Peeking into the Stingers: A Comprehensive SWATH-MS Study of the European Hornet Vespa crabro (Linnaeus, 1758) (Hymenoptera: Vespidae) Venom Sac Extracts.

This study aimed to investigate the venom sac extracts (VSEs) of the European hornet (EH) Vespa crabro (Linnaeus, 1758) (Hymenoptera: Vespidae), focusing on the differences between stinging females, gynes (G), and workers (W), at the protein level. Using a quantitative "Sequential Window Acquisition of all Theoretical Fragment Ion Mass Spectra" (SWATH-MS) analysis, we identified and quantified a total of 240 proteins. Notably, within the group, 45.8% (n = 110) showed significant differential expression between VSE-G and VSE-W. In this set, 57.3% (n = 63) were upregulated and 42.7% (n = 47) downregulated in the G. Additionally, the two-hundred quantified proteins from the class Insecta belong to sixteen different species, six of them to the Hymenoptera/Apidae lineage, comprising seven proteins with known potential allergenicity. Thus, phospholipase A1 (Vesp v 1), phospholipase A1 verutoxin 2b (VT-2b), hyaluronidase A (Vesp v 2A), hyaluronidase B (Vesp v 2B), and venom allergen 5 (Vesp v 5) were significantly downregulated in the G, and vitellogenin (Vesp v 6) was upregulated. Overall, 46% of the VSE proteins showed differential expression, with a majority being upregulated in G. Data are available via ProteomeXchange with identifier PXD047955. These findings shed light on the proteomic differences in VSE between EH castes, potentially contributing to our understanding of their behavior and offering insights for allergy research.

JAK/STAT mediated insulin resistance in muscles is essential for effective immune response.

BACKGROUND: The metabolically demanding nature of immune response requires nutrients to be preferentially directed towards the immune system at the expense of peripheral tissues. We study the mechanisms by which this metabolic reprograming occurs using the parasitoid infection of Drosophila larvae. To overcome such an immune challenge hemocytes differentiate into lamellocytes, which encapsulate and melanize the parasitoid egg. Hemocytes acquire the energy for this process by expressing JAK/STAT ligands upd2 and upd3, which activates JAK/STAT signaling in muscles and redirects carbohydrates away from muscles in favor of immune cells. METHODS: Immune response of Drosophila larvae was induced by parasitoid wasp infestation. Carbohydrate levels, larval locomotion and gene expression of key proteins were compared between control and infected animals. Efficacy of lamellocyte production and resistance to wasp infection was observed for RNAi and mutant animals. RESULTS: Absence of upd/JAK/STAT signaling leads to an impaired immune response and increased mortality. We demonstrate how JAK/STAT signaling in muscles leads to suppression of insulin signaling through activation of ImpL2, the inhibitor of Drosophila insulin like peptides. CONCLUSIONS: Our findings reveal cross-talk between immune cells and muscles mediates a metabolic shift, redirecting carbohydrates towards immune cells. We emphasize the crucial function of muscles during immune response and show the benefits of insulin resistance as an adaptive mechanism that is necessary for survival.

VespAI: a deep learning-based system for the detection of invasive hornets.

The invasive hornet Vespa velutina nigrithorax is a rapidly proliferating threat to pollinators in Europe and East Asia. To effectively limit its spread, colonies must be detected and destroyed early in the invasion curve, however the current reliance upon visual alerts by the public yields low accuracy. Advances in deep learning offer a potential solution to this, but the application of such technology remains challenging. Here we present VespAI, an automated system for the rapid detection of V. velutina. We leverage a hardware-assisted AI approach, combining a standardised monitoring station with deep YOLOv5s architecture and a ResNet backbone, trained on a bespoke end-to-end pipeline. This enables the system to detect hornets in real-time-achieving a mean precision-recall score of ≥0.99-and send associated image alerts via a compact remote processor. We demonstrate the successful operation of a prototype system in the field, and confirm its suitability for large-scale deployment in future use cases. As such, VespAI has the potential to transform the way that invasive hornets are managed, providing a robust early warning system to prevent ingressions into new regions.

Wasp sting-induced bilateral thalamic and midbrain infarction.

