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Nanosized plastics (nanoplastics) releasing into the wastewater may pose a potential threat to biological nitrogen removal. Constructed wetland (CW), a wastewater treatment or shore buffer system, is an important sink of nanoplastics, while it is unclear how nitrogen removal in CWs occurs in response to nanoplastics. Here, we investigated the effects of polystyrene (PS) nanoplastics (0, 10, and 1000 μg/L) on nitrogen removal for 180 days in CWs. The results revealed that total nitrogen removal efficiency decreased by 29.5–40.6%. We found that PS penetrated the cell membrane and destroyed both membrane integrity and reactive oxygen species balance. Furthermore, PS inhibited microbial activity in vivo, including enzyme (ammonia monooxygenase, nitrate reductase, and nitrite reductase) activities and electron transport system activity (ETSA). These adverse effects, accompanied by a decline in the relative abundance of nitrifiers (e.g., Nitrosomonas and Nitrospira) and denitrifiers (e.g., Thauera and Zoogloea), directly accounted for the strong deterioration observed in nitrogen removal. The decline in leaf and root activities decreased nitrogen uptake by plants, which is an important factor of deterioration in nitrogen removal. Overall, our results imply that the presence of nanoplastics in the aquatic environment is a hidden danger to the global nitrogen cycle and should receive more attention.

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