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Certain species of true fruit flies (Diptera: Tephritidae) cause tremendous damage to commercially important fruits and vegetables, and many countries operate continuous trapping programs which rely on male-specific lures such as trimedlure (TML), methyl eugenol (ME), and cue-lure (CL). Traditionally, these attractants have been applied as liquids to cotton wicks inside traps, although this results in high evaporative loss of the lure. Slow-release, polymeric plugs have been widely adopted for TML, but such devices are not widely used for ME or CL. Recent data, however, suggest that ME and CL plugs may be attractive for as long as 12 wk in the field. The present study investigates whether ME and CL plugs weathered for 18 or 24 wk are effective in capturing males of Bactrocera dorsalis (Hendel) and Zeugodacus cucurbitae (Coquillett), respectively. For B. dorsalis, 6 g ME plugs were as effective as the control treatment (fresh liquid on a wick) after 12 wk of weathering but not after 18 or 24 wk. For Z. cucurbitae, 3 g CL plugs were as effective as the control treatment (fresh CL plugs) after 12 and 18 wk of weathering but not after 24 wk. The residual content and release rate of the 2 lures were also measured over time, but, with the exception of the residual content of ME, we did not find a direct correlation between these parameters and numbers of flies captured., (Published by Oxford University Press on behalf of Entomological Society of America 2024.)

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.)

Pyrethroid are the primary insecticides used for controlling of Bactricera dorsalis, a highly destructive and invasive fruit pest. Field populations have developed serious resistance, especially to β-cypermethrin. While mutations in the voltage-gated sodium channel (Vgsc) are a common mechanism of pyrethroid resistance, variations in BdVgsc associated with β-cypermethrin resistance remain unclear. Here, we reported the resistance levels of five field populations from China, with resistance ratio ranging from 1.54 to 21.34-fold. Cloning the full length of BdVgsc revealed no specific or known amino acid mutations between the most resistant population and the susceptible strain. However, three types of partial intron retention (IRE4-5, IRE19-f and IREL-24) were identified in BdVgsc transcripts, with these intron retentions containing stop codons. The expression of IRE4-5 transcripts and total BdVgsc showed different trends across developmental stages and tissues. Exposure to β-cypermethrin led to increased expression of IRE4-5. Comparison of genomic and transcriptional sequences reveled that IRE4-5 transcripts had two types (IRE4-5.5 T and IRE4-5.6 T) caused by genomic variations. Both field and congenic strains indicated that homozygotes for IRE4-5.5 T had lower IRE4-5 transcript levels than homozygotes for IRE4-5.6 T. However, congenic and field strains exhibited inconsistent results about the association of expression levels of IRE4-5 transcripts with sensitivity to β-cypermethrin. In summary, this study is the first to identify intron retention transcripts in the Vgsc gene from B. dorsalis and to examine their expression patterns across different developmental stages, tissues, and strains with varying sensitivities to β-cypermethrin. The potential role of the intron retentions of BdVgsc in insecticide toxicity is also discussed., Competing Interests: Declaration of competing interest The authors involved in this work declare that they do not have any type of competing for financial interests or personal relationships that have influenced the work reported in this paper., (Copyright © 2024. Published by Elsevier Inc.)

Background: Fopius arisanus (Sonan) is an important egg-pupal endoparasitic wasp of Bactrocera dorsalis (Hendel). As traditional method of sorting nonparasitic B. dorsalis from parasitic wasps is labor-intensive, requires specific equipment and poses the risk of spreading fertile hosts, the development of a more convenient, economical and safe sorting procedure is important., Results: The optimal cyromazine emergence inhibition procedure (CEIP) involved facilitating the pupation of B. dorsalis mature larvae (Bdml) in 3 mg kg -1 cyromazine sand substrate (CSS) for 48 h. When the Bdml that had been exposed to F. arisanus during the egg stage were treated with 3-7 mg kg -1 CSS for 48 h, no negative effects on the emergence parameters of parasitoids were observed. Treatment with 3-4 mg kg -1 CSS had insignificant effects on the biological and behavioral parameters of F. arisanus. However, treatment with 5-6 mg kg -1 CSS adversely affected the fecundity and antennating activity of the wasps; specifically, 6 mg kg -1 CSS negatively affected the lifespan and flight ability of wasps. Fortunately, no transgenerational effects on these parameters were observed in the progeny. Regarding the nutrient reserves of both sexes of F. arisanus, significant dose-dependent effects were observed. Moreover, 5-6 mg kg -1 CSS significantly reduced the protein and carbohydrate content in F. arisanus; in particular, 6 mg kg -1 CSS notably reduced the lipid content., Conclusion: CEIP provides a more flexible, economical and safe mass-release program for F. arisanus. In addition, it has profound implications for the biological control of other dipteran pests. © 2024 Society of Chemical Industry., (© 2024 Society of Chemical Industry.)

Background: Insecticide resistance among invasive tephritid fruit flies poses a great risk to national food security and has the potential to disrupt quarantine and eradication programs, which rely on the efficacy of Spinosad to prevent widespread establishment in North America. During 2022 to 2023 we surveyed the extent of Spinosad resistance of two key species, oriental fruit fly Bactrocera dorsalis, and melon fly Zeugodacus cucurbitae, from 20 sites across five Hawaiian Islands including Kaua'i, O'ahu, Maui, Molokai and the "Big Island" (Hawai'i)., Results: We used topical thoracic applications of eight concentrations of Spinosad ranging from 0.028 to 3.6 mg/mL to evaluate the lethal concentration (LC 50 and LC 99 ) required to kill wild-caught males. Resistance ratios (RR) were calculated by comparing the LC 50 of wild flies to laboratory susceptible lines maintained in colony. Our results identified at least two new sites of concern for melon fly resistance on the Big Island, and at least four sites of concern for oriental fruit fly, all of which were located on the Big Island. At these locations RRs were >5. On O'ahu, melon fly RRs were >10., Conclusions: The persistence of Spinosad resistance is concerning, yet it is a reduction compared to the values reported previously and before changes to Spinosad use recommendations by local extension agents beginning in 2017. For oriental fruit fly, these RR values are the highest levels that have been detected in wild Hawai'i populations. These data suggest that expanded Spinosad reduction and replacement programs are warranted given the ongoing issues with Spinosad resistance in Hawai'i and expansion in the number of species affected. © 2024 The Author(s). Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA., (© 2024 The Author(s). Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.)

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.)

Chitosan is a naturally derived polymer that has significant potential for use as a bioinsecticide. Despite this, there is a lack of research as to the efficacy of chitosan for many insect pest species. The apple maggot fly, Rhagoletis pomonella (Walsh) (Diptera: Tephritidae), is one such pest for which chitosan toxicity has not been explored. In this study, the toxicity of chitosan for R. pomonella adults was tested via no-choice feeding assays. An aging trial was further used to test the mortality of flies provided dried chitosan-sucrose treatments (CST), which were aged for 0 or 3 days in greenhouse conditions. This study found that the CST is toxic for R. pomonella adults when ingested, leading to a significant increase in the rate of mortality compared to control groups. The use of dried chitosan, however, did not change the mortality of flies, suggesting this will not be an effective delivery mechanism. Effective biopesticide delivery systems have not been defined for chitosan use outside of a laboratory, indicating the need for further research testing delivery mechanisms. It is suggested that an effective method of delivery can be as a food-based bait in attract-and-kill traps, as chitosan must be ingested to kill flies. Forming a viscous solution, chitosan may be best suited to these lure systems., (© The Author(s) 2024. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

