In this study, the inhibitory effects of acid orange 7 (AO7), a common azo dye, on nitrification in biofilms were investigated in situ using microelectrodes. Biofilms were obtained from laboratory rotating drum biofilm reactor after the nitrification process reached a pseudo-steady state. Dissolved oxygen, pH, NH4+, NO3, and redox potential microelectrodes, with tip diameters ranging from 3–15 μm, were used to monitor the spatial distribution and change of microbial activities within nitrifying biofilms. It was found that at lower concentration (1 mg/L), AO7 had only a slight impact on the NH4+-N concentration profiles. The ammonium consumption rate decreased as higher AO7 concentrations (15 mg/L and 25 mg/L) were exposed to the biofilms. A similar trend was observed for the NO3-N microprofiles. The nitrate production rate decreased as the AO7 concentration in the bulk solution increased. The dissolved oxygen and pH microprofiles also showed oxygen and alkalinity utilization, but at lower rates throughout the biofilms when the nitrification process was inhibited. No significant redox potential differences were observed in the biofilms after AO7 was applied.

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