CO2 perturbation experiments: similarities and differences between dissolved inorganic carbon and total alkalinity manipulations

Increasing atmospheric carbon dioxide (CO2) through human activities and invasion of anthropogenic CO2 into the surface ocean alters the seawater carbonate chemistry, increasing CO2 and bicarbonate (HCO3−) at the expense of carbonate ion (CO32−) concentrations. This redistribution in the dissolved inorganic carbon (DIC) pool decreases pH and carbonate saturation state (Ω). Several components of the carbonate system are considered potential key variables influencing for instance calcium carbonate precipitation in marine calcifiers such as coccolithophores, foraminifera, corals, mollusks and echinoderms. Unravelling the sensitivities of marine organisms and ecosystems to CO2 induced ocean acidification (OA) requires well-controlled experimental setups and accurate carbonate system manipulations. Here we describe and analyse the chemical changes involved in the two basic approaches for carbonate chemistry manipulation, i.e. changing DIC at constant total alkalinity (TA) and changing TA at constant DIC. Furthermore, we briefly introduce several methods to experimentally manipulate DIC and TA. Finally, we examine responses obtained with both approaches using published results for the coccolithophore Emiliania huxleyi. We conclude that under most experimental conditions in the context of ocean acidification DIC and TA manipulations yield similar changes in all parameters of the carbonate system, which implies direct comparability of data obtained with the two basic approaches for CO2 perturbation

Reviews and Syntheses: Responses of coccolithophores to ocean acidification: a meta-analysis

Concerning their sensitivity to ocean acidification, coccolithophores, a group of calcifying single-celled phytoplankton, are one of the best-studied groups of marine organisms. However, in spite of the large number of studies investigating coccolithophore physiological responses to ocean acidification, uncertainties still remain due to variable and partly contradictory results. In the present study we have used all existing data in a meta-analysis to estimate the effect size of future pCO2 changes on the rates of calcification and photosynthesis and the ratio of particulate inorganic to organic carbon (PIC / POC) in different coccolithophore species. Our results indicate that ocean acidification has a negative effect on calcification and the cellular PIC / POC ratio in the two most abundant coccolithophore species: Emiliania huxleyi and Gephyrocapsa oceanica. In contrast, the more heavily calcified species Coccolithus braarudii did not show a distinct response when exposed to elevated pCO2/reduced pH. Photosynthesis in Gephyrocapsa oceanica was positively affected by high CO2, while no effect was observed for the other coccolithophore species. There was no indication that the method of carbonate chemistry manipulation was responsible for the inconsistent results regarding observed responses in calcification and the PIC / POC ratio. The perturbation method, however, appears to affect photosynthesis, as responses varied significantly between total alkalinity (TA) and dissolved inorganic carbon (DIC) manipulations. These results emphasize that coccolithophore species respond differently to ocean acidification, both in terms of calcification and photosynthesis. Where negative effects occur, they become evident at CO2 levels in the range projected for this century in the case of unabated CO2 emissions. As the data sets used in this meta-analysis do not account for adaptive responses, ecological fitness and ecosystem interactions, the question remains as to how these physiological responses play out in the natural environment

Effects of ocean acidification on marine dissolved organic matter are not detectable over the succession of phytoplankton blooms

Marine dissolved organic matter (DOM) is one of the largest active organic carbon reservoirs on Earth, and changes in its pool size or composition could have a major impact on the global carbon cycle. Ocean acidification is a potential driver for these changes because it influences marine primary production and heterotrophic respiration. We simulated ocean acidification as expected for a “business-as-usual” emission scenario in the year 2100 in an unprecedented long-term mesocosm study. The large-scale experiments (50 m3 each) covered a full seasonal cycle of marine production in a Swedish Fjord. Five mesocosms were artificially enriched in CO2 to the partial pressure expected in the year 2100 (900 μatm), and five more served as controls (400 μatm). We applied ultrahigh-resolution mass spectrometry to monitor the succession of 7360 distinct DOM formulae over the course of the experiment. Plankton blooms had a clear effect on DOM concentration and molecular composition. This succession was reproducible across all 10 mesocosms, independent of CO2 treatment. In contrast to the temporal trend, there were no significant differences in DOM concentration and composition between present-day and year 2100 CO2 levels at any time point of the experiment. On the basis of our results, ocean acidification alone is unlikely to affect the seasonal accumulation of DOM in productive coastal environments

Primary production during nutrient-induced blooms at elevated CO2 concentrations

Mesocosms experiments (PeECE II and PeECE III) were carried out in 9 transparent mesocosms. Prior to the experimental period, the seawater carbonate system was manipulated to achieve three different levels of CO2. At the onset of the experimental period, nutrients were added to all mesocosms in order to initiate phytoplankton blooms. Rates of primary production were measured by in-situ incubations using 14C-incorporation and oxygen production/consumption. Particulate primary production by 14C was also size fractionated and compared with phytoplankton species composition. Nutrient supply increased the primary production rates, and a net autotrophic phase with 14C-fixation rates up to 4 times higher than initial was observed midway through the 24 days experiment before net community production returned to near-zero and 14C-fixation rates relaxed back to lower than initial. We found a trend in the 14C-based measurements towards higher cumulative primary production at higher pCO2, consistent with recently published results for DIC removal (Riebesell et al., 2007). There where found differences to the size fractionated primary production response to CO2 treatments. The plankton composition changes throughout the bloom, however, resulted in no significant response until the final phase of the experiment where phytoplankton growth became nutrient limited, and phytoplankton community changed from diatom to flagellate dominance. This opens for the two alternative hypotheses that such an effect is associated with mineral nutrient limited growth, and/or with phytoplankton species composition. The lack of a clear net heterotrophic phase in the last part of the experiment supports the idea that a substantial part of production in the upper layer was not degraded locally, but either accumulated there or was exported vertically