The Queensland fruit fly, Bactrocera tryoni (Froggatt) (Diptera: Tephritidae), is a crop pest of global economic importance because of its wide range of hosts and its invasiveness capacities. To develop a novel integrated and sustainable crop protection, we have investigated the insecticidal properties of different varieties of kava (Piper methysticum [Frost]) extracted by two methods and the attractive effects of six plant volatiles identified from B. tryoni host plants to female, mated or not. We did not identify any significant insecticidal effect of the traditional Pacific kava plant at the tested concentrations. Among mated females, ethyl acetate compared to the no odor control elicited the highest attraction (87%, of which 60% for this odor), while ethyl butyrate was preferred compared with ethyl acetate in dual choice assays. Flies' preferences for specific odors depended on their mating status and the odor landscape they were confronted with. Combination with the commercial ingestion insecticide (Success 4: spinosad, 480 g/l, Dow AgroSciences, Valbonne, France) with the plant volatiles were tested to detect an increase in mortality related to the addition of an attractant. The 2-heptanone slightly showed a tend to increase the attractiveness of mated females within 4-6 h to the food bait, but the results were not statistically significant after 8 h. Further tests should be performed with other concentrations or mixtures of the identified host plant volatiles to develop a strong lure and kill strategy., (© The Author(s) 2024. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

Zeugodacus cucurbitae Coquillett (Diptera: Tephritidae) is an agriculturally and economically important pest worldwide that has developed resistance to β-cypermethrin. Glutathione S-transferases (GSTs) have been reported to be involved in the detoxification of insecticides in insects. We have found that both ZcGSTd6 and ZcGSTd10 were up-regulated by β-cypermethrin induction in our previous study, so we aimed to explore their potential relationship with β-cypermethrin tolerance in this study. The heterologous expression of ZcGSTd6 and ZcGSTd10 in Escherichia coli showed significantly high activities against 1-chloro-2,4-dinitrobenzene (CDNB). The kinetic parameters of ZcGSTd6 and ZcGSTd10 were determined by Lineweaver-Burk. The V max and K m of ZcGSTd6 were 0.50 μmol/min·mg and 0.3 mM, respectively. The V max and K m of ZcGSTd10 were 1.82 μmol/min·mg and 0.53 mM. The 3D modelling and molecular docking results revealed that β-cypermethrin exhibited a stronger bounding to the active site SER-9 of ZcGSTd10. The sensitivity to β-cypermethrin was significantly increased by 18.73% and 27.21%, respectively, after the knockdown of ZcGSTd6 and ZcGSTd10 by using RNA interference. In addition, the inhibition of CDNB at 50% (IC 50 ) and the inhibition constants (Ki) of β-cypermethrin against ZcGSTd10 were determined as 0.41 and 0.33 mM, respectively. The Ki and IC 50 of β-cypermethrin against ZcSGTd6 were not analysed. These results suggested that ZcGSTd10 could be an essential regulator involved in the tolerance of Z. cucurbitae to β-cypermethrin., (© 2024 Royal Entomological Society.)

G-protein coupled receptors (GPCRs) are the largest and most diverse transmembrane receptor family in the cell. They are involved in regulating a wide range of biological processes, including behavior, reproduction, and development. However, GPCRs have not yet been identified in Zeugodacus cucurbitae. The current study focuses on the GPCRs identification, classification, distribution, and their expression analysis under β-cypermethrin stress to uncover novel targets for pest management and assist in the development of effective strategies for controlling the melon fly population. We identified 80 GPCRs genes including 50 GPCRs identified in family A, 17 GPCRs identified in family B, 8 identified in family C, and 5 identified in family F. Z. cucurbitae GPCRs showed significant differences in both the number of genes in families or subfamilies, as well as the sequencing of the genes. Interestingly, newly identified GPCRs genes are expressed differently at various developmental stages of Z. cucurbitae. Further, we evaluated these 80 GPCRs using Realtime quantitative PCR to confirm their expression between β-cypermethrin-resistant (RS) strain and susceptible strain (SS) of Z. cucurbitae. We identified 50 GPCR genes were highly overexpressed in a RS. Among these genes, eight genes were strongly induced by the 30% lethal concentration (LC) while two genes were significantly increased by the 50% LC of β-cypermethrin. This first genome-wide profiling and characterization of GPCRs could lay foundation for unraveling detoxification mechanism and target site modifications which may improve the insect resistance and could be effective insecticide targets for Z. cucurbitae management., Competing Interests: Declaration of competing interest None., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Phenolic compounds are the secondary metabolites (SMs) present in plants carrying different bioactivities. In the present study, we explored the influence of a phenolic compound namely phloroglucinol on oviposition behaviour and different biochemical entities of an insect pest Zeugodacus cucurbitae (Coquillett) (Diptera: Tephritidae) using artificial diet. Phloroglucinol (IUPAC name: benzene-1,3,5-triol) affected the activity of antioxidant and detoxifying enzymes viz. superoxide dismutases (SOD), catalase (CAT), ascorbate peroxidases (APOX). dehydroascorbate reductase (DHAR), peroxidases (POX), phenol oxidase (PO), glutathione peroxidase (GPOX), glutathione S-transferase peroxidase (GSTpox), glutathione reductase (GR), glutathione S-transferase (GST) and esterases (EST) as well as the biological antioxidants viz. ascorbate content and glutathione. The lipid peroxide content (LP) and hydrogen peroxide content (H 2 O 2 ) were significantly enhanced in the treated larvae indicating oxidative stress in the insect. Significant inhibition in oviposition was observed and effective repellency percentage increased with phloroglucinol treatment as compared to control. The oviposition deterrent activity and toxic effects of phloroglucinol on various biochemical parameters of Z. cucurbitae larvae revealed in the present study clearly confirms its suitability for use in pest management., (Copyright © 2022 Elsevier Inc. All rights reserved.)

The present study analyzed the chemical composition of Juniperus foetidissima Willd. essential oils (EOs) and evaluated their attractancy and toxicity to two agriculturally important tephritid fruit flies. The composition of hydrodistilled EOs obtained from leaves (JFLEO) and fruits (JFFEO) of J. foetidissima was analyzed by GC-FID and GC-MS. The main compounds were α -pinene (45%) and cedrol (18%) in the JFLEO and α -pinene (42%), α -thujone (12%), and β -thujone (25%) in the JFFEO. In behavioral bioassays of the male Mediterranean fruit fly, Ceratitis capitata (Wiedemann), both JFLEO and JFFEO showed strong attraction comparable to that observed with two positive controls, Melaleuca alternifolia and Tetradenia riparia EOs. In topical bioassays of the female Caribbean fruit fly, Anastrepha suspensa (Loew), the toxicity of JFFEO was two-fold higher than that of JFLEO, with the LD 50 values being 10.46 and 22.07 µg/µL, respectively. This could be due to differences in chemical components between JFLEO and JFFEO. The JFFEO was dominated by 48% monoterpene hydrocarbons (MH) and 46% oxygenated monoterpenes (OM), while JFLEO consisted of 57% MH, 18% OM, and 20% oxygenated sesquiterpenes (OS). This is the first study to evaluate the attractancy and toxicity of J. foetidissima EOs to tephritid fruit flies. Our results indicate that JFFEO has the potential for application to the management of pest tephritid species, and further investigation is warranted.

A series of 3-oxygenated α-ionone analogs have been developed as highly specific male lures for the solanaceous fruit fly Bactrocera latifrons, a pest of solanaceous fruits. We compared the attractant and phagostimulant activities of analogs with or without (i) unsaturations at the 4,5- and/or 7,8-positions and (ii) oxygen moieties at the 3- and/or 9-positions of the ionone molecule. Since naturally occurring vomifoliol (V2) was found to induce a highly potent phagostimulant activity in B. latifrons males, related analogs including dehydrovomifoliol (V1), 6-hydroxy-α-ionone (U1), and 6-hydroxy-α-ionol (U2) were synthesized to evaluate their attractant and phagostimulant activities. Synthetic V1, V2, U1, and U2 exhibited low attractant activity, but their phagostimulant activity was relatively high. Optical isomers of 3-oxo-7,8-dihydro-α-ionone (P3) and V1 were prepared to examine the stereochemical specificity of attractants. (+)-(6R)-P3 and (+)-(6S)-V1 exhibited the corresponding activities, while their respective antipodal enantiomers were found entirely inactive., (© The Author(s) 2021. Published by Oxford University Press on behalf of Japan Society for Bioscience, Biotechnology, and Agrochemistry.)

