in the male adult stage, and its expression level gradually increased with the increase in age and then remained stable at the sexually matured stage. The distribution among tissues demonstrated the specific expression of Zctakeout1 in the MAGs, and fluorescence immunohistochemical results confirmed the presence of Zctakeout1 in close proximity to binuclear cells of the mesoderm epidermal MAGs. In continuation, CRISPR/Cas9-mediated genome editing was employed, resulting in successfully generating a homozygous strain with an +8 bp insertion. The mating experiments with the Zctakeout1 -/- males resulted in significant reductions in both the mating rate and egg production of females., Conclusion: These findings prove that the MAGs-specific Zctakeout1 is essential in regulating fecundity in female Z. cucurbitae fruit flies. Our data suggests its utilization in future essential insect-specific gene-directed sterility insect technique (SIT) by the genetic manipulation to keep these important Tephritidae populations under control. © 2024 Society of Chemical Industry., (© 2024 Society of Chemical Industry.)

In Bactrocera dorsalis , both males and females release chemical signals to attract mates. In our previous study, we identified ethyl laurate, ethyl myristate, and ethyl palmitate as potent female-derived pheromones that contribute to mate attraction. However, the mechanisms underlying the olfactory recognition remain unclear. In this study, we observed strong antennal and behavioral responses in male B. dorsalis to these female-derived pheromones, and further investigation revealed significant upregulation of OBP49a and OBP83b following exposure to these compounds. Through fluorescence competitive binding assays and RNA interference techniques, we demonstrated the crucial roles of OBP49a and OBP83b in detecting female-derived pheromones. Finally, molecular docking analysis identified key residues, including His134 in OBP83b and a lysine residue in OBP49a, which formed hydrogen bonds with female-derived pheromones, facilitating their binding. These findings not only advance our understanding of olfactory recognition of pheromones in B. dorsalis but also offer potential targets for developing olfaction-interfering techniques for pest control.

Confronting environments with low relative humidity is one of the main challenges faced by insects with expanding distribution ranges. Anastrepha ludens (the Mexican fruit fly) has evolved to cope with the variable conditions encountered during its lifetime, which allows it to colonise a wide range of environments. However, our understanding of the mechanisms underpinning the ability of this species to confront environments with low relative humidity is incomplete. In this sense, omic approaches such as transcriptomics can be helpful for advancing our knowledge on how this species copes with desiccation stress. Considering this, in this study, we performed transcriptomic analyses to compare the molecular responses of laboratory-reared A. ludens exposed and unexposed to desiccation. Data from the transcriptome analyses indicated that the responses to desiccation are shared by both sexes. We identified the up-regulation of transcripts encoding proteins involved in lipid metabolism and membrane remodelling, as well as proteases and cuticular proteins. Our results provide a framework for understanding the response to desiccation stress in one of the most invasive fruit fly species in the world.

Two fruit fly (Diptera: Tephritidae) species of economic importance: Ceratitis rosa Karsch and Ceratitis quilicii De Meyer, Mwatawala & Virgilio are present in South Africa. The two species were considered as one species prior to 2016, but were subsequently separated. In this study, the distribution and abundance of the two species were quantified in seven provinces in South Africa through trapping with Enriched Ginger Oil as an attractant. Trapping was conducted over three seasons across two years (2020 and 2021): late summer, autumn-winter, and spring-early summer. Host ranges of the two species were investigated by fruit sampling in and outside of trapping sites. Ceratitis quilicii was more widely distributed than C. rosa with the latter being recorded in only three north-eastern provinces. There were geographical limits for both species with no records of them in Northern Cape Province. Catches of C. quilicii were higher in summer with average temperatures varying from 15 to 27°C while for C. rosa , catches remained low and consistent between seasons. Ceratitis quilicii catches decreased at lower rates than those of C. rosa at temperatures below 15°C. The two species were reared from 13 plant species from nine families. Four of these hosts were infested by both C. quilicii and C. rosa in the same province where they occurred. Preferred hosts of the two species belonged to the Myrtaceae family. The characterisation of the distribution, abundance and host ranges of these pests will provide a baseline for pest status determination and implementation of management actions.

Determining movement parameters for pest insects such as tephritid fruit flies is critical to developing models which can be used to increase the effectiveness of surveillance and control strategies. In this study, harmonic radar was used to track wild-caught male Queensland fruit flies (Qflies), Bactrocera tryoni, in papaya fields. Experiment 1 continuously tracked single flies which were prodded to induce movement. Qfly movements from this experiment showed greater mean squared displacement than predicted by both a simple random walk (RW) or a correlated random walk (CRW) model, suggesting that movement parameters derived from the entire data set do not adequately describe the movement of individual Qfly at all spatial scales or for all behavioral states. This conclusion is supported by both fractal and hidden Markov model (HMM) analysis. Lower fractal dimensions (straighter movement paths) were observed at larger spatial scales (> 2.5 m) suggesting that Qflies have qualitatively distinct movement at different scales. Further, a two-state HMM fit the observed movement data better than the CRW or RW models. Experiment 2 identified individual landing locations, twice a day, for groups of released Qflies, demonstrating that flies could be tracked over longer periods of time., (© 2024. This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply.)

Invasive fruit flies (Diptera: Tephritidae) pose a serious threat to the production and export of many commercially important fruits and vegetables. Detection of the agricultural pests Bactrocera dorsalis (Hendel) and Zeugodacus cucurbitae (Coquillett) relies heavily on traps baited with male-specific attractants. For B. dorsalis, traps are typically baited with liquid methyl eugenol (ME), and for Z. cucurbitae, traps are baited with liquid cue-lure (CL). Operating large-scale trapping networks is costly, consequently, there is much interest in identifying ways to maintain network sensitivity while reducing costs. One cost-cutting approach is the possibility of combining different male lures in the same dispenser, thus reducing the number of traps requiring servicing. The chief objective of this study was to compare captures of B. dorsalis and Z. cucurbitae males in Jackson traps baited with polymeric wafers impregnated with both ME and raspberry ketone (RK, a hydrolyzed form of CL) versus traps baited with liquid ME or CL freshly applied to cotton wicks. Captures were measured when the ME/RK wafers had been weathered for 12, 18, or 24 wk. Captures of B. dorsalis and Z. cucurbitae males were similar between fresh lure and weathered wafers over all trapping periods, with a single exception apparently due to the lessened potency of the associated killing agent. The residual amount and release rate of ME and RK from the wafers were also measured to examine possible relationships between wafer chemistry and trap catch. The possible implications of the present results to area-wide trapping programs are discussed., (Published by Oxford University Press on behalf of Entomological Society of America 2024.)

