Zinc Oxide nanoparticles (ZnO NPs) are being increasingly applied in the industry, which results inevitably in the release of these materials into the hydrosphere. In this study, simulated waste-activated sludge experiments were conducted to investigate the effects of Zinc Oxide NPs and to compare it with its ionic counterpart (as ZnSO). It was found that even 1 mg/L of ZnO NPs could have a small impact on COD and ammonia removal. Under 1, 10 and 50 mg/L of ZnO NP exposure, the Chemical Oxygen Demand (COD) removal efficiencies decreased from 79.8% to 78.9%, 72.7% and 65.7%, respectively. The corresponding ammonium (NH N) concentration in the effluent significantly (P < 0.05) increased from 11.9 mg/L (control) to 15.3, 20.9 and 28.5 mg/L, respectively. Under equal Zn concentration, zinc ions were more toxic towards microorganisms compared to ZnO NPs. Under 50 mg/L exposure, the effluent Zn level was 5.69 mg/L, implying that ZnO NPs have a strong affinity for activated sludge. The capacity for adsorption of ZnO NPs onto activated sludge was found to be 2.3, 6.3, and 13.9 mg/g MLSS at influent ZnO NP concentrations of 1.0, 10 and 50 mg/L respectively, which were 1.74-, 2.13- and 2.05-fold more than under Zn ion exposure.