More than stoichiometry: the molecular composition of inorganic and organic substrates controls ammonium regeneration by bacteria

Abstract

The mineralization of nitrogen (N) and especially the regeneration of ammonium are critical processes performed by bacteria in aquatic ecosystems. Quantifying these processes is complicated because bacteria simultaneously consume and produce ammonium. Here we use experimental data on the effects of the molecular composition of the supplied substrates, combined with a classical stoichiometric model of ammonium regeneration, to demonstrate how the quantification of these processes can be improved. We manipulated a batch culture experiment with an isolated bacterial community by adding three different types of N substrates: dissolved inorganic nitrogen (DIN, nitrate), dissolved organic nitrogen (DON, amino acid) and a mixture of DIN and DON. With such experiment set-up, the ammonium regeneration per se could be easily tracked without using complicated methods (e.g. isotope dilution). We compared the experimental data with the predictions of Goldman et al model (1987) as well as with a revised version, using the measured consumption carbon:nitrogen ratio (C:N ratio), rather than an estimated consumption ratio. We found that, for all substrates, and in particular, mixed substrates where C and N are partially dissociated between different molecules, estimates of ammonium regeneration rates can be improved by measuring the actual consumption C: N ratio. ImportanceMeasuring bacterial ammonium regeneration in natural aquatic ecosystem is difficult because bacteria in the field simultaneously consume and produce ammonium. In our experimental design, we used nitrate as the inorganic nitrogen substrate. This way, we could measure separately the uptake and excretion of inorganic nitrogen by bacteria without incorporating cumbersome methods such as isotope dilution. Our experiment allowed us to evaluate the accuracy of various stoichiometric models for the estimation of net bacterial nitrogen regeneration. We found that: O_LIThe exact distribution of C and N among the various molecules that make the bulk of DOM is a crucial factor to consider for bacterial net nitrogen regeneration. C_LIO_LIFor all substrates, and in particular, mixed substrates where C and N are partially dissociated between different molecules, estimates of net nitrogen regeneration rates can be improved by measuring the actual C: N ratio of bacterial consumption. C_LI