Quantitative effect of adding percentages of anammox granules on the start-up process and microbial community analysis

The anaerobic ammonium oxidation (anammox) process is challenging due to its long start-up duration and high demand for mature anammox seed sludge. However, adding a small amount of anammox sludge to the inoculum can be a reasonable solution. This study investigated the effect of adding percentage of anammox granules (0, 1, 2, 4, and 8%) in the seed sludge on the anammox start-up process. The anammox process was achieved in all five reactors after 55, 6, 5, 3 and 0 days. Increasing the adding percentage effectively shortened the duration of lag phase and cell lysis, but had little effect on the final nitrogen removal performance, except for 4% adding percentage. Families of Brocadiaceae, Burkholderiaceae, Ignavibacteriaceae, SJA-28, and Rhodocyclaceae were dominant, with a core microbiota of eight operational taxonomic unites (OTUs), and Candidatus Brocadia fulgida became the dominant anammox species. Seven synergistic members with anammox bacteria were identified by correlation network analysis. Major potential functional groups involved in C and N cycle were also observed by FAPROTAX. Together with the qPCR and sequencing results, it was suggested that more than 2% of adding percentages would result in a short lag phase, rapid growth rate in elevation stage, high final performances, and anammox bacteria abundance comparable to that in the anammox seed sludge. This crucial finding indicated the feasibility of economical and rapid start-up of the anammox process with a minimum amount of anammox seed sludge.