Direct and latent effects of ocean acidification on the transition of a sea urchin from planktonic larva to benthic juvenile.

Ongoing ocean acidification is expected to affect marine organisms and ecosystems. While sea urchins can tolerate a wide range of pH, this comes at a high energetic cost, and early life stages are particularly vulnerable. Information on how ocean acidification affects transitions between life-history stages is scarce. We evaluated the direct and indirect effects of pH (pHT 8.0, 7.6 and 7.2) on the development and transition between life-history stages of the sea urchin Strongylocentrotus droebachiensis, from fertilization to early juvenile. Continuous exposure to low pH negatively affected larval mortality and growth. At pH 7.2, formation of the rudiment (the primordial juvenile) was delayed by two days. Larvae raised at pH 8.0 and transferred to 7.2 after competency had mortality rates five to six times lower than those kept at 8.0, indicating that pH also has a direct effect on older, competent larvae. Latent effects were visible on the larvae raised at pH 7.6: they were more successful in settling (45% at day 40 post-fertilization) and metamorphosing (30%) than larvae raised at 8.0 (17 and 1% respectively). These direct and indirect effects of ocean acidification on settlement and metamorphosis have important implications for population survival.

Broad Thermal Tolerance in the Cold-Water Coral Lophelia pertusa From Arctic and Boreal Reefs.

Along the Norwegian coasts and margins, extensive reefs of the stony coral Lophelia pertusa act as hotspots for local biodiversity. Climate models project that the temperature of Atlantic deep waters could rise by 1-3°C by 2100. In this context, understanding the effects of temperature on the physiology of cold-water species will help in evaluating their resilience to future oceanic changes. We investigated the response of L. pertusa to stepwise short-term increases in temperature. We sampled corals from four reefs, two located north of the Arctic circle and two at the mid-Norwegian shelf (boreal). In on-board experiments (one per reef), the sampled fragments were exposed to increasing temperatures from 5 to 15°C over 58 h. Respiration increased linearly by threefold for a 10°C increase. The short-term temperature increase did not induce mortality, cellular (neutral red assay for lysosome membrane stability; but one exception) or oxidative stress (lipid peroxidation assay) - to a few exceptions. However, the variability of the respiration responses depended on the experiment (i.e., reef location), possibly linked to the genetic structure of the individuals that we sampled (e.g., clones or siblings). The corals from the Arctic and boreal regions appear to have a high tolerance to the rapid temperature fluctuations they experience in the field. Over extended periods of time however, an increased metabolism could deplete the energy stored by the corals, if not met by an increased food availability and/or uptake. Empirical data on organisms' thermal performance curves, such as the one presented in this study for L. pertusa, will be useful to implement predictive models on the responses of species and populations to climate change.

Development of the sea urchin Heliocidaris crassispina from Hong Kong is robust to ocean acidification and copper contamination.

Metallic pollution is of particular concern in coastal cities. In the Asian megacity of Hong Kong, despite water qualities have improved over the past decade, some local zones are still particularly affected and could represent sinks for remobilization of labile toxic species such as copper. Ocean acidification is expected to increase the fraction of the most toxic form of copper (Cu2+) by 2.3-folds by 2100 (pH ≈7.7), increasing its bioavailability to marine organisms. Additionally, multiple stressors are likely to exert concomitant effects (additive, synergic or antagonist) on the organisms living in the sea. Here, we tested the hypothesis that copper-contaminated waters are more toxic to sea urchin larvae under future pH conditions. We exposed sea urchin embryos and larvae to two low-pH and two copper treatments (0.1 and 1.0 µM) in three separate experiments. Over the short time typically used for toxicity tests (up to 4-arm plutei, i.e. 3 days), larvae of the sea urchin Heliocidaris crassispina were robust and survived the copper levels present in Hong Kong waters today (≤0.19 µM) as well as the average pH projected for 2100. We, however, observed significant mortality with lowering pH in the longer, single-stressor experiment (Expt A: 8-arm plutei, i.e. 9 days). Abnormality and arm asymmetry were significantly increased by pH or/and by copper presence (depending on the experiment and copper level). Body size (d3; but not body growth rates in Expt A) was significantly reduced by both lowered pH and added copper. Larval respiration (Expt A) was doubled by a decrease at pHT from 8.0 to 7.3 on d6. In Expt B1.0 and B0.1, larval morphology (relative arm lengths and stomach volume) were affected by at least one of the two investigated factors. Although the larvae appeared robust, these sub-lethal effects may have indirect consequences on feeding, swimming and ultimately survival. The complex relationship between pH and metal speciation/uptake is not well-characterized and further investigations are urgently needed to detangle the mechanisms involved and to identify possible caveats in routinely used toxicity tests.

