Insight into biofilm formation of wastewater treatment processes: Nitrogen removal performance and biological mechanisms

Biofilm formation affects biological nitrogen (N) removal, and a sequencing batch biofilm reactor (SBBR) was set up to evaluate the changes in N removal and microbial characteristics during biofilm formation. The results indicated that the average effluent concentration of chemical oxygen demand (COD), ammonia nitrogen (NH₄⁺-N) and total nitrogen (TN) in the SBBR were 27.48, 1.41, and 13.52 mg L⁻¹, respectively after biofilm formation. Furthermore, this process increased microbial richness, but reduced microbial diversity. Patescibacteria, Proteobacteria, and Bacteroides were the dominant phyla that did not change after biofilm formation. After biofilm formation, Firmicutes was eliminated while Spirochaetes involved in the interspecies relationship. Biofilm increased the nitrification and denitrification relating coding genes abundance (hao, narG, narZ, nxrA, narH, narY, nxrB, napA, napB, norB, norC and nosZ), and enhanced the processes of N respiration and denitrification, carbohydrate metabolism, amino acid metabolism and membrane transport. Meanwhile, correlation analysis between genera and transcriptome reflected that Zooglea, Micropruina, Aeromonas and Tessaracoccus played essential roles in biofilm formation and N removal. The key enzyme abundance of EC:1.7.99.1, EC:1.7.2.4, and EC:1.1.1.42 of N and tricarboxylic acid (TCA) cycle increased after biofilm formation. This study can reveal the effect of biofilm formation on biological N removal and provide a theoretical foundation for the application of biofilm process.