Biochemical, metabolic and morphological responses of the intertidal gastropod Littorina littorea to ocean acidification and increase temperature

Future changes to the pH and temperature of the oceans are predicted to impact the biodiversity of marine ecosystems, particularly those animals that rely on the process of calcification. The marine intertidal gastropod Littorina littorea can be used as a model of intertidal organism for investigating the effects of ocean acidification and high temperature, alone and in combination because its ability to be quickly adapt against environmental stressor. In the first study a single species population of L. littorea was used to test for physiological and biochemical effects underpinning organismal responses to climate change and ocean acidification. Compared with control conditions, snails decreased metabolic rates by 31% in response to elevated pCO2 while by 15% in response to combined pCO2 and temperature. Decreased metabolic rates were associated with metabolic depression, a strategy to match oxygen demand and availability, and an increase in end-product metabolites in the tissue under acidified treatments, indicating an increased reliance on anaerobic metabolism. This study also showed that anthropogenic alteration of CO2 and temperature may also lead to plastic responses, a fundamental mechanism of many marine gastropods to cope environmental variability. At low pH and elevated temperature in isolation or combined showing lower shell growth than individuals kept under control conditions. Percentage change in shell length and thicknesses was also lower under acidified and temperature in isolation or combined than control condition, making shells were more globular and desiccation rates were higher. Further studies to broader latitudinal ranges for six populations of L. littorea showed that shell growth decreased in all six populations under elevated pCO2 compared to control snails particularly those at range edges. Elevated pCO2 also affected to the reduction of shell length and width that causing shell aspect ratio to increase across latitudinal gradients except individuals from Millport, UK. Percentage changes of aperture width and aperture area were also decrease under elevated pCO2 with greater reduction of aperture area were found at populations in the mid-ranges which is assumed this response might be linked to local adaptation of the individual to microclimatic conditions. This study also showed that metabolic rates were negatively affected by high pCO2 and show non-linear trend across latitudinal gradients in compared to individual kept under normal pCO2 conditions. Metabolomic analysis showed that two northern populations of Trondheim and TromsØ were distinct from other populations when exposed to low temperature (15 °C) with elevated pCO2 due to, in part, high concentrations of thymine, uracil, valine and lysine. A similar separation also occurred under medium (25 °C) and high (35 °C) temperature exposure in which one of northern population (Trondheim) was distinct from other populations and had lower concentrations of alanine, betaine and taurine while higher of valine. These results suggest that populations at northern latitudes may apply different ionic transport mechanisms under elevated pCO2 and elevated temperatures and those populations are likely to vary in terms of their physiological responses to this environmental challenge.

Response of two marine bacterial isolates to high CO2 concentration

Experimental results related to the effects of ocean acidification on planktonic marine microbes are still rather inconsistent and occasionally contradictory. Moreover, laboratory or field experiments that address the effects of changes in CO₂ concentrations on heterotrophic microbes are very scarce, despite the major role of these organisms in the marine carbon cycle. We tested the direct effect of an elevated CO₂ concentration (1000 ppmv) on the biomass and metabolic rates (leucine incorporation, CO₂ fixation and respiration) of 2 isolates belonging to 2 relevant marine bacterial families, Rhodobacteraceae (strain MED165) and Flavobacteriaceae (strain MED217). Our results demonstrate that, contrary to some expectations, high pCO₂ did not negatively affect bacterial growth but increased growth efficiency in the case of MED217. The elevated partial pressure of CO₂ ( pCO₂) caused, in both cases, higher rates of CO₂ fixation in the dissolved fraction and, in the case of MED217, lower respiration rates. Both responses would tend to increase the pH of seawater acting as a negative feedback between elevated atmospheric CO₂ concentrations and ocean acidification.

