The biochemical response of wild sticklebacks collected in Autumn 2005 at seven stations in the North of France was studied using a set of complementary biomarkers. Here, data on biotransformation of xenobiotics, oxidative stress exposure and damages, neurotoxicity and endocrine disruption are provided. All the sites are characterized by a specific response pattern that allows distinguishing sampling locations. Moreover, these responses are in accordance with data on existing environmental pressures and the chemical analysis of metals performed in surface water. The assessment of individual responses is completed by fish population disturbance monitoring. Based on these measurements, the investigated sites are characterized by different levels of disturbance. This study argues for a multi-parametric approach of aquatic ecosystem contamination based on association between chemical, biochemical and ecological endpoints and provides a testimony of the usefulness of stickleback for this purpose.