Abstract. The hydrogen isotopic composition (δ²H) of lipid biomarkers has diverse applications in the fields of paleoclimatology, biogeochemistry, and microbial community dynamics. Large changes in hydrogen isotope fractionation have been observed among microbes with differing core metabolisms, while environmental factors including temperature and nutrient availability can affect isotope fractionation by photoautotrophs. Much effort has gone into studying these effects under laboratory conditions with single species cultures. Moving beyond controlled environments and quantifying the natural extent of these changes in freshwater lacustrine settings and identifying their causes is essential for robust application of δ²H values of common short-chain fatty acids as a proxy of net community metabolism and of phytoplankton-specific biomarkers as a paleohydrologic proxy.

This work targets the effect of community dynamics, temperature, and productivity on ²H∕¹H fractionation in lipid biomarkers through a comparative time series in two central Swiss lakes: eutrophic Lake Greifen and oligotrophic Lake Lucerne. Particulate organic matter was collected from surface waters at six time points throughout the spring and summer of 2015, and δ²H values of short-chain fatty acids, as well as chlorophyll-derived phytol and the diatom biomarker brassicasterol, were measured. We paired these measurements with in situ incubations conducted with NaH¹³CO₃, which were used to calculate the production rates of individual lipids in lake surface water. As algal productivity increased from April to June, net discrimination against ²H in Lake Greifen increased by as much as 148 ‰ for individual fatty acids. During the same time period in Lake Lucerne, net discrimination against ²H increased by as much as 58 ‰ for individual fatty acids. A large portion of this signal is likely due to a greater proportion of heterotrophically derived fatty acids in the winter and early spring, which are displaced by more ²H-depleted fatty acids as phytoplankton productivity increases. Smaller increases in ²H discrimination for phytol and brassicasterol suggest that a portion of the signal is due to changes in net photoautotrophic ²H fractionation, which may be caused by increasing temperatures, a shift from maintenance to high growth, or changes in the community assemblage. The fractionation factors for brassicasterol were significantly different between the two lakes, suggesting that its hydrogen isotope composition may be more sensitive to nutrient regime than is the case for fatty acids or phytol.