ALBERT

Description: Community approaches to investigating ocean acidification (OA) effects suggest a high tolerance of micro- and mesozooplankton to carbonate chemistry changes expected to occur within this century. Plankton communities in the coastal areas of the Baltic Sea frequently experience pH variations partly exceeding projections for the near future both on a diurnal and seasonal basis. We conducted a large-scale mesocosm CO2 enrichment experiment ( ∼  55 m3) enclosing the natural plankton community in Tvärminne–Storfjärden for 8 weeks during June–August 2012 and studied community and species–taxon response of ciliates and mesozooplankton to CO2 elevations expected for this century. In addition to the response to fCO2, we also considered temperature and chlorophyll a variations in our analyses. Shannon diversity of ciliates significantly decreased with fCO2 and temperature with a greater dominance of smaller species. The mixotrophic Myrionecta rubra seemed to indirectly and directly benefit from higher CO2 concentrations in the post-bloom phase through increased occurrence of picoeukaryotes (most likely Cryptophytes) and Dinophyta at higher CO2 levels. With respect to mesozooplankton, we did not detect significant effects for either total abundance or for Shannon diversity. The cladocera Bosmina sp. occurred at distinctly higher abundance for a short time period during the second half of the experiment in three of the CO2-enriched mesocosms except for the highest CO2 level. The ratio of Bosmina sp. with empty to embryo- or resting-egg-bearing brood chambers, however, was significantly affected by CO2, temperature, and chlorophyll a. An indirect CO2 effect via increased food availability (Cyanobacteria) stimulating Bosmina sp. reproduction cannot be ruled out. Although increased regenerated primary production diminishes trophic transfer in general, the presence of organisms able to graze on bacteria such as cladocerans may positively impact organic matter transfer to higher trophic levels. Thus, under increasing OA in cladoceran-dominated mesozooplankton communities, the importance of the microbial loop in the pelagic zone may be temporarily enhanced and carbon transfer to higher trophic levels may be stimulated.

Description: The oceans absorb about a quarter of the annually produced anthropogenic atmospheric carbon dioxide (CO2), resulting in a decrease in surface water pH, a process termed ocean acidification (OA). Surprisingly little is known about how OA affects the physiology of heterotrophic bacteria or the coupling of heterotrophic bacteria to phytoplankton when nutrients are limited. Previous experiments were, for the most part, undertaken during productive phases or following nutrient additions designed to stimulate algal blooms. Therefore, we performed an in situ large-volume mesocosm ( ∼  55 m3) experiment in the Baltic Sea by simulating different fugacities of CO2 (fCO2) extending from present to future conditions. The study was conducted in July–August after the nominal spring bloom, in order to maintain low-nutrient conditions throughout the experiment. This resulted in phytoplankton communities dominated by small-sized functional groups (picophytoplankton). There was no consistent fCO2-induced effect on bacterial protein production (BPP), cell-specific BPP (csBPP) or biovolumes (BVs) of either free-living (FL) or particle-associated (PA) heterotrophic bacteria, when considered as individual components (univariate analyses). Permutational Multivariate Analysis of Variance (PERMANOVA) revealed a significant effect of the fCO2 treatment on entire assemblages of dissolved and particulate nutrients, metabolic parameters and the bacteria–phytoplankton community. However, distance-based linear modelling only identified fCO2 as a factor explaining the variability observed amongst the microbial community composition, but not for explaining variability within the metabolic parameters. This suggests that fCO2 impacts on microbial metabolic parameters occurred indirectly through varying physicochemical parameters and microbial species composition. Cluster analyses examining the co-occurrence of different functional groups of bacteria and phytoplankton further revealed a separation of the four fCO2-treated mesocosms from both control mesocosms, indicating that complex trophic interactions might be altered in a future acidified ocean. Possible consequences for nutrient cycling and carbon export are still largely unknown, in particular in a nutrient-limited ocean.