A male patient in his early 40s presented to the emergency department with an acute onset of respiratory distress and facial oedema, indicative of anaphylaxis. These symptoms emerged 2 hours subsequent to a wasp sting on the left side of his face. Despite initial stabilisation, the patient's state deteriorated into somnolence and disorientation. Notably, he denied any history of seizures, sensory or motor deficits, or bowel/bladder complications. Physical examination unveiled no focal neurological deficits. Routine laboratory tests and drug screening yielded no significant findings. Subsequent brain MRI with angiography exposed bilateral thalami diffusion restriction, strongly implying an acute infarction within the artery of Percheron territory, an atypical vascular variant. The sequence of events, alongside the absence of other conclusive aetiologies, indicated a wasp sting-induced thalamic infarction driven by vasogenic and thrombogenic effects of inflammatory substances.

Subtle morphological changes in the visual and antennal sensory system of bees and wasps across an urbanisation gradient.

Increased temperature and fragmentation of green spaces in urban areas could drive variations in functional traits of insects. Such morphological shifts may occur for sensory systems, which were previously reported to be prone to change with habitat characteristics in non-urban contexts. Here, we measured traits related to the visual and antennal sensory systems in the bees Halictus scabiosae and Osmia cornuta and the wasp Polistes dominula along an urbanisation gradient within Milan (Italy). We hypothesised that fragmentation could filter for better visual properties, and that higher temperature could filter for fewer thermoreceptors and more olfactory hairs. While controlling for body size, results show subtle but appreciable responses to urbanisation in one or more traits in all species, though not always supporting our hypotheses. O. cornuta shows marginally higher ommatidia density and smaller ommatidia diameter (associated with better visual resolution) in more fragmented sites, as well as marginally fewer thermoreceptors in hotter sites, in agreement with our two predictions. On the other hand, H. scabiosae has marginally smaller antennae and P. dominula has smaller eyes at warmer locations, and the wasp also has smaller antennae and 9th flagellomeres in more fragmented areas. Perhaps higher temperatures accelerate development of sensory system at higher speed than the rest of body in these two species. Our results represent the first evidence of urbanisation effects on the visual and antennal sensory systems of bees and wasps and underline how such effects may involve a much broader bouquet of traits then previously observed.

Giant honeybees (Apis dorsata) trade off defensiveness against periodic mass flight activity.

The giant honeybee Apis dorsata (Fabricius, 1793) is an evolutionarily ancient species that builds its nests in the open. The nest consists of a single honeycomb covered with the bee curtain which are several layers of worker bees that remain almost motionless with their heads up and abdomens down on the nest surface, except for the mouth area, the hub between inner- and outer-nest activities. A colony may change this semi-quiescence several times a day, depending on its reproductive state and ambient temperature, to enter the state of mass flight activity (MFA), in which nest organisation is restructured and defense ability is likely to be suppressed (predicted by the mass-flight-suspend-defensiveness hypothesis). For this study, three episode of MFA (mfa1-3) of a selected experimental nest were analysed in a case study with sequences of >60 000 images at 50 Hz, each comprise a short pre-MFA session, the MFA and the post-MFA phase of further 10 min. To test colony defensiveness under normative conditions, a dummy wasp was cyclically presented with a standardised motion programme (Pd) with intervening sessions without such a presentation (nPd). Motion activity at five selected surveillance zones (sz1-5) on the nest were analysed. In contrast to mfa1,2, in mfa3 the experimental regime started with the cyclic presentation of the dummy wasp only after the MFA had subsided. As a result, the MFA intensity in mfa3 was significantly lower than in mfa1-2, suggesting that a colony is able to perceive external threats during the MFA. Characteristic ripples appear in the motion profiles, which can be interpreted as a start signal for the transition to MFA. Because they are strongest in the mouth zone and shift to higher frequencies on their way to the nest periphery, it can be concluded that MFA starts earlier in the mouth zone than in the peripheral zones, also suggesting that the mouth zone is a control centre for the scheduling of MFA. In Pd phases of pre- and postMFA, the histogram-based motion spectra are biphasic, suggesting two cohorts in the process, one remaining at quiescence and the other involved in shimmering. Under MFA, nPd and Pd spectra were typically Gaussian, suggesting that the nest mates with a uniform workload shifted to higher motion activity. At the end of the MFA, the spectra shift back to the lower motion activities and the Pd spectra form a biphasic again. This happens a few minutes earlier in the peripheral zones than in the mouth zone. Using time profiles of the skewness of the Pd motion spectra, the mass-flight-suspend-defensiveness hypothesis is confirmed, whereby the inhibition of defense ability was found to increase progressively during the MFA. These sawtooth-like time profiles of skewness during MFA show that defense capability is recovered again quite quickly at the end of MFA. Finally, with the help of the Pd motion spectra, clear indications can be obtained that the giant honeybees engage in a decision in the sense of a tradeoff between MFA and collective defensiveness, especially in the regions in the periphery to the mouth zone.

