Biological nutrient removal (BNR) is an effective process for removing nitrogen from wastewater using microorganisms under different environmental conditions. However, these processes can result in nitrous oxide (N2O) emission. Dissolved oxygen (DO) levels shape bacterial communities and affect N2O emissions in BNR. Ammonia-oxidizing bacteria (AOB), the main N2O producers in the BNR process, are significantly affected by DO. The bacterial communities responding to DO at the DNA level in BNR have been elucidated. However, the metabolically active microbes are truly participant in the biodegradation process. Thus, it is worth exploring how metabolically active bulk and AOB microbial communities and their N2O production respond to the DO levels. In this study, N2O emission were measured in lab-scale sequencing batch reactors at varying DO levels. High-throughput sequencing of the metabolically active bacterial 16S rRNA and amoA genes was performed. The results indicate that increasing DO levels positively influence metabolically active bacterial biodiversity and AOB relative abundance without increasing N2O emissions. The carbon source significantly affected denitrification and nitrification in activated sludge. Optimal DO levels and adequate carbon sources can effectively reduce N2O accumulation. However, excessive aeration reduces the abundance of Zoogloea, leading to compromised sludge settling. This study revealed the effect of DO on N2O producing microorganisms based on cDNA levels, which provided an important basis for mitigating N2O emission in BNR.
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