Growth of cultured giant clams (Tridacna spp.) in low pH, high-nutrient seawater: species-specific effects of substrate and supplemental feeding under acidification

Four species of giant clams, Tridacna maxima, T. squamosa, T. derasa and T. crocea, were cultured in outdoor raceways for 364 days at the Waikīkī Aquarium and the Oceanic Institute on the island of O‘ahu, Hawai‘i, USA.  Growth of each species was compared among individuals grown with and without supplemental phytoplankton feeding, and directly on the substrate or mounted on concrete plugs in low pH, high nutrient seawater.  Among clams cultured with and without supplemental phytoplankton (Chaetoceros spp.), feeding resulted in significantly lower mortality in all species but T. deresa, whereas growth was significantly higher among fed clams for all species except T. squamosa. Tridacna derasa showed roughly a three-fold increase in growth when fed (88.5 g ± 4.4 SD) than when unfed (26.0 g ± 2.1 SD), whereas T. maxima growth was substantially lower, but nearly 10-fold greater in response to feeding (9.0 g ± 1.9 SD). The overall mortality rate of juvenile clams was significantly lower in the fed (44.4 ± 10.0%) than the unfed (71.8 ± 9.6%) trials, with the greatest effect observed in mortality of T. maxima (fed 15% versus unfed 80%) and T. squamosa (fed 65% versus unfed 95%). None of the T. squamosa remained on concrete plugs for the duration of the experiment. Among the remaining three species, there was no difference in either wet weight or shell length for T. maxima and for wet weight only in T. derasa on (186.5 g ± 16.1 SD) and off (147.0 g ± 6.0 SD) the concrete plugs.  In contrast, T. crocea had significantly greater shell growth off the plugs (14.3 mm ± 1.0 SD versus 8.5 mm ± 1.7 SD) but significantly greater gain in wet weight on the concrete plugs (26.3 g ± 1.5 SD versus 58.5 g ± 2.5 SD).  The seawater wells used for this study are well characterized with elevated levels of inorganic nutrients and higher pCO2 relative to tropical ocean waters, roughly approximating predictions for future oceanic conditions under IPCC IS92a emission scenarios. In comparison to previous studies in natural seawater, T. derasa had a significantly higher shell growth rate in the high-nutrient, low-pH well water.  In contrast, T. maxima and T. squamosa had significantly lower growth rates in low pH, whereas growth of T. crocea was not significantly different between low pH and ambient seawater.  These experiments demonstrate species-specific differences with each treatment, which cautions against making broad generalizations regarding the effects of substrate type, feeding effects, nutrient enrichment, and ocean acidification on tridacnid culture and survival.

Effect of carbonate chemistry manipulations on calcification, respiration, and excretion of a Mediterranean pteropod

Although shelled pteropods are expected to be particularly sensitive to ocean acidification, the few available studies have mostly focused on polar species and have not allowed determining which parameter of the carbonate system controls their calcification. Specimens of the temperate Mediterranean species Creseis acicula were maintained under seven different conditions of the carbonate chemistry, obtained by manipulating pH and total alkalinity, with the goal to disentangle the effects of the pH and the saturation state with respect to aragonite (Ωa). Our results tend to show that respiration, excretion as well as rates of net and gross calcification were not directly affected by a decrease in pH but decreased significantly with a decrease in Ωa. Due to the difficulties in maintaining pteropods in the laboratory and the important variability in their abundances in our study site, long-term acclimation as well as replication of the experiment was not possible. However, we strongly believe that these results represent an important step in the mechanistic understanding of the effect of ocean acidification on pteropods physiology.

Marine bivalve shell geochemistry and ultrastructure from modern low pH environments: environmental effect versus experimental bias (update)

Bivalve shells can provide excellent archives of past environmental change but have not been used to interpret ocean acidification events. We investigated carbon, oxygen and trace element records from different shell layers in the mussels Mytilus galloprovincialis combined with detailed investigations of the shell ultrastructure. Mussels from the harbour of Ischia (Mediterranean, Italy) were transplanted and grown in water with mean pHT 7.3 and mean pHT 8.1 near CO2 vents on the east coast of the island. Most prominently, the shells recorded the shock of transplantation, both in their shell ultrastructure, textural and geochemical record. Shell calcite, precipitated subsequently under acidified seawater responded to the pH gradient by an in part disturbed ultrastructure. Geochemical data from all test sites show a strong metabolic effect that exceeds the influence of the low-pH environment. These field experiments showed that care is needed when interpreting potential ocean acidification signals because various parameters affect shell chemistry and ultrastructure. Besides metabolic processes, seawater pH, factors such as salinity, water temperature, food availability and population density all affect the biogenic carbonate shell archive.

