Aggregation enhances the activity and growth rate of anammox bacteria and its mechanisms.

The aggregation of anaerobic ammonium oxidation (anammox) bacteria is important for the start-up and biomass retention of anammox processes. However, it is unclear whether it is beneficial to the activity, growth and reproduction of anammox bacteria. In this study, four reactor systems were developed to explore the effects of aggregation on anammox activity, growth and reproduction, after excluding the contribution of aggregation to sludge settling and retention. Results demonstrated that (i) compared with free-living planktonic bacteria, the aggregated bacteria had a higher volumetric nitrogen removal rate (0.75 kg-N/(m³·d)) and specific nitrogen removal activity (1.097 kg-N/VSS/d). And after 67 days cultivation, it had the higher sludge concentration and relative abundance (92.4%); (ii) compared with acidic polysaccharides and α-d-glucopyranose polysaccharides, β-d-glucopyranose polysaccharide play more essential roles of anammox aggregation; (iii) norspermidine triggered the secretion of α-d-glucopyranose polysaccharides to combat the toxicity, and inhibited biomass growth rate; (iv) immobilization in polyvinyl alcohol (10%) or sodium alginate (2%) gel beads was better than sodium alginate-chitosan gel beads and norspermidine (biofilm inhibitor) for the cultivation of free-living planktonic anammox bacteria. This is the first comparative study of three methods for cultivating free-living anammox bacteria. In conclusion, we found that the aggregation of anammox sludge not only facilitates biomass retention but also enhances the bioactivity, relative abundance, growth, and reproduction rate of anammox bacteria. The work is helpful to understand the formation of anammox granular sludge and contribute to the fast start-up and stable operation in anammox application.