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ABSTRACT
Quantitative evaluation of denitrification by the dual isotope approach, which measures the stable isotope ratios of nitrogen (δ15N) and oxygen (δ18O) in nitrate, has been hampered by the wide range of values reported for the ratio of enrichment factors for 15N and 18O (15ε and 18ε, respectively) during denitrification. The objectives of this study were to determine 15ε and 18ε values at different denitrification rates under controlled conditions, and to infer possible mechanisms by which the 18ε/15ε ratio is influenced under different conditions. Column experiments were conducted at 25, 15, and 10°C, which enabled determination of 15ε and 18ε at different denitrification rates, in the absence of nitrate replenishment from ammonium oxidation and other sources. The values of 15ε and 18ε ranged from −11.8 to −14.9‰ and from −8.4 to −15.9‰, respectively, with 15ε less sensitive to changes in the denitrification rates. The resultant 18ε/15ε ratio, ranging from 0.70 to 1.17, was close to the values reported for sediment incubations, and larger than those for groundwater systems. These results are consistent with the explanations that 18ε/15ε value itself is close to unity during denitrification, and that at smaller denitrification rates, concurrent reactions including re-oxidation of nitrite to nitrate lead to smaller apparent fractionation of 18O and smaller 18ε/15ε ratios. This suggests that while linear relationships between δ18O and δ15N give a strong evidence of denitrification, apparent 18ε/15ε values are site specific and depend on the ambient conditions. In evaluating denitrification in such systems, we suggest the use of 15ε in preference to 18ε because 15ε is less sensitive to denitrification rates.
Acknowledgments
We thank Drs. Satoshi Ohno and Tetsuo Sekiguchi of the Central Region Agricultural Research Center, who kindly provided the soil samples used in this study, and Mrs. Hiromi Gouhara and Mrs. Emiko Sakurai for their assistance with the chemical analysis. We also acknowledge the support and suggestions provided by the members of the Institute for Agro-Environmental Sciences, National Agriculture and Food Research Organization.