Description: Although coccolithophore physiological responses to CO2-induced changes in seawater carbonate chemistry have been widely studied in the past, there is limited knowledge on the variability of physiological responses between populations from different areas. In the present study, we investigated the specific responses of growth, particulate organic (POC) and inorganic carbon (PIC) production rates of three populations of the coccolithophore Emiliania huxleyi from three regions in the North Atlantic Ocean (Azores: six strains, Canary Islands: five strains, and Norwegian coast near Bergen: six strains) to a CO2 partial pressure (pCO2) range from 120 to 2630 µatm. Physiological rates of each population and individual strain increased with rising pCO2 levels, reached a maximum and declined thereafter. Optimal pCO2 for growth, POC production rates, and tolerance to low pH (i.e., high proton concentration) was significantly higher in an E. huxleyi population isolated from the Norwegian coast than in those isolated near the Azores and Canary Islands. This may be due to the large environmental variability including large pCO2 and pH fluctuations in coastal waters off Bergen compared to the rather stable oceanic conditions at the other two sites. Maximum growth and POC production rates of the Azores and Bergen populations were similar and significantly higher than that of the Canary Islands population. This pattern could be driven by temperature–CO2 interactions where the chosen incubation temperature (16 ∘C) was slightly below what strains isolated near the Canary Islands normally experience. Our results indicate adaptation of E. huxleyi to their local environmental conditions and the existence of distinct E. huxleyi populations. Within each population, different growth, POC, and PIC production rates at different pCO2 levels indicated strain-specific phenotypic plasticity. Accounting for this variability is important to understand how or whether E. huxleyi might adapt to rising CO2 levels.

Description: The Cretaceous ocean witnessed intervals of profound perturbations such as volcanic input of large amounts of CO2, anoxia, eutrophication, and introduction of biologically relevant metals. Some of these extreme events were characterized by size reduction and/or morphological changes of a few calcareous nannofossil species. The correspondence between intervals of high trace metal concentrations and coccolith dwarfism suggests a negative effect of these elements on nannoplankton biocalcification process in past oceans. In order to verify this hypothesis, we explored the potential effect of a mixture of trace metals on growth and morphology of four living coccolithophore species, namely Emiliania huxleyi, Gephyrocapsa oceanica, Pleurochrysis carterae and Coccolithus pelagicus. These taxa are phylogenetically linked to the Mesozoic species showing dwarfism under excess metal concentrations. The trace metals tested were chosen to simulate the environmental stress identified in the geological record and upon known trace metal interaction with living coccolithophores algae. Our laboratory experiments demonstrated that elevated trace metal concentrations not only affect coccolithophore algae production but, similarly to the fossil record, coccolith size and/or weight. Smaller coccoliths were detected in E. huxleyi and C. pelagicus, while coccoliths of G. oceanica showed a decrease in size only at the highest trace metal concentrations. P. carterae coccolith size was unresponsive for changing trace metal amounts. These differences among species allow to discriminate most- (P. carterae), intermediate- (E. huxleyi), and least- (C. pelagicus and G. oceanica) tolerant taxa. The fossil record and the experimental results converge on a selective response of coccolithophores to metal availability. These species-specific differences must be considered before morphological features of coccoliths are used to reconstruct paleo-chemical conditions.

Description: The Cretaceous ocean witnessed intervals of profound perturbations such as volcanic input of large amounts of CO2, anoxia, eutrophication and introduction of biologically relevant metals. Some of these extreme events were characterized by size reduction and/or morphological changes of a few calcareous nannofossil species. The correspondence between intervals of high trace metal concentrations and coccolith dwarfism suggests a negative effect of these elements on nannoplankton biocalcification processes in past oceans. In order to test this hypothesis, we explored the potential effect of a mixture of trace metals on growth and morphology of four living coccolithophore species, namely Emiliania huxleyi, Gephyrocapsa oceanica, Pleurochrysis carterae and Coccolithus pelagicus. The phylogenetic history of coccolithophores shows that the selected living species are linked to Mesozoic species showing dwarfism under excess metal concentrations. The trace metals tested were chosen to simulate the environmental stress identified in the geological record and upon known trace metal interactions with living coccolithophore algae.Our laboratory experiments demonstrated that elevated trace metal concentrations, similarly to the fossil record, affect coccolithophore algae size and/or weight. Smaller coccoliths were detected in E. huxleyi and C. pelagicus, while coccoliths of G. oceanica showed a decrease in size only at the highest trace metal concentrations. P. carterae coccolith size was unresponsive to changing trace metal concentrations. These differences among species allow discriminating the most- (P. carterae), intermediate- (E. huxleyi and G. oceanica) and least-tolerant (C. pelagicus) taxa. The fossil record and the experimental results converge on a selective response of coccolithophores to metal availability.These species-specific differences must be considered before morphological features of coccoliths are used to reconstruct paleo-chemical conditions.

