Deciphering the key role of biofilm and mechanisms in high-strength nitrogen removal within the anammox coupled partial S0-driven autotrophic denitrification system

Anammox coupled partial S 0 -driven autotrophic denitrification (PS 0 AD) technology represents an innovative approach for removing nitrogen from wastewater. The research highlighted the crucial role of biofilm on sulfur particles in the nitrogen removal process. Further analysis revealed that sulfur-oxidizing bacteria (SOB) are primarily distributed in the inner layer of the biofilm, while anammox bacteria (AnAOB) are relatively evenly distributed in inner and outer layers, with Thiobacillus and Candidatus Brocadia being the dominant species, respectively. Except for anammox and PS 0 AD processes, 15 N isotope labeling tests determined that sulfur reshaped nitrogen metabolism pathways, providing solid evidence for the occurrence of sulfammox process. SOB and AnAOB collaborate in nitrogen and sulfur conversion, with SOB-drived PS 0 AD processes reducing nitrate to nitrite for AnAOB to remove ammonia. Conversely, the nitrate produced from anammox process can be reused by SOB. Metagenomic analyses verified that SOB drove the PS 0 AD process through encoding soxBYZ gene, while AnAOB might play an important role in simultaneously driving the anammox and sulfammox processes. These findings underscore the importance of biofilm and clarify the nitrogen-sulfur cycle mechanisms within the coupled system.