Identification of pesticide exposure-induced metabolic changes in mosquito larvae

Highlights: • Pesticides affect ecosystems at concentrations below regulatory thresholds. • We observed metabolic changes at concentrations considered safe in risk assessment. • The insecticide clothianidin induced energy-related metabolic changes in mosquitoes. • Effects were weaker but lasted longer after exposure to very low concentrations. • Low concentrations may induce environmental effects through increased energy demand. The European regulatory framework for pesticides generally applies an assessment factor of up to 100 below the acute median lethal concentration (LC50) in laboratory tests to predict the regulatory acceptable concentrations (RACs). However, long-term detrimental effects of pesticides in the environment occur far below the RACs. Here, we explored the metabolic changes induced by exposure to the neonicotinoid insecticide clothianidin in larvae of the mosquito Culex pipiens. We exposed the test organisms to the insecticide for 24 h and then measured the levels of 184 metabolites immediately and 48 h after the pulse contamination. We established a link between the exposure to clothianidin and changes in the level of three specific classes of metabolites involved in energy metabolism, namely, glycerophospholipids, acylcarnitines and biogenic amines. Remarkably, exposure to concentrations considered to be safe according to the regulatory framework (2–4 orders of magnitude lower than the acute LC50), induced longer-term effects than exposure to the highest concentration. These results suggest that a specific detoxification mechanism was only triggered by the highest concentration. We conclude that even very low insecticide concentrations increase the energy demands of exposed organisms, which potentially translates into a decline in sensitive species in the field.