Description: Coccolithophores are a group of phytoplankton species which cover themselves with small scales (coccoliths) made of calcium carbonate (CaCO3). The reason why coccolithophores form these calcite platelets has been a matter of debate since decades but has remained elusive so far. One hypothesis is that they serve a role in light/UV protection, especially in surface dwelling species like Emiliania huxleyi which can tolerate exceptionally high levels of solar radiation. In this study, we tested this hypothesis by culturing a calcifying and a non-calcifying strain under different light conditions with and without UV radiation. The coccoliths of E. huxleyi reduced the transmission of visible radiation (400–700 nm) by 7.5 %, UV-A (315–400 nm) by 14.1 % and UVB (280–315 nm) by 18.4 %. Growth rates of the calcifying strain (PML B92/11) were about 2 times higher than those of the non-calcifying strain (CCMP 2090) under indoor constant light levels in the absence of UV radiation. When exposed to outdoor conditions (fluctuating sunlight with UV radiation), growth rates of calcified cells were almost 3.5 times higher compared to naked cells. Furthermore, relative electron transport rate was 114 % higher and non-photochemical quenching (NPQ) 281 % higher in the calcifying compared to the non-calcifying strain, implying higher energy transfer associated with higher NPQ in the presence of calcification. When exposed to natural solar radiation including UV radiation, maximal quantum yield of photosystem II was only slightly reduced in the calcifying but strongly reduced in the non-calcifying strain. Our results reveal an important role of coccoliths in mitigating light and UV stress in E. huxleyi.

Description: Annually, the oceans absorb about one fourth of the anthropogenically produced atmospheric carbon dioxide (CO2) resulting in a drop in surface water pH, a process termed ocean acidification (OA). Surprisingly little is known about how OA affects physiology as well as food web interactions of heterotrophic bacteria when essential nutrients are limited, since most previous experiments were carried out during productive phases or even after nutrient additions to stimulate algal blooms. Therefore, we conducted an in situ large-volume mesocosm (~55 m3) experiment in the Baltic Sea by simulating different fugacities of CO2 (fCO2) extending from present to future conditions. The study was carried out after the spring-bloom in July–August to maintain low-nutrient conditions throughout the experiment, which resulted in a small-sized phytoplankton community dominated by picophytoplankton. Several positive as well as negative effects on free-living (FL) and particle-associated (PA) bacterial protein production (BPP) and biovolume (BV) could be related to fCO2-induced differences in phytoplankton composition and subsequent the availability of phytoplankton-derived organic matter. However, dynamics of BV and cell-specific BPP (csBPP) of FL heterotrophic bacteria could not be explained exclusively by the availability of phytoplankton-derived organic carbon. The dynamics were also related to enhanced grazing on DNA rich (HDNA) bacterial cells at higher fCO2 as revealed by flow cytometry. Additionally, a decoupling of autotrophic production and heterotrophic consumption during the last third of the experiment resulted in low, but significantly higher accumulation of DOC at enhanced fCO2. Interestingly we could not detect any consistent and direct fCO2-induced effect on BPP, csBPP nor BV of either FL or PA heterotrophic bacteria. In contrast, our results reveal several indirect fCO2-induced effects on BPP and bacterial BV with potential consequences for oceanic carbon cycling, in particular in a low nutrient and high fCO2 future ocean.

Description: The Baltic Sea is a unique environment as the largest body of brackish water in the world. Acidification of the surface oceans due to absorption of anthropogenic CO2 emissions is an additional stressor facing the pelagic community of the already challenging Baltic Sea. To investigate its impact on trace gas biogeochemistry, a large-scale mesocosm experiment was performed off Tvärminne Research Station, Finland, in summer 2012. During the second half of the experiment, dimethylsulfide (DMS) concentrations in the highest-fCO2 mesocosms (1075–1333 µatm) were 34 % lower than at ambient CO2 (350 µatm). However, the net production (as measured by concentration change) of seven halocarbons analysed was not significantly affected by even the highest CO2 levels after 5 weeks' exposure. Methyl iodide (CH3I) and diiodomethane (CH2I2) showed 15 and 57 % increases in mean mesocosm concentration (3.8 ± 0.6 increasing to 4.3 ± 0.4 pmol L−1 and 87.4 ± 14.9 increasing to 134.4 ± 24.1 pmol L−1 respectively) during Phase II of the experiment, which were unrelated to CO2 and corresponded to 30 % lower Chl a concentrations compared to Phase I. No other iodocarbons increased or showed a peak, with mean chloroiodomethane (CH2ClI) concentrations measured at 5.3 (±0.9) pmol L−1 and iodoethane (C2H5I) at 0.5 (±0.1) pmol L−1. Of the concentrations of bromoform (CHBr3; mean 88.1 ± 13.2 pmol L−1), dibromomethane (CH2Br2; mean 5.3 ± 0.8 pmol L−1), and dibromochloromethane (CHBr2Cl, mean 3.0 ± 0.5 pmol L−1), only CH2Br2 showed a decrease of 17 % between Phases I and II, with CHBr3 and CHBr2Cl showing similar mean concentrations in both phases. Outside the mesocosms, an upwelling event was responsible for bringing colder, high-CO2, low-pH water to the surface starting on day t16 of the experiment; this variable CO2 system with frequent upwelling events implies that the community of the Baltic Sea is acclimated to regular significant declines in pH caused by up to 800 µatm fCO2. After this upwelling, DMS concentrations declined, but halocarbon concentrations remained similar or increased compared to measurements prior to the change in conditions. Based on our findings, with future acidification of Baltic Sea waters, biogenic halocarbon emissions are likely to remain at similar values to today; however, emissions of biogenic sulfur could significantly decrease in this region.

