Analyzing the effect of storage conditions on anammox recovery performance from the perspectives of time, temperature and biomass form

In this study, the effects of different storage conditions, such as temperature, storage time and biomass form, on the properties of anaerobic ammonia oxidation (anammox) were investigated along with the identification of the process mechanism. The results showed that the influence of storage time on anammox properties was stronger than that of storage temperature and biomass form. Also, the anammox recovery activity at 15 °C was better than that at 4 °C, and the anammox recovery activity of immobilized filler was better than that of anammox granular sludge (AnGS). Although cryogenic storage severely damaged anammox activity, lower loss of extracellular polymeric substances maintained the AnGS structure. The maximum recovery of specific anammox activity at 15 °C for the immobilized filler was observed to be 109%. In addition, intermittent substrate supplementation weakened the adverse effect of long-term storage on anammox activity, and was conducive to maintaining stable flora composition and promoting regeneration of anammox bacteria (AnAOB). High-throughput sequencing analysis showed that starvation resulted in increased community diversity, and the functional bacteria Candidatus Brocadia was observed to be more tolerant to starvation than Candidatus Kuenenia. Finally, principal component analysis was used to explain the complex relationship between process performance and preservation conditions. Based on the results of this work, it is recommended to preserve AnAOB in the form of immobilized filler at 15 °C and supplement substrate intermittently during long term storage. This study provides an economical and robust strategy for the short-term and long-term preservation of AnAOB. [Display omitted] •The relationship between storage conditions and anammox performance was analyzed.•Preservation at 15 °C was beneficial to maintain AnAOB metabolism.•Preservation at 4 °C reduced EPS loss and effectively maintained structure of AnGS.•Intermittent substrate supplementation was beneficial for AnAOB regeneration.•The starved samples contained higher number of Candidatus Brocadia.