Melon fly ( Bactrocera cucurbitae ) is the most common pest of cucurbits, and it directly causes damage to cucurbit fruits in the early developmental stage. The infection of fruit tissues induces oxidative damage through increased generation of cellular reactive oxygen species. The effects of melon fly infestation on the production of defensive enzymes and antioxidant capabilities in five cucurbit species, namely, bottle gourd, chayote, cucumber, snake gourd, and bitter gourd, were investigated in this study. The total phenolic and flavonoid content was considerably higher in melon fly infestation tissues compared to healthy and apparently healthy tissues. The chayote and bottle gourd tissues expressed almost 1.5- to 2-fold higher phenolic and flavonoid contents compared to the tissues of bitter gourd, snake gourd, and cucumber upon infestation. Defensive enzymes, such as peroxidase (POD), superoxide dismutase (SOD), polyphenol oxidase (PPO), and catalase (CAT), were high in healthy and infected tissues of chayote and bottle gourd compared to bitter gourd, snake gourd, and cucumber. The activity of POD (60-80%), SOD (30-35%), PPO (70-75%), and CAT (40-50%) were high in infected chayote and bottle gourd tissue, representing resistance against infestation, while bitter gourd, snake gourd, and cucumber exhibited comparatively lower activity suggesting susceptibility to melon fly infection. The antioxidant properties were also high in the resistant cucurbits compared to the susceptible cucurbits. The current research has enlightened the importance of redox-regulatory pathways involving ROS neutralization through infection-induced antioxidative enzymes in host cucurbit resistance. The melon fly infestation depicts the possible induction of pathways that upregulate the production of defensive enzymes and antioxidants as a defensive strategy against melon fly infestation in resistant cucurbits.

The family Tephritidae (Diptera) includes species that are highly invasive and harmful to crops. Due to globalization, international trade, and human displacement, their spread is continuously increasing. Unfortunately, the control of tephritid flies is still closely linked to the use of synthetic insecticides, which are responsible for detrimental effects on the environment and human health. Recently, research is looking for alternative and more eco-friendly tools to be adopted in Integrated Pest Management (IPM) programs. In this regard, essential oils (EOs) and their main compounds represent a promising alternative to chemical insecticides. EOs are made up of phytoconstituents formed from the secondary metabolism of many plants and can act as attractants or toxics, depending on the dose. Because of this unique characteristic, EOs and their main constituents are promising tools that can be used both in Sterile Insect Technique (SIT) programs and in the "lure and kill" technique, exploiting the attractiveness of the product in the former case and its toxicity in the latter. In this article, current knowledge on the biological and behavioral effects of EOs and their main constituents on tephritid fruit flies is reviewed, mainly focusing on species belonging to the Anastrepha , Bactrocera , Ceratitis , and Zeugodacus genera. The mechanisms of action of EOs, their real-world applications, and challenges related to their use in IPM are critically discussed.

The Oriental fruit fly, Bactrocera dorsalis (Hendel) is an economically devastating pest of fruit crops across the globe with stringent quarantine restrictions to limit its further spread. The current management programs increasingly depend on male annihilation but trapping female flies is equally important to reduce fruit damage. Considering the importance of kairomones in courtship and oviposition site selection behavior of B. dorsalis, the aim of this work was to isolate and identify potential cues from the volatiles of arils of jackfruit, Artocarpus heterophyllus. Using olfactometer assays and gas-chromatography linked to electroantennographic detection, attraction of both female and male B. dorsalis to specific jackfruit volatiles was demonstrated. Ethyl 3-methylbutanoate, ethyl hexanoate, pentyl butanote, 2-methylbutyl 3-methylbutanoate, 2-methylpropyl hexanoate, (Z)-3-hexenyl 3-methylbutanoate and dodecanal were found to attract female B. dorsalis specifically. Butyl acetate, 2 phenylethanol and pentyl 3-methylbutanoate elicited attraction in male B. dorsalis only. Synthetic blends of these compounds were found to attract female and male B. dorsalis in laboratory as well as field conditions. Using specific cues common to each set, a blend of methyl 3-methylbutanoate, butyl acetate, 3-methylbutyl acetate and hexyl acetate attracted both sexes of B dorsalis. This study demonstrates the use of kairomone-based lures for sex-specific as well as bisexual attraction for the first time.

The Chinese citrus fly, Bactrocera minax, is a notorious univoltine pest that causes damage to citrus. B. minax enters obligatory pupal diapause in each generation to resist harsh environmental conditions in winter. Despite the enormous efforts that have been made in the past decade, the understanding of pupal diapause of B. minax is currently still fragmentary. In this study, the 20-hydroxyecdysone solution and ethanol solvent was injected into newly-formed pupae to obtain non-diapause- (ND) and diapause-destined (D) pupae, respectively, and a comparative proteomics analysis between ND and D pupae was performed 1 and 15 d after injection. A total of 3,255 proteins were identified, of which 190 and 463 were found to be differentially abundant proteins (DAPs) in ND1 vs D1 and ND15 vs D15 comparisons, respectively. The reliability and accuracy of LFQ method was validated by qRT-PCR. Functional analyses of DAPs, including Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment, and protein-protein interaction (PPI) network construction, were conducted. The results revealed that the diapause program of B. minax is closely associated with several physiological activities, such as phosphorylation, chitin biosynthesis, autophagy, signaling pathways, endocytosis, skeletal muscle formation, protein metabolism, and core metabolic pathways of carbohydrate, amino acid, and lipid conversion. The findings of this study provide insights into diapause program of B. minax and lay a basis for further investigation into its underlying molecular mechanisms., Competing Interests: The authors have declared that no competing interests exist.

Anastrepha ludens is a key pest of mangoes and citrus from Texas to Costa Rica but the mechanisms of odorant perception in this species are poorly understood. Detection of volatiles in insects occurs mainly in the antenna, where molecules penetrate sensillum pores and link to soluble proteins in the hemolymph until reaching specific odor receptors that trigger signal transduction and lead to behavioral responses. Scrutinizing the molecular foundation of odorant perception in A. ludens is necessary to improve biorational management strategies against this pest. After exposing adults of three maturity stages to a proteinaceous attractant, we studied antennal morphology and comparative proteomic profiles using nano-LC-MS/MS with tandem mass tags combined with synchronous precursor selection (SPS)-MS3. Antennas from newly emerged flies exhibited dense agglomerations of olfactory sensory neurons. We discovered 4618 unique proteins in the antennas of A. ludens and identified some associated with odor signaling, including odorant-binding and calcium signaling related proteins, the odorant receptor co-receptor (Orco), and putative odorant-degrading enzymes. Antennas of sexually immature flies exhibited the most upregulation of odor perception proteins compared to mature flies exposed to the attractant. This is the first report where critical molecular players are linked to the odor perception mechanism of A. ludens .