Background: The potential of copper nanoparticles (Cu-NPs) to be used as an alternative control strategy against olive fruit flies (Bactrocera oleae) with reduced sensitivity to the pyrethroid deltamethrin and the impact of both nanosized and bulk copper hydroxide (Cu(OH) 2 ) on the insect's reproductive and endosymbiotic parameters were investigated., Results: The application of nanosized and bulk copper applied by feeding resulted in significant levels of adult mortality, comparable to or surpassing those achieved with deltamethrin at recommended doses. Combinations of Cu-NPs or copper oxide nanoparticles (CuO-NPs) with deltamethrin significantly enhanced the insecticide's efficacy against B. oleae adults. When combined with deltamethrin, Cu-NPs significantly reduced the mean total number of offspring compared with the control, and the number of stings, pupae, female and total number of offspring compared with the insecticide alone. Both bulk and nanosized copper negatively affected the abundance of the endosymbiotic bacterium Candidatus Erwinia dacicola which is crucial for the survival of B. oleae larvae., Conclusion: The Cu-NPs can aid the control of B. oleae both by reducing larval survival and by enhancing deltamethrin performance in terms of toxicity and reduced fecundity, providing an effective anti-resistance tool and minimizing the environmental footprint of synthetic pesticides by reducing the required doses for the control of the pest. © 2024 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry., (© 2024 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.)

Implementation of augmentative biological control requires estimates of parasitoid dispersal from the release point to determine appropriate release density, spacing, and timing. This study evaluated the movement patterns of Fopius arisanus Sonan (Hymenoptera: Braconidae) parasitoids, which have historically been used to control invasive tephritid fruit flies. The wasps were released from the central point, and dispersal was monitored over time using parasitism in sentinel fruit and trap captures at 40 points radiating out from the center (15-240 m). The releases were conducted 4 times during June, July, September, and November 2006. The data showed that there were large declines in dispersal by distance. Parasitism was greatest closest to the release point, within 30 m. Parasitism was also greatest within the first 24 h of the release. After 1 wk, parasitism decreased from 41% to 1.5% within 30 m. These data correlated strongly with trap capture data, which also showed that parasitoid movement favored the SE region of our release site, roughly corresponding to the overall prevailing winds. Wind speed, relative humidity, and temperature all affected parasitoid movement during our trial, indicating the complex environmental factors that can affect release success. This is the first report of dispersal metrics for F. arisanus. Our findings are in agreement with other similar studies on braconid movement generally and suggest that frequent, high-density releases are most effective since the dispersal of F. arisanus is limited and retention in the environment is low. We discuss our results in the context of international augmentative biological control release programs., (Published by Oxford University Press on behalf of Entomological Society of America 2024.)

Insects rely heavily on their olfactory system for various behaviors, including foraging, mating, and oviposition. Numerous studies have demonstrated that insects can adjust their olfactory behaviors in response to different physiological states and environmental conditions. This flexibility allows them to perceive and process odorants according to different conditions. The Oriental fruit fly, Bactrocera dorsalis, is a highly destructive and invasive pest causing significant economic losses to fruit and vegetable crops worldwide. The olfactory behavior of B. dorsalis exhibits strong plasticity, resulting in its successful invasion. To enhance our understanding of B. dorsalis' olfactory behavior and explore potential strategies for behavior control, we have reviewed recent literature on its olfactory plasticity and potential molecular mechanisms., Competing Interests: Declaration of Competing Interest The authors declare no competing interests existed., (Copyright © 2024 Elsevier Inc. All rights reserved.)

AbstractPhysiological time is important for understanding the development and seasonal timing of ectothermic animals but has largely been applied to developmental processes that occur during spring and summer, such as morphogenesis. There is a substantial knowledge gap in the relationship between temperature and development during winter, a season that is increasingly impacted by climate change. Most temperate insects overwinter in diapause, a developmental process with little obvious morphological change. We used principles from the physiological time literature to measure and model the thermal sensitivity of diapause development rate in the apple maggot fly Rhagoletis pomonella , a univoltine fly whose diapause duration varies substantially within and among populations. We show that diapause duration can be predicted by modeling a relationship between temperature and development rate that is shifted toward lower temperatures compared with typical models of morphogenic, nondiapause development. However, incorporating interindividual variation and ontogenetic variation in the temperature-to-development rate relationship was critical for accurately predicting fly emergence, as diapause development proceeded more quickly at high temperatures later in diapause. We conclude that the conceptual framework may be flexibly applied to other insects and discuss possible mechanisms of diapause timers and implications for phenology with warming winters.

Photic entrainment is an essential function of the circadian clock, which enables organisms to set the appropriate timing of daily behavioral and physiological events. Recent studies have shown that the mechanisms of the circadian clock and photic entrainment vary among insect species. This study aimed to elucidate the circadian photoreceptors necessary for photic entrainment in firebrats Thermobia domestica, one of the most primitive apterygote insects. A homology search of publicly available RNA sequence (RNA-seq) data from T. domestica exhibited a cryptochrome 2 (cry2) gene and three opsin genes, opsin long wavelength 1 (opLW1), opLW2, and opUV, as candidate circadian photoreceptors. We examined the possible involvement of these genes in photic entrainment of firebrat locomotor rhythms. Firebrats had the highest entrainability to the light-dark cycle of green light. Treatment with dsRNA of the candidate genes strongly downregulated the respective targeted genes, and in the case of opsin genes, other untargeted genes were occasionally downregulated to various degrees. Under constant light, most control firebrats became arrhythmic, whereas a fraction of those treated with double RNAi of the two opLWs remained rhythmic. Behavioral experiments revealed that the transient cycles necessary for re-entrainment to shifted light cycles were lengthened when opLW2 expression was reduced. These results suggest that opLW2 is involved in the photic entrainment of circadian rhythm in firebrats., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Background: Gut bacteria, which serve as essential modulators, exert a significant impact on insect physiology and behavior and have substantial application potential in pest management. The dynamics of gut bacteria and their impact on Phortica okadai behavior remain unclear., Methods: In this study, the dynamics of gut bacteria at different developmental stages in P. okadai were analyzed using 16S ribosomal RNA (rRNA) gene sequencing, and the species and abundance of gut bacteria that affect host behavior were examined via behavioral experiments., Results: A total of 19 phyla, 29 classes, 74 orders, 101 species, and 169 genera were identified. The results of the behavioral experiments indicated that the species Lactiplantibacillus argentoratensis, Acetobacter tropicalis, Leuconostoc citreum, and Levilactobacillus brevis effectively influenced the feeding preference of P. okadai, and the single-bacterium-seeded P. okadai exhibited feeding preferences distinct from those of the germ-free (GF) and wild-type P. okadai., Conclusions: The species and relative abundance of gut bacteria together positively impact P. okadai behavior. Lactiplantibacillus argentoratensis, as the most attractive bacteria to P. okadai, presents opportunities for novel pest control strategies targeting this vector and agricultural pest., (© 2024. The Author(s).)

