Unveiling the effect of acetate on the interactions of functional bacteria in an anammox biofilm system

Biodegradable organics make an important impact on anaerobic ammonium oxidation (anammox) system. In this study, acetate was selected as a typical biodegradable organic, and its effect on the anammox biofilm system was comprehensively discussed from the macro and micro perspectives. Under a low influent concentration of acetate (<240 ± 10 mg/L), the best total nitrogen (TN) removal performance was 96%, but it decreased to 83% when the acetate concentration increased to 350 ± 20 mg/L. With the addition of acetate, the relative abundance of the family Brocadiaceae, which contains all known anammox bacteria, gradually increased from 7.97% to 12.79%, indicating that the presence of acetate promoted enrichment of anammox bacteria in the biofilm. Metagenomic analysis further demonstrated that an appropriate concentration of acetate helps to increase the abundances of the key enzymes related to nitrogen removal and enhance the metabolism of anammox and denitrification, thereby promoting the synergy of anammox and denitrifying bacteria. Hydrazine synthase (hzs), which is unique to the anammox process, was detected in association with the genera Candidatus Kuenenia, Candidatus Jettenia and Candidatus Brocadia, with its abundance increasing from 13268 (with no addition of acetate) to 19186 (with acetate addition of 240 ± 10 mg/L). This work provides a deeper understanding of the intrinsic interactions between functional bacteria in an anammox biofilm system.