Metabolomics analysis of larval secretions reveals a caste-driven nutritional shift in a social wasp colony.

Social wasps exhibit a unique nutritional cycle in which adults feed larvae with prey, and larvae provide adults with larval secretions (LS). LS serves as a vital nutritional source for adults, contributing to the colony's health and reproductive success. The LS nutrient composition has been previously reported in various wasp species, yet these analyses focused solely on worker-destined larvae, overlooking the potential caste designation effects on LS composition. Using metabolomics techniques, we analysed and compared the metabolite and nutrient composition in LS of queen- and worker-destined larvae of the Oriental hornet. We found that queen-destined LS (QLS) contain greater amounts of most metabolites, including amino acids, and smaller amounts of sugars compared to worker-destined LS (WLS). The amino acid-to-sugar ratio in QLS was approximately tenfold higher than in WLS. Thus, as the colony transitions from the production of workers to the production of reproductives, it gradually experiences a nutritional shift that may influence the behaviour and physiology of the adult nest population. This caste-specific metabolite profile and nutrient composition of LS reflect the differences in the diet and physiological requirements of worker- and queen-destined larvae and may play a critical role in caste determination in social wasps.

Passive heat diffusion in nests with downward-facing cells: Implications for early colony development in social wasps.

Social insects employ a variety of active and passive mechanisms for nest thermoregulation. Many social wasp species exhibit a particular nest-architecture by building their nests with cells facing downward. By using thermal imaging to characterize the heat diffusion throughout Oriental hornet nests from different angular positions, we show that the heat diffusion along the vertical gradient of nests is more efficient when the cell openings face downward than when facing sideways or upward, demonstrating the efficiency of this specific architecture in increasing the nest temperature. This passive thermoregulation mechanism could be especially important during the initial stage of the colony, when the queen is alone to rear her first brood. Among the social insects that build cells to raise their brood, we suggest that wasps can take advantage of the thermal benefits of this particular architecture of their cells as, unlike bees, they do not usually store food in them.

Continuous exchange of nectar nutrients in an Oriental hornet colony.

Nutritional exchanges play a fundamental role in the evolution of animal societies. In higher animal societies, while adult individuals can be both food donors and receivers, the offspring usually only receive food from the adults. Hornets and wasps are fierce insect hunters that feed their larvae with prey. However, although the adults also consume floral nectar, the role of nectar in vespid nutrition has remained largely unknown. We provided experimental colonies of the Oriental hornet with artificial nectar enriched with a 13C-labeled amino acid, and found that a continuous cycle of nutrition took place, in which nectar nutrients were used and exchanged back and forth between adults and larvae. We posit that this continuous cycle of nutrients constitutes a mechanism contributing to social cohesion. In an additional experiment, we found that nectar consumption was essential for adult and larval survival, suggesting the importance of wasps and hornets as pollinators in natural ecosystems.

The effects of dietary proline, ß-alanine, and γ-aminobutyric acid (GABA) on the nest construction behavior in the Oriental hornet (Vespa orientalis).

Adult wasps primary food resource is larval saliva. This liquid secretion consists mainly of amino acids and carbohydrates processed from the prey brought to the colony by the foragers. However, adults also regularly consume floral nectar. The nectar's most abundant proteinogenic amino acid is proline, and the two most abundant non-proteinogenic amino acids are ß-alanine and GABA. These three amino acids are also common in larval saliva. Here, we study the effect of these dietary amino acids on the physiology and nest construction behavior of the Oriental hornet. Our results reveal their deleterious effects, especially at high concentrations: ß-alanine and GABA consumption reduced the hornets' lifespan and completely inhibited their construction behavior; while proline induced a similar but more moderate effect. At low concentrations, these amino acids had no effect on hornet survival but did slow down the nest construction process. Using carbon isotopically labeled amino acids, we show that, unlike proline, ß-alanine is stored in most body tissues (brain, muscles, and fat body), suggesting that it is rapidly metabolized after consumption. Our findings demonstrate how a single amino acid can impact the fitness of a nectarivore insect.

