The Water Framework Directive requires the development of biological tools which can act as early-warning indicators of a sudden increase (accidental pollution) or decrease (recovery due to prevention) of the chemical status of aquatic systems. River biofilms, which respond quickly to modifications of environmental parameters and also play a key part in the functioning of aquatic ecosystems, are therefore good candidates to monitor an increase or a decrease of water pollution. In the present study, we investigated the biological response of biofilms transplanted either upstream (recovery) or downstream (deterioration of exposure levels) the urban area of Paris (France). Both modifications of Cu community tolerance levels and of global bacterial and eukaryotic community structure using automated ribosomal intergenic spacer analysis (ARISA) fingerprints were examined 15 and 30 days after the transplantation. Cu tolerance levels of the heterotrophic component of biofilms were assessed using a short-term toxicity test based on β-glucosidase (heterotrophic) activity. Cu tolerance increased for biofilms transplanted upstream to downstream Paris (5-fold increase on day 30) and conversely decreased for biofilms transplanted downstream to upstream (8-fold decrease on day 30). ARISA fingerprints revealed that bacterial and eukaryotic community structures of transplanted biofilms were closer to the structures of biofilms from the transplantation sites (or sites with similar contamination levels) than to biofilms from their sites of origin. Statistical analysis of the data confirmed that the key factor explaining biofilm Cu tolerance levels is the sampling site and not the site of origin. It also showed that Cu tolerance levels are related to the global urban contamination (both metals and nutrients). The study shows that biofilms adapt fast to modifications of their surroundings. In particular, community tolerance varies quickly and reflects the new exposure levels only 15 days after transplantation. Those results support the use of biofilms as reliable early-warning indicators of diffuse urban contamination.