The long multiplication time and extremely demanding enrichment environment requirements of Anammox bacteria (AAOB) have led to difficult reactor start-ups and hindered its practical dissemination. Few feasibility studies have been reported on the recovery of AAOB activity initiation after inlet substrate disconnection caused by an unfavorable condition, and few factors, such as indicators of the recovery process, have been explored. Therefore, in this experiment, two modified expanded granular sludge bed reactors (EGSB) were inoculated with 1.5 L anaerobic granular sludge (AGS) + 1 L Anammox sludge (AMS) (R1) and 2.5 L anaerobic granular sludge (AGS) (R2), respectively. After a long-term (140 days) starvation shock at a high temperature (38 °C), the bacteria population activity recovery experiments were conducted. After 160 days, both reactors were successfully started up, and the total nitrogen removal rates exceeded 87%. Due to the experimental period, the total nitrogen removal rate of R2 was slightly higher than that of R1 in the final stage. However, it is undeniable that R2 had a relatively long activity delay during startup, while R1 had no significant activity delay during startup. The sludge obtained from R1 had a higher specific anammox activity (SAA). Analysis of the extracellular polymer substances (EPS) results showed that the extracellular polymer content in R1 was higher than that in R2 throughout the recovery process, indicating that R1 had higher sludge stability and denitrification performance. Scanning electron microscopy (SEM) analysis showed that more extracellular filamentous bacteria could be seen in the R1 reactor with better morphology of Anammox bacteria. In contrast, the R2 reactor had fewer extracellular hyphae and micropores as a percentage and higher filamentous bacteria content. The results of microbial 16SrDNA analysis showed that R1 used AAOB as inoculum to initiate Anammox, and the reactor was enriched with Anammox bacteria earlier and in much greater abundance than R2. The experimental results indicated that inoculating mixed anaerobic granular sludge and Anammox sludge to initiate an anammox reactor was more effective.
Keywords: anaerobic granular sludge; anammox; start-up; starvation shock.