Experimental growth characteristics, cell size, yessotoxin values and bioassay data of Protoceratium reticulatum (Dinophyceae)

Harmful algal blooms are mainly caused by marine dinoflagellates and are known to produce potent toxins that may affect the ecosystem, human activities and health. Such events have increased in frequency and intensity worldwide in the past decades. Numerous processes involved in Global Change are amplified in the Arctic, but little is known about species specific responses of arctic dinoflagellates. The aim of this work was to perform an exhaustive morphological, phylogenetical and toxinological characterization of Greenland Protoceratium reticulatum and, in addition, to test the effect of temperature on growth and production of bioactive secondary metabolites. Seven clonal isolates, the first isolates of P. reticulatum available from arctic waters, were phylogenetically characterized by analysis of the LSU rDNA. Six isolates were further characterized morphologically and were shown to produce both yessotoxins (YTX) and lytic compounds, representing the first report of allelochemical activity in P. reticulatum. As shown for one of the isolates, growth was strongly affected by temperature with a maximum growth rate at 15 °C, a significant but slow growth at 1 °C, and cell death at 25 °C, suggesting an adaptation of P. reticulatum to temperate waters. Temperature had no major effect on total YTX cell quota or lytic activity but both were affected by the growth phase with a significant increase at stationary phase. A comparison of six isolates at a fixed temperature of 10 °C showed high intraspecific variability for all three physiological parameters tested. Growth rate varied from 0.06 to 0.19 per day, and total YTX concentration ranged from 0.3 to 15.0 pg YTX/cell and from 0.5 to 31.0 pg YTX/cell at exponential and stationary phase, respectively. All six isolates performed lytic activity; however, for two isolates lytic activity was only detectable at higher cell densities in stationary phase.

(Table T1) Initial water quality conditions for bioassay experiments during Nathaniel B. Palmer cruise NBP09-01 in the Amundsen Sea

Nutrient addition experiments were performed during the austral summer in the Amundsen Sea (Southern Ocean) to investigate the availability of organically bound iron (Fe) to the phytoplankton communities, as well as assess their response to Fe amendment. Changes in autotrophic biomass, pigment concentration, maximum photochemical efficiency of photosystem II, and nutrient concentration were recorded in response to the addition of dissolved free Fe (DFe) and Fe bound to different model ligands. Analysis of pigment concentrations indicated that the autotrophic community was dominated by the prymnesiophyte Phaeocystis antarctica throughout most of the Amundsen Sea, although diatoms dominated in two experiments conducted in the marginal ice zone. Few significant differences in bulk community biomass (particulate organic carbon, nitrogen, and chlorophyll a) were observed, relative to the controls, in treatments with Fe added alone or bound to the ligand phytic acid. In contrast, when Fe was bound to the ligand desferrioxamine B (DFB), decreases in the bulk biomass indices were observed. The concentration of the diatom accessory pigment fucoxanthin showed little response to Fe additions, while the concentration of the P. antarctica-specific pigment, 19'-hexanoyloxyfucoxanthin (19'-hex), decreased when Fe was added alone or bound to the model ligands. Lastly, differences in the nitrate:phosphate (NO3- :PO4**3-) utilization ratio were observed between the Fe-amended treatments, with Fe bound to DFB resulting in the lowest NO3- :PO4**3- uptake ratios (~ 10) and the remaining Fe treatments having higher NO3- :PO4**3- uptake ratios (~ 17). The data are discussed with respect to glacial inputs of Fe in the Amundsen Sea and the bioavailability of Fe. We suggest that the previously observed high NO3- :PO4**3- utilization ratio of P. antarctica is a consequence of its production of dissolved organic matter that acts as ligands and increases the bioavailability of Fe, thereby stimulating the uptake of NO3-.

Effect of enhanced pCO2 levels on the production of dissolved organic carbon and transparent exopolymer particles in short-term bioassay experiments

It has been proposed that increasing levels of pCO2 in the surface ocean will lead to more partitioning of the organic carbon fixed by marine primary production into the dissolved rather than the particulate fraction. This process may result in enhanced accumulation of dissolved organic carbon (DOC) in the surface ocean and/or concurrent accumulation of transparent exopolymer particles (TEPs), with important implications for the functioning of the marine carbon cycle. We investigated this in shipboard bioassay experiments that considered the effect of four different pCO2 scenarios (ambient, 550, 750 and 1000 µatm) on unamended natural phytoplankton communities from a range of locations in the northwest European shelf seas. The environmental settings, in terms of nutrient availability, phytoplankton community structure and growth conditions, varied considerably between locations. We did not observe any strong or consistent effect of pCO2 on DOC production. There was a significant but highly variable effect of pCO2 on the production of TEPs. In three of the five experiments, variation of TEP production between pCO2 treatments was caused by the effect of pCO2 on phytoplankton growth rather than a direct effect on TEP production. In one of the five experiments, there was evidence of enhanced TEP production at high pCO2 (twice as much production over the 96 h incubation period in the 750 ?atm treatment compared with the ambient treatment) independent of indirect effects, as hypothesised by previous studies. Our results suggest that the environmental setting of experiments (community structure, nutrient availability and occurrence of phytoplankton growth) is a key factor determining the TEP response to pCO2 perturbations.