Effects of long-term high CO₂ exposure on two species of coccolithophores

The physiological performance of two coccolithophore species, Emiliania huxleyi and Coccolithus braarudii, was investigated during long-term exposure to elevated pCO2 levels. Mono-specific cultures were grown over 152 (E. huxleyi) and 65 (C. braarudii) generations while pCO2 was gradually increased to maximum levels of 1150 μatm (E. huxleyi) and 930 μatm (C. braarudii) and kept constant thereafter. Rates of cell growth and cell quotas of particulate organic carbon (POC), particulate inorganic carbon (PIC) and total particulate nitrogen (TPN) were determined repeatedly throughout the incubation period. Increasing pCO2 caused a decrease in cell growth rate of 9% and 29% in E. huxleyi and C. braarudii, respectively. In both species cellular PIC:TPN and PIC:POC ratios decreased in response to rising pCO2, whereas no change was observed in the POC:TPN ratios of E. huxleyi and C. braarudii. These results are consistent with those obtained in shorter-term high CO2 exposure experiments following abrupt pertubations of the seawater carbonate system and indicate that for the strains tested here a gradual CO2 increase does not alleviate CO2/pH sensitivity

CO2 and HCO3- uptake in marine diatoms acclimated to different CO2 concentrations.

Rates of cellular uptake of CO2 and HCO3- during steady-state photosynthesis were measured in the marine diatoms Thalassiosira weissflogii and Phaeodactylum tricornutum, acclimated to CO2 partial pressures of 36, 180, 360, and 1,800 ppmv. In addition, in vivo activity of extracellular (eCA) and intracellular (iCA) carbonic anhydrase was determined in relation to CO2 availability. Both species responded to diminishing CO2 supply with an increase in eCA and iCA activity. In P. tricornutum, eCA activity was close to the detection limit at higher CO2 concentrations. Simultaneous uptake of CO2 and HCO3- was observed in both diatoms. At air-equilibrated CO2 levels (360 ppmv), T. weissflogii took up CO2 and HCO3- at approximately the same rate, whereas CO2 uptake exceeded HCO3- uptake by a factor of two in P. tricornutum. In both diatoms, CO2 :HCO3- uptake ratios progressively decreased with decreasing CO2 concentration, whereas substrate affinities of CO2 and HCO3- uptake increased. Half-saturation concentrations were always <=5 mM CO2 for CO2 uptake and <700 mM HCO3- for HCO3- uptake. Our results indicate the presence of highly efficient uptake systems for CO2 and HCO3- in both diatoms at concentrations typically encountered in ocean surface waters and the ability to adjust uptake rates to a wide range of inorganic carbon supply

Environmental control on coccolithophore morphology : do modern species yield information that is transferable to the geological past?

It is generally assumed that calcareous nannofossils conserve palaeoenvironmental information from the time of their formation. Changes in coccolith morphology can result from physiological responses to environmental drivers. Temperature, salinity, nutrient concentration, light and carbonate chemistry are among the environmental drivers that impact extant coccolithophores and may alter coccolith size, as well as coccosphere size and morphology. Many palaeoreconstruction studies have assessed the biological responses of living coccolithophore species to environmental drivers with the expectation that it is possible to use this information for calibrating the biomineralisation responses of ancient coccolithophores. However, there is a large uncertainty concerning whether the morphological responses of living coccolithophores to environmental changes are similar to the morphological responses of fossil species, when you consider the fact that millions of years of evolutionary adaptation lie between the extant species and their fossilised ancestors. In order to test this caveat, we examined four extant species (Emiliania huxleyi, Gephyrocapsa oceanica , Coccolithus pelagicus subsp. braarudii and Pleurochrysis carterae), which have been evolutionarily distinct for millions of years. We cultured them under changing environmental conditions in order to evaluate any changes in coccolith morphology. Our underlying hypothesis was that if the species showed a uniform reaction to any of the tested environmental drivers, then this would suggests that the same response may well occur over geological timescales, and that coccolith morphological changes could serve as a palaeo-proxy for that particular driver. Our experiments demonstrated that the four species had no common response to changing light intensity, Mg/Ca, nutrient content or temperature with respect to coccolith size. These results revealed the difficulties in using coccolith size as a proxy for environmental drivers. One exception was an increase in malformations when coccolithophores were grown under excess CO2 , and these data provided evidence that this response variable can be used as a palaeo-proxy for episodes of acute carbonate chemistry perturbations