Serratia montpellierensis sp. nov., Isolated from Laboratory-Reared Parasitic Wasps Psyttalia lounsburyii Silvestri and Psyttalia ponerophaga Silvestri (Hymenoptera: Braconidae).

Two strains of bacteria, PsyLou2AT and PsyPon4B, were isolated from adult braconid wasps Psyttalia lounsburyii and Psyttalia ponerophaga, respectively. These laboratory-reared wasps were investigated as agents for biological control of the olive fruit fly, Bactrocera oleae. Analysis of 16S rRNA genes of the two isolates demonstrated that they were highly related and belonged to the genus Serratia. Genomic sequencing of these isolates revealed genomes of 5,152,551 bp and 5,154,385 bp for PsyLou2AT and PsyPon4B, respectively, and both genomes had a mol% G+C content of 59.6%. Phylogenetic analyses using BLAST-based average nucleotide identity (ANIb), and digital DNA-DNA hybridization methods indicated that PsyLou2AT was most closely related to Serratia nevei S15T, producing ANIb and dDDH values of 96.11% and 70.2%, respectively. Since these values were literally on the species cutoff threshold, additional S. nevei genome assemblies were analyzed using ANIb and dDDH calculations. This revealed that among assemblies that were clearly identifiable as S. nevei, S. nevei S15T was the most closely related to PsyLou2AT, and that a majority of assemblies produced dDDH values of 68.3-68.7% relative to PsyLou2AT. Additionally, PsyLou2AT differed biochemically from S. nevei S15T in that it produced positive Voges Proskauer tests, produced protease, lacked arginine dihydrolase, and did not utilize D-lactose. Hence, PsyLou2AT represents a novel taxon within the Serratia, for which we propose the name Serratia montpellierensis sp. nov. The type strain is PsyLou2AT (=LMG 32817T =NRRL B-65689T).

New records of three parasitoids, Pteroptrix chinensis, Aphytis hispanicus, and Marlattiella prima (Hymenoptera: Aphelinidae) associated with an exotic scale, Lopholeucaspis japonica (Hemiptera: Diaspididae) in Tennessee.

A survey for parasitoids of Lopholeucaspis japonica Cockerell (Hemiptera: Diaspididae), an exotic scale of woody ornamentals, resulted in the discovery of 3 species of aphelinid parasitoid wasps, Pteroptrix chinensis (Howard), Aphytis hispanicus (Mercet), and Marlattiella prima Howard. This serves as the first report of these parasitoids reared from a host in the state of Tennessee, USA. Despite routine pesticide applications in the surveyed nursery and directed treatments of the infested plants to control the scale outbreak, the percentage of parasitized scale in privet and euonymus shrubs averaged 7.0% and 7.9%, respectively. These parasitoids may be useful in the natural or managed control of this pest in the United States, but additional research is needed to understand how these parasitoids contribute to the control of L. japonica in the landscape and how nursery production practices can be modified to promote parasitoid populations.

Effect of egg production dynamics on the functional response of two parasitoids.