Bactrocera tryoni (Froggatt) and Bactrocera neohumeralis (Hardy) are sibling fruit fly species that are sympatric over much of their ranges. Premating isolation of these close relatives is thought to be maintained in part by allochrony-mating activity in B. tryoni peaks at dusk, whereas in B. neohumeralis, it peaks earlier in the day. To ascertain whether differences in pheromone composition may also contribute to premating isolation between them, this study used solid-phase microextraction and gas chromatography-mass spectrometry to characterize the rectal gland volatiles of a recently collected and a more domesticated strain of each species. These glands are typical production sites and reservoirs of pheromones in bactrocerans. A total of 120 peaks were detected and 50 were identified. Differences were found in the composition of the rectal gland emissions between the sexes, species, and recently collected versus domesticated strains of each species. The compositional variation included several presence/absence and many quantitative differences. Species and strain differences in males included several relatively small alcohols, esters, and aliphatic amides. Species and strain differences in females also included some of the amides but additionally involved many fatty acid esters and 3 spiroacetals. While the strain differences indicate there is also heritable variation in rectal gland emissions within each species, the species differences imply that compositional differences in pheromones emitted from rectal glands could contribute to the premating isolation between B. tryoni and B. neohumeralis. The changes during domestication could also have significant implications for the efficacy of Sterile Insect Technique control programs., (© The Author(s) 2024. Published by Oxford University Press on behalf of Entomological Society of America.)

The maxillary palp is an auxiliary olfactory organ in insects, which, different from the antennae, is equipped with only a few olfactory sensory neuron (OSN) types. We postulated that these derived mouthpart structures, positioned at the base of the proboscis, may be particularly important in mediating feeding behaviors. As feeding is spatio-temporally segregated from oviposition in most Tephritidae, this taxonomic group appears quite suitable to parse out sensory breadth and potential functional divergence of palps and antennae. Scanning electron microscopy and anterograde staining underlined the limited palpal olfactory circuit in Tephritidae: only three morphological subtypes of basiconic sensilla were found, each with two neurons, and project to a total of six antennal lobe glomeruli in Bactrocera dorsalis. Accordingly, the palps detected only few volatiles from the headspace of food (fermentation and protein lures) and fruit (guava and mango) compared to the antennae (17 over 77, using gas-chromatography coupled electrophysiology). Interestingly, functionally the antennae were more tuned to fruit volatiles, detecting eight times more fruit than food volatiles (63 over 8), whereas the number of fruit and food volatile detection was more comparable in the palps (14 over 8). As tephritids diverge in oviposition preferences, but converge on food substrates, we postulated that the receptive ranges of palpal circuits would be more conserved compared to the antennae. However, palpal responses of three tephritid species that differed in phylogenetic relatedness and ecologically niche, diverged across ecological rather than phylogenetic rifts. Two species with strongly overlapping ecology, B. dorsalis and Ceratitis capitata, showed inseparable response profiles, whereas the cucurbit specialist Zeugodacus cucurbitae strongly diverged. As Z. cucurbitae is phylogenetically placed between B. dorsalis and C. capitata, the results indicate that ecology overrides phylogeny in the evolution of palpal tuning, in spite of being predisposed to detecting food volatiles., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

This study was carried out in 3 types of biotopes where vegetable crops are not grown to highlight their contribution to the dynamics of vegetable-infesting flies. To this end, a trapping system based on a sexual attractant, the Cuelure associated with an insecticide was set up in 18 biotopes (6 natural areas, 6 mango orchards, and 6 agroforestry parks) in the regions of Hauts Bassins and Cascades in the South-West of Burkina Faso. During the trapping monitoring, which was done every 2 wk to collect insects captured, fruits present in 3 types of biotopes were sampled and incubated for insect emergence. Ten Dacus (Fabricius) [Diptera: Tephritidae] species and Zeugodacus cucurbitae (Coquillett) [Diptera: Tephritidae] were trapped in the study area. The predominant species captured was Z. cucurbitae (52.93%) followed by Dacus punctatifrons (Karsch) [Diptera: Tephritidae] (29.89%) and Dacus humeralis (Bezzi) (12.71%). Six tephritid species were emerged from 6 wild fruit species belonging to Cucurbitaceae, Apocynaceae, and Passifloraceae families. Fruit flies were more abundant from Jul to Nov with peaks observed in Aug or Oct depending on the species. Citrullus colocynthis L. (Cucurbitaceae), Lagenaria sp. (Cucurbitaceae), Passiflora foetida L. (Passifloraceae), and Passiflora sp. acted as reservoir host plants of Dacus ciliatus (Loew), Dacus bivittatus (Bigot), Dacus vertebratus (Bezzi) [Diptera: Tephritidae], D. punctatifrons, and Z. cucurbitae, the major vegetable insect pests in West Africa. The 3 types of biotopes acted as suitable refuge areas of vegetable crop-infesting fruit flies either for the favorable microclimate or for the alternative host plants., (© The Author(s) 2024. Published by Oxford University Press on behalf of Entomological Society of America.)

The ability to recognize a host plant is crucial for insects to meet their nutritional needs and locate suitable sites for laying eggs. Bactrocera dorsalis is a highly destructive pest in fruit crops. Benzothiazole has been found to induce oviposition behavior in the gravid B. dorsalis . However, the ecological roles and the olfactory receptor responsible for benzothiazole are not yet fully understood. In this study, we found that adults were attracted to benzothiazole, which was an effective oviposition stimulant. In vitro experiments showed that BdorOR49b was narrowly tuned to benzothiazole. The electroantennogram results showed that knocking out BdorOR49b significantly reduced the antennal electrophysiological response to benzothiazole. Compared with wild-type flies, the attractiveness of benzothiazole to BdorOR49b knockout adult was significantly attenuated, and mutant females exhibited a severe decrease in oviposition behavior. Altogether, our work provides valuable insights into chemical communications and potential strategies for the control of this pest.

The oriental fruit fly, Bactrocera dorsalis (Hendel), is a notorious pest of fruit crops, causing severe damage to fleshy fruits during oviposition and larval feeding. Gravid females locate suitable oviposition sites by detecting the host volatiles. Here, the oviposition preference of antenna-removed females and the electrophysiological response of ovipositors to benzothiazole indicated that both antennae and ovipositors are involved in perceiving benzothiazole. Subsequently, odorant receptors (ORs) expressed in both antennae and ovipositors were screened, and BdorOR43a-1 was further identified to respond to benzothiazole using voltage-clamp recording. Furthermore, BdorOR43a-1 -/- mutants were obtained using the CRISPR/Cas9 system and their oviposition preference to benzothiazole was found to be significantly altered compared to WT females, suggesting that BdorOR43a-1 is one of the important ORs for benzothiazole perception. Our results not only demonstrate the important role of antennae and ovipositors in benzothiazole-induced oviposition but also elucidate on the OR responsible for benzothiazole perception in B. dorsalis .