Tephritid fruit flies are commonly reared on artificial larval diets for laboratory studies and for sterile insect technique pest management programs. While significant effort has been invested in developing artificial larval diets, surprisingly little is known about the specific nutritional requirements of tephritid flies. Recently developed gel larval diets have provided new opportunities for nutritional studies in Queensland fruit fly, Bactrocera tryoni ('Q-fly'). Wheat germ oil (WGO) is the main source of fatty acids and vitamin E in this diet, and is key for production of high-quality adults. To identify the importance of nutritional components of WGO for Q-fly productivity and quality, linoleic, linolenic, oleic and palmitic fatty acids as well as α-tocopherol (vitamin E) were included in the diet individually and in combination. Diets that included all of the tested fatty acids or just unsaturated fatty acids performed as well as diets containing WGO in most quality control parameters except fecundity, and addition of vitamin E reduced the pupal productivity. Considering individual fatty acids, larval diets containing only linolenic acid produced adults with higher percentage of fliers than did larval diets containing only palmitic acid or oleic acid. Compared with diets containing WGO, nutritional requirements for egg production in Q-fly were not entirely met by either grouped fatty acids or individual polyunsaturated, monounsaturated or saturated fatty acids, however, diets containing linoleic acid alone produced more eggs than any other fatty acid. The present study is a significant advance in understanding of the role of fatty acids as a component of WGO in larval diet in meeting the needs of developing Q-fly for somatic performance, but highlight also that other, untested, components of WGO appear to be important for reproduction., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Plant-derived compounds can be an environmentally friendly alternative to synthetic pesticide use for pest management. Essential oils (EOs) in several plant families have been found to be toxic to various pest species of insects through topical application, ingestion, and as fumigants. Previous studies revealed that, among various environmentally friendly insecticides, the EOs of Baccharis dracunculifolia and Pinus elliottii and an ethanol extract of Solanum granulosoleprosum plus Ricinus communis, were toxic to Ceratitis capitata and Anastrepha fraterculus (Diptera: Tephritidae) when applied topically to pupae or when ingested by adults. Here, we aimed to examine the potentially toxic effects of these plant-derived compounds when these two pestiferous fruit fly species were exposed to their vapors. We also examined their fumigant effect on female fecundity and fertility and compared it with water and ethanol controls. Exposure of C. capitata and A. fraterculus sexually mature adults to volatiles and vapors of both B. dracunculifolia and P. elliottii EOs resulted in lower longevity (half-life), survivorship, and female fecundity than the water vapor control. Toxicity of C. capitata was greater for P. elliottii than for B. dracunculifolia while the reverse was true for A. fraterculus. Exposure to vapors of S. granulosoleprosum + R. communis (S + R) had no effect on longevity but reduced survivorship of adults of both species. Interestingly, exposure to vapors of S + R, 50% (v/v) and pure ethanol resulted in greater fecundity of females of both frugivorous fly species than the water control. By contrast, fertility (% egg hatch) was in all cases high (>85%) and not different than the water control. Exposure to ethanol vapors appears to have similar effects on frugivorous tephritids as those reported on saprophagous and frugivorous species of Drosophila, a novel finding that may have important practical implications.

Queensland fruit fly [Bactrocera tryoni (Froggatt), Diptera, Tephritidae] is the most devastating insect pest impacting Australian horticulture. The Sterile Insect Technique (SIT) is an important component of tephritid pest management programs. However, mass-rearing and irradiation (to render insects sterile) may reduce the fitness and performance of the insect, including the ability of sterile males to successfully compete for wild females. Manipulation of the gut microbiome, including the supplementation with bacterial probiotics shows promise for enhancing the quality of mass-reared sterile flies, however there are fewer published studies targeting the larval stage. In this study, we supplemented the larval stage of mass-reared B. tryoni with bacterial probiotics. We tested several individual bacteria that had been previously isolated and characterized from the gut of wild B. tryoni larvae including Asaia sp., Enterobacter sp., Lactobacillus sp., Leuconostoc sp. We also tested a consortium of all four of these bacterial isolates. The fitness parameters tested included adult survival in field cages, laboratory mate selection of bacteria supplemented males by bacteria nonsupplemented females, and laboratory locomotor activity of adult flies. None of the bacterial probiotic treatments in the current study was significantly different to the control for field survival, mate selection or locomotor activity of adult B. tryoni, which agree with some of the other studies regarding bacterial probiotics fed to the larval stage of tephritids. Future work is needed to determine if feeding the same, and/or other probiotics to adults, as opposed to larvae can positively impact survival, mating performance, mating competitiveness and locomotor activity of B. tryoni. The bacterial group(s) and function of bacterial species that increase fitness and competitiveness is also of interest to tephritid mass-rearing programs., (© 2019 Institute of Zoology, Chinese Academy of Sciences.)

Insect glutathione S-transferases (GSTs) are important in insecticide detoxification and Insect-specific GSTs, Epsilon and Delta, have largely expanded in insects. In this study, we functionally expressed and characterized an epsilon class GST gene (BdGSTe8), predominant in the adult Malpighian tubules of Bactrocera dorsalis. This gene may be associated with malathion resistance based on transcriptional studies of resistant and susceptible strains. RNA interference-mediated knockdown of this gene significantly recovered malathion susceptibility in the adults of a malathion-resistant strain, and overexpression of BdGSTe8 enhanced resistance in transgenic Drosophila. Analysis of BdGSTe8 polymorphism showed that several point mutations may be associated with metabolic resistance to malathion. A cytotoxicity assay in Escherichia coli indicated that both of the recombinant BdGSTe8 proteins may play a functional role in protecting cells from toxicity. The allele of BdGSTe8-B conferred higher levels of malathion detoxification capability. Liquid chromatography and ultra-performance liquid chromatography-tandem mass spectrometry analysis showed that the BdGSTe8-A allele did not metabolize malathion directly. However, the BdGSTe8-B allele was involved in the direct metabolism of malathion, which was caused by a mutation in V128A. Further analysis of the sequence suggests that BdGSTe8 evolved rapidly. It maybe play the role of a backup gene and could become a new gene in the future in order to retain the ability of detoxification of malathion, which was driven by positive selection. These results suggest that divergent molecular evolution in BdGSTe8 has played a role in metabolic resistance to malathion in B. dorsalis., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Azadirachtin exhibits excellent bioactivities against several hundred arthropods. However, current knowlege of its biochemical effect on B. dorsalis larvae is not deep enough. In this study, integrated LC-MS and GC-MS-based untargeted metabolomics were used to analyze the changes of endogenous metabolites and the biochemical effects of azadirachtin on B. dorsalis larvae. Azadirachtin has excellent bioactivities against B. dorsalis larvae in this study, leading to a longer developmental duration, lower survival rate, and low pupa weight. The effect of azadirachtin was investigated on a total of 22 and 13 differentially abundant metabolites in the LC-MS and GC-MS-based metabolomics results, are selected respectively. Pathway analysis indicated that 14 differentially enriched metabolic pathways, including seven influential pathways, are worthy of attention. Further integrated key metabolic pathway analysis showed that histidine metabolism, D-glutamine and D-glutamate metabolism, biotin metabolism, ascorbate and aldarate metabolism, pentose and glucuronate interconversions, and alanine, aspartate and glutamate metabolism in B. dorsalis larvae are significantly relevant pathways affected by azadirachtin. Although extrapolating the bioactivity results in this study to the practical project of B. dorsalis pest management in the field has limitations, it was found that azadirachtin has a significant effect on the primary metabolism of B. dorsalis larvae.

