Multidimensional Reveal of Nitrogen Regulation on Comammox

Abstract Background The discovery of complete ammonia oxidizer (comammox) was groundbreaking. Comammox can use ammonia as the sole nitrogen source and turn it to nitrate. Moreover, genomic data indicated that comammox contained genes which can metabolize urea and nitrite. However, the feasibility of enriching comammox with urea and nitrite in long term has not been proved. This study enriched comammox’s culture by using nitrite in reactor SA and urea in reactor SB. Results The nitrification rate of reactor SB (1.29 mg N·g -1 biofilm · d -1 ) was higher than that in reactor SA (0.6 mg N · g -1 biofilm · d -1 ) at the 390 th day. Comammox outnumbered ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB) in both reactor SA (9.04 × 10 9 copies / g biofilm) and reactor SB (5.34×10 10 copies/ g biofilm). In reactor SA, comammox’s amoA accounted for 92% of the total amoA, which was higher than that in reactor SB (85%). However, the percentage of comammox (4%) in total bacteria was much lower than reactor SB (14%). The results of metagenomic sequencing showed that all the pathways of nitrogen cycle including nitrification, nitrogen fixation, denitrification, assimilation nitrate reduction, and dissimilation nitrate reduction can be detected in both reactor SA and reactor SB except the anammox pathway. The genes related to nitrite oxidation and nitrate reduction in reactor SA (TPM = 5099; TPM = 3329) was higher than that of in reactor SB (TPM = 4071; TPM = 2984), presumably due to the demand of turning nitrite to nitrate and turning nitrate to ammonia. While genes related to ammonia oxidation and urea metabolism in reactor SB (TPM = 3915; TPM = 3638) was higher than that in reactor SA (TPM = 2708; TPM = 3002). Conclusion Nitrite and urea can regulate the enrichment culture of comammox by converting its metabolic pathway. Using nitrite as sole nitrogen source can improve the proportion comammox’s amoA in total amoA while using urea as the sole nitrogen source may increase comammox’s proportion in total bacteria. These results can accelerate the enrichment of comammox and facilitate the promotion of comammox’s engineering operation.