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Hardwater lakes are common in human-dominated regions of the world and often experience pollution due to agricultural and urban effluent inputs of inorganic and organic nitrogen (N). Although these lakes are landscape hotspots for CO2 exchange and food web carbon (C) cycling, the effect of N enrichment on hardwater lake food web functioning and C cycling patterns remains unclear. Specifically, it is unknown if different eutrophication scenarios (e.g., modest non point vs. extreme point sources) yield consistent effects on auto- and heterotrophic C cycling, or how biotic responses interact with the inorganic C system to shape responses of air-water CO2 exchange. To address this uncertainty, we induced large metabolic gradients in the plankton community of a hypereutrophic hardwater Canadian prairie lake by adding N as urea (the most widely applied agricultural fertilizer) at loading rates of 0, 1, 3, 8 or 18 mg N L-1 week-1 to 3240-L, in-situ mesocosms. Over three separate 21-day experiments, all treatments of N dramatically increased phytoplankton biomass and gross primary production (GPP) two- to six-fold, but the effects of N on autotrophs plateaued at ~3 mg N L-1. Conversely, heterotrophic metabolism increased linearly with N fertilization over the full treatment range. In nearly all cases, N enhanced net planktonic uptake of dissolved inorganic carbon (DIC), and increased the rate of CO2 influx, while planktonic heterotrophy and CO2 production only occurred in the highest N treatments late in each experiment, and even in these cases, enclosures continued to in-gas CO2. Chemical effects on CO2 through calcite precipitation were also observed, but similarly did not change the direction of net CO2 flux. Taken together, these results demonstrate that atmospheric exchange of CO2 in eutrophic hardwater lakes remains sensitive to increasing N loading and eutrophication, and that even modest levels of N pollution are capable of enhancing autotrophy and CO2 in-gassing in P-rich lake ecosystems.

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Limnological theory posits that phosphorus (P) limits primary production in freshwater lakes, in part because fixation of atmospheric nitrogen (N2) can compensate for limitations in nitrogen (N) supply to phytoplankton. However, quantitative estimates of the degree to which N2 fixation satisfies planktonic N demand are rare. Here we used biweekly sampling during summer in seven lakes over 2 decades to estimate both planktonic N2 fixation and phytoplankton N demand. We further assessed the ability of biologically fixed N to satisfy N needs of primary producers in productive hardwater lakes. Phytoplankton N requirements, derived from estimates of phytoplankton productivity and N content, were moderately synchronous (S = 0.41) among lakes (ca. 0.1–9.2 mg N m–3 hr–1). In contrast, rates of N2 fixation determined using isotopic natural abundance method (NAM; 0.002–3.2 mg N m–3 hr–1), or heterocyte‐based calculations (0.10–1.78 mg N m–3 hr–1), varied asynchronously (SNAM = –0.03 and SHeterocyte = –0.11) among basins, accounted for a median of 3.5% (mean 11.3% ± 21.6) of phytoplankton demand, and were correlated to the abundance of Nostocales cyanobacteria when analysed using generalised additive models. Overall, the total mass of fixed N accounted for a median of only 3.0% of the spring standing stock of total dissolved N in study lakes (mean 7.5 ± 12.1%), with higher relative importance of fixed N in highly productive downstream lakes. Thus, while fixed N helps sustain primary productivity, particularly in years with high rates of N2‐fixation, it does not appear to eliminate N limitation of phytoplankton growth in these P‐rich hardwater lakes.