Essential (Mg, Fe, Zn and Cu) and non-essential (Cd and Pb) elements in predatory insects (Vespa crabro and Vespa velutina): a molecular perspective

The recent introduction of the Asian hornet, Vespa velutina, into Europe has raised concern regarding the threat to honeybees and the competition with the European hornet, Vespa crabro. The aim of this study was to investigated essential (Mg, Fe, Zn, Cu) and non-essential (Cd and Pb) elements in these two species. Element concentrations were determined in the whole body and separately in the head, thorax and abdomen using atomic absorption spectrometry (AAS). The changes in essential element concentration and speciation during metamorphosis were also studied using gel filtration chromatography followed by AAS and proteomic analysis. In both species, the essential elements were more concentrated in the abdomen due to the presence of fat bodies. Magnesium, Fe and Zn concentrations were significantly higher in V. crabro than in V. velutina and could have been related to a higher aerobic energy demand of the former species required to sustain foraging flight at higher speeds and to cover longer distances. Low concentrations of Cd and Pb were indicative of low environmental exposure. The concentration and speciation of essential elements, particularly Fe, varied among the developmental stages, indicating a modification of ligand preferences during metamorphosis. Overall, the results in the present study provide a better understanding of the hornet metal metabolism and a foundation for additional studies

Combined exposure to sublethal concentrations of an insecticide and a fungicide affect feeding, ovary development and longevity in a solitary bee

Pollinators in agroecosystems are often exposed to pesticide mixtures. Even at low concentrations, the effects of these mixtures on bee populations are difficult to predict due to potential synergistic interactions. In this paper, we orally exposed newly emerged females of the solitary bee Osmia bicornis to environmentally realistic levels of clothianidin (neonicotinoid insecticide) and propiconazole (fungicide), singly and in combination. The amount of feeding solution consumed was highest in bees exposed to the neonicotinoid, and lowest in bees exposed to the pesticide mixture. Ovary maturation and longevity of bees of the neonicotinoid and the fungicide treatments did not differ from those of control bees. By contrast, bees exposed to the pesticide mixture showed slow ovary maturation and decreased longevity. We found a synergistic interaction between the neonicotinoid and the fungicide on survival probability. We also found an interaction between treatment and emergence time (an indicator of physiological condition) on longevity. Longevity was negatively correlated to physiological condition only in the fungicide and the mixture treatments. Delayed ovary maturation and premature death imply a shortened nesting period (highly correlated to fecundity in Osmia). Our findings provide a mechanism to explain the observed dynamics of solitary bee populations exposed to multiple chemical residues in agricultural environments

A holistic approach for the investigation of bee health

Despite current evidence is in strong disagreement with an emergency for the conservation of Apis mellifera, great concern is related to profitability of beekeeping operations. A growing involvement of veterinary science in addressing bee health topics will therefore be fundamental to preserve and protect the entire sector. The experiments in this thesis focused on two different and interdependent levels related to bee health: the biochemical level and the parasitological level. At the biochemical level the impact of plant protection products on bee physiology and survival was studied, elucidating synergistic interactions between poor nutrition and pesticide exposure in A. mellifera and between an insecticide and a fungicide in Osmia bicornis. Moreover, an innovative fingerprinting approach on honey bee haemolymph was applied to detect population imbalances in the hive. The control of Varroa infestations was studied both at the biochemical and parasitological level. A panel of biomarkers in honey bee haemolymph was applied to compare different mite control protocols. This resulted in relevant indications for beekeeping operations pursuing the least impact on nutritional status of the colonies. To guide the decision making of beekeepers, a new formic acid evaporator was tested in comparison with a more established one. Considering its widespread distribution in the country, efforts were directed also towards N. ceranae. In particular, the pivotal aspect of diagnosis was studied, proposing a new qPCR method to overcome some limits of the existing ones. In conclusion, this works fills some of the knowledge gaps of the beekeeping sector. However, many of them still need to be addressed and the upcoming menaces of climate change and dispersal of pathogens via globalization should be targeted by research efforts in the near future. Therefore, a multifaceted vision of bee health is of capital importance, aware of the complementarity of reductionist and holistic approaches

