Description: A method is described to simultaneously determine the neutral, amino, and acidic sugar content of combined carbohydrates in high molecular weight (HMW, 〉 1 kDa) dissolved organic matter and in particles from seawater samples. Monomeric sugars are determined after acid hydrolysis and neutralization through acid evaporation using high performance anion exchange chromatography (HPAEC) coupled with pulsed amperometric detection (PAD). The separation of sugars during chromatography is achieved in two steps, an isocratic elution (18 mM NaOH) followed by a gradient course of two mobile eluent phases (NaOH and CH3COONa). HPAEC-PAD has previously been applied to measure neutral and amino sugars in marine samples. Since salt anions interfere with the measurement, some of the earlier studies used ion exchange resins for seawater desalting. Thereby, variable losses of neutral and amino sugars, and the complete removal of acidic sugars have been reported. Here, we show that desalting by membrane dialysis (1 kDa) is an efficient alternative to ion exchange resins and yields recoveries of 〉 90% for HMW carbohydrates. We conducted several tests to determine the accuracy and reproducibility of the method. Sugar concentrations determined with our protocol were compared to results obtained with the colorimetric TPTZ-method, and with earlier HPAEC-PAD protocols using cation/ anion exchange resins. Applications of our protocol to field samples indicated that acidic sugars can comprise a substantial fraction (30-50%) of HMW dissolved carbohydrates in seawater. The simultaneous analysis of the three classes of sugars appears promising to detect a larger fraction of marine combined carbohydrates, and thus to improve our understanding of organic matter cycling in the ocean.

Description: With the accumulation of anthropogenic carbon dioxide (CO2), a proceeding decline in seawater pH has been induced that is referred to as ocean acidification. The ocean's capacity for CO2 storage is strongly affected by biological processes, whose feedback potential is difficult to evaluate. The main source of CO2 in the ocean is the decomposition and subsequent respiration of organic molecules by heterotrophic bacteria. However, very little is known about potential effects of ocean acidification on bacterial degradation activity. This study reveals that the degradation of polysaccharides, a major component of marine organic matter, by bacterial extracellular enzymes was significantly accelerated during experimental simulation of ocean acidification. Results were obtained from pH perturbation experiments, where rates of extracellular α- and β-glucosidase were measured and the loss of neutral and acidic sugars from phytoplankton-derived polysaccharides was determined. Our study suggests that a faster bacterial turnover of polysaccharides at lowered ocean pH has the potential to reduce carbon export and to enhance the respiratory CO2 production in the future ocean.

Description: Since the 1990s the AWI conducts studies on phytoplankton ecology at various locations in the central Arctic Ocean, the Greenland Sea and the Fram Strait onboard the ice going research vessel Polarstern. These studies provide valuable insight into the pelagic Arctic ecosystem. However, plankton abundance and composition were determined only sporadically and only few biogeochemical components were analysed. Considering the rapid changes of environmental conditions due to increasing temperatures, sea ice loss and ocean acidification, a comprehensive view of their impact on pelagic biological processes and the consequences for organic matter cycling is essential for our understanding of Arctic ecosystems. Thus a new group - Plankton Ecology and Biogeochemistry in a Changing Arctic Ocean (PEPBCAO) - established at the AWI studying the pelagic system in greater detail. The aim of this group is to complement the measurements of bulk variables and phyto- and zooplankton abundances by molecular assessment of the phytoplankton diversity including pico- and nanoplankton, allowing to better quantifying the intrusion of invading species into the polar habitat. Point measurements during cruises will serve as ground-truthing data to create basin wide satellite images, focussing on the quantitative estimation of phytoplankton functional types, which can serve as an input for modelling approaches. Changes in the composition of organic matter are investigated by molecular analyses and together with abundance and activity of zooplankton key species this will improve our ability of estimating carbon export under climate change. Beside the Central Arctic, one focus of the group will be the ‘AWI Hausgarten Deep Sea Monitoring Station’ in the Fram Strait off Svalbard. These data will be integrated in the existing data sets of the Arctic Ocean and this will provide a more detailed view of the changing pelagic system in the transition region between the central Arctic Ocean and the North Atlantic Ocean and first results will be presented.

