Macroalgae δ15N values in well-mixed estuaries: Indicator of anthropogenic nitrogen input or macroalgae metabolism?

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Although nitrogen stable isotope ratio (δ15N) in macroalgae is widely used as a bioindicator of anthropogenic nitrogen inputs to the coastal zone, recent studies suggest the possible role of macroalgae metabolism in δ15N variability. Simultaneous determinations of δ15N of dissolved inorganic nitrogen (DIN) along the land–sea continuum, inter-species variability of δ15N and its sensitivity to environmental factors are necessary to confirm the efficiency of macroalgae δ15N in monitoring nitrogen origin in mixed-use watersheds. In this study, δ15N of annual and perennial macroalgae (Ulva sp., Enteromorpha sp., Fucus vesiculosus and Fucus serratus) are compared to δ15N-DIN along the Charente Estuary, after characterizing δ15N of the three main DIN sources (i.e. cultivated area, pasture, sewage treatment plant outlet). During late winter and spring, when human activities produce high DIN inputs, DIN sources exhibit distinct δ15N signals in nitrate (NO3−) and ammonium (NH4+): cultivated area (+6.5 ± 0.6‰ and +9.0 ± 11.0‰), pasture (+9.2 ± 1.8‰ and +12.4‰) and sewage treatment plant discharge (+16.9 ± 8.7‰ and +25.4 ± 5.9‰). While sources show distinct δ15N-NO3− in this multiple source catchment, the overall mixture of NO3− sources – generally >95% DIN – leads to low variations of δ15N-NO3− at the mouth of the estuary (+7.7 to +8.4‰). Even if estuarine δ15N-NO3− values are not significantly different from pristine continental and oceanic site (+7.3‰ and +7.4‰), macroalgae δ15N values are generally higher at the mouth of the estuary. This highlights high anthropogenic DIN inputs in the estuary, and enhanced contribution of 15N-depleted NH4+ in oceanic waters. Although seasonal variations in δ15N-NO3− are low, the same temporal trends in macroalgae δ15N values at estuarine and oceanic sites, and inter-species differences in δ15N values, suggest that macroalgae δ15N values might be modified by the metabolic response of macroalgae to environmental parameters (e.g., temperature, light, DIN concentrations). Differences between annual and perennial macroalgae indicate both a higher integration time of perennial compared to annual macroalgae and the possible role of passive versus active uptake mechanisms. Further studies are required to characterize the sensitivity of macroalgae fractionation to variable environmental conditions and uptake mechanisms.