Unveiling the interaction mechanisms of key functional microorganisms in the partial denitrification-anammox process induced by COD

● The availability of PD-anammox was investigated with higher NO₃^––N concentration. ● NO₃^––N concentration affects NO₃^––N accumulation during denitrification. ● COD concentration is determinant for N removal pathways in PD-anammox process. ● The synergy/competition mechanisms between denitrifiers and anammox was explored. Partial denitrification-anammox (PD-anammox) is an innovative process to remove nitrate (NO₃^––N) and ammonia (NH₄⁺–N) simultaneously from wastewater. Stable operation of the PD-anammox process relies on the synergy and competition between anammox bacteria and denitrifiers. However, the mechanism of metabolic between the functional bacteria in the PD-anammox system remains unclear, especially in the treatment of high-strength wastewater. The kinetics of nitrite (NO₂^––N) accumulation during denitrification was investigated using the Michaelis-Menten equation, and it was found that low concentrations of NO₃^––N had a more significant effect on the accumulation of NO₂^––N during denitrification. Organic matter was a key factor to regulate the synergy of anammox and denitrification, and altered the nitrogen removal pathways. The competition for NO₂^––N caused by high COD concentration was a crucial factor that affecting the system stability. Illumina sequencing techniques demonstrated that excess organic matter promoted the relative abundance of the Denitratesoma genus and the nitrite reductase gene nirS, causing the denitrifying bacteria Denitratisoma to compete with Cadidatus Kuenenia for NO₂^––N, thereby affecting the stability of the system. Synergistic carbon and nitrogen removal between partial denitrifiers and anammox bacteria can be effectively achieved by controlling the COD and COD/NO₃^––N.