Predominance of heavily calcified coccolithophores at low CaCO3 saturation during winter in the Bay of Biscay

Coccolithophores are an important component of the Earth system, and, as calcifiers, their possible susceptibility to ocean acidification is of major concern. Laboratory studies at enhanced pCO₂ levels have produced divergent results without overall consensus. However, it has been predicted from these studies that, although calcification may not be depressed in all species, acidification will produce “a transition in dominance from more to less heavily calcified coccolithophores” [Ridgwell A, et al., (2009) Biogeosciences 6:2611–2623]. A recent observational study [Beaufort L, et al., (2011) Nature 476:80–83] also suggested that coccolithophores are less calcified in more acidic conditions. We present the results of a large observational study of coccolithophore morphology in the Bay of Biscay. Samples were collected once a month for over a year, along a 1,000-km-long transect. Our data clearly show that there is a pronounced seasonality in the morphotypes of Emiliania huxleyi, the most abundant coccolithophore species. Whereas pH and CaCO₃ saturation are lowest in winter, the E. huxleyi population shifts from <10% (summer) to >90% (winter) of the heavily calcified form. However, it is unlikely that the shifts in carbonate chemistry alone caused the morphotype shift. Our finding that the most heavily calcified morphotype dominates when conditions are most acidic is contrary to the earlier predictions and raises further questions about the fate of coccolithophores in a high-CO₂ world.

Before ocean acidification: calcifier chemistry lessons

Ocean Acidification (OA) has been an important research topic for a decade. Scientists have focused on how the predicted 56% decline in the seawater carbonate ion () concentration will dramatically impair the ability of calcifiers, ranging from coccolithophores to shellfish, to form calcium carbonate (CaCO₃) structures, and the implications of the reduced carbonate saturation state (Ω) for increased dissolution of such structures. However, most published OA studies have overlooked a fundamental issue: most calcifying organisms do not rely on carbonate from seawater to calcify; they use either bicarbonate () or metabolically-produced CO₂. The ability of important primary (corals, coralline seaweeds and coccolithophores) and secondary (mollusks) producers to modify their local carbonate chemistry suggests that the primary threat to them from OA is by dissolution rather than impaired calcification. Here, we draw on calcification research from an era before OA and combine it with recent studies that question the source of the carbonate ion, to provide new insights into how OA might affect calcifying organisms. Organismal modification of local carbonate chemistry may enable calcifiers to successfully form calcareous structures despite OA.

Impacts of seawater acidification on mantle gene expression patterns of the Baltic Sea blue mussel: implications for shell formation and energy metabolism

Marine organisms have to cope with increasing CO₂ partial pressures and decreasing pH in the oceans. We elucidated the impacts of an 8-week acclimation period to four seawater pCO₂ treatments (39, 113, 243 and 405 Pa/385, 1,120, 2,400 and 4,000 μatm) on mantle gene expression patterns in the blue mussel Mytilus edulis from the Baltic Sea. Based on the M. edulis mantle tissue transcriptome, the expression of several genes involved in metabolism, calcification and stress responses was assessed in the outer (marginal and pallial zone) and the inner mantle tissues (central zone) using quantitative real-time PCR. The expression of genes involved in energy and protein metabolism (F-ATPase, hexokinase and elongation factor alpha) was strongly affected by acclimation to moderately elevated CO₂ partial pressures. Expression of a chitinase, potentially important for the calcification process, was strongly depressed (maximum ninefold), correlating with a linear decrease in shell growth observed in the experimental animals. Interestingly, shell matrix protein candidate genes were less affected by CO₂ in both tissues. A compensatory process toward enhanced shell protection is indicated by a massive increase in the expression of tyrosinase, a gene involved in periostracum formation (maximum 220-fold). Using correlation matrices and a force-directed layout network graph, we were able to uncover possible underlying regulatory networks and the connections between different pathways, thereby providing a molecular basis of observed changes in animal physiology in response to ocean acidification.