Functional response describes the number of hosts attacked by a parasitoid in relation to host densities and plays an important role by connecting behavioral-level processes with community-level processes. Most functional response studies were carried out using simple experimental designs where the insects were confined to a plain and small arena with different host densities during a fixed period of time. With these designs, other factors that might affect the functional response of parasitoids were not analyzed, such as fecundity, age, and experience. We proposed a series of latent-variables Markovian models that comprised an integrated approach of functional response and egg production models to estimate the realized lifetime reproductive success of parasitoids. As a case study, we used the parasitoids Anagyrus cachamai and A. lapachosus (Hymenoptera: Encyrtidae), two candidate agents for neoclassical biocontrol of the Puerto Rican cactus pest mealybug, Hypogeococcus sp. (Hemiptera: Pseudococcidae). The tested species were assessed according to their physiology and prior experience. We estimated the number of mature eggs after emergence, egg production on the first day, egg production rate, the proportion of eggs resorbed, egg resorption threshold, and egg storage capacity. Anagyrus cachamai and A. lapachosus both presented a type III functional response. However, the two parasitoids behaved differently; for A. cachamai, the number of parasitized hosts decreased with female age and depended on the number of mature eggs that were available for oviposition, whereas A. lapachosus host parasitism increased with female age and was modulated by its daily egg load and previous experience. The methodology presented may have large applicability in pest control, invasive species management, and conservation biology, as it has the potential to increase our understanding of the reproductive biology of a wide variety of species, ultimately leading to improved management strategies.

Identification of Parthenogenesis-Inducing Effector Proteins in Wolbachia.

Bacteria in the genus Wolbachia have evolved numerous strategies to manipulate arthropod sex, including the conversion of would-be male offspring to asexually reproducing females. This so-called "parthenogenesis induction" phenotype can be found in a number of Wolbachia strains that infect arthropods with haplodiploid sex determination systems, including parasitoid wasps. Despite the discovery of microbe-mediated parthenogenesis more than 30 yr ago, the underlying genetic mechanisms have remained elusive. We used a suite of genomic, computational, and molecular tools to identify and characterize two proteins that are uniquely found in parthenogenesis-inducing Wolbachia and have strong signatures of host-associated bacterial effector proteins. These putative parthenogenesis-inducing proteins have structural homology to eukaryotic protein domains including nucleoporins, the key insect sex determining factor Transformer, and a eukaryotic-like serine-threonine kinase with leucine-rich repeats. Furthermore, these proteins significantly impact eukaryotic cell biology in the model Saccharomyces cerevisiae. We suggest that these proteins are parthenogenesis-inducing factors and our results indicate that this would be made possible by a novel mechanism of bacterial-host interaction.

Functional Characterization of the Niemann-Pick C2 Protein BdioNPC2b in the Parasitic Wasp Baryscapus dioryctriae (Chalcidodea: Eulophidae).

Reverse chemical ecology has been widely applied for the functional characterization of olfactory proteins in various arthropods, but few related studies have focused on parasitic wasps. Here, the odorant carrier Niemann-Pick C2 protein of Baryscapus dioryctriae (BdioNPC2b) was studied in vitro and in vivo. Ligand binding analysis revealed that BdioNPC2b most strongly bound to 2-butyl-2-octenal and which compound could elicit an EAG response and attracted B. dioryctriae adults. Moreover, this odorant attractant significantly improved the reproductive efficiency of B. dioryctriae compared to that of the control. Then, the relationship between BdioNPC2b and 2-butyl-2-octenal was validated by RNAi, and site-directed mutagenesis revealed the involvement of three key residues of BdioNPC2b in binding to 2-butyl-2-octenal through hydrogen bonding. Our findings provide not only a deeper understanding of the olfactory function of NPC2 in wasps but also useful information for improving the performance of the parasitoid B. dioryctriae as a biological control agent.

Small wasps, big muscles: Fore and hind leg modifications in chalcidoid wasps (Hymenoptera: Chalcidoidea).