Prolonged exposure to elevated CO2 promotes growth of the algal symbiont Symbiodinium muscatinei in the intertidal sea anemone Anthopleura elegantissima

Some photosynthetic organisms benefit from elevated levels of carbon dioxide, but studies on the effects of elevated PCO₂ on the algal symbionts of animals are very few. This study investigated the impact of hypercapnia on a photosynthetic symbiosis between the anemone Anthopleura elegantissima and its zooxanthella Symbiodinium muscatinei. Anemones were maintained in the laboratory for 1 week at 37 Pa PCO₂ and pH 8.1. Clonal pairs were then divided into two groups and maintained for 6 weeks under conditions naturally experienced in their intertidal environment, 45 Pa PCO₂, pH 8.1 and 231 Pa PCO₂, pH 7.3. Respiration and photosynthesis were measured after the 1-week acclimation period and after 6 weeks in experimental conditions. Density of zooxanthellal cells, zooxanthellal cell size, mitotic index and chlorophyll content were compared between non-clonemate anemones after the 1-week acclimation period and clonal anemones at the end of the experiment. Anemones thrived in hypercapnia. After 6 weeks, A. elegantissima exhibited higher rates of photosynthesis at 45 Pa (4.2 µmol O₂ g⁻¹ h⁻¹) and 231 Pa (3.30 µmol O₂ g⁻¹ h⁻¹) than at the initial 37 Pa (1.53 µmol O₂ g⁻¹ h⁻¹). Likewise, anemones at 231 Pa received more of their respiratory carbon from zooxanthellae (CZAR  = 78.2%) than those at 37 Pa (CZAR  = 66.6%) but less than anemones at 45 Pa (CZAR  = 137.3%). The mitotic index of zooxanthellae was significantly greater in the hypercapnic anemones than in anemones at lower PCO₂. Excess zooxanthellae were expelled by their hosts, and cell densities, cell diameters and chlorophyll contents were not significantly different between the groups. The response of A. elegantissima to hypercapnic acidification reveals the potential adaptation of an intertidal, photosynthetic symbiosis for high PCO₂.

Productivity gains do not compensate for reduced calcification under near-future ocean acidification in the photosynthetic benthic foraminifera Marginopora vertebralis

Changes in the seawater carbonate chemistry (ocean acidification) from increasing atmospheric carbon dioxide (CO₂) concentrations negatively affect many marine calcifying organisms, but may benefit primary producers under dissolved inorganic carbon (DIC) limitation. To improve predictions of the ecological effects of ocean acidification, the net gains and losses between the processes of photosynthesis and calcification need to be studied jointly on physiological and population levels. We studied productivity, respiration, and abundances of the symbiont-bearing foraminifera Marginopora vertebralis on natural CO₂ seeps in Papua New Guinea and conducted additional studies on production and calcification on the Great Barrier Reef (GBR) using artificially enhanced pCO₂. Net oxygen production increased up to 90% with increasing pCO₂, with temperature, light and pH together explaining 61% of the variance in production. Production increased with increasing light and increasing pCO₂, and declined at higher temperatures. Respiration was also significantly elevated (~25%), while calcification was reduced (16-39%) at low pH/high pCO₂ compared to present day conditions. In the field, M. vertebralis was absent at three CO₂ seep sites at pH_Total levels below ~7.9 (pCO₂ ~700 μatm), but it was found in densities of over 1000 m⁻² at all three control sites. The study showed that endosymbiotic algae in foraminifera benefit from increased DIC availability, and may be naturally carbon limited. The observed reduction in calcification may have been caused either by increased energy demands for proton pumping (measured as elevated rates of respiration), or by stronger competition for DIC from the more productive symbionts. The net outcome of these two competing processes is that M. vertebralis cannot maintain populations under pCO₂ exceeding 700 μatm, thus are likely to be extinct in the next century.