Reduced calcification decreases photoprotective capability in the coccolithophorid Emiliania huxleyi

Intracellular calcification of coccolithophores generate CO₂ and consumes additional energy for acquisition of calcium and bicarbonate ions, therefore, it may correlate with photoprotective processes by influencing the energetics. To address this hypothesis, a calcifying Emiliania huxleyi strain (CS-369) was grown semi-continuously at reduced (0.1 mM, LCa) and ambient Ca²⁺ concentrations (10 mM, HCa) for 150 days (>200 generations). The HCa-grown cells had higher photosynthetic and calcification rates and higher contents of chl a and carotenoids compared to the naked (bearing no coccoliths) LCa-grown cells. When exposed to stress-full levels of PAR, LCa-grown cells displayed lower photochemical yield and less efficient non-photochemical quenching (NPQ). When the LCa or HCa-grown cells were inversely shifted to their counterpart medium, LCa to HCa transfer increased photosynthetic carbon fixation (P), calcification rate (C), C/P ratio, NPQ and pigments contents, while those shifted from HCa to LCa exhibited the opposite performance. Increased NPQ, carotenoids and quantum yield clearly linked with increased or sustained calcification in E. huxleyi. The calcification must have played a role in dissipating excessive energy or as an additional drainage of electrons absorbed by the photosynthetic antennae. This phenomenon was further supported by testing two noncalcifying strains, which showed insignificant changes photosynthetic carbon fixation and NPQ when transferred to LCa condition.

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.

Interactive effects of ocean acidification and temperature on two scleractinian corals from Moorea, French Polynesia

This study tested the hypothesis that the response of corals to temperature and pCO2 is consistent between taxa. Juvenile massive Porites spp. and branches of P. rus from the back reef of Moorea were incubated for 1 month under combinations of temperature (29.3 °C and 25.6 °C) and pCO2 (41.6 Pa and 81.5 Pa) at an irradiance of 599 μmol quanta m−2 s−1. Using microcosms and CO2 gas mixing technology, treatments were created in a partly nested design (tanks) with two between-plot factors (temperature and pCO2), and one within-plot factor (taxon); calcification was used as a dependent variable. pCO2 and temperature independently affected calcification, but the response differed between taxa. Massive Porites spp. was largely unaffected by the treatments, but P. rus grew 50% faster at 29.3 °C compared with 25.6 °C, and 28% slower at 81.5 Pa vs. 41.6 Pa CO2. A compilation of studies placed the present results in a broader context and tested the hypothesis that calcification for individual coral genera is independent of pH, [HCO3−], and [CO32−]. Unlike recent reviews, this analysis was restricted to studies reporting calcification in units that could be converted to nmol CaCO3 cm−2 h−1. The compilation revealed a high degree of variation in calcification as a function of pH, [HCO3−], and [CO32−], and supported three conclusions: (1) studies of the effects of ocean acidification on corals need to pay closer attention to reducing variance in experimental outcomes to achieve stronger synthetic capacity, (2) coral genera respond in dissimilar ways to pH, [HCO3−], and [CO32−], and (3) calcification of massive Porites spp. is relatively resistant to short exposures of increased pCO2, similar to that expected within 100 y.

Effects of diurnally oscillating pCO2 on the calcification and survival of coral recruits

Manipulative studies have demonstrated that ocean acidification (OA) is a threat to coral reefs, yet no experiments have employed diurnal variations in pCO₂ that are ecologically relevant to many shallow reefs. Two experiments were conducted to test the response of coral recruits (less than 6 days old) to diurnally oscillating pCO₂; one exposing recruits for 3 days to ambient (440 µatm), high (663 µatm) and diurnally oscillating pCO₂ on a natural phase (420–596 µatm), and another exposing recruits for 6 days to ambient (456 µatm), high (837 µatm) and diurnally oscillating pCO₂ on either a natural or a reverse phase (448–845 µatm). In experiment I, recruits exposed to natural-phased diurnally oscillating pCO₂ grew 6–19% larger than those in ambient or high pCO₂. In experiment II, recruits in both high and natural-phased diurnally oscillating pCO₂ grew 16 per cent larger than those at ambient pCO₂, and this was accompanied by 13–18% higher survivorship; the stimulatory effect on growth of oscillatory pCO₂ was diminished by administering high pCO₂ during the day (i.e. reverse-phased). These results demonstrate that coral recruits can benefit from ecologically relevant fluctuations in pCO₂ and we hypothesize that the mechanism underlying this response is highly pCO₂-mediated, night-time storage of dissolved inorganic carbon that fuels daytime calcification.