This study discovered one nitrate-calcium-based anammox start-up pathway. Compared with control, the start-up time of anammox was saved by 33.3%, and the average total nitrogen removal efficiency increased from 29.6% to 53.7% during the start-up. Besides, the continuous nitrite accumulation (1.18 mg/L) and a marked increase in the relative abundance of denitrifying and anammox bacteria were observed in the only Ca(NO)-added group. These results suggested that calcium nitrate induced partial denitrification to provide nitrite for anammox. Additionally, the role of dissimilatory nitrate reduction to ammonium (DNRA) in the Ca(NO)-added systems also deserved attention, for the contribution of DNRA to nitrate removal as well as the relative abundance of DNRA bacteria were both increased for the Ca(NO)-added groups. These results suggested that a mutualistic symbiosis among denitrification, DNRA and anammox exists in the calcium nitrate-added systems, which may explain the reason for acceleration of anammox start-up by adding calcium nitrate.

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