Azadirachtin, as the most promising and effective botanical insecticide, exhibits significant growth inhibition activity against agricultural and forestry pests. However, its biochemical effects at the metabolic level compared with those of other insect growth regulators have not been studied. Therefore, in this study, a GC-MS based untargeted metabolomics approach was applied to compare azadirachtin with pyriproxyfen (a juvenile hormone analog) and tebufenozide (a molting hormone analog) in terms of their metabolic effects on Bactrocera dorsalis larvae. The bioactivity of azadirachtin against B. dorsalis larvae was significantly different than those of pyriproxyfen and tebufenozide. A total of 693 mass features were recognized, and 112 metabolites were identified in this study. The results showed that a total of 16, 13 and 10 differentially regulated metabolites corresponding to 12, 5 and 8 pathways occur in Aza versus CK, Pyr versus CK and Teb versus CK group, respectively. Further analysis showed that 6 differentially regulated metabolites corresponding to 5 key pathways could be the primary differential metabolic response of B. dorsalis larvae to the three insect growth regulators. The pathways were myo-inositol corresponding to ascorbate and aldarate metabolism as the specific response of B. dorsalis larvae to azadirachtin; xylitol, xylulose and 3-aminopropionitrile corresponding to pentose and glucuronate interconversions, and cyanoamino acid metabolism as the common responses to azadirachtin and pyriproxyfen; and 3-hydroxypropionic acid and beta-alanine corresponding to propanoate metabolism and beta-alanine metabolism as the specific responses to tebufenozide. The results showed that the metabolic response of B. dorsalis larvae to azadirachitin is closer to that of pyriproxyfen than tebufenozide. The differentially regulated metabolites and pathways responsible for this difference are discussed., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Background: Ammoides verticillata Briq is an aromatic and medicinal herb that has been widely used in folk medicine for treatment of several types of pathologies such as respiratory problems, colds, fever, headache, gastric disorders and renal infections., Objective: Essential oils have been subjected of numerous works for their antimicrobial and insecticidal properties which have been valued by hundreds of publications and patents. The main objective of this work was to evaluate the chemical composition, and the in vitro and in vivo antifungal, and insecticidal properties of essential oil of Ammoides verticillata., Methods: In this work, the air-dried aerial parts of Ammoides verticillata were hydrodistilled in a Clevenger-type apparatus. The essential oil isolated was analyzed using gas chromatography (GC) and mass spectrometry (GC/MS). The in vitro antifungal activity of the essential oil was investigated against four plant fungi using radial growth technique. The effect of the essential oil on disease development of olive caused by Penicillium crustosum and Aspergillus niger in the in vivo conditions. The fumigation test of the essential oil was evaluated against L3 stages, pupas and adults of Bactrocera oleae known as the olive fly., Results: The essential oil of A. verticillata was characterized mainly by carvacrol (44.3%), limonene (19.3%) and p-cymene (19.2%). The result of in vitro antifungal activity of essential oil showed an interesting antifungal inhibition against Alternaria alternata and Fusarium solani strains with percentage inhibition of 89%. Furthermore, oil of A. verticillata has demonstrated promising in vivo antifungal activity to control infections of olives caused by Aspergillus niger and Penicillium crustosum. In addition, the essential oil exhibited fumigation toxicity against Bactrocera oleae with mortality percentage of 100% at 2 μL/L air., Conclusion: The results showed that essential oil of A. verticillata has interesting biological activities, which suggests that oil have the potential to be used as biocide to protect olives of pathogenic fungi and pests., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Bactrocera dorsalis (Hendel) is a notorious insect pest that attacks diverse vegetables and fruits worldwide. The sterile insect technique has been developed as an environmentally friendly and effective control method that depends on the mass production of target flies. Because dietary yeast (protein) and sucrose (carbohydrate) are important in adult diets, yeast:sucrose (Y:S) mixtures are crucial for the mass-rearing of B. dorsalis. In this study, we found adult diets with different ratios of yeast to sucrose-influenced fecundity, and an extremely high or low Y:S ratios significantly decreased egg production of B. dorsalis. Additionally, the maximum oviposition efficiency was realized at dietary yeast to sucrose ratios of 1:1 and 1:3, suggesting their potential use to produce more eggs for the mass production of B. dorsalis. Here, new gel diets having different yeast concentrations (g/L water) were also assessed for rearing B. dorsalis larvae. Gel diets containing 20 g/L yeast led to a higher pupation, pupal weight and adult eclosion rate, and a shorter developmental time than other yeast concentrations. Moreover, the present gel diet also resulted in greater pupal production and adult emergence rates than previously used liquid and solid artificial diets, revealing that it is suitable for rearing B. dorsalis larvae. This research provides a useful reference on artificial diets mixtures for mass rearing B. dorsalis, which is critical for employing the sterile insect technique., (© The Author(s) 2020. Published by Oxford University Press on behalf of Entomological Society of America.)

Pesticides are necessary to fight agricultural pests, yet they are often nonspecific, and their widespread use is a hazard to the environment and human health. The genomic era allows for new approaches to specifically target agricultural pests, based on analysis of their genome and their microbiome. We present such an approach, to combat Bactrocera oleae, a widespread pest whose impact is devastating on olive production. To date, there is no specific pesticide to control it. Herein, we propose a novel strategy to manage this pest via identifying novel pharmacological targets on the genome of its obligate endosymbiotic bacterium Candidatus Erwinia dacicola. Three genes were selected as pharmacological targets. The 3D models of the Helicase, Polymerase, and Protease-C gene products were designed and subsequently optimized by means of molecular dynamics simulations. Successively, a series of structure-based pharmacophore models were elucidated in an effort to pave the way for the efficient high-throughput virtual screening of libraries of low molecular weight compounds and thus the discovery of novel modulating agents. Our methodology provides the means to design, test, and identify highly specific pest control substances that minimize the impact of toxic chemicals on health, economy, and the environment.

Background: The interaction between gut bacterial symbionts and Tephritidae became the focus of several studies that showed that bacteria contributed to the nutritional status and the reproductive potential of its fruit fly hosts. Anastrepha fraterculus is an economically important fruit pest in South America. This pest is currently controlled by insecticides, which prompt the development of environmentally friendly methods such as the sterile insect technique (SIT). For SIT to be effective, a deep understanding of the biology and sexual behavior of the target species is needed. Although many studies have contributed in this direction, little is known about the composition and role of A. fraterculus symbiotic bacteria. In this study we tested the hypothesis that gut bacteria contribute to nutritional status and reproductive success of A. fraterculus males., Results: AB affected the bacterial community of the digestive tract of A. fraterculus, in particular bacteria belonging to the Enterobacteriaceae family, which was the dominant bacterial group in the control flies (i.e., non-treated with AB). AB negatively affected parameters directly related to the mating success of laboratory males and their nutritional status. AB also affected males' survival under starvation conditions. The effect of AB on the behaviour and nutritional status of the males depended on two additional factors: the origin of the males and the presence of a proteinaceous source in the diet., Conclusions: Our results suggest that A. fraterculus males gut contain symbiotic organisms that are able to exert a positive contribution on A. fraterculus males' fitness, although the physiological mechanisms still need further studies.

Tephritid fruit flies are amongst the most significant horticultural pests globally and male chemical lures are important for monitoring and control. Zingerone has emerged as a unique male fruit fly lure that can attract dacine fruit flies that are weakly or non-responsive to methyl eugenol and cuelure. However, the key features of zingerone that mediate this attraction are unknown. As Jarvis's fruit fly, Bactrocera jarvisi (Tryon), is strongly attracted to zingerone, we evaluated the response of B. jarvisi to 37 zingerone analogues in a series of field trials to elucidate the functional groups involved in attraction. The most attractive analogues were alkoxy derivatives, with isopropoxy being the most attractive, followed by ethoxy and trifluoromethoxy analogues. All of the phenolic esters tested were also attractive with the response typically decreasing with increasing size of the ester. Results indicate that the carbonyl group, methoxy group, and phenol of zingerone are key sites for the attraction of B. jarvisi and identify some constraints on the range of structural modifications that can be made to zingerone without compromising attraction. These findings are important for future work in developing and optimising novel male chemical lures for fruit flies.

Peptidase inhibitors (PIs) are defense proteins of plants which are active against gut peptidases of different insects. Sapindus mukorossi was identified as a source of bioactive PIs which could confer resistance against Bactrocera cucurbitae, a most devastating pest of several economically important crops. In the present study, a trypsin inhibitor was purified from mature dry seeds of S. mukorossi and characterized for its biochemical properties as well as its potential for bio control of B. cucurbitae. The purified fractions from RP- HPLC through SDS-PAGE gave an apparent molecular weight of ~29 kDa. S. mukorossi trypsin inhibitor (SMTI) was found to be a non-competitive inhibitor which was active over a broad range of temperature (10-100 °C) and pH (6-11). SMTI when incorporated in artificial diet inhibited the growth and development of B. cucurbitae larvae. Gene expression analysis of trypsin and chymotrypsin genes via qRT-PCR indicated that their mRNA expression was down-regulated while that of other genes namely, Catalase, Elastase, Superoxide Dismutase, Glutathione -S-transferase and Alkaline Phosphatase was up regulated. SMTI also showed deleterious effects against different bacterial strains. The results of this study indicated that S. mukorossi trypsin inhibitor has potential to be used as a bio control agent that can reduce the harm caused by melon fruit fly and other devastating pests.

