Denitrifying phosphorus removal (DPR) performance and metabolic mechanisms of Tetrasphaera at long-term nitrite exposure

Abstract

Tetrasphaera has been identified as the main polyphosphate-accumulating organism (PAOs) in enhanced biological phosphate removal (EBPR), but the ability of Tetrasphaera to use nitrite as electron acceptor for denitrifying phosphorus removal (DPR) is still unclear. This study investigated the DPR performance and mechanism, microbial structures, and FNA inhibition of Tetrasphaera-dominated culture in the long-term acclimatization at different nitrite concentrations (10, 20, 40 and 50 mgN/L). The results showed that the most optimal influent nitrite concentration was 20 mgN/L (C/N = 6), and the removal efficiency of nitrite and phosphorus reached above 90 % and 95 %, respectively. Fluorescence In Situ Hybridization-Flow Cytometry (FISH-FCM) showed that the population abundance of Tetrasphaera was 80.95 %, in which Tet2-892 clade was the most abundant and mainly responsible for DPR with nitrite as electron acceptor. When the nitrite concentration reached 50 mgN/L (FNA = 12.80 µgHNO2-N/L, pH = 7.0 ± 0.1), electron supply deficiency and FNA inhibition resulted in only 43.0 % and 68.4 % of nitrite and phosphorus removal efficiency, respectively. The intracellular amino acid and glycogen metabolism were inhibited by FNA under this condition. A significant accumulation of N2O was observed, resulting from a combination of the insufficient electron supply, absence of nos-Z gene, and long-term FNA inhibition. DPR dominated by Tetrasphaera with appropriate nitrite as electron acceptor provides a new way to achieve nitrogen and phosphorus simultaneous removal. In addition, this study found that Tet2-892 clade of Tetrasphaera was more tolerant to FNA compared to Accumulibacter and GAOs, which provides a strategy for inhibiting GAOs activity to maintain the stability of the EBPR system.