NOx and ammonium isotopic fingerprints of anammox in natural and engineered systems: Implications for N isotope budgets and the use of NOx isotopes to diagnose process stability in wastewater treatment

The anaerobic oxidation of ammonium with nitrite (anammox) has been identified as a very important fixed nitrogen (N) sink, accounting for a large fraction of global fixed N loss in marine, freshwater, and semi-terrestrial environments. In engineered systems, combined nitritation-anammox is an efficient process to remove N from ammonium-rich wastewater, with nitrite as the central intermediate. During the anammox process, nitrate is being produced, providing reducing equivalents for carbon fixation. Measuring the N isotope ratios in fixed N species (i.e., ammonium, nitrite, nitrate) has proven to be a valuable tool to track N cycling in freshwater and marine ecosystems, yet its application in wastewater treatment as a tool to diagnose nitrate production pathways is novel. In this presentation we will elucidate, and compare, the N isotope effects associated with anammox 1) in vitro, 2) in a lacustrine setting, and 3) in a small-scale batch reactor for wastewater treatment. We demonstrate that the anammox nitrite/nitrate isotopic signatures are modulated by the superposition of strong kinetic (normal and inverse) and equilibrium (nitrite-nitrate) N isotope fractionation. The ammonium N isotope effect is driven by kinetic N isotope fractionation, and is similar to that of nitrification. We will discuss the possible controls on the expression of the anammox N isotope effects in the natural environment. We will also evaluate the use of nitrate/nitrite N (and O) isotope signatures to distinguish between nitrate production by anammox versus nitrite oxidation, which is important for optimizing process efficiency during wastewater treatment.