Bombus terrestris (L.) is one of the most important native and commercial pollinator species worldwide. Along with other pollinators their populations are in decline due to a multifactorial phenomenon that includes the extensive use of neonicotinoid insecticides. Thus, the characterisation and understanding of neonicotinoid effects on bees at the molecular level is essential to mitigate the risks of their use in the environment. This study initially characterised the brain proteomes of bumblebees in response to aging prior to assessing changes at the behavioural, cellular and molecular level as a response to neonicotinoid exposure. We demonstrated the highly catalytic nature of the developing bumblebee brain and how energy and carbohydrate metabolism increase in response to aging, while genetic information processes are downregulated. By considering differences in mode of action and mode of exposure to the neonicotinoids clothianidin and imidacloprid, the effects of acute and chronic oral exposure on bumblebee workers were determined. Neonicotinoids differentially impair energy metabolism and structural processes in the brain suggesting possible divergence of insecticide mode of action. Clothianidin and imidacloprid triggered different behavioural responses and toxicity in bees, with the former causing hyperactivity and the latter, temporal paralysis. Imidacloprid is less toxic to bumblebees and the brain physiology is differentially affected depending on chemical, dose or mode of exposure selected. The levels of the synapse associated protein synapsin increased in bumblebee brains for imidacloprid-exposed bees only, and functional annotation analysis of differential expressed proteins indicated impairment of intracellular transport, energy metabolism, translational activity, purines and pyrimidines metabolism, endocytic and exocytic activity and synaptic functioning as a whole. The pathways affected by neonicotinoid exposure vary depending on chemical and mode of exposure, which complicates the identification of biomarkers of neonicotinoid exposure in bumblebees. In addition, neonicotinoid metabolism in bees is poorly understood and these chemicals can accumulate in the bee body, which potentially contributes to long term toxicity. Overall the results presented in this thesis demonstrate individual and distinct ways by which neonicotinoids influence neuronal communication and provide novel insights into molecular aspects of bee health, through highlighting the pathways affected by aging and pesticide use on this important pollinator species