Ocean acidification modulates the incorporation of radio-labeled heavy metals in the larvae of the Mediterranean sea urchin Paracentrotus lividus.

The marine organisms which inhabit the coastline are exposed to a number of anthropogenic pressures that may interact. For instance, the accumulation of toxic metals present in coastal waters is expected to be modified by ocean acidification through e.g. changes in physiological performance and/or elements availability. Changes in bioaccumulation due to lowering pH are likely to be differently affected depending on the nature (essential vs. non-essential) and speciation of each element. The Mediterranean is of high concern for possible cumulative effects due to strong human influences on the coastline. The aim of this study was to determine the effect of ocean acidification (from pH 8.1 down to -1.0 pH units) on the incorporation kinetics of six trace metals (Mn, Co, Zn, Se, Ag, Cd, Cs) and one radionuclide (241Am) in the larvae of an economically- and ecologically-relevant sea urchin of the Mediterranean coastline: Paracentrotus lividus. The radiolabelled metals and radionuclides added in trace concentrations allowed precise tracing of their incorporation in larvae during the first 74 h of their development. Independently of the expected indirect effect of pH on larval size/developmental rates, Paracentrotus lividus larvae exposed to decreasing pHs incorporated significantly more Mn and Ag and slightly less Cd. The incorporation of Co, Cs and 241Am was unchanged, and Zn and Se exhibited complex incorporation behaviors. Studies such as this are necessary prerequisites to the implementation of metal toxicity mitigation policies for the future ocean. We discuss possible reasons and mechanisms for the specific effect of pH on each metals.

Ocean acidification increases larval swimming speed and has limited effects on spawning and settlement of a robust fouling bryozoan, Bugula neritina.

Few studies to date have investigated the effects of ocean acidification on non-reef forming marine invertebrates with non-feeding larvae. Here, we exposed adults of the bryozoan Bugula neritina and their larvae to lowered pH. We monitored spawning, larval swimming, settlement, and post-settlement individual sizes at two pHs (7.9 vs. 7.6) and settlement dynamics alone over a broader pH range (8.0 down to 6.5). Our results show that spawning was not affected by adult exposure (48h at pH7.6), larvae swam 32% faster and the newly-settled individuals grew significantly larger (5%) at pH7.6 than in the control. Although larvae required more time to settle when pH was lowered, reduced pH was not lethal, even down to pH6.5. Overall, this fouling species appeared to be robust to acidification, and yet, indirect effects such as prolonging the pelagic larval duration could increase predation risk, and might negatively impact population dynamics.

Genomic Characterization of the Evolutionary Potential of the Sea Urchin Strongylocentrotus droebachiensis Facing Ocean Acidification.

