Constructed wetlands (CWs) have been proven to effectively immobilize plastic particles. However, little is known about the differences in the impact of varying sized plastic particles on nitrous oxide (N₂O) release, as well as the intervention mechanisms in CWs. Here, we built a lab-scale wetland model and introduced plastic particles of macro-, micro-, and nano-size at 100 μg/L for 370 days. The results showed that plastic particles of all sizes reduced N₂O release in CWs, with the degrees being the strongest for the Nano group, followed by Micro and Macro groups. Meanwhile, ¹⁵N- and ¹⁸O-tracing experiment revealed that the ammoxidation process contributed the most N₂O production, followed by denitrification. While for every N₂O-releasing process, the contributing proportion of N₂O in nitrification-coupled denitrification were most significantly cut down under exposing to macro-sized plastics and had an obvious increase in nitrifier denitrification in all groups, respectively. Finally, we revealed the three mechanism pathways of N₂O release reduction with macro-, micro-, and nano-sized plastics by impacting carbon assimilation (RubisCO activity), ammonia oxidation (gene amo abundance and HAO activity), and N-ion transmembrane and reductase activities, respectively. Our findings thus provided novel insights into the potential effects of plastic particles in CWs as an eco-technology.

Ammonia-oxidizing microorganisms (AOMs, archaea (AOA) and bacteria (AOB)) are primarily responsible for the ammoxidation in constructed wetlands (CWs). However, little is known about evaluating the response of AOA and AOB to engineered nanomaterials (ENMs) and quantifying the shift of their contribution to ammoxidation. Herein, we operated a series of CWs exposing to silver nanoparticles (Ag-NPs), single-walled carbon nanotubes (SWCNTs), and polystyrene nano-sized plastics (PS-NPs) with the wastewater-accumulating concentration of ENMs for 180 days. The results showed that the abundance of AOA amoA gene in situ was far lower than that of AOB, while the abundance ratio of AOA to AOB increased by 15 folds after 180-day experiment. Using DNA stable isotope probing (DNA-SIP) experiment, we found that the active AOB microbiota varied substantially but the AOA was more stable across different groups. Furthermore, the co-occurrence analysis proved that ENMs stress increased the negative coexistence pattern of AOA and AOB; predictive functional profiling showed that the ENMs enhanced the functional advantage of AOA by inhibiting AOB (mainly hydroxylamine oxidation process). Finally, the contribution of AOA increased under exposing to SWCNTs (18.35%), PS-NPs (24.92%), and Ag-NPs (32.14%) compared with control group (0.03%) for 180 days. Despite this, AOB was still the primary executant of ammoxidation in CWs. Overall, in our study, the differences in activities and contributions of AOMs were quantified in CWs, and a significantly negative coexistence relationship between AOA and AOB was revealed when exposed to emerging nanomaterials.

The extracellular polymeric substances (EPS) from activated sludge played significant roles in the removal of nanoparticles from wastewater. A series of batch experiments were carried out to determine the adsorption mechanism of three nano-Ag by activated sludge, as well as the contributions of EPS fractions including dissolved EPS (DEPS), loosely bound EPS (LB-EPS) and tightly bound EPS (TB-EPS). The results demonstrated that the adsorption of nano-Ag by sludge biomass agreed with pseudo-second-order kinetic reaction model and Freundlich isotherm model. About 26.0-41.2% of nano-Ag was trapped by the bound EPS (BEPS) matrix of activated sludge (especially LB-EPS) and 42.5-52.6% of them was adsorbed onto the inner cells after the adsorption. Moreover, the interaction energy contributions of EPS fractions followed the order of EDE > 0 > ETB > ELB, suggesting DEPS in wastewater went against the removal of nano-Ag due to steric repulsion while LB-EPS and TB-EPS were positive to nano-Ag adsorption by modifying biomass surface and providing extensive binding sites. Besides, EPS fractions played significant roles in the adsorption of nano-Ag with low initial concentrations but had limited effect at high concentrations. Overall, this study investigated the effect of EPS fractions on the adsorption behaviors of nano-Ag by activated sludge biomass, which is meaningful to understand the removal mechanism of nanoparticles in sewage and the potential role of EPS fractions.

Dissolved black carbon (DBC), widely found in soil and water environments is likely to affect the transport of nanoplastics in aquatic environments. The aggregation and deposition behaviors of fresh and aged polystyrene nanoplastics (PSs) with and without DBC in NaCl solution were investigated by time-resolved dynamic light scattering (DLS) and quartz crystal microbalance with dissipation monitoring equipment (QCM-D) techniques. The results suggest that DBC can screen the surface charges of PSs by interacting with PSs through hydrogen bonding, hydrophobic interactions and π-π interactions, although they were negatively charged. DBC promoted the aggregation of PSs under relatively low ionic strengths, and it minimally affected the stability of PSs under high ionic strength. Deposition experiments showed that both DBC in salt solution and DBC adsorption on silica surface facilitated the deposition of fresh PSs while HA inhibited both deposition processes. After aging, PSs were more stable, and the effects of DBC and HA were weakened. This study investigated the influence mechanism of DBC on the aggregation and deposition behaviors, which provides new insights into the stability and transport of PSs in complex aquatic environments.