The effects of CeO2 nanoparticles (CeO2 NPs) exposure on biological nitrogen removal in a sequencing batch biofilm reactor (SBBR) were investigated. At low concentration (1 mg/L), no significant effect was observed on total nitrogen (TN) removal. However, at high concentrations (10 and 50 mg/L), the TN removal efficiency reduced from 74.09% to 64.26% and 55.17%, respectively. Scanning electron microscope imaging showed large amounts of CeO2 NPs adsorbed on the biofilm, which increased the production of reactive oxygen species. The exposure at only 50 mg/L CeO2 NPs measurably affected the lactate dehydrogenase release. Confocal laser scanning microscopy showed that high concentrations of CeO2 NPs reduced bacterial viability. Moreover, after a short-term exposure, extracellular polymeric substances (EPS) were observed to increase, forming a compact matrix to protect the bacteria. The activities of nitrate reductase and ammonia monooxygenase were inhibited, but there was no significant impact on the activity of nitrite oxidoreductase. (C) 2015 Elsevier Ltd. All rights reserved.