Our results underscore the importance to test pesticide combinations likely to occur in agricultural environments, and to include several bee species in environmental risk assessment schemes. © 2016 Society of Chemical Industry.

-Pollen diet is essential to nurse bees to develop hypopharyngeal glands (HPGs) and produce royal jelly to feed larvae. The effect of different diet regimes combined with pesticide exposure on the development of HPGs in honeybees was studied. In particular, a high-quality pollen mixture (high pollen diversity and protein content) and a low-quality diet (low pollen diversity and protein content) were combined with the chronic administration of thiamethoxam (10 and 40 μg/L). The impact of these stressors was evaluated by measuring the size of HPG acini and the total protein content of bee heads in 8-and 12-day-old bees. Bees fed with sub-lethal concentrations of thiamethoxam and with the low-quality diet exhibited smaller and irregularly shaped acini. The total protein content in bee head was also affected by pesticide administration, while the pollen quality did not cause significant modifications. In conclusion, these results highlighted the importance of the food quality, considered as nutritional value and pesticide contamination, for the physiological development of honeybees.HPGs / honeybee / neonicotinoid / pollen diet / chronic toxicity

In the agricultural environment, honey bees may be exposed to combinations of pesticides. Until now, the effects of these combinations on honey bee health have been poorly investigated. In this study, we assessed the impacts of biological and chemical insecticides, combining low dietary concentrations of Bacillus thuringiensis (Bt) spores (100 and 1000µg/L) with the chemical insecticide fipronil (1µg/L). In order to assess the possible effects of Cry toxins, the Bt kurstaki strain (Btk) was compared with a Bt strain devoid of toxin-encoding plasmids (Bt Cry(-)). The oral exposure to fipronil and Bt spores from both strains for 10 days did not elicit significant effects on the feeding behavior and survival after 25 days. Local and systemic physiological effects were investigated by measuring the activities of enzymes involved in the intermediary and detoxication metabolisms at two sampling dates (day 10 and day 20). Attention was focused on head and midgut glutathione-S-transferase (GST), midgut alkaline phosphatase (ALP), abdomen glyceraldehyde-3-phosphate dehydrogenase (GAPD) and glucose-6-phosphate dehydrogenase (G6PD). We found that Bt Cry(-) and Btk spores induced physiological modifications by differentially modulating enzyme activities. Fipronil influenced the enzyme activities differently at days 10 and 20 and, when combined with Bt spores, elicited modulations of some spore-induced physiological responses. These results show that an apparent absence of toxicity may hide physiological disruptions that could be potentially damaging for the bees, especially in the case of combined exposures to other environmental stressors.

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