Consumption

Consumption datase

Hemolymph

Hemolymph datase

Looking for biomarkers of health status in the hemolymph of Apis mellifera

The demographic decline of honeybees (Apis mellifera) determines negative effects not only for agriculture and apiculture, but also for the ecosystems. For this reason, it is mandatory to understand the determinants causing the decline and to establish objective criteria to evaluate their health status. Proteins in the hemolymph may represent a source of valuables biomarkers to assess the health status of the hive; however, they are still poorly investigated. The aim of this study is to separate, to identify the most abundant proteins in the hemolymph, and to identify biomarkers of health status that could help beekeepers and veterinarians in the management and treatment of the hives. Four apiaries were chosen for this research, characterized by different management and location (A= hill, B and D= plain, C= apiary subjected to nomadism). In each apiary, three hives were selected and bees were collected in June, July, August, September, and October. Then 2 µL of hemolymph were withdrawn from each bee and the total proteins were determined by the Bradford method. The proteins of the hemolymph were separated using the SDS-PAGE electrophoresis and identified using mass spectrometry. Colony-level traits (areas covered with bees, brood, sealed brood and pollen) were also estimated. In the hemolymph of honeybees, 20 proteins were identified and among these, five proteins showed relevant variations. These are vitellogenin, apolipophorin, hexamerin, transferrin and prophenoloxidase. In particular, in October, the apiary C presented worst general conditions and a lower population associated with lower concentrations of total proteins, apolipoforin and vitellogenin. The apiary D, on the contrary, had the best trophic and health conditions, associated with higher concentrations of all the aforementioned five proteins. Thereby, it is possible to hypothesize that these proteins may be valuables biomarkers of health status, and it is worthy to continue their study

Data from: Neonicotinoid pesticides and nutritional stress synergistically reduce survival in honey bees

The honey bee (Apis mellifera L.) is a major pollinator whose health is of global concern. Declines in honey bee health are related to multiple factors, including resource quality and pesticide contamination. Intensive agricultural areas with crop monocultures potentially reduce the quality and quantity of available nutrients and expose bee foragers to pesticides. However, there is, to date, no evidence for synergistic effects between pesticides and nutritional stress in animals. The neonicotinoids clothianidin (CLO) and thiamethoxam (TMX) are common systemic pesticides that are used worldwide and found in nectar and pollen. We therefore tested if nutritional stress (limited access to nectar, and access to nectar with low sugar concentrations) and sublethal, field-realistic acute exposures to two neonicotinoids (CLO and TMX at 1/5 and 1/25 of LD50) could alter bee survival, food consumption, and hemolymph sugar levels. Bee survival was synergistically reduced by the combination of poor nutrition and pesticide exposure (-50%). Nutritional and pesticide stressors reduced also food consumption (-48%) and hemolymph levels of glucose (-60%) and trehalose (-27%). Our results provide the first demonstration that field-realistic nutritional stress and pesticide exposure can synergistically interact and cause significant harm to animal survival. These findings have implications for current pesticide risk-assessment procedures and pollinator protection

Survival

Survival datase

SDS-PAGE-Based Quantitative Assay of Hemolymph Proteins in Honeybees: Progress and Prospects for Field Application

In human and veterinary medicine, serum proteins are considered to be useful biomarkers for assessing the health and nutritional status of the organism. Honeybee hemolymph has a unique proteome that could represent a source of valuable biomarkers. Therefore, the aims of this study were to separate and identify the most abundant proteins in the hemolymph of worker honeybees to suggest a panel of these proteins that could represent useful biomarkers for assessing the nutritional and health status of the colonies and, finally, to analyze them in different periods of the year. Four apiaries were selected in the province of Bologna, and the bees were analyzed in April, May, July, and November. Thirty specimens from three hives of each apiary were sampled and their hemolymph was collected. The most represented bands obtained after 1D sodium-dodecyl-sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) were cut from the gel, and the proteins were identified using an LC-ESI-Q-MS/MS System. A total of twelve proteins were unmistakably identified; the two most abundant proteins were apolipophorin and vitellogenin, which are known biomarkers of bee trophic and health status. The two other proteins identified were transferrin and hexamerin 70a, the first being involved in iron homeostasis and the second being a storage protein. Most of these proteins showed an increase from April to November, mirroring the physiological changes of honeybees during the productive season. The current study suggests a panel of biomarkers from honeybee hemolymph worth testing under different physiological and pathological field conditions