Anastrepha fraterculus (Wiedemann) and A. obliqua (Macquart) are important pests of fruit crops. In Brazil, these species cause damage to fruit growing in the South (annual average temperature of 20.9 °C) and Northeast (average yearly temperature of 24 °C). We evaluated the effect of temperature on the viability and development time of A. fraterculus and A. obliqua immature stages in their respective preferred hosts, guava (Psidium guajava L., Myrtaceae) and mango (Mangifera indica L., Anacardiaceae). The duration of egg and pupal stages, egg to pre-pupa, and viability of egg and pupal stages under different temperatures (15, 20, 25, 30, and 35 °C) were assessed. For both species, development time decreased with increasing temperature. Viability in the evaluated stages was only observed between 15 and 30 °C. However, the species responded differently to the exposure temperatures (15 and 30 °C), especially in the pupal stage and from egg to pre-pupa. Anastrepha fraterculus showed a lower tolerance to high temperatures, especially in the pupal stage and from egg to pre-pupa, which may explain its lower importance and economic impact in warmer Brazilian regions. Anastrepha obliqua had a lower tolerance at 15 °C, indicating greater adequacy for temperatures above 20 °C, characteristic of Northeast Brazil, suggesting the capacity to spread to cooler areas with rising temperatures., (© 2023. Sociedade Entomológica do Brasil.)

Calling males of Anastrepha obliqua release volatile compounds to attract conspecific males to form leks and females to mate. Male volatiles from Mexican and Brazilian populations of A . obliqua have been previously identified. However, there are differences in the number and identity of volatile compounds between the populations. These differences in volatile profiles may be due to male origin (e.g. wild or mass-reared flies) or methodological issues (e.g. sampling techniques). In this study, we evaluated the attractiveness of wild, laboratory non-irradiated, and laboratory-irradiated flies under semi-field conditions. Male volatiles were collected using dynamic headspace sampling (DHS) and solid-phase microextraction (SPME) techniques, and identified using gas chromatography-coupled mass spectrometry. The results showed no difference in the attractiveness of wild, laboratory non-irradiated, and irradiated males to females. However, the number of captured females differed according to the origin; wild and non-irradiated females were captured more frequently than the irradiated flies. A total of 21 compounds were found using SPME, whereas only 12 were collected using DHS, although the relative amounts of these compounds were higher than those obtained using the former sampling technique. In addition, only laboratory non-irradiated males released α-pinene and menthol, which have not been previously reported in this fruit fly species. Additionally, we identified novel compounds in A . obliqua ; however, certain compounds previously reported were not detected. This study suggests that despite the qualitative and quantitative variations in the volatile profiles of A . obliqua males, their attractiveness was unaffected.

Olfaction plays indispensable roles in insect behavior such as host location, foraging, oviposition, and avoiding predators. Chemosensory proteins (CSPs) can discriminate the hydrophobic odorants and transfer them to the odorant receptors. Presently, CSPs have been identified in many insect species. However, their presence and functions remain unknown in Bactrocera dorsalis , a destructive and invasive insect pest in the fruit and vegetable industry. Here, we annotated eight CSP genes in the genome of B. dorsalis . The results of quantitative real-time polymerase chain reaction (RT-qPCR) showed that BdorCSP3 was highly expressed in the antennae. Molecular docking and in vitro binding assays showed that BdorCSP3 had a good binding ability to host volatiles methyl eugenol (ME, male-specific attractant) and β-caryophyllene (potential female attractant). Subsequently, CRISPR/Cas9 was used to generate BdorCSP3 -/- mutants. Electroantennograms (EAGs) and behavioral assays revealed that male mutants significantly reduced the preference for ME, while female mutants lost their oviposition preference to β-caryophyllene. Our data indicated that BdorCSP3 played important roles in the perception of ME and β-caryophyllene. The results not only expanded our knowledge of the olfaction perception mechanism of insect CSPs but also provided a potential molecular target for the control of B. dorsalis .

The carambola fruit fly Bactrocera carambolae Drew and Hancock (Diptera: Tephritidae) is an invasive fruit fly reported in North Brazil that threatens Brazilian fruit culture. Assessing the potential risk of establishing this pest is necessary to reduce the threat of B. carambolae dispersion to other countries and Brazilian regions and to avoid damage to the fruit trade. In this study, the CLIMEX model was used to understand the response of B. carambolae to climate change and to determine its potential global distribution with and without irrigation practices. Based on ecophysiological parameters, the model simulates factors limiting species distribution concerning the climate. To assess the seasonal variation in the density of B. carambolae, monitoring data in Uiramutã municipality, Roraima, from 2013 to 2019 was used. According to the CLIMEX forecast, large parts of America, Africa, and Asia, mainly in areas closest to the equator, are highly suitable for the survival of B. carambolae. Brazil is a good part of its territory with high suitability for B. carambolae, especially the North, South, and Southeast regions and the entire coastal area. The periods of the highest climatic suitability in the five Brazilian regions were January-May and October-December. The potential distribution area expands under irrigation and is highly suitable for most areas without cold stress. The CLIMEX model for B. carambolae generated in the present study provides important information for the Brazilian eradication program and other surveillance activities established in pest-free areas., (© 2023. Sociedade Entomológica do Brasil.)

The species distributions migration poleward and into higher altitudes in a warming climate is especially concerning for economically important insect pest species, as their introduction can potentially occur in places previously considered unsuitable for year-round survival. We explore the expansion of the climatically suitable areas for a horticultural pest, the Mediterranean fruit fly (medfly) Ceratitis capitata (Diptera, Tephritidae), with an emphasis on Europe and California. We reviewed and refined a published CLIMEX model for C. capitata, taking into consideration new records in marginal locations, with a particular focus on Europe. To assess the model fit and to aid in interpreting the meaning of the new European distribution records, we used a time series climate dataset to explore the temporal patterns of climate suitability for C. capitata from 1970 to 2019. At selected bellwether sites in Europe, we found statistically significant trends in increasing climate suitability, as well as a substantial northward expansion in the modelled potential range. In California, we also found a significant trend of northward and altitudinal expansion of areas suitable for C. capitata establishment. These results provide further evidence of climate change impacts on species distributions and the need for innovative responses to increased invasion threats., (© 2024. The Author(s).)

