The short- and long-term effects of nitrite on denitrifying anaerobic methane oxidation (DAMO) organisms

The denitrifying anaerobic methane oxidation (DAMO) process can achieve methane oxidation and denitrification at the same time by using nitrate or nitrite as an electron acceptor. The short- and long-term effects of nitrite on DAMO organisms were studied from macro (such as denitrification) to micro (such as microbial structure and community) based on two types of DAMO microbial systems. The results showed that the inhibitory effects of nitrite on the two DAMO microbial systems increased with rising concentration and prolonged time. In the short-term inhibitory phase, nitrite with concentrations below 100 mg N L^-1 did not inhibit the two distinct DAMO enrichments. When nitrite concentration was increased to 950 mg N L^-1, the nitrogen removal performance was completely inhibited. However, in the long-term inhibition experiment, when nitrite concentration was increased to 650 mg N L^-1, the nitrogen removal performance was completely inhibited. In addition, in acidic conditions, the real inhibitor of nitrite is FNA (free nitrous acid), while in alkaline conditions, the real inhibitor is the ionized form of nitrite. By using high-throughput sequencing technology, the species abundance and diversity of the two DAMO microbial systems showed an apparent decrease after long-term inhibition, and the community structure also changed significantly. For the enrichment culture dominated by DAMO bacteria, the substantial drop of Methylomonas may be the internal cause of the decreased nitrogen removal rate, and for the coexistence system that hosted both DAMO bacteria and archaea, the reduction of Nitrospirae may be an internal reason for the decline of the denitrification rate.