Activity of ammonia-oxidizing bacteria in enriched cultures exposed to 3,4-dimethyl-1H-pyrazole dihydrogen phosphate nitrification inhibitor

The use of nitrification inhibitors is an interesting tool to achieve a higher N efficiency in plants while decreasing the environmental impact of N fertilization. However, an integrated evaluation of the efficiency of nitrification inhibitors over time, understood as the period in which the nitrifying activity is inhibited or slows down, is necessary to assess whether their use is ecofriendly and sustainable. To test the direct efficiency of 3,4-dimethyl-1H-pyrazole dihydrogen phosphate (DMPP) on nitrification, a study has been carried out in two cultures enriched with ammonia-oxidizing bacteria (AOB) obtained from a soil with continuous N fertilization (80 kg N ha−1 year−1 as NH4NO3) and from soil without N fertilization. In addition, Cu has been evaluated as a cofactor of ammonia monoxygenase, a key enzyme in the nitrifying activity of AOBs. On the other hand, the stability of DMP has been studied both in the cultivation system enriched in AOBs and in soil to assess the efficiency of the inhibitor due to its persistence over time. Our work reveals that nitrification rates observed in cultures enriched in AOBs from genus Nitrosospira isolated from soils with continuous N fertilization were not higher than those of cultures without N fertilization. In AOB cultures, DMPP was a very efficient inhibitor of nitrification (> 50 % inhibition of integrated AMO activity), mainly due to the stability of DMP (3,4-dimethyl-1 H-pyrazole) in the cultures. However, DMP stability was significantly lower under soil conditions (> 90 % of DMP was degraded in the first 30 days of incubation). Other metals are suggested as cofactors of the enzyme ammonia monooxygenase alternatively to Cu.This work was supported by Spanish Government-Ministry of Science, Innovation and Universities (RTI2018-094623-B-C22), Spain. MCIN/AEI/10.130039/501100011033/FEDER, EU. J.M.R. was supported by a doctoral fellowship from Coordination for the Improvement of Higher Education Personnel (CAPES), of Brazilian Government (1I11903/13-9), Brazil. Open access funding provided by the Public University of Navarra