Essential roles of eDNA and AI-2 in aerobic granulation in sequencing batch reactors operated at different settling times

Settling time has been considered as one of the most effective selection pressures for aerobic granulation in sequencing batch reactors (SBRs), i.e., poorly settleable bioparticles would be washed out from SBRs, and the heavy and good settling ones would be retained at a shorter setting time. However, its biological implication remains unclear. This study investigated the microbiological mechanisms of aerobic granulation at different settling times. It provided experimental evidence for the first time showing that a shorter settling time could enhance release of extracellular DNA through cell lysis, which in turn initiated microbial aggregation leading to increased biomass size and density, while AI-2-mediated quorum sensing was found not to be involved in initial aggregation. It was further shown that the AI-2-mediated quorum sensing system was activated to regulate the growth and maturation of aerobic granules when the biomass density reached a threshold of 1.025 g ml⁻¹. It appears from this study that a short settling time of SBR would induce microbiological and physiological responses of bacteria which are required at different stages of aerobic granulation and provide new insights into biological mechanisms of settling time-triggered aerobic granulation.