The striped fruit fly, Zeugodacus scutellata is a significant pest in East and Southeast Asia by damaging Cucurbitaceae blossoms and fruits. To control this pest, a novel strategy to suppress the gene(s) associated with sexually dimorphic phenotypes has been devised and implemented in a laboratory scale. However, comprehensive transcriptomic analysis related to this sex differentiation of Z. scutellata was necessary to determine effective target genes for the genetic control. We performed de novo assembly of the transcript obtained by paired-end sequencing using an Illumina HiSeq platform and let to 217,967 unigenes (i.e., unique genes) with a minimum length of 200 bp. The female produced 31, 604, 442 reads with 97.93% of Q20, 94.76% of Q30, and the male produced 130, 592, 828 reads with 97.93% of Q20 and 94.76 of Q30%. The differentially expressed genes were used to predict genetic factors associated with sex differentiation, which included Rho1, extra-macrochaetae (emc), hopscotch (hop), doublesex (dsx), sex-lethal (sxl), transformer-2 (tra-2), testis-specific serine/threonine-protein kinase (tssk1), tektin1 (tkt1) and 2 (tkt2), odorant binding proteins (OBPs), fruitless (fru), vitellogenin receptor, and hormone receptors in Z. scutellata. In addition, this transcriptome analysis provides the additional gene associated with sex determination and mating behaviors, which would be applied to develop a novel sterile insect technique against Z. scutellata., (© 2023 Wiley Periodicals LLC.)

Insect response to cold stress is often associated with adaptive strategies and chemical variation. However, low-temperature domestication to promote the cold tolerance potential of Bactrocera dorsalis and transformation of main internal substances are not clear. Here, we use a series of low-temperature exposure experiments, supercooling point (SCP) measurement, physiological substances and cryoprotectants detection to reveal that pre-cooling with milder low temperatures (5 and 10°C) for several hours (rapid cold hardening) and days (cold acclimation) can dramatically improve the survival rate of adults and pupae under an extremely low temperature (-6.5°C). Besides, the effect of rapid cold hardening for adults could be maintained even 4 h later with 25°C exposures, and SCP was significantly declined after cold acclimation. Furthermore, content of water, fat, protein, glycogen, sorbitol, glycerol and trehalose in bodies were measured. Results showed that water content was reduced and increased content of proteins, glycogen, glycerol and trehalose after two cold domestications. Our findings suggest that rapid cold hardening and cold acclimation could enhance cold tolerance of B. dorsalis by increasing proteins, glycerol, trehalose and decreasing water content. Conclusively, identifying a physiological variation will be useful for predicting the occurrence and migration trend of B. dorsalis populations.

Oviposition in a generalist parasitoid is a complex process that involves interactions among its host, host plant, and spatiotemporal environment. Our study was aimed at exploring the ability of a generalist parasitic wasp, Diachasmimorpha longicaudata (Ashmead) (Hymenoptera: Braconidae), in perceiving odor cues from its host insect [Oriental fruit fly Bactrocera dorsalis (Hendel, Diptera: Tephritidae)] in infested mango fruits. A series of experiments involving behavioral and electrophysiological studies suggest that D. longicaudata is able to distinguish host infested fruit volatiles from uninfested. Additionally, we found that D. longicaudata can distinguish between high and low-level infestations using specific cues emanating from fruits. Coupled gas chromatography-electroantennographic detection (GC-EAD) and gas chromatography-mass spectrometry (GC-MS) analysis of infested mango fruit head space volatiles identified compounds such as ethyl butanoate, ethyl 3-hydroxybutyrate, β-myrcene, trans-β-ocimene, allo-ocimene, ethyl octanoate, ethyl-α-toulate, β-elemene, β-caryophyllene, humulene, caryophyllene oxide, ethyl dodecanoate, ethyl hexadecanoate, 2-furancarboxylic acid, 3-hydroxy-2-methyl-4-pyrone, and phenethyl alcohol emanated from three different host treatments with different levels of larval infestation which elicited antennal responses in D. longicaudata. We suggest that these cues might play a significant role in attracting D. longicaudata in the field., (© The Author(s) 2023. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

The Mediterranean fruit fly, Ceratitis capitata (Diptera: Tephritidae), holds an impressive record of successful invasion events promoted by globalization in fruit trade and human mobility. In addition, C. capitata is gradually expanding its geographic distribution to cooler temperate areas of the Northern Hemisphere. Cold tolerance of C. capitata seems to be a crucial feature that promotes population establishment and hence invasion success. To elucidate the interplay between the invasion process in the northern hemisphere and cold tolerance of geographically isolated populations of C. capitata, we determined (a) the response to acute cold stress survival of adults, and (b) the supercooling capacity (SCP) of immature stages and adults. To assess the phenotypic plasticity in these populations, the effect of acclimation to low temperatures on acute cold stress survival in adults was also examined. The results revealed that survival after acute cold stress was positively related to low temperature acclimation, except for females originating from Thessaloniki (northern Greece). Adults from the warmer environment of South Arava (Israel) were less tolerant after acute cold stress compared with those from Heraklion (Crete, Greece) and Thessaloniki. Plastic responses to cold acclimation were population specific, with the South Arava population being more plastic compared to the two Greek populations. For SCP, the results revealed that there is little to no correlation between SCP and climate variables of the areas where C. capitata populations originated. SCP was much lower than the lowest temperature individuals are likely to experience in their respective habitats. These results set the stage for asking questions regarding the evolutionary adaptive processes that facilitate range expansions of C. capitata into cooler temperate areas of Europe., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Nikos T. Papadopoulos reports financial support was provided by the European Union’s Horizon 2020 Program for Research and Innovation the FF-IPM Project (HORIZON 2020, GA818184). The remaining authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Bactrocera dorsalis (Hendel) (Diptera: Tephritidae), is a major global pest of fruits. Currently, the sequential male annihilation technique, followed by the sterile insect technique has been used to significantly reduce the population of feral males in this species. However, issues with sterile males being killed by going to male annihilation traps have reduced the efficacy of this approach. The availability of males that are non-methyl eugenol-responding would minimize this issue and increase the efficacy of both approaches. For this, we recently established two separate lines of non-methyl eugenol-responding males. These lines were reared for 10 generations and in this paper, we report on the assessment of males from these lines in terms of methyl eugenol response and mating ability. We saw a gradual decrease in non-responders from ca. 35 to 10% after the 7th generation. Despite that, there were still significant differences until the 10th generation in numbers of non-responders over controls using laboratory strain males. We did not attain pure isolines of non-methyl eugenol-responding males, so we used non-responders from the 10th generation of those lines as sires to initiate two reduced-responder lines. Using these reduced responder flies, we found that there was no significant difference in mating competitiveness when compared with control males. Overall, we suggest that it may be possible to establish lines of low or reduced responder males to be used for sterile release programs, that could be applied until the 10th generation of rearing. Our information will contribute to the further development of an increasingly successful management technique incorporating the use of SIT alongside MAT to contain wild populations of B. dorsalis .

