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While many studies on the toxicity of pesticides looked at the effects of a higher mean temperature, effects of the realistic scenario of daily temperature variation are understudied. Moreover, despite the increasing interest for the toxicity of pesticide mixtures how this is influenced by temperature has been largely ignored. We tested whether daily temperature variation (DTV) magnifies the toxicity of two pesticides with a different mode of action, the organophosphate pesticide chlorpyrifos (CPF) and the biopesticide Bacillus thuringiensis var. israelensis (Bti), and of their mixture in the vector mosquito Culex pipiens. Single exposure to CPF and Bti increased mortality and reduced female development time, and exposure to CPF also increased female wing length. DTV was not lethal and did not change the toxicity of the individual pesticides. Yet, a key novel finding was that high DTV increased the mortality of the mixture by changing the interaction between both pesticides from additive to synergistic. Given that in nature daily temperature variation is omnipresent, this is important both for vector control and for ecological risk assessment. The higher toxicity of the mixture at high DTV compared to the typically used constant test temperatures in the laboratory urges caution when evaluating the environmental impact of pesticide mixtures. (c) 2018 Elsevier B.V. All rights reserved.