Description: The bacterial turnover of organic matter is the major CO2-regenerating process in the ocean. About 75-95% of freshly produced organic carbon gets remineralized within the surface ocean, the zone that is most strongly affected by ocean acidification. Thus, rates of organic matter turnover changing in response to elevated seawater pCO2 would comprise a high feedback potential to climate change. Bacterial activity in the ocean is mainly fuelled by labile organic matter, a fraction of the chemically complex organic matter pool that provides high carbon, nitrogen and energy yields to bacterioplankton metabolism and growth. We investigated the impact of ocean acidification on the bacterial processing of polysaccharides and proteins, two important components of bioreactive organic matter, at a full marine site (Gulf of Biscay, North Atlantic) and in a brackish coastal ecosystem (Baltic Sea). During in situ studies and experimental simulations of present-day and future-ocean seawater pCO2, we determined rates of bacterial extracellular enzyme activities and of glucose and amino acid uptake, as well as bacterial biomass production using fluorescent and radioactive labelled substrate analogues. Furthermore, concentration and composition of organic matter were determined by different analytical tools. Our results reveal that the bacterial turnover of organic matter is sensitive to future changes in seawater pCO2. Effects of ocean acidification on enzymatic processes in coincidence with effects on the organic matter supply to bacterioplankton can potentially alter carbon fluxes in the future ocean and may feed back to rising pCO2 in the atmosphere

Description: The present study investigates the combined effect of phosphorous limitation, elevated partial pressure of CO2 (pCO2) and temperature on a calcifying strain of Emiliania huxleyi (PML B92/11) by means of a fully controlled continuous culture facility. Two levels of phosphorous limitation were consecutively applied by renewal of culture media (N:P=26) at dilution rates (D) of 0.3 d−1 and 0.1 d−1. CO2 and temperature conditions were 300, 550 and 900 μatm pCO2 at 14 °C and 900 μatm pCO2 at 18 °C. In general, the steady state cell density and particulate organic carbon (POC) production increased with pCO2, yielding significantly higher concentrations in cultures grown at 900 μatm pCO2 compared to 300 and 550 μatm pCO2. At 900 μatm pCO2, elevation of temperature as expected for a greenhouse ocean, further increased cell densities and POC concentrations. In contrast to POC concentration, C-quotas (pmol C cell−1) were similar at D=0.3 d−1 in all cultures. At D=0.1 d−1, a reduction of C-quotas by up to 15% was observed in the 900 μatm pCO2 at 18 °C culture. As a result of growth rate reduction, POC:PON:POP ratios deviated strongly from the Redfield ratio, primarily due to an increase in POC. Ratios of particulate inorganic and organic carbon (PIC:POC) ranged from 0.14 to 0.18 at D=0.3 d−1, and from 0.11 to 0.17 at D=0.1 d−1, with variations primarily induced by the changes in POC. At D=0.1 d−1, cell volume was reduced by up to 22% in cultures grown at 900 μatm pCO2. Our results indicate that changes in pCO2, temperature and phosphorus supply affect cell density, POC concentration and size of E. huxleyi (PML B92/11) to varying degrees, and will likely impact bloom development as well as biogeochemical cycling in a greenhouse ocean.

Description: An experimental study was conducted to test the effects of projected sea surface warming (according to the IPPC scenarios) on the accumulation and composition of dissolved organic matter (DOM) during marine phytoplankton blooms in cold seas (〈4°C). Eight mesocosms (∼1400 L) were filled with natural seawater, and two replicate mesocosms each were incubated by raising temperature by +0, +2, +4 and +6°C, respectively. The enclosed water was initially fertilized with inorganic nutrients to induce the development of phytoplankton blooms, which were then dominated by diatoms. Over a 4-week period, dissolved combined carbohydrates (DCCHO) and dissolved amino acids (DAA) were determined as major components of freshly produced, labile to semi-labile DOM. In all mesocosms, the increase in DCCHO concentration occurred sharply after the peak of chlorophyll a concentration, when nutrients became depleted. Rising temperature resulted in an earlier, faster and higher accumulation of DCCHO and of combined glucose predominantly. DCCHO yielded a maximum percentage of 35, 40, 49 and 59% of total combined carbohydrates in the +0, +2, +4 and +6°C treatments, respectively. Accumulation of DAA occurred more continuously and at an average rate of 0.79 ± 0.20 nmol L−1 h−1, but was not affected by rising temperature. Owing to the higher accumulation of DCCHO, the C:N ratio of DOM increased strongly during the course of the bloom, with higher ratios in the warmer treatments. Our study suggests that warming increases the extracellular release of carbohydrates from phytoplankton and, therefore, may affect the bottom-up control of the microbial loop in cold seas in the future.