Methoprene, a juvenile hormone analog, is used to accelerate sexual maturation in males of species of economic importance in support to the sterile insect technique (SIT). In the SIT, mass-reared sterile males are released into the field and need to survive until they reach sexual maturation, find a wild female, mate with her and then induce female sexual refractoriness, so she will not remate with a wild counterpart. The use of methoprene shortens the time between release and copulation. However, in South American fruit flies, Anastrepha fraterculus , the ability of methoprene-treated males to inhibit female remating has been shown to be lower than wild males, when methoprene was applied by pupal immersion or topical application. Here we evaluated the possibility of incorporating methoprene into the male diet at different doses and the ability of those males to inhibit female remating, as well as the effect of methoprene on male reproductive organ size, due to the possible correlation between male accessory gland size and their content, and the role of male accessory gland proteins in female inhibition. We found that A. fraterculus males fed with methoprene in the adult protein diet at doses as high as 1% were less likely to inhibit female remating, however, at all other lower doses males had the same ability as untreated males to inhibit female remating. Males fed with methoprene had bigger male accessory glands and testes compared to methoprene-deprived males. We demonstrate that the incorporation of methoprene in adult male diets is possible in this species and potentially useful as a post-teneral, pre-release supplement at doses as low as 0.01%. Even at higher doses, the percentage of females remating after 48 h from the first copulation is sufficiently low in this species so as not compromise the efficiency of the SIT.

Steroid hormone 20-hydroxyecdysone (20E) plays critical roles in reproductive development in dipterans and several other insect species. Ecdysteroidogenesis in the glands of larval or nymphal insects and other arthropods has been extensively studied, but that in the adult gonads remains largely unknown. Here we identified a proteasome β3 subunit (PSMB3) from a highly invasive pest Bactrocera dorsalis, and found that this gene was crucial for ecdysone production during female reproduction. PSMB3 was enriched in the ovary, and it was upregulated during sexual maturation. RNAi-mediated depletion of PSMB3 resulted in retarded ovarian development and decreased fecundity. Additionally, knockdown of PSMB3 reduced 20E titer in hemolymph of B. dorsalis. Molecularly, RNA sequencing and qPCR validation revealed that PSMB3 depletion suppressed the expression of 20E biosynthetic genes in the ovary and 20E responsive genes in the ovary and fat body. Furthermore, exogenous 20E rescued the inhibition of the ovarian development caused by PSMB3 depletion. Taken together, this study provides new insights into the adult reproductive development-related biological processes controlled by PSMB3, and proposed a potential eco-friendly control strategy against this notorious agricultural pest., Competing Interests: Declaration of competing interest The authors declare that they have no conflicts of interest with the contents of this article., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Anastrepha obliqua Macquart (Diptera: Tephritidae) is a polyphagous species with hog plums (Spondias spp.) (Sapindales: Anacardiaceae) and mangoes (Mangifera indica L.) (Sapindales: Anacardiaceae) as primary host fruits. In this study, the olfactory preference of A. obliqua for three stages of ripeness of two mango cultivars ('Coche' and 'Ataulfo') was investigated. The female flies were more attracted to ripe 'Coche' fruits compared to those ripe 'Ataulfo'. Further, they were more attracted to the 'Coche' half-ripe and ripe fruits than to the unripe ones, but they did not discriminate among the stages of ripeness of 'Ataulfo' fruits. The male flies did not show preference for any specific mango cultivars or ripeness stage tested. Four compounds from ripe 'Coche' mangoes, and two from ripe 'Ataulfo' fruits were identified using coupled gas chromatography-electroantennographic (GC-EAD) recording and gas chromatography-mass spectrometry (GC-MS) analysis. 'Coche' mango volatiles eliciting responses from the female antennae were ethyl butyrate, ethyl hexanoate, ethyl heptanoate, and ethyl octanoate. The two 'Ataulfo' mango volatiles were identified as 3-carene and ethyl octanoate. These compounds were absent in unripe mangoes of both cultivars. Synthetic blends of these compounds were attractive to females as mango extracts in field cage tests. Our results suggest that the olfactory preference of A. obliqua for attractive hosts is based on the presence or absence of the compounds associated with fruit maturity., (© The Author(s) 2023. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

The oriental fruit fly, Bactrocera dorsalis (Hendel) (Diptera: Tephritidae), is an invasive and polyphagous pest of horticultural crops, and it can cause huge economic losses in agricultural production. The rapid development of CRISPR/Cas9 gene editing technology has provided new opportunities for the scientific control of agricultural pests. Here, we explore the applicability of the B. dorsalis sex peptide receptor (Bdspr) as a target gene for the CRISPR/Cas9-based sterile insect technique (SIT) in B. dorsalis. We screened two high-efficient single guide RNAs (sgRNAs) for gene editing. The results showed that both mutation efficiency and germline transmission rate were 100% in the surviving G0 females (8/8) from injected embryos, and that 75% of mosaically mutated G0 females (6/8) were sterile. The 50% of heterozygous G1 females (4/8) could not lay eggs; 100% of eggs laid by them could not survive; and 62.5% of individual females (5/8) had abnormal ovaries. These results indicate that Bdspr plays an important role in regulating fertility, egg viability, and ovary development in female B. dorsalis, suggesting that the spr gene can be used for CRISPR/Cas9-based SIT in B. dorsalis., (© The Author(s) 2023. Published by Oxford University Press on behalf of Entomological Society of America.)

Tephritid fruit flies are major horticultural pests of fruit and vegetable crops. Developing an odour lure that can attract mated female flies under field conditions has presented a major challenge to chemical ecologists around the globe. We have designed a new lure to attract female Queensland fruit fly, Bactrocera tryoni, based on the knowledge that this pest tephritid is attracted not only to odours from ripening fruits, but also to odours of symbiotic (gut-associated) yeasts on which the larvae feed. Initial field trials screened four volatile mixtures, each containing a base mix of three, short-chain "fruit ripening" esters and differing in the addition of long-chain "fruity" esters and / or alcohols produced by live yeasts. Results demonstrated that the lure formulated from short-chain esters and live yeast alcohols attracted the most female flies, with significantly higher catches than an odourless control trap, and a protein-baited trap. As electrophysiological studies exploring peripheral responses in tephritids to alcohols associated with fermentation have not been extensively investigated, we conducted this work on both the antennae and maxillary palps, and discovered that three yeast alcohols were detected by both sensory organs. Performance trials conducted in pome fruit and stone fruit orchards evaluated the six-component blend of short-chain ester and live yeast alcohol lure against a commercially available synthetic fruit-odour lure and a protein-based lure. In the apple orchard, the new lure caught significantly more female flies, and in particular mated females, compared to the commercially available lures. In the stone fruit orchard, while the new lure again caught the most mated female flies compared to the other lures, differences were not significant. The visual stimulus (trap design) used had a significant effect on capture, with a Ladd trap (red sphere on a yellow panel) being considerably more effective than the other traps. We discuss the implications of this study, including why the degree of effectiveness of the new lure might have differed among the test orchards (citrus, apple, stonefruit), and why yeast odours might increase attractiveness to "ripening fruit" volatile blends under field conditions., (© 2022. Crown.)