A particularly conspicuous morphological feature in chalcidoid wasps are strikingly modified legs present in both males and females. It evolved convergently multiple times on either fore or hind legs implying strong evolutionary pressure and a prominent function in the wasps' life history. We investigate the external and internal morphology of the modified legs of five species of chalcidoid wasps representing four families (Ooderidae, Heydeniidae, Chalcididae, and Leucospidae), using light microscopy, scanning electron microscopy and micro computed tomography. We aimed to identify shared characteristics as well as differences between genera/species, leg pairs and sexes and to draw first conclusions about the shared or different functions. All species and sexes share the same general leg morphology, with enlarged femur, curved tibia and a huge flexor tibiae muscle. However, there are also genus/species-specific differences such as distinctive spine-like setae on the femur of Oodera spp., or leg pair-specific differences in the position of the extensor tibiae muscle. Shared characteristics imply a common primary function in which strong forces are required to pull the tibia against the femur while differences imply different secondary functions. Both primary and secondary functions have yet to be revealed beyond informed speculations.

An odorant receptor mediates the avoidance of Plutella xylostella against parasitoid.

BACKGROUND: Ecosystems are brimming with myriad compounds, including some at very low concentrations that are indispensable for insect survival and reproduction. Screening strategies for identifying active compounds are typically based on bioassay-guided approaches. RESULTS: Here, we selected two candidate odorant receptors from a major pest of cruciferous plants-the diamondback moth Plutella xylostella-as targets to screen for active semiochemicals. One of these ORs, PxylOR16, exhibited a specific, sensitive response to heptanal, with both larvae and adult P. xylostella displaying heptanal avoidance behavior. Gene knockout studies based on CRISPR/Cas9 experimentally confirmed that PxylOR16 mediates this avoidance. Intriguingly, rather than being involved in P. xylostella-host plant interaction, we discovered that P. xylostella recognizes heptanal from the cuticular volatiles of the parasitoid wasp Cotesia vestalis, possibly to avoid parasitization. CONCLUSIONS: Our study thus showcases how the deorphanization of odorant receptors can drive discoveries about their complex functions in mediating insect survival. We also demonstrate that the use of odorant receptors as a screening platform could be efficient in identifying new behavioral regulators for application in pest management.

Mitochondrial perturbation in immune cells enhances cell-mediated innate immunity in Drosophila.

BACKGROUND: Mitochondria participate in various cellular processes including energy metabolism, apoptosis, autophagy, production of reactive oxygen species, stress responses, inflammation and immunity. However, the role of mitochondrial metabolism in immune cells and tissues shaping the innate immune responses are not yet fully understood. We investigated the effects of tissue-specific mitochondrial perturbation on the immune responses at the organismal level. Genes for oxidative phosphorylation (OXPHOS) complexes cI-cV were knocked down in the fruit fly Drosophila melanogaster, targeting the two main immune tissues, the fat body and the immune cells (hemocytes). RESULTS: While OXPHOS perturbation in the fat body was detrimental, hemocyte-specific perturbation led to an enhanced immunocompetence. This was accompanied by the formation of melanized hemocyte aggregates (melanotic nodules), a sign of activation of cell-mediated innate immunity. Furthermore, the hemocyte-specific OXPHOS perturbation induced immune activation of hemocytes, resulting in an infection-like hemocyte profile and an enhanced immune response against parasitoid wasp infection. In addition, OXPHOS perturbation in hemocytes resulted in mitochondrial membrane depolarization and upregulation of genes associated with the mitochondrial unfolded protein response. CONCLUSIONS: Overall, we show that while the effects of mitochondrial perturbation on immune responses are highly tissue-specific, mild mitochondrial dysfunction can be beneficial in immune-challenged individuals and contributes to variation in infection outcomes among individuals.

The paper wasps (Hymenoptera: Vespidae: Polistinae) of Sri Lanka recorded from recent investigations.

Paper wasps of subfamily Polistinae Lepeletier have been studied in many countries of the world due to their importance as pest species, predators, model organisms in research and medical significance. Seven species have been well documented in Sri Lanka, of these five species represent genus Ropalidia Gurin-Mneville, and two species genus Polistes Latrielle. However, the species have not been studied systematically for many years and recent records are not available. In the present study investigations for wasps (Vespidae) were conducted in 28 locations of all provinces and climatic zones of the country. Five species of paper wasps were found in 15 of the locations investigated, four in the genus Ropalidia and one in the genus Polistes. Ropalidia marginata Lepeletier was the most abundant and widely distributed species, while the other species had more limited distribution. Polistes (Gyrostoma) olivaceus De Geer, previously recorded from Sri Lanka, was not recorded during the present study. All the species of paper wasps encountered in the present study showed changes in distribution from their historical locations, decline in distributional ranges and occurrence in new locations.

