The Jucar River, in a typical Mediterranean Basin, is expected to suffer a decline in water quality and quantity as a consequence of the climate change. This study is focused on the presence and distribution of pesticides in water and fish, using the first extensive optimization and application of the QuEChERS method to determine pesticides in freshwater fish. Majority pesticides in water - in terms of presence and concentration - were dichlofenthion, chlorfenvinphos, imazalil, pyriproxyfen and prochloraz (associated with a frequent use in farming activities), as well as buprofezin, chlorpyriphos and hexythiazox. In fish, the main compounds were azinphos-ethyl, chlorpyriphos, diazinon, dimethoate and ethion. The analysis of bio-concentration in fish indicated differences by species. The maximum average concentration was detected in European eel (a critically endangered fish species). The wide presence of pesticides in water and fish suggests potential severe effects on fish populations and other biota in future scenarios of climate change, in a river basin with several endemic and endangered fish species. The potential effects of pesticides in combination with multiple stressors require further research to prioritize the management of specific chemicals and suggest effective restoration actions at the basin scale. (C) 2013 Elsevier B.V. All rights reserved.