Ocean acidification (OA) is increasing due to anthropogenic CO2 emissions and poses a threat to marine species and communities worldwide. To better project the effects of acidification on organisms' health and persistence, an understanding is needed of the 1) mechanisms underlying developmental and physiological tolerance and 2) potential populations have for rapid evolutionary adaptation. This is especially challenging in nonmodel species where targeted assays of metabolism and stress physiology may not be available or economical for large-scale assessments of genetic constraints. We used mRNA sequencing and a quantitative genetics breeding design to study mechanisms underlying genetic variability and tolerance to decreased seawater pH (-0.4 pH units) in larvae of the sea urchin Strongylocentrotus droebachiensis. We used a gene ontology-based approach to integrate expression profiles into indirect measures of cellular and biochemical traits underlying variation in larval performance (i.e., growth rates). Molecular responses to OA were complex, involving changes to several functions such as growth rates, cell division, metabolism, and immune activities. Surprisingly, the magnitude of pH effects on molecular traits tended to be small relative to variation attributable to segregating functional genetic variation in this species. We discuss how the application of transcriptomics and quantitative genetics approaches across diverse species can enrich our understanding of the biological impacts of climate change.

Assessing physiological tipping point of sea urchin larvae exposed to a broad range of pH.

Our ability to project the impact of global change on marine ecosystem is limited by our poor understanding on how to predict species sensitivity. For example, the impact of ocean acidification is highly species-specific, even in closely related taxa. The aim of this study was to test the hypothesis that the tolerance range of a given species to decreased pH corresponds to their natural range of exposure. Larvae of the green sea urchin Strongylocentrotus droebachiensis were cultured from fertilization to metamorphic competence (29 days) under a wide range of pH (from pHT  = 8.0/pCO2  ≈ 480 µatm to pHT  = 6.5/pCO2  ≈ 20 000 µatm) covering present (from pHT 8.7 to 7.6), projected near-future variability (from pHT 8.3 to 7.2) and beyond. Decreasing pH impacted all tested parameters (mortality, symmetry, growth, morphometry and respiration). Development of normal, although showing morphological plasticity, swimming larvae was possible as low as pHT  ≥ 7.0. Within that range, decreasing pH increased mortality and asymmetry and decreased body length (BL) growth rate. Larvae raised at lowered pH and with similar BL had shorter arms and a wider body. Relative to a given BL, respiration rates and stomach volume both increased with decreasing pH suggesting changes in energy budget. At the lowest pHs (pHT  ≤ 6.5), all the tested parameters were strongly negatively affected and no larva survived past 13 days post fertilization. In conclusion, sea urchin larvae appeared to be highly plastic when exposed to decreased pH until a physiological tipping point at pHT  = 7.0. However, this plasticity was associated with direct (increased mortality) and indirect (decreased growth) consequences for fitness.

Acidified seawater impacts sea urchin larvae pH regulatory systems relevant for calcification.

Calcifying echinoid larvae respond to changes in seawater carbonate chemistry with reduced growth and developmental delay. To date, no information exists on how ocean acidification acts on pH homeostasis in echinoderm larvae. Understanding acid-base regulatory capacities is important because intracellular formation and maintenance of the calcium carbonate skeleton is dependent on pH homeostasis. Using H(+)-selective microelectrodes and the pH-sensitive fluorescent dye BCECF, we conducted in vivo measurements of extracellular and intracellular pH (pH(e) and pH(i)) in echinoderm larvae. We exposed pluteus larvae to a range of seawater CO(2) conditions and demonstrated that the extracellular compartment surrounding the calcifying primary mesenchyme cells (PMCs) conforms to the surrounding seawater with respect to pH during exposure to elevated seawater pCO(2). Using FITC dextran conjugates, we demonstrate that sea urchin larvae have a leaky integument. PMCs and spicules are therefore directly exposed to strong changes in pH(e) whenever seawater pH changes. However, measurements of pH(i) demonstrated that PMCs are able to fully compensate an induced intracellular acidosis. This was highly dependent on Na(+) and HCO(3)(-), suggesting a bicarbonate buffer mechanism involving secondary active Na(+)-dependent membrane transport proteins. We suggest that, under ocean acidification, maintained pH(i) enables calcification to proceed despite decreased pH(e). However, this probably causes enhanced costs. Increased costs for calcification or cellular homeostasis can be one of the main factors leading to modifications in energy partitioning, which then impacts growth and, ultimately, results in increased mortality of echinoid larvae during the pelagic life stage.