Two new species of Epirhyssa (Hymenoptera: Ichneumonidae: Rhyssinae) from Costa Rica.

As a result of continuing research on the ichneumonid wasps of Costa Rica, two new species of Epirhyssa (Rhyssinae) are described: E. leonoreae and E. lewisi. A key is provided for the identification of all 14 species now known to occur in the country.

Review of the subgenus Tarpheion of the genus Blacus Nees (Hymenoptera, Braconidae, Brachistinae) from China, with description of nine new species and eight new records.

An updated key to the currently known species of the subgenus Tarpheion van Achterberg, 1976 (Hymenoptera, Braconidae, Blacus) in China is provided. Nine new species are proposed, B. (T.) adelphius sp. nov., B. (T.) frontalis sp. nov., B. (T.) gilvus sp. nov., B. (T.) hainanensis sp. nov., B. (T.) parilis sp. nov., B. (T.) reticulatus sp. nov., B. (T.) sculptilis sp. nov., B. (T.) tanae sp. nov., and B. (T.) wuyishanensis sp. nov. Eight species, B. (T.) achterbergi Haeselbarth, 1976, B. (T.) albiventris van Achterberg, 1988, B. (T.) angichorus van Achterberg, 1988, B. (T.) antennalis van Achterberg, 1988, B. (T.) apicalis van Achterberg, 1976, B. (T.) artomandibularis van Achterberg, 1976, B. (T.) bicolor van Achterberg, 1988, and B. (T.) soror van Achterberg, 1988, are newly recorded from China.

Taxonomic notes on some neglected doryctine wasps (Hymenoptera: Braconidae) from Argentina described by J. Brthes and E. Blanchard.

The type specimens of Austrodolops eremitae Blanchard, Doryctes ridiaschinae Brthes, and Hormiopius ptericoptophagus Blanchard were studied and reclassified according to the current classification of the subfamily Doryctinae. The monotypic genera Austrodolops Blanchard and Hormiopius Blanchard are treated as junior synonyms of Allorhogas Gahan (syn. nov.) and Heterospilus Haliday (syn. nov.) respectively. Examination of the gall associated species D. ridiaschinae shows that it belongs in the genus Allorhogas. All species are redescribed and illustrated and the following new combinations are proposed: Allorhogas eremitae (Blanchard) (comb. nov.), Allorhogas ridiaschinae (Brthes) (comb. nov.) and Heterospilus ptericoptophagus (Blanchard) (comb. nov.). Lectotypes are designated for all studied species and the genus Doryctes is excluded from the currently known Argentine fauna.

Revision of the genus Cyrtolabulus van der Vecht in Madagascar (Hymenoptera: Vespidae: Eumeninae).

The species of the genus Cyrtolabulus van der Vecht, 1969 occurring in Madagascar are revised, with the description of three new species: Cyrtolabulus flavorufus sp. nov., Cyrtolabulus parvulus sp. nov. and Cyrtolabulus scrobalis sp. nov. Pseudonortonia madacassa Gusenleitner, 2012 is synonymized under Cyrtolabulus suboscurus (Giordani Soika, 1941) syn. nov., and a neotype is designated for Labus bekilyensis Giordani Soika, 1941. A comparison with the closely related genus Cyrteumenes Giordani Soika, 1991 and a key to the species are provided.

Review of the spider wasp genus Episyron Schidte, 1837 (Hymenoptera: Pompilidae) from India, with description of three new species.

The spider wasp genus Episyron Schidte, 1837 is reviewed, with five species recorded from various parts of India along with three new species described and illustrated from Kerala: E. keralaensis Anju, Girish Kumar & Thejass, sp. nov., E. nigrocalcarius Anju, Girish Kumar & Thejass, sp. nov., and E. rufotibius Anju, Girish Kumar & Thejass, sp. nov. Diagnosis of species occurring in India, comparisons of new species with congeners and a key to the Indian species also are provided.