Insights into integrated glycerol-driven partial denitrification-anaerobic ammonium oxidation system using bioinformatic analysis: The dominance of Bacillus spp. and the potential of nitrite producing via assimilatory nitrate reduction.

Partial denitrification-anaerobic ammonium oxidation (PD/A) was considered a novel technology for biological nitrogen removal. In this study, a glycerol-driven PD/A granular sludge reactor was constructed, and its nitrogen removal efficiency and microbial mechanisms were investigated systematically. After optimization, the PD/A reactor achieved 92.3 % of the nitrogen removal (~90 % by anammox) with the influent COD/NO3--N ratio of 2.6, and approximate 1.36 mol NO3--N was required for removing 1 mol NH4+-N. Granular sludge with layered structure (anaerobic ammonium oxidizing bacteria (AnAOB) was wrapped by the heterotrophic bacteria) was successfully developed, which resulted in the sludge floating. Bacillus was firstly found to be the dominant genus in PD/A system with an abundance of 46.1 %, whereas the AnAOB only accounted for 0.2-2.8 %. Metatranscriptomic analysis showed that the metabolic characteristics obviously changed during the operation, and the differential expressing genes mainly belonged to ABC transport and quorum sensing pathway. Further analysis about the expressing patterns of nitrogen metabolism related genes indicated that the anammox related genes (mainly from Candidatus Brocadia and Candidatus Jettenia) exhibited a much higher expressing level than other genes. Interestingly, the assimilatory nitrate reduction process in Bacillus showed great NO2--N producing potential, so it was considered to be an essential pathway participating in PD/A process. This study provided a comprehensive insight into the glycerol-driven PD/A system.