Insights into N2O turnovers under polyethylene terephthalate microplastics stress in mainstream biological nitrogen removal process

The ubiquitous microplastics in wastewater have raised growing concerns due to their unintended effects on microbial activities. However, whether and how microplastics affect nitrous oxide (N₂O) (a potent greenhouse gas) turnovers in mainstream biological nitrogen removal (BNR) process remain unclear. This work therefore aimed to fill such knowledge gap by conducting both long-term and batch tests. After over 100 days of feeding with wastewater containing polyethylene terephthalate (PET) microplastics (0–500 μg/L), the long-term results showed that both production and reduction of N₂O during denitrification were reduced, as well as the N₂O production during nitrification. Accordingly, 60% reduction in N₂O accumulation and 70% reduction in N₂O production were observed in the denitrification and nitrification batch tests, respectively. Nevertheless, the long-term N₂O emission factors under PET microplastics stress were comparable to that in the control reactor, mainly because PET microplastics led to more nitrite accumulation in anoxic period. With the aid of online N₂O sensors and site-preference analysis, it was demonstrated that the heterotrophic bacteria pathway and ammonia oxidizing bacteria denitrification pathway for N₂O production were negatively affected by PET microplastics, whereas a clear increase in the contribution of hydroxylamine pathway (+ 22.9%) was observed. Further investigation revealed that PET microplastics even at environmental level (i.e. 10 μg/L) significantly reshaped the BNR sludge characteristics (e.g. much larger particle size) and microbial communities (e.g. Thauera, Rhodobacte and Nitrospira) as well as the nitrogen metabolism pathways, which were chiefly responsible for the changes of N₂O turnovers and N₂O production pathways under the PET microplastics stress.