Humans have used weaver ants, Oecophylla smaragdina, as biological control agents to control insect pests in orchards for many centuries. Over recent decades, the effectiveness of weaver ants as biological control agents has been attributed in part to deterrent and oviposition inhibiting effects of kairomones produced by the ants, but the chemical identity of these kairomones has remained unknown. We have identified the kairomone responsible for deterrence and oviposition inhibition by O. smaragdina, providing a significant advance in understanding the chemical basis of their predator/prey interactions. Olfactometer assays with extracts from weaver ants demonstrated headspace volatiles to be highly repellent to Queensland fruit fly, Bactrocera tryoni. Using electrophysiology and bioassays, we demonstrate that this repellence is induced by a single compound, 1-octanol. Of 16 compounds identified in O. smaragdina headspace, only 1-octanol evoked an electrophysiological response from B. tryoni antennae. Flies had greatly reduced oviposition and spent significantly less time in an olfactometer arm in the presence of 1-octanol or a synthetic blend of headspace volatiles containing 1-octanol than in the presence of a synthetic blend of headspace volatiles without 1-octanol, or clean air. Taken together, our results demonstrate that 1-octanol is the functional kairomone component of O. smaragdina headspace that explains repellence and oviposition deterrence, and is hence an important contributor to the effectiveness of these ants as biological control agents., (© 2022. The Author(s).)

Nutrition is a central factor influencing immunity and resistance to infection, but the extent to which nutrition during development affects adult responses to infections is poorly understood. Our study investigated how the nutritional composition of the larval diet affects the survival, pathogen load and food intake of adult fruit flies, Bactrocera tryoni, after septic bacterial infection. We found a sex-specific effect of larval diet composition on survival post-infection: survival rate was higher and bacterial load was lower for infected females raised on a sugar-rich larval diet than for females raised on a protein-rich larval diet, an effect that was absent in males. Both males and females were heavier when fed a balanced larval diet compared with a protein- or sugar-rich diet, while body lipid reserves were higher for those that had consumed the sugar-rich larval diet compared with other diets. Body protein reserves were lower for flies that had been raised on the sugar-rich larval diet compared with other diets in males, but not females. Both females and males shifted their nutrient intake to ingest a sugar-rich diet when infected compared with sham-infected flies without any effect of the larval diet, suggesting that sugar-rich diets can be beneficial to fight off bacterial infection as shown in previous literature. Overall, our findings show that nutrition during early life can shape individual fitness in adulthood., Competing Interests: Competing interests The authors declare no competing or financial interests., (© 2022. Published by The Company of Biologists Ltd.)

Background: Olfaction has an indispensable role in insect behavior, enabling location of suitable host plants and oviposition sites, finding mates and evasion of natural enemies. Chemosensory proteins (CSPs) function to screen external odorants and transport them to olfactory receptor neurons, thereby increasing the sensitivity of the olfactory system. At present, CSP genes have been identified in many insect species, but there are relatively few studies on the function of CSP, especially in Tephritidae., Results: In this study, we sequenced and analyzed 12 transcriptomes of Bactrocera minax and identified five CSP genes. The results of polymerase chain reactions with reverse transcription showed that BminCSP3 was highly expressed only in antennae. Results from competitive binding experiments showed that BminCSP3 has good binding ability to citral compared with 23 other volatile organic compounds. The docking model with citral showed hydrogen bond formation with residues (ARG97); however, no hydrogen bonds were formed in the docking of five other ligands (furfuryl alcohol, linalool, cis-3-hexenyl acetate, (R)-(+)-limonene and (+)-carvone). Electroantennogram (EAG) analyses revealed that citral was active in B. minax at the antennal level, and the EAG response value of female adults was significantly higher than that of male adults. Furthermore, the results of behavioral bioassays showed that females were significantly attracted to citral., Conclusion: Our results suggest that BminCSP3 plays an important role in the recognition of citral by B. minax adults. © 2022 Society of Chemical Industry., (© 2022 Society of Chemical Industry.)

Seasonal temperatures select for eclosion timing of temperate insects and their parasitoids. In western North America, the fruit fly Rhagoletis zephyria Snow (Diptera: Tephritidae) is parasitized by the hymenopterous wasps Utetes lectoides (Gahan), an egg parasite, and Opius downesi Gahan, a larval parasite (both Braconidae). Eclosion of wasps should be timed with the presence of susceptible fly stages, but reports indicate U. lectoides ecloses in the absence of flies under no-chill conditions. Based on this, we tested the hypotheses that chill durations and no-chill temperatures both differentially regulate eclosion times of R. zephyria and its parasitic wasps. When fly puparia were chilled at ~3°C for 130-180 d, U. lectoides and O. downesi always eclosed on average later than flies. However, after 180-d chill, flies eclosed on average earlier than after 130- and 150-d chill, whereas eclosion times of U. lectoides and O. downesi were less or not affected by chill duration. When fly puparia were exposed to 20-22°C (no chill), U. lectoides eclosed before flies, with 88.9% of U. lectoides versus only 0.61% of flies eclosing. Taken together, findings show that eclosion times of flies are more sensitive to changes in chill duration than those of wasps. Flies are less sensitive than wasps to no-chill in that most flies do not respond by eclosing after no-chill while most wasps do. Our results suggest that shorter winters and longer summers due to climate change could cause mismatches in eclosion times of flies and wasps, with potentially significant evolutionary consequences., (Published by Oxford University Press on behalf of Entomological Society of America 2022.)