Nutrient Utilization during Male Maturation and Protein Digestion in the Oriental Hornet.

Males of social Hymenoptera spend the first days following eclosion inside the nest before dispersing to find a young queen to mate with. During this period, they must acquire enough nutrients to enable their sexual maturation and store energy to sustain them through their nuptial journey. It was previously argued that adult hornets are unable to process dietary proteins and rely on the larvae to supply them with free amino acids and carbohydrates that they secrete via trophallaxis. Using isotopically enriched diets, we examined nutrient allocation and protein turnover in newly-emerged males of the Oriental hornet during their maturation period and tested the protein digestion capability in the presence and absence of larvae in both males and worker hornets. The results indicated that protein turnover in males occurs during the first days following eclosion, while carbohydrates are incorporated into body tissues at higher rates towards the end of the maturation period. Additionally, we found that males cannot digest protein and depend on larval secretions as a source of nutrition, while workers, in contrast to previous reports, can metabolize protein independently. Our findings demonstrate the contribution of adult male nutrition and larval secretions to colony fitness.

Allocation and metabolism of naturally occurring dietary amino acids in the Oriental hornet.

Oriental hornet (Vespa orientalis) foragers are strong, long-distance flyers exhibiting a high metabolic rate. Accordingly, they feed on carbohydrate-rich diets, such as floral nectar and larval secretions. These nutritional sources, in addition to carbohydrates, also contain free amino acids (AAs). Leucine, glycine, and proline are three common AAs in the diet of social wasps. Using diets enriched with carbon-specific (13C1) isotopically labeled leucine, glycine, and proline, and a cavity ring-down spectroscope (CRDS) stable carbon isotope analyzer, we examined the metabolism of these AAs, their allocation in the hornets' respiration during rest and activity, and their incorporation into the body tissues. In hornets that consumed 13C proline, we detected the heavy isotope only in the exhaled CO2, suggesting that proline was utilized solely as a metabolic fuel and was not incorporated into their body (i.e., as protein). Labeled carbons from glycine and leucine, in contrast, were found in all the examined tissues (i.e., muscles, brain, fat bodies, ovaries, and venom glands), and were also utilized as a metabolic fuel, but mostly during rest. Using AAs labeled with a specific stable carbon isotope, we demonstrate the compatibility between the hornet's metabolic requirements and AA use, in both the living organism as a whole and in its different body tissues.

Morphological and Molecular Revision of the Genus Ozirhincus (Diptera: Cecidomyiidae)-Long-Snouted Seed-Feeding Gall Midges on Asteraceae.

The Palaearctic gall-midge genus Ozirhincus is unique among the Cecidomyiidae for its morphology and biology. Unlike most other phytophagous gall midges, species in this genus do not induce galls but develop inside achenes of Asteraceae plants. The heads of adults are characterized by an unusually elongate proboscis, the function of which is unclear. Despite a lot of attention from taxonomists in the 19th and early 20th century, a proper revision of the genus has been hindered by complex host associations, the loss of most relevant type material, and the lack of a thorough comparative study of all life stages. The present revision integrated morphological, molecular, and life-history data to clearly define species boundaries within Ozirhincus, and delimit host-plant ranges for each of them. A phylogenetic analysis based on the mitochondrial COI and 16S genes confirmed the validity of four distinct species but did not resolve the relationships among them. All species are oligophages, and some may occur together on the same host plant. Species with wider host-plant ranges have wider European and circum-Mediterranean distribution ranges, whereas species with narrower host ranges are limited to Europe and the Russian Far East. As part of the present work, O. hungaricus is reinstated from synonymy, O. tanaceti is synonymized under O. longicollis, neotypes are designated for O. longicollis and O. millefolii, and a lectotype is designated for O. anthemidis.