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The project OLCAPP was aimed at exploring the response of a natural, eutrophic system to ocean liming by slaked lime (calcium hydroxide) dispersal in the wake of ships. The main objectives were:- to monitor and model the slaked lime dissolution kinetics and the carbonate equilibrium (alkalinity, pH spikes/alteration, Dissolved Inorganic Carbon);- to assess Dissolved Organic Matter changes in quantity and composition;- to assess possible changes in primary production, photosynthetic efficiency and phytoplankton abundance and associations;- to assess the short and medium-term response of planktonic and benthic calcareous primary producers (calcareous red algae = maerl) to alkalinization and potential precipitation of carbonate crystals induced by the treatments.In the framework of the Transnational Access provided by AQUACOSM-plus, at the ECIMAT-UVIGO facility (Vigo, Spain) we had the opportunity to test ocean liming in nine mesocosm tanks. A sediment trap and a basket of calcareous algae (maerl) were positioned at the bottom of each tank. Mearl was previously collected in the Ria de Vigo at 7m depth and prepared for the experiment. Successively, each tank was filled with natural coastal seawater (~1m3).Out of the nine mesocosms, three tanks were treated with calcium hydroxide 0.02g/L (High) and three tanks with 0.006g/L (Low) per treatment, repeated on days 1, 3 and 5 (multiple exposure). The remaining three tanks were kept as control. A record of pH, O2, salinity, temperature and PAR was performed in the mesocosms during the experiment with a ten-minutes frequency.Nutrient concentration is monitored on a long-term basis in the Ria de Vigo, and was also tested on days 1, 3 and 5 for all treatments and controls. Seawater samples were collected from the mesocosms before (pre-treatment), and after 1h, 4h, and 24h from each treatment, with a 5L Niskin bottle. Dissolved Organic Matter as Dissolved Organic Carbon (DOC) and Chromophoric Dissolved Organic Matter (CDOM) were analysed, along with the Dissolved Inorganic Carbon (DIC). Moreover, samples collected after 1h from treatments were used for assessing also the bacterial association, the size-fractionated Chl-a concentration and the plankton primary production and photosynthetic efficiency. Gross primary production, community respiration and net community production were measured by changes in oxygen concentrations after 24 h light-dark bottle incubations. Dissolved oxygen was measured by Winkler titration. A total of 165 samples were obtained from filtering 2L of mesocosm water from pre-treatments, 4h and 24h samples, for the collection of the phytoplankton community, to be analysed under optical and scanning electron microscope.Preliminary observations during the experiment and the first data on the plankton community suggest that the High treatment leads to important flocculation and sedimentation affecting both the transparency of the water and the bottom environment, with significant and stable pH increase and decrease in phytoplankton production and efficiency. The mineralogical nature of the flocculation, the response of benthic calcareous algae and phytoplankton community, in term of composition and abundance of the major components, is here discussed.

We experimentally evaluated the temporal (interday and interseason) and spatial variability in microbial plankton responses to vitamin B12 and/or B1 supply (solely or in combination with inorganic nutrients) in coastal and oceanic waters of the northeast Atlantic Ocean. Phytoplankton and, to a lesser extent, prokaryotes were strongly limited by inorganic nutrients. Interday variability in microbial plankton responses to B vitamins was limited compared to interseason variability, suggesting that B-vitamin availability might be partially controlled by factors operating at seasonal scale. Chlorophyll a (Chl a) concentration and prokaryote biomass (PB) significantly increased after B-vitamin amendments in 13 % and 21 %, respectively, of the 216 cases (36 experiments × 6 treatments). Most of these positive responses were produced by treatments containing either B12 solely or B12 combined with B1 in oceanic waters, which was consistent with the significantly lower average vitamin-B12 ambient concentrations compared to that in the coastal station. Negative responses, implying a decrease in Chl a or PB, represented 21 % for phytoplankton and 26 % for prokaryotes. Growth stimulation by B1 addition was more frequent on prokaryotes than in phytoplankton, suggesting that B1 auxotrophy in the sampling area could be more widespread in prokaryotes than in phytoplankton. Negative responses to B vitamins were generalized in coastal surface waters in summer and were associated with a high contribution of Flavobacteriales to the prokaryote community. This observation suggests that the external supply of B12 and/or B1 may promote negative interactions between microbial components when B-vitamin auxotrophs are abundant. The microbial response patterns to B12 and/or B1 amendments were significantly correlated with changes in the prokaryotic community composition, highlighting the pivotal role of prokaryotes in B-vitamin cycling in marine ecosystems.

Atmospheric nutrient deposition into the open ocean increased over the past decades as a result of human activity and water-soluble organic nitrogen accounts for up to 30% of the total nitrogen inputs. The effects of inorganic and/or organic nutrient inputs on phytoplankton and heterotrophic bacteria have never been concurrently assessed in open ocean oligotrophic communities over a wide spatial gradient. We studied the effects of potentially limiting inorganic (nitrate, ammonium, phosphate, silica) and organic nutrient (glucose, aminoacids) inputs on microbial plankton biomass, community structure and metabolism in five microcosm experiments conducted along a latitudinal transect in the Atlantic Ocean (from 26° N to 29° S). Primary production rates increased up to 1.8-fold. Bacterial respiration and microbial community respiration increased up to 14.3 and 12.7-fold, respectively. Bacterial production and bacterial growth efficiency increased up to 58.8-fold and 2.5-fold, respectively. The largest increases were measured after mixed inorganic-organic nutrients additions. Changes in microbial plankton biomass were small as compared with those in metabolic rates. A north to south increase in the response of heterotrophic bacteria was observed, which could be related to a latitudinal gradient in phosphorus availability. Our results suggest that organic matter inputs associated with atmospheric deposition into the Atlantic Ocean will result in a predominantly heterotrophic versus autotrophic response and in increases in bacterial growth efficiency, particularly in the Southern Hemisphere. Subtle differences in the initial environmental and biological conditions are likely to result in differential microbial responses to inorganic and organic matter inputs.