Insect responses to chemical attractants are often measured using olfactory bioassays prior to testing in field experiments. The attraction of sexually mature male Bactrocera dorsalis to methyl eugenol (ME) and the loss of attraction by ME pre-fed males have been demonstrated in laboratory bioassays and field trapping studies. It has been suggested that ME nonresponsiveness can be exploited to improve the effectiveness of B. dorsalis management programs by protecting sterile males from ME-based control measures. Currently, work is underway to identify alternatives that reduce or eliminate ME response. To support the development of compounds and evaluation of their effect on B. dorsalis attraction to ME, we compared the effectiveness of three common bioassay methods that have been used to measure lure response in Bactrocera flies under controlled conditions (choice assays using Y-tube [Y], small-cage arena [SC], and rotating carousel field-cage [RC]) to determine which bioassay method is efficient and reliable. A series of bioassays comparing ME-exposed and ME-naïve wild-type and genetic sexing strain males showed that the RC and SC were effective at both observing attraction to ME and detecting a significant reduction in ME response from ME-exposed males. However, the male attraction to ME and a significant decrease in response to ME after ME feeding was not observed in our Y-tube assays. These suggest that RC and SC are preferable options to evaluate ME non-responsiveness in B. dorsalis, and that Y-tube tests are difficult to administer correctly., (Published by Oxford University Press on behalf of Entomological Society of America 2022.)

Bactrocera dorsalis is an invasive polyphagous pest causing considerable ecological and economic damage worldwide. We report a high-quality chromosome-level genome assembly and combine various transcriptome data to explore the molecular mechanisms of its rapid adaptation to new environments. The expansions of the DDE transposase superfamily and key gene families related to environmental adaptation and enrichment of the expanded and unique gene families in metabolism and defence response pathways explain its environmental adaptability. The relatively high but not significantly different expression of heat-shock proteins, regardless of the environmental conditions, suggests an intrinsic mechanism underlying its adaptation to high temperatures. The mitogen-activated protein kinase pathway plays a key role in adaptation to new environments. The prevalence of duplicated genes in its genome explains the diversity in the B. dorsalis complex. These findings provide insights into the genetic basis of the invasiveness and diversity of B. dorsalis, explaining its rapid adaptation and expansion., (© 2022. The Author(s).)

Insect mitogenome organisation is highly conserved, yet, some insects, especially with parasitic life cycles, have rearranged mitogenomes. Furthermore, intraspecific mitochondrial diversity can be reduced by fitness-affecting bacterial endosymbionts like Wolbachia due to their maternal coinheritance with mitochondria. We have sequenced mitogenomes of the Wolbachia-infected endoparasitoid Dipterophagus daci (Strepsiptera: Halictophagidae) and four of its 22 known tephritid fruit fly host species using total genomic extracts of parasitised flies collected across > 700 km in Australia. This halictophagid mitogenome revealed extensive rearrangements relative to the four fly mitogenomes which exhibited the ancestral insect mitogenome pattern. Compared to the only four available other strepsipteran mitogenomes, the D. daci mitogenome had additional transpositions of one rRNA and two tRNA genes, and a single nucleotide frameshift deletion in nad5 requiring translational frameshifting or, alternatively, resulting in a large protein truncation. Dipterophagus daci displays an almost completely endoparasitic life cycle when compared to Strepsiptera that have maintained the ancestral state of free-living adults. Our results support the hypothesis that the transition to extreme endoparasitism evolved together with increased levels of mitogenome changes. Furthermore, intraspecific mitogenome diversity was substantially smaller in D. daci than the parasitised flies suggesting Wolbachia reduced mitochondrial diversity because of a role in D. daci fitness., (© 2022. The Author(s).)

Females of many insect species are unreceptive to remating for a period following their first mating. This inhibitory effect may be mediated by either the female or her first mate, or both, and often reflects the complex interplay of reproductive strategies between the sexes. Natural variation in remating inhibition and how this phenotype responds to captive breeding are largely unexplored in insects, including many pest species. We investigated genetic variation in remating propensity in the Queensland fruit fly, Bactrocera tryoni, using strains differing in source locality and degree of domestication. We found up to threefold inherited variation between strains from different localities in the level of intra-strain remating inhibition. The level of inhibition also declined significantly during domestication, which implied the existence of genetic variation for this trait within the starting populations as well. Inter-strain mating and remating trials showed that the strain differences were mainly due to the genotypes of the female and, to a lesser extent, the second male, with little effect of the initial male genotype. Implications for our understanding of fruit fly reproductive biology and population genetics and the design of Sterile Insect Technique pest management programs are discussed., (© 2022. The Author(s).)

The short neuropeptide F (sNPF) signaling system, consisting of sNPF and its receptor (sNPFR), influences many physiological processes in insects, including feeding, growth and olfactory memory. We previously showed that sNPF regulates olfactory sensitivity in the oriental fruit fly Bactrocera dorsalis (Hendel) during starvation. However, the functional analysis of sNPFR is constrained by the failure of RNA interference in this species. Here, we generated a null sNPFR mutant using the CRISPR/Cas9 system to investigate the physiological roles of this receptor in more detail. G0 adults were produced at a frequency of 60.8%, and sNPFR -/- mutants were obtained after several generations of backcrossing followed by self-crossing among heterozygous flies. We found that the mutants were significantly less successful at foraging for certain foods and showed increased foraging latency. Electroantennogram (EAG) assays indicated that the mutants had significantly lower electrophysiological responses to three tested odorants. Furthermore, qPCR data revealed the inhibition of several olfactory receptor genes, including Orco. Immunohistochemistry showed that BdsNPFR was localized in cells under the sensillum on the antennae. Based on their shape and size, the BdsNPFR + cells differ from odorant receptor neurons (ORNs), which were labeled using a Drosophila melanogaster Orco antibody. Our data suggest that sNPFR regulates olfaction-mediated foraging behavior by mediating interactions between BdsNPFR + cells and selected ORNs., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Olfaction underpins many insect behaviors, such as foraging, host location, mating, and predator avoidance. In the first step of insect olfaction, odorant-binding proteins (OBPs) bind hydrophobic odorants and transport them to odorant receptors. Methyl eugenol (ME) is a powerful attractant for mature males of the oriental fruit fly Bactrocera dorsalis (Hendel), one of the most destructive fruit pests. The underlying molecular mechanism is unclear, but there is in vitro evidence that BdorOBP56f-2 is involved in ME perception. We used microscale thermophoresis to confirm that BdorOBP56f-2 directly binds ME with strong affinity in vitro. We then used CRISPR/Cas9 to knock out the BdorOBP56f-2 gene, allowing us to establish a homozygous mutant B. dorsalis line. The electroantennogram response and behavioral attraction to ME were significantly reduced in the mutant, providing in vivo evidence that BdorOBP56f-2 is necessary for efficient ME perception. Our results offer insight into the molecular mechanism of ME perception in B. dorsalis and provide a theoretical basis for the functional analysis of other OBPs., (Copyright © 2021 Elsevier Ltd. All rights reserved.)