Major cellular and physiological impacts of ocean acidification on a reef building coral

As atmospheric levels of CO₂ increase, reef-building corals are under greater stress from both increased sea surface temperatures and declining sea water pH. To date, most studies have focused on either coral bleaching due to warming oceans or declining calcification due to decreasing oceanic carbonate ion concentrations. Here, through the use of physiology measurements and cDNA microarrays, we show that changes in pH and ocean chemistry consistent with two scenarios put forward by the Intergovernmental Panel on Climate Change (IPCC) drive major changes in gene expression, respiration, photosynthesis and symbiosis of the coral, Acropora millepora, before affects on biomineralisation are apparent at the phenotype level. Under high CO₂ conditions corals at the phenotype level lost over half their Symbiodinium populations, and had a decrease in both photosynthesis and respiration. Changes in gene expression were consistent with metabolic suppression, an increase in oxidative stress, apoptosis and symbiont loss. Other expression patterns demonstrate upregulation of membrane transporters, as well as the regulation of genes involved in membrane cytoskeletal interactions and cytoskeletal remodeling. These widespread changes in gene expression emphasize the need to expand future studies of ocean acidification to include a wider spectrum of cellular processes, many of which may occur before impacts on calcification.

Exposure to Elevated Temperature and pCO2 Reduces Respiration Rate and Energy Status in the Periwinkle Littorina littorea

In the future, marine organisms will face the challenge of coping with multiple environmental changes associated with increased levels of atmospheric Pco2, such as ocean warming and acidification. To predict how organisms may or may not meet these challenges, an in-depth understanding of the physiological and biochemical mechanisms underpinning organismal responses to climate change is needed. Here, we investigate the effects of elevated Pco2 and temperature on the whole-organism and cellular physiology of the periwinkle Littorina littorea. Metabolic rates (measured as respiration rates), adenylate energy nucleotide concentrations and indexes, and end-product metabolite concentrations were measured. Compared with values for control conditions, snails decreased their respiration rate by 31% in response to elevated Pco2 and by 15% in response to a combination of increased Pco2 and temperature. Decreased respiration rates were associated with metabolic reduction and an increase in end-product metabolites in acidified treatments, indicating an increased reliance on anaerobic metabolism. There was also an interactive effect of elevated Pco2 and temperature on total adenylate nucleotides, which was apparently compensated for by the maintenance of adenylate energy charge via AMP deaminase activity. Our findings suggest that marine intertidal organisms are likely to exhibit complex physiological responses to future environmental drivers, with likely negative effects on growth, population dynamics, and, ultimately, ecosystem processes.

Effects of increased CO2 and nutrients on seagrass (Cymodocea nodosa) metabolism

Continuous global change leads to acidify ocean through increasing of atmospheric CO2 level which is major issue for our ecosysem now-a-days. Addressing this ocean acidification and ongoing anthropogenic problems of eutrophication with ocean temperature increase and teir cumulative impacts or interactive effects are still demanding a lot in research arena of oceanic environment. In this connection, this experiment conducted to investigate the effect of both nutrient and CO2 enrichment on the net community production (NCP) of Cymodocea nodosa beds collected from the western sector of the highly dynamic coastal lagoon Ria Formosa (south Portugal: 37° 01´ N, 7° 50´ W) in a mesocosm set up situated in Ramalhete Marine Station of University of Algarve where the open circulation of seawater exiss. To address the interaction with seagrass metabolism; two types of CO2 concentration (enriched: 700 ppm with pH 7.84 and control: existing 370 ppm with pH 8.12) and two types of nutrient concentration (enriched and control) were used with seawater. However, four types of different combinations from CO2 and nutrient concentration can explain effects of net community production for to complementary methods performed: light incubation and dark incubation. To estimate seagrass community metabolism, I measured change in calculated concentration of dissolved inorganic carbon (DIC) throughout photosynthesis and respiration from conducted twelve light incubations and nine dark incubations respectively in diferent days and times in order to catch possible wider range of underwater irradiances in case of light incubations. There were mild different trends suggesting increased production (± 38000 µmol C h-1 m-2) at underwater irradiance of ± 900 PAR µmol m-2 s-1 in the treatment of enriched nutrients and control CO2 concentration while decreased production (± 30000 µmol C h-1 m-2) found in the treatment with control CO2 and control nutrient at same irradiance. However, in consider to daytime, the net community production in afternoon found to differ a little bit after photoinhibition (observed at 13.30 h with ±1100 PAR µmol m-2 s-1) where maximum increased of NCP (± 35000 µmol C h-1 m-2) found at 17.00 h in the enriched (both in CO2 and nutrient) treatment. In all cases, average positive NCP values (from light) are found lower than the average negative NCP values (from dark) suggesting more community respiration in the equal day-night dates though the treatment with control CO2 and enriched nitrogen showed maximum net community production (around 60000 µmol C h-1 m-2) in the study place of south Portugal in the month of April-May when the daylight existed around 14 hours in a day. However, both CO2 and Nitrogen contents of seawater were not significantly affected yet in Cymodocea nodosa beds in generally even thouh there was significant difference (p = 0.002) among the daily average net community production of the four treatments. Further study should be carried out in order to better understand the underlying metabolic activities of C. nodosa leading to net community production in elevated CO2 and nutrients concentration to meet the upcoming global change.

