To determine the relationship between biofilm characteristics and organic nutrient content, we investigated the development of hyporheic biofilms on artificial substrata (glass slides) in the early phase of bacterial colonization (one week) in contrasting river habitats with significantly different concentrations of dissolved organic carbon (DOC) and biodegradable and refractory DOC (BDOC and RDOC). Results suggest that the quality of DOC (i.e. BDOC versus RDOC content) controlled electron -transport system (ETS)-active bacterial abundances early in colonization, while DOC concentrations limit biofilm development onlyin the latter phase of colonization.