Azadirachtin, a botanical insecticide with high potential, has been widely used in pest control. Azadirachtin has shown strong biological activity against Bactrocera dorsalis in toxicological reports, but its mechanism remains unclear. This study finds that azadirachtin A inhibits the growth and development of Bactrocera dorsalis larvae. The larval weights and body sizes of the azadirachtin-treated group were significantly less than those of the control group in a concentration-dependent manner. Further, pathological sections revealed that azadirachtin destroyed the midgut cell structure and intestinal walls, while TUNEL staining showed that azadirachtin could induce apoptosis of midgut cells, and Western blot analysis indicated that Bcl-XL expression was inhibited and cytochrome c (CytC) released into the cytoplasm. The results also imply azadirachtin-induced structural alterations in the Bactrocera dorsalis larvae midgut by activation of apoptosis. RNA-seq analysis of midgut cells found that 482 and 708 unique genes were upregulated and downregulated, respectively. These differentially expressed genes (DEGs) were enriched in apoptotic and lysosomal signaling pathways and included 26 genes of the cathepsin family. qRT-PCR verified the expression patterns of some DEGs, indicating that Cathepsin F was upregulated by 278.47-fold and that Cathepsin L and Cathepsin D were upregulated by 28.06- and 8.97-fold, respectively. Finally, association analysis between DEGs and DEMs (differentially expressed metabolites) revealed that azadirachtin significantly reduced the digestion and absorption of carbohydrates, proteins, fats, vitamins and minerals in the midgut. In conclusion, azadirachtin induces the release of cathepsin from lysosomes, causing apoptosis in the midgut. Ultimately, this leads to reduced digestion and absorption of nutrient metabolites in the midgut and inhibition of the growth and development of Bactrocera dorsalis larvae., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Certain male fruit flies from the genera Bactrocera and Zeugodacus (Diptera: Tephritidae) actively forage for floral semiochemicals produced by some endemic Bulbophyllum orchids found in tropical and subtropical forests. These floral semiochemicals are largely classified as either phenylbutanoids (e.g., raspberry ketone (RK)) or phenylpropanoids (e.g., methyl eugenol (ME)). Zingerone (ZN) is a phenylbutanoid recently found that structurally resembles ME and RK, both of which are phytochemicals commonly used as male attractants in fruit fly control programmes. It was previously shown that feeding on ME and RK increased the mating success of certain tephritid fruit flies, specifically in B. dorsalis and B. tryoni males, respectively, through enhancement of sexual signaling. However, ZN, which acts as a metabolic enhancer to increase male courtship activity in B. tryoni, did not show the same effect. As fruit fly-phytochemical lure interactions are unique and species-specific phenomena, this study seeks to elucidate the ecological significance of ZN feeding to Zeugodacus tau in terms of sexual signaling. We demonstrate here that ZN feeding by Z. tau males enhanced female attraction and subsequent mating success by increasing male courtship, and the attractiveness of the sexual signals in both wind tunnel and semi-field cage bioassays. In addition, we also demonstrated temporal effects on male behaviour in relation to the amount of lure intake. However, feeding on ZN did not appear to affect the total time spent in copula for Z. tau. This is the first report showing an important role of ZN in increasing courtship activity as well as enhancement of sexual signaling in Z. tau males., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

A trypsin inhibitor was purified from the seeds of Psoralea corylifolia by ion-exchange and affinity chromatography. The purified fractions were subjected to RP- HPLC which resolved into a single peak. SDS-PAGE analysis gave an apparent molecular weight of 18 kDa. P. corylifolia trypsin inhibitor (PCTI) was found to be a competitive inhibitor and was active over a broad temperature (10-100 °C) and pH (6-11) range. It was shown to have a deleterious effect on growth and development of larvae of the melon fruit fly, Bactrocera cucurbitae, when incorporated in artificial diet using various concentrations. The larval, pupal, total development period and larval mortality significantly increased during the treatment. Inhibitory effects were also observed on percentage emergence which was significantly reduced. Nutritional indices namely food assimilated (FA) and mean relative growth rate (MRGR) also decreased significantly with increase in concentration of PCTI. qRT-PCR results indicated that the expression of trypsin and chymotrypsin genes were down-regulated while elastase, catalase, GST, SOD and AP were up-regulated. PCTI was also effective against certain bacterial strains. These results indicated that the peptidase inhibitor from P. corylifolia may be a potential bio-control agent which can decrease the damage caused by B. cucurbitae and other related destructive pests., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The melon fruit fly, Bactrocera cucurbitae (Coquillett), is a serious agricultural pest which has defied the various control measures employed against it. Protease inhibitors present in plants which have the potential to inhibit the growth and development of associated insect pests can be a possible alternative which can be manipulated for developing resistance in plants to the pest. In the present study, winged bean (Psophocarpus tetragonolobus) protease inhibitor isolated through affinity chromatography was explored for its potential to disrupt the development of melon fruit fly, B. cucurbitae. Different concentrations (12.5, 25, 50, 100, 200, and 400 µg ml-1) of the winged bean protease inhibitor (WBPI) were incorporated into the artificial diet of the second instar (64-72 h old) larvae of B. cucurbitae. The WBPI significantly delayed the larval, pupal, and total development period. The percentage pupation and adult emergence of the treated larvae was reduced as compared with control. The activities of major digestive enzymes (trypsin, chymotrypsin, leucine aminopeptidase, and elastase) decreased significantly in the larvae treated with different concentrations (50, 100, 200, and 400 µg ml-1) of WBPI. The findings reveal that the inhibitor holds considerable promise for the management of the melon fruit fly.

Nutrition and infection are closely linked. While it is now well established that hosts can modulate their nutrition after being infected, the extent to which this change in foraging provides the host with a greater fitness remains to be fully understood. Our study explored the relationships between dietary choice, macronutrients intake [i.e., protein (P) and carbohydrate (C)], infection, survival rate and growth of pathogenic bacterial population in the true fruit fly Bactrocera tryoni. Results showed that flies injected with the bacterium Serratia marcescens decreased their macronutrient intake and shifted their diet choice to carbohydrate-biased diet compared to naïve individuals. Interestingly, flies injected with either PBS (i.e., sham-infected) or heat-killed bacteria also reduced food intake and modulated diet choice but only for a day after injection. When infected flies were restricted to the diet they selected (i.e., PC 1:8), they survived better the infection than those restricted to a protein-biased diet (i.e., PC 1:5). In addition, we did not observe any growth of pathogen load in infected flies fed PC 1:8 for the first 3 days post-infection. Finally, a decrease in lipid body reserves was found in flies injected with live bacteria and, interestingly, this loss of body lipid also occurred in flies injected with heat-killed bacteria, but in a diet-dependent manner. Our results indicated that B. tryoni flies modulated their macronutrient intake and decreased the negative effects of the infection on their survival ("nutritional self-medication") the first days following the infection., (Copyright © 2019. Published by Elsevier Ltd.)

Background: Tephritid fruit flies are recognized as the most economically important insect pest group, causing significant losses to horticultural crops globally. The sterile insect technique (SIT) is used to suppress or eradicate pest fruit flies in many countries. The provisioning of adult dietary or olfactory supplementation pre-release is commonly used to improve the mating performance of sterile male flies in SIT. This study on a major pest species, Queensland fruit fly, Bactrocera tryoni (Froggatt), focused on improving mating performance by providing a semiochemical, raspberry ketone (RK), in the pre-release adult diet., Results: Survival was numerically higher for RK-supplemented males. Sexual maturity occurred 1 day earlier (from 7 to 6 days) in RK-supplemented sterile males. The mating latency period decreased with maturation age and was lower for RK-fed males. RK-supplemented sterile males increasingly mated with fertile females as they aged (10-19 days). The mating competitiveness of both RK-supplemented sterile males and RK-denied sterile males was greater than that of wild males., Conclusion: The early sexual maturity and increased mating performance of RK-supplemented sterile males indicate that the effectiveness of SIT programmes can be increased through dietary supplementation with RK during the pre-release period. © 2018 Society of Chemical Industry., (© 2018 Society of Chemical Industry.)