Effect of marine acidification on calcification and respiration of Chlamys farreri

Marine acidification will be an important environmental problem in the near future as a result of persistent emissions of CO₂ and dissolution into seawater. In this study, we found that calcification and respiration of the Zhikong scallop (Chlamys farreri) are likely to be severely affected by increasing acidification. Calcification and respiration significantly declined as pH decreased. The calcification rate decreased by 33% when the pH of water was 7.9 compared with a pH of 8.1, and decreased close to 0 when the pH was reduced to 7.3. CO₂ and O₂ respiratory rates were reduced by 14% and 11%, respectively, when pH decreased from 7.9 to 7.3. Increasing acidification also led to changes in the metabolic pathways of C. farreri. Under acidic conditions, proteins may replace carbohydrates as the metabolic substrate. The survival of C. farreri is likely to be severely threatened in the next few centuries.

Physiological energetics of juvenile clams Ruditapes decussatus in a high CO2 coastal ocean

Effects of coastal ocean acidification, other than calcification, were tested on juvenile clams (Ruditapes decussatus) during a controlled CO2 perturbation experiment. The carbonate chemistry of natural (control) seawater was manipulated by injecting CO2 to attain two reduced pH levels: -0.4 and -0.7 pH units as compared with the control seawater. After 87 days of exposure, we found that the acidification conditions tested in this experiment significantly reduced the clearance, ingestion and respiration rates, and increased the ammonia excretion rate of R. decussatus seeds. Reduced ingestion combined with increased excretion is generally associated with a reduced energy input, which will likely contribute to a slower growth of the clams in a future high CO2 coastal ocean. These results emphasize the need for management policies to mitigate the adverse effects of global change on aquaculture, which is an economically relevant activity in most coastal areas worldwide.

Response of larval barnacle proteome to CO2-driven seawater acidification

The majority of benthic marine invertebrates have a complex life cycle, during which the pelagic larvae select a suitable substrate, attach to it, and then metamorphose into benthic adults. Anthropogenic ocean acidification (OA) is postulated to affect larval metamorphic success through an altered protein expression pattern (proteome structure) and post-translational modifications. To test this hypothesis, larvae of an economically and ecologically important barnacle species Balanus amphitrite, were cultured from nauplius to the cyprid stage in the present (control) and in the projected elevated concentrations of CO2 for the year 2100 (the OA treatment). Cyprid response to OA was analyzed at the total proteome level as well as two protein post-translational modification (phosphorylation and glycosylation) levels using a 2-DE based proteomic approach. The cyprid proteome showed OA-driven changes. Proteins that were differentially up or down regulated by OA come from three major groups, namely those related to energy-metabolism, respiration, and molecular chaperones, illustrating a potential strategy that the barnacle larvae may employ to tolerate OA stress. The differentially expressed proteins were tentatively identified as OA-responsive, effectively creating unique protein expression signatures for OA scenario of 2100. This study showed the promise of using a sentinel and non-model species to examine the impact of OA at the proteome level.