Behaviour links physiological function with ecological processes and can be very sensitive towards environmental stimuli and chemical exposure. As such, behavioural indicators of toxicity are well suited for assessing impacts of pesticides at sublethal concentrations found in the environment. Recent developments in video-tracking technologies offer the possibility of quantifying behavioural patterns, particularly locomotion, which in general has not been studied and understood very well for aquatic macroinvertebrates to date. In this study, we aim to determine the potential effects of exposure to two neurotoxic pesticides with different modes of action at different concentrations (chlorpyrifos and imidacloprid) on the locomotion behaviour of the water louse Asellus aquaticus. We compare the effects of the different exposure regimes on the behaviour of Asellus with the effects that the presence of food and shelter exhibit to estimate the ecological relevance of behavioural changes. We found that sublethal pesticide exposure reduced dispersal distances compared to controls, whereby exposure to chlorpyrifos affected not only animal activity but also step lengths while imidacloprid only slightly affected step lengths. The presence of natural cues such as food or shelter induced only minor changes in behaviour, which hardly translated to changes in dispersal potential. These findings illustrate that behaviour can serve as a sensitive endpoint in toxicity assessments. However, under natural conditions, depending on the exposure concentration, the actual impacts might be outweighed by environmental conditions that an organism is subjected to. It is, therefore, of importance that the assessment of toxicity on behaviour is done under relevant environmental conditions.