Exposure to plant compounds and analogues of juvenile hormone (JH) increase male mating success in several species of tephritid fruit flies. Most of these species exhibit a lek mating system, characterized by active female choice. Although the pattern of enhanced male mating success is evident, few studies have investigated what benefits, if any, females gain via choice of exposed males in the lek mating system. In the South American fruit fly, Anastrepha fraterculus, females mate preferentially with males that were exposed to volatiles released by guava fruit or treated with methoprene (a JH analogue). Here, we tested the hypothesis that female choice confers direct fitness benefits in terms of fecundity and fertility. We first carried out mate choice experiments presenting females with males treated and non-treated with guava volatiles or, alternatively, treated and non-treated with methoprene. After we confirmed female preference for treated males, we compared the fecundity and fertility between females mated with treated males and non-treated ones. We found that A. fraterculus females that mated with males exposed to guava volatiles showed higher fecundity than females mated to non-exposed males. On the other hand, females that mated methoprene-treated males showed no evidence of direct benefits. Our findings represent the first evidence of a direct benefit associated to female preference for males that were exposed to host fruit odors in tephritid fruit flies. Differences between the two treatments are discussed in evolutionary and pest management terms., Competing Interests: The authors have declared that no competing interests exist.

Phenoloxidase (PO) plays a key role in melanin biosynthesis during insect development. Here, we isolated the 2310-bp full-length cDNA of PPO1 from Zeugodacus tau, a destructive horticultural pest. qRT-polymerase chain reaction showed that the ZtPPO1 transcripts were highly expressed during larval-prepupal transition and in the haemolymph. When the larvae were fed a 1.66% kojic acid (KA)-containing diet, the levels of the ZtPPO1 transcripts significantly increased by 2.79- and 3.39-fold in the whole larvae and cuticles, respectively, while the corresponding PO activity was significantly reduced; in addition, the larval and pupal durations were significantly prolonged; pupal weights were lowered; and abnormal phenotypes were observed. An in vitro inhibition experiment indicated that KA was an effective competitive inhibitor of PO in Z. tau. Additionally, the functional analysis showed that 20E could significantly up-regulate the expression of ZtPPO1, induce lower pupal weight, and advance pupation. Knockdown of the ZtPPO1 gene by RNAi significantly decreased mRNA levels after 24 h and led to low pupation rates and incomplete pupae with abnormal phenotypes during the larval-pupal interim period. These results proved that PO is important for the normal growth of Z. tau and that KA can disrupt the development of this pest insect.

Male Annihilation Technique (MAT) is a key tool to suppress or eradicate pestiferous tephritid fruit flies for which there exist powerful male lures. In the case of Bactrocera dorsalis (Hendel), a highly invasive and destructive species, current implementations of MAT utilize a combination of the male attractant methyl eugenol (ME) and a toxicant such as spinosad ("SPLAT-MAT-ME") applied at a high density with the goal of attracting and killing an extremely high proportion of males. We conducted direct comparisons of trap captures of marked B. dorsalis males released under three experimental SPLAT-MAT-ME site densities (110, 220, and 440 per km2) near Hilo, Hawaii using both fresh and aged traps to evaluate the effectiveness of varying densities and how weathering of the SPLAT-MAT-ME formulation influenced any density effects observed. Counterintuitively, we observed decreasing effectiveness (percent kill) with increasing application density. We also estimated slightly higher average kill for any given density for weathered grids compared with fresh. Spatial analysis of the recapture patterns of the first trap service per replicate x treatment reveals similar positional effects for all grid densities despite differences in overall percent kill. This study suggests that benefits for control and eradication programs would result from reducing the application density of MAT against B. dorsalis through reduced material use, labor costs, and higher effectiveness. Additional research in areas where MAT programs are currently undertaken would be helpful to corroborate this study's findings., Competing Interests: The authors have declared that no competing interests exist.

The Oriental fruit fly, Bactrocera dorsalis, is a highly destructive pest of various fruits. The reproductive and host-finding behaviors of this species are affected by several plant semiochemicals that are perceived through chemosensory receptors. However, the chemosensory mechanisms by which this perception occurs have not been fully elucidated. We conducted RNA sequencing analysis of the chemosensory organs of B. dorsalis to identify the genes coding for chemosensory receptors. We identified 60 olfactory receptors (ORs), 17 gustatory receptors and 23 ionotropic receptors-including their homologs and variants-from the transcriptome of male antennae and proboscises. We functionally analyzed ten ORs co-expressed with the obligatory co-receptor ORCO in Xenopus oocytes to identify their ligands. We tested 24 compounds including attractants for several Bactrocera species and volatiles from the host fruits of B. dorsalis. We found that BdorOR13a co-expressed with ORCO responded robustly to 1-octen-3-ol. BdorOR82a co-expressed with ORCO responded significantly to geranyl acetate, but responded weakly to farnesenes (a mixture of isomers) and linalyl acetate. These four compounds were subsequently subjected to behavioral bioassays. When each of the aforementioned compound was presented in combination with a sphere model as a visual cue to adult flies, 1-octen-3-ol, geranyl acetate, and farnesenes significantly enhanced landing behavior in mated females, but not in unmated females or males. These results suggest that the ORs characterized in the present study are involved in the perception of plant volatiles that affect host-finding behavior in B. dorsalis., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

The use of insecticides has been a primary tool to manage Bactrocera dorsalis in Pakistan; however, recent reports of field control failures necessitate mapping out the insecticide resistance problem. Therefore, eight field strains from Pakistan, were evaluated for their resistance against trichlorfon and spinosad. Compared with a reference strain, six field strains showed high levels of resistance to trichlorfon, while two field strains expressed intermediate resistance. In case of spinosad, five field strains fell in the susceptible range, whereas, the rest of the strains represented minor resistance. Correlation analysis between LD 50 values of trichlorfon and spinosad of all the field strains revealed non-significant association, suggesting the possibility of lack of cross-resistance between both insecticides. Synergism bioassays implementing S,S,S-tributylphosphorotrithioate (DEF) and piperonyl butoxide (PBO) revealed that the LD 50 values of trichlorfon in the presence of either DEF or PBO in seven field strains were significantly reduced. However, DEF and PBO had a non-significant effect on synergizing spinosad toxicity. The results revealed resistance to trichlorfon in field strains of B. dorsalis, which might be metabolic-based. Absence or minor resistance to spinosad and lack of cross-resistance to trichlorfon, suggest that spinosad could be a potential candidate for managing B. dorsalis.

The objective of this study was to evaluate the toxicity of flavor enhancers to the oriental fruit fly Bactrocera dorsalis (Hendel). The flavor enhancers glycine, disodium guanylate, succinic acid disodium salt, monosodium glutamate (MSG), disodium inosinate, and L-alanine significantly increased the mortality of B. dorsalis flies. The mortality of flies that fed on glycine, disodium guanylate, succinic acid disodium salt, and MSG was greater than 90%. Additionally, fruit fly mortality increased with increases in both time and concentration. Glycine not only reduced the climbing ability of B. dorsalis but also affected the duration and frequency of its behavioral patterns (flight, walking, grooming and inactivity). Compared with adult flies in the control group, adult B. dorsalis flies that fed on glycine exhibited a significantly increased duration and frequency of inactivity and a decreased duration and frequency of both flight and walking. However, the effect of glycine on grooming activity was not significant. These findings demonstrate the toxic effects of flavor enhancers on B. dorsalis. Glycine also affected the behavior of adult flies at a low dose. Therefore, glycine has potentially toxic to insects and also likely to have a negative impact at sublethal concentrations.