Description: Nitrogen fixation by filamentous cyanobacteria supplies significant amounts of new nitrogen (N) to the Baltic Sea. This balances N loss processes such as denitrification and anammox, and forms an important N source supporting primary and secondary production in N-limited post-spring bloom plankton communities. Laboratory studies suggest that filamentous diazotrophic cyanobacteria growth and N2-fixation rates are sensitive to ocean acidification, with potential implications for new N supply to the Baltic Sea. In this study, our aim was to assess the effect of ocean acidification on diazotroph growth and activity as well as the contribution of diazotrophically fixed N to N supply in a natural plankton assemblage. We enclosed a natural plankton community in a summer season in the Baltic Sea near the entrance to the Gulf of Finland in six large-scale mesocosms (volume ∼ 55 m3) and manipulated fCO2 over a range relevant for projected ocean acidification by the end of this century (average treatment fCO2: 365–1231 µatm). The direct response of diazotroph growth and activity was followed in the mesocosms over a 47 day study period during N-limited growth in the summer plankton community. Diazotrophic filamentous cyanobacteria abundance throughout the study period and N2-fixation rates (determined only until day 21 due to subsequent use of contaminated commercial 15N-N2 gas stocks) remained low. Thus estimated new N inputs from diazotrophy were too low to relieve N limitation and stimulate a summer phytoplankton bloom. Instead, regeneration of organic N sources likely sustained growth in the plankton community. We could not detect significant CO2-related differences in neither inorganic nor organic N pool sizes, or particulate matter N : P stoichiometry. Additionally, no significant effect of elevated CO2 on diazotroph activity was observed. Therefore, ocean acidification had no observable impact on N cycling or biogeochemistry in this N-limited, post-spring bloom plankton assemblage in the Baltic Sea.

Description: Sediment traps are the most common tool to investigate vertical particle flux in the marine realm. However, the spatial and temporal decoupling between particle formation in the surface ocean and particle collection in sediment traps at depth often handicaps reconciliation of production and sedimentation even within the euphotic zone. Pelagic mesocosms are restricted to the surface ocean, but have the advantage of being closed systems and are therefore ideally suited to studying how processes in natural plankton communities influence particle formation and settling in the ocean's surface. We therefore developed a protocol for efficient sample recovery and processing of quantitatively collected pelagic mesocosm sediment trap samples for biogeochemical analysis. Sedimented material was recovered by pumping it under gentle vacuum through a silicon tube to the sea surface. The particulate matter of these samples was subsequently separated from bulk seawater by passive settling, centrifugation or flocculation with ferric chloride, and we discuss the advantages and efficiencies of each approach. After concentration, samples were freeze-dried and ground with an easy to adapt procedure using standard lab equipment. Grain size of the finely ground samples ranged from fine to coarse silt (2–63 µm), which guarantees homogeneity for representative subsampling, a widespread problem in sediment trap research. Subsamples of the ground material were perfectly suitable for a variety of biogeochemical measurements, and even at very low particle fluxes we were able to get a detailed insight into various parameters characterizing the sinking particles. The methods and recommendations described here are a key improvement for sediment trap applications in mesocosms, as they facilitate the processing of large amounts of samples and allow for high-quality biogeochemical flux data.