Long-term exposure to zinc oxide nanoparticles improves PAOs function in enhanced biological phosphorus removal

As the most widely applied process for biological phosphorus removal, enhanced biological phosphorus removal (EBPR) relies on phosphorus accumulating organisms (PAOs) and glycogen accumulating organisms (GAOs), whose function is crucial for the removal of phosphorus. In this study, the effect of zinc oxide nanoparticles (ZnO NPs, 0-50 mg/L) on EBPR performance was investigated in both long-term reactors and batch experiments. It was found that the performance of biological phosphorus removal was recovered from 0% (day 0) to >99% (day 70) after long-term exposure of ZnO NPs (50 mg/L). Further studies revealed that ZnO NPs treatment caused no significant effects on the morphology and settleability of activated sludge, but enhanced the release and uptake of phosphorus as well as the transformations of polyhydroxyalkanoates and glycogen in activated sludge, which suggested that PAOs were re-activated during long-term exposure to ZnO NPs. Fluorescence in-situ hybridization (FISH) analysis showed that the relative abundance of PAOs was increased after long-term exposure. Meanwhile, the enzymatic activities of PPX and PPK were also enhanced. These results indicated that compared with short-term exposure, long-term exposure to ZnO NPs favours PAOs function and thus led to the recovery of biological phosphorus removal.