Dissolved inorganic N fluxes, including nitrification and denitrification rates, were measured in situ under dark and light conditions using epilithic biofilms from a 6th to 8th Strahler-order reach of the River Garonne, France. A broad range of epilithic biofilm biomasses (6–51 g ash-free dry mass [AFDM]/m2) was assessed in 39 benthic chamber experiments. Nitrification was detected in 31 cases (0.1–7.7 mg N m2 h1) and caused a net release of NO3 – in spite of algal assimilation in light conditions in 5 cases. Denitrification (Dw) (0–8.1 mg N m2 h1), which accounted for 83% of gross NO3 – removal in the dark, was a significant, permanent N sink. Calculation of mean (61 SE) N budgets indicated epilithon release of inorganic N in the dark (1.5 6 0.3 mg N m2 h1) that was significantly related to biofilm mass. Calculation of mean N budgets under light indicated net uptake (1.8 6 0.4 mg N m2 h1). However, a net release was observed when biomass was .40 g AFDM/m2. A calculated daily budget showed evidence for a threshold between photoautotrophy (N assimilation) and heterotrophy (N mineralization) of 23 g AFDM/m2. This threshold approximated mean biomass at low water and may correspond to an equilibrium value for epilithic biomass in the river. A conceptual scheme of the epilithic biofilm function and the consequences on the water-column N content in rivers is proposed.