Effects of sterilization techniques on chemodenitrification and N₂O production in tropical peat soil microcosms

Chemodenitrification – the non-enzymatic process of nitrite reduction – may be an important sink for fixed nitrogen in tropical peatlands. Rates and products of chemodenitrification are dependent on O₂, pH, Fe²⁺ concentration, and organic matter composition, which are variable across peat soils. Assessing abiotic reaction pathways is difficult because sterilization and inhibition agents can alter the availability of reactants by changing iron speciation and organic matter composition. We compared six commonly used soil sterilization techniques – γ irradiation, chloroform, autoclaving, and the use of three different chemical inhibitors (mercury, zinc, and azide) – for their compatibility with chemodenitrification assays for tropical peatland soils (organic-rich, low-pH soil from the eastern Amazon). Out of the six techniques, γ irradiation resulted in soil treatments with the lowest cell viability and denitrification activity and the least effect on pH, iron speciation, and organic matter composition. Nitrite depletion rates in γ-irradiated soils were highly similar to untreated (live) soils, whereas other sterilization techniques showed deviations. Chemodenitrification was a dominant process of nitrite consumption in tropical peatland soils assayed in this study. Nitrous oxide (N₂O) is one possible product of chemodenitrification reactions. Abiotic N₂O production was low to moderate (3 %–16 % of converted nitrite), and different sterilization techniques lead to significant variations on production rates due to inherent processes or potential artifacts. Our work represents the first methodological basis for testing the abiotic denitrification and N₂O production potential in tropical peatland soil.