Α reduction of pyrethroid efficacy has been recently recorded in Bactrocera oleae, the most destructive insect of olives. The resistance levels of field populations collected from Crete-Greece scaled up to 22-folds, compared to reference laboratory strains. Sequence analysis of the IIS4-IIS6 region of para sodium channel gene in a large number of resistant flies indicated that resistance may not be associated with target site mutations, in line with previous studies in other Tephritidae species. We analyzed the transcriptomic differences between two resistant populations versus an almost susceptible field population and two laboratory strains. A large number of genes was found to be significantly differentially transcribed across the pairwise comparisons. Interestingly, gene set analysis revealed that genes of the 'electron carrier activity' GO group were enriched in one specific comparison, which might suggest a P450-mediated resistance mechanism. The up-regulation of several transcripts encoding detoxification enzymes was qPCR validated, focusing on transcripts coding for P450s. Of note, the expression of contig00436 and contig02103, encoding CYP6 P450s, was significantly higher in all resistant populations, compared to susceptible ones. These results suggest that an increase in the amount of the CYP6 P450s might be an important mechanism of pyrethroid resistance in B. oleae., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Males of Bactrocera dorsalis (Hendel) (Diptera: Tephritidae) are strongly attracted to methyl eugenol (ME) (1,2-dimethoxy-4-(2-propenyl)benzene), a phenylpropanoid compound occurring in many plant species. Feeding on ME is known to enhance male B. dorsalis mating competitiveness, which can increase the effectiveness of the sterile insect technique (SIT) manifold. However, currently used systems for holding the mass-reared males in fly emergence and release facilities before release, do not allow for application of ME through feeding. Therefore, the current study was designed to evaluate different delivery systems of ME that would be applicable for large-scale application to sterile males held in such facilities. Males of a genetic sexing strain (GSS) of B. dorsalis treated by ME-aromatherapy or ME-airblown-aromatherapy that were competing with ME-fed males achieved a similar level of mating success in walk-in field cages, but the mating success was significantly higher when compared to untreated males. The results confirm the feasibility of developing ME-airblown-aromatherapy as a practical way of large scale ME delivery to enhance the mating competitiveness of sterile B. dorsalis males.

Male fruit fly attractants, cue-lure (CL) and methyl eugenol (ME), are important in the monitoring and control of pest fruit fly species. Species respond to CL or ME but not both, and there are conflicting reports on whether combining CL (or its hydroxy analogue raspberry ketone) and ME decreases their attractiveness to different species. Fruit fly monitoring programs expend significant effort using separate CL and ME traps and avoiding lure cross-contamination, and combining the two lures in one trap would create substantial savings. To determine if combining lures has an inhibitory effect on trap catch, CL and ME wicks placed in the same Steiner trap were field tested in comparison to CL alone and ME alone in Australia and Papua New Guinea (PNG). In Australia, 24 out of 27 species trapped were significantly more attracted to CL or ME alone than the combination ME/CL lure, including the pests Bactrocera bryoniae (Tryon), B. frauenfeldi (Schiner), B. kraussi (Hardy), B. neohumeralis (Hardy), B. tryoni (Froggatt) (CL-responsive), and B. musae (Tryon) (ME-responsive). In PNG, 13 out of 16 species trapped were significantly more attracted to CL or ME alone than the ME/CL combination, including the pests B. bryoniae, B. frauenfeldi, B. neohumeralis, B. trivialis (Drew), Zeugodacus cucurbitae (Coquillett) (CL-responsive) and B. dorsalis (Hendel), B. musae, and B. umbrosa (Fabricius) (ME-responsive). This study shows that combining CL and ME in the one trap in equal parts significantly reduces catches of most species of Dacini fruit flies in Australia and PNG., (© The Author(s) 2017. 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 Artocarpus fruit fly, Bactrocera umbrosa (Fabricius) (Diptera: Tephritidae), is an oligophagous fruit pest infesting Moraceae fruits, including jackfruit (Artocarpus heterophyllus Lamarck), a fruit commodity of high value in Malaysia. The scarcity of fundamental biological, physiological and ecological information on this pest, particularly in relation to behavioural response to phytochemical lures, which are instrumental to the success of many area-wide fruit fly control and management programmes, underpins the need for studies on this much-underrated pest. The positive response of B. umbrosa males to methyl eugenol (ME), a highly potent phytochemical lure, which attracts mainly males of many Bactrocera species, was shown to increase with increasing age. As early as 7 days after emergence (DAE), ca. 22% of males had responded to ME and over 50% by 10 DAE, despite no occurrence of matings (i.e. the males were still sexually immature). Male attraction to ME peaked from 10 to 27 DAE, which corresponded with the flies' attainment of sexual maturity. In wind-tunnel assays during the dusk courtship period, ME-fed males exhibited earlier calling activity and attracted a significantly higher percentage of virgin females compared with ME-deprived males. ME-fed males enjoyed a higher mating success than ME-deprived males at 1-day post ME feeding in semi-field assays. ME consumption also promotes aggregation behaviour in B. umbrosa males, as demonstrated in wind-tunnel and semi-field assays. We suggest that ME plays a prominent role in promoting sexual communication and enhancing mating performance of the Artocarpus fruit fly, a finding that is congruent with previous reports on the consequences of ME acquisition by other economically important Bactrocera species.

Field and lab trials took place in Crete (July to September 2016), concerning the residual degradation and toxicity of seven active ingredients applied as bait sprays against the olive fruit fly. Highest residues were recorded in olive leaves for dimethoate and phosmet (~ 60 mg/kg) immediately after application (day 1+), while a threefold and fivefold reduction was observed 1 week later, respectively. Residues of pyrethroids were determined at lower levels (< 10 mg/kg) but remained almost stable for a longer period of time. Finally, thiacloprid and spinosad residues were determined at 5.81 and 0.19 mg/kg respectively (day 1+), and rapidly decreased below the LOQ. Highest toxicity against the olive fruit fly was observed just right after the application of dimethoate (100%), a-cypermethrin (80%), and L-cyhalothrin (72.92%). Although the toxicity of dimethoate was significantly reduced 1 week after the application (80%) and then minimized, toxicity of pyrethroids remained almost stable (> 60%) for the first 2 weeks and then decreased to 30-40%, which remained stable up to the end of the study (8 weeks). Concerning phosmet, its toxicity ranged from 35 to 56% for 3 weeks with no significant reduction, while spinosad presented a lower toxicity profile (50% only for 1 week). The benefits of these results in the knowledge of insecticide residues and their toxicity against olive fruit fly can be used for improving olive fruit fly control.

Symbionts are associated with many insects and play several multifunctional roles in insect-microorganism mutualistic relationships. The trichlorphon-degrading symbiont Citrobacter freundii (CF-BD) of the oriental fruit fly Bactrocera dorsalis was recently discovered; however, its intraspecies transmission pathway among flies remains unknown. Here, we use fluorescence in situ hybridization (FISH), PCR detection, and a series of ingenious experiments to reveal that CF-BD was aggregated in rectal pads associated with the female ovipositor, and the CF-BD symbiont was vertically transmitted via egg surface contamination. Although CF-BD was not detected in ovaries, it was found in deposited eggs. In addition, CF-BD was readily acquired horizontally between larvae or adults via oral uptake, although it was not transferred via mating behavior. Surface sterilization of eggs had a negative effect on the insects, which exhibited a lower body weight and a sharp decrease in fecundity, suggesting important biological roles of CF-BD in the fitness of the host insects. Our findings may also help to explain the high pesticide resistance levels of B. dorsalis. Furthermore, identifying a clear transmission pathway of this organophosphorus-degrading symbiont will be useful for pesticide resistance management and future pest control technologies.

Bactrocera dorsalis (Hendel) is a fruit-eating pest that causes substantial economic damage to the fresh produce industry in tropical and sub-tropical countries. Methyl eugenol (ME) is a powerful attractant for mature males of B. dorsalis, and has been widely used for detecting, luring and eradicating B. dorsalis populations worldwide. However, the molecular mechanism underlying the olfactory perception of ME remains largely unknown. Here, we analyzed the differential proteomics profiling of the antennae between ME-responsive and ME-non-responsive males by using isobaric tags for relative and absolute quantitation (iTRAQ). In total, 4622 proteins were identified, of which 277 proteins were significant differentially expressed, with 192 up-regulated and 85 down-regulated in responsive male antennae. Quantitative real-time PCR (qRT-PCR) analysis confirmed the authenticity and accuracy of the proteomic analysis. Based on the iTRAQ and qRT-PCR results, we found that the odorant-binding protein 2 (BdorOBP2) was abundantly expressed in responsive male antennae. Moreover, BdorOBP2 was significantly up-regulated by ME in male antennae. Mature males showed significantly greater taxis toward ME than did mature females. Silencing BdorOBP2 reduced mature males' responsiveness to ME. These results indicate that BdorOBP2 may play an essential role in the molecular mechanism underlying B. dorsalis olfactory perception of ME.