Measurement of the Potential Rates of Dissimilatory Nitrate Reduction to Ammonium Based on 14NH4 +/15NH4 + Analyses...

The importance of understanding the fate of nitrate (NO3^-), which is the dominant N species transferred from terrestrial to aquatic ecosystems, has been increasing because global nitrogen loads have dramatically increased following industrialization. Dissimilatory nitrate reduction to ammonium (DNRA) and denitrification are both microbial processes that use NO3^- for respiration. Compared to denitrification, quantitative determinations of the DNRA activity have been carried out only to a limited extent. This has led to an insufficient understanding of the importance of DNRA in NO3^- transformations and the regulating factors of this process. The objective of this paper is to provide a detailed procedure for the measurement of the potential DNRA rate in environmental samples. In brief, the potential DNRA rate can be calculated from the ¹⁵N-labeled ammonium (¹⁵NH4⁺) accumulation rate in ¹⁵NO3^- added incubation. The determination of the ¹⁴NH4⁺ and ¹⁵NH4⁺ concentrations described in this paper is comprised of the following steps. First, the NH4⁺ in the sample is extracted and trapped on an acidified glass filter as ammonium salt. Second, the trapped ammonium is eluted and oxidized to NO3^- via persulfate oxidation. Third, the NO3^- is converted to N2O via an N2O reductase deficient denitrifier. Finally, the converted N2O is analyzed using a previously developed quadrupole gas chromatography-mass spectrometry system. We applied this method to salt marsh sediments and calculated their potential DNRA rates, demonstrating that the proposed procedures allow a simple and more rapid determination compared to previously described methods.