AHL-mediated quorum sensing drives microbial community succession and metabolic pathway in algal-bacterial biofilm system.

Microalgae, ammonia-oxidizing bacteria (AOB), and anaerobic ammonium-oxidizing bacteria (AnAOB) have been proven to form an integrated algal-bacterial biofilm system with over 93 % of total nitrogen removal. Compared to conventional nitrification-denitrification process, this system operated without additional organic carbon or aeration. In order to understand the interaction mechanisms between bacteria and algae, this study investigated microbial community succession, the changes in metabolic pathways and the potential role of acyl-homoserine lactone (AHL)-mediated quorum sensing (QS) during the formation of the algae/partial nitrification/anammox biofilm system. Within this algal-bacterial symbiotic biofilm, the dominant genera identified were Candidatus_Brocadia (AnAOB), Nitrosomonas (AOB), and Geitlerinema (microalgae), with relative abundances of 13.86 %, 6.37 %, and 2.88 %, respectively. Compared with the first two stages, the abundance of genes related to nitrogen metabolism pathways (anaerobic ammonium oxidation, denitrification, and ammonia assimilation) increased, indicating enhanced nitrogen transformation capacity in the algal-bacterial symbiotic stage. Co-occurrence network analysis also revealed enhanced microbial interactions, with increased negative correlations (from 36.07 % to 39.38 %), high average standard betweenness centrality (from 0.193 to 0.304), and reduced community vulnerability (from 0.037 to 0.028), contributing to biofilm stability and resilience. The variations in AHLs provided direct evidence for more frequent interspecies communication, facilitating the ecological reconfiguration in the biofilm. Overall, the close synergistic relationship between microalgae and bacteria supports stable biofilm development and high nitrogen removal efficiency.