Céline Ridame

More than inorganic copper is bioavailable to aquatic mosses at environmentally relevant concentrations

The present study investigates how dissolved organic matter (DOM) alters copper bioavailability at environmentally relevant concentrations (1-5 microg/L of dissolved copper, 1-4 mg/L of dissolved organic copper). A methodology combining two biological endpoints (short-term and steady-state bioaccumulation of copper by the aquatic moss Fontinalis antipyretica) and a sampling of labile copper with diffusion gradient in thin films (DGT) is proposed for batch experiments conducted with mineral water and various DOM, ethylenediaminetetra-acetic acid (EDTA), humic acid, and natural Seine River (France) extracts (hydrophobic and transphilic fractions). All types of DOM reduce the bioavailability of copper to aquatic mosses, and this reduction was more pronounced for the short-term biological endpoint, which was taken as being representative for environmental exposure. Labile copper sampled with DGT made it possible to estimate short-term bioaccumulation in the case of EDTA and natural Seine River extracts. With humic acid solutions, however, labile copper was lower than bioavailable copper. This result suggests that at realistic metal concentrations and with certain types of natural DOM, bioavailable copper might comprise not only inorganic copper but also some weak organic complexes. Hence, labile copper, in situ sampled with DGT, might not systematically overestimate bioavailable copper, as suggested previously on the basis of in vitro toxicity studies.

Strong stimulation of N 2 fixation in oligotrophic Mediterranean Sea: results from dust addition in large in situ mesocosms

The response of N 2 (dinitrogen) fixation to contrasted (wet and dry) Saharan dust deposition was studied in the framework of the DUNE project (a DUst experiment in a low-Nutrient, low-chlorophyll Ecosystem) during which realistic simulations of dust deposition (10 g m -2 ) into large mesocosms (52 m 3 ) were performed. Three distinct experimental dust additions were conducted in June 2008 (DUNE-1-P: simulation of a wet deposition, DUNE-1-Q: simulation of a dry deposition) and 2010 (DUNE-2-R: simulation of 2 successive wet depositions) in the northwestern oligotrophic Mediterranean Sea. Here we show that wet and dry dust deposition induced a rapid (24 h or 48 h after dust additions), strong (from 2- to 5.3-fold) and long (at least 4-6 days duration) increase in N 2 fixation, indicating that both wet and dry Saharan dust deposition was able to relieve efficiently the nutrient limitation(s) of N 2 fixation. This means in particular that N 2 fixation activity was not inhibited by the significant input of nitrate associated with the simulated wet deposition (~ 9 mmol NO 3 - m -2 ). The input of new nitrogen associated with N 2 fixation was negligible relative to the atmospheric NO 3 - input associated with the dust. The contribution of N 2 fixation to primary production was negligible (≤ 1%) before and after dust addition in all experiments, indicating that N 2 fixation was a poor contributor to the nitrogen demand for primary production. Despite the stimulation of N 2 fixation by dust addition, the rates remained low, and did not significantly change the contribution of N 2 fixation to new production since only a maximum contribution of 10% was observed. The response of N 2 fixation by diazotrophs and CO 2 fixation by the whole phytoplankton community suggests that these metabolic processes were limited or co-limited by different nutrients. With this novel approach, which allows us to study processes as a function of time while atmospheric particles are sinking, we show that new atmospheric nutrients associated with Saharan dust pulses do significantly stimulate N 2 fixation in the Mediterranean Sea and that N 2 fixation is not a key process in the carbon cycle in such oligotrophic environments.

Response of the Unicellular Diazotrophic Cyanobacterium Crocosphaera watsonii to Iron Limitation

Iron (Fe) is widely suspected as a key controlling factor of N2 fixation due to the high Fe content of nitrogenase and photosynthetic enzymes complex, and to its low concentrations in oceanic surface seawaters. The influence of Fe limitation on the recently discovered unicellular diazotrophic cyanobacteria (UCYN) is poorly understood despite their biogeochemical importance in the carbon and nitrogen cycles. To address this knowledge gap, we conducted culture experiments on Crocosphaera watsonii WH8501 growing under a range of dissolved Fe concentrations (from 3.3 to 403 nM). Overall, severe Fe limitation led to significant decreases in growth rate (2.6-fold), C, N and chlorophyll a contents per cell (up to 4.1-fold), N2 and CO2 fixation rates per cell (17- and 7-fold) as well as biovolume (2.2-fold). We highlighted a two phased response depending on the degree of limitation: (i) under a moderate Fe limitation, the biovolume of C. watsonii was strongly reduced, allowing the cells to keep sufficient energy to maintain an optimal growth, volume-normalized contents and N2 and CO2 fixation rates; (ii) with increasing Fe deprivation, biovolume remained unchanged but the entire cell metabolism was affected, as shown by a strong decrease in the growth rate, volume-normalized contents and N2 and CO2 fixation rates. The half-saturation constant for growth of C. watsonii with respect to Fe is twice as low as that of the filamentous Trichodesmium indicating a better adaptation of C. watsonii to poor Fe environments than filamentous diazotrophs. The physiological response of C. watsonii to Fe limitation was different from that previously shown on the UCYN Cyanothece sp, suggesting potential differences in Fe requirements and/or Fe acquisition within the UCYN community. These results contribute to a better understanding of how Fe bioavailability can control the activity of UCYN and explain the biogeography of diverse N2 fixers in ocean.