Reprint: The Importance of Natural Acidified Systems in the Study of Ocean Acidification: What Have We Learned?

Human activity is generating an excess of atmospheric CO2, resulting in what we know as ocean acidification, which produces changes in marine ecosystems. Until recently, most of the research in this area had been done under small-scale, laboratory conditions, using few variables, few species and few life cycle stages. These limitations raise questions about the reproducibility of the environment and about the importance of indirect effects and synergies in the final results of these experiments. One way to address these experimental problems is by conducting studies in situ, in natural areas where expected future pH conditions already occur, such as CO2 vent systems. In the present work, we compile and discuss the latest research carried out in these natural laboratories, with the objective to summarize their advantages and disadvantages for research to improve these investigations so they can better help us understand how the oceans of the future will change.

Gonadal rematuration and sex-specific reproductive impairment in Manila clams under ocean acidification.

Ocean acidification (OA) can affect marine bivalves at various levels of biological organization. Yet, little effort has been devoted to understanding how OA affects the reproductive events of marine bivalves during multiple cycles of maturation. Here, we tested sex-specific reproductive responses of Manila clams (Ruditapes philippinarum) to OA during gonadal rematuration. Under acidified conditions, both male and female clams exhibited delayed gonadal rematuration following spawning and impairments in gonadal tissues, which can be likely ascribed to lowered concentrations of hormones and vitellogenin. The findings indicate that marine bivalves experience significant declines in reproductive capacity as a result of OA during their reproductive cycles, with clear sex-specific differences. Consequently, it is essential to consider sex-specific reproduction responses of marine bivalves to OA when developing conservation strategies and forecasting population sustainability in a rapidly acidifying marine environment.

Investigating intraspecific variability in the biological responses of sea urchins (Paracentrotus lividus) to seawater acidification.

Alterations in seawater chemistry posed by acidification may lead to immunological and antioxidant defence impairment in sea urchins, with differences among local populations. Here, we analyzed the effects of reduced pH on Paracentrotus lividus, with a multibiomarker approach, and the possible intraspecific variations in sea urchin responses. Two groups of animals with different ecological histories (i.e., the pattern of environmental characteristics and pressures experienced throughout the organism's lifetime) were maintained at ambient pH and pH reduced of 0.4 units for 8 months. Changes in gonadosomatic index (GSI), immunological, and oxidative stress biomarkers were assessed in coelomic fluid, gonads, and digestive tract. Animals maintained at reduced pH showed limited impact of seawater acidification compared to the ambient pH condition. However, sea urchins from the two sites were differently influenced by the seawater pH (as shown by multivariate analyses). GSI and immunological and antioxidant status were differentially modulated between the two sexes, with generally higher values in females, but differences between sexes in relation to the pH of exposure were limited. Overall, our findings highlight that the impact of environmental stressors may differ in sea urchins from different locations. This has implications for the maintenance of P. lividus wild populations under future global change scenarios.

Mitigation of hypoxia and ocean acidification on the inner East China Sea shelf impacted by the 2023 summer drought.

Hypoxia and acidification are universal environmental issues in coastal seas, especially in large river dominated shelves, and the East China Sea shelf is a typical case among them. However, the responses of status of hypoxia and acidification in coastal seas to the extremes of river discharges are still to be revealed. This study surveyed the influences of a summer drought on the status of hypoxia and acidification on the inner East China Sea shelf off the Changjiang estuary. In August of 2023 during a summer drought, carbonate system parameters and dissolved oxygen (DO) were surveyed on the East China Sea shelf off the Changjiang estuary. As expected, dissolved inorganic carbon (DIC) removal (up to >40 µmol kg-1) and DO over-saturation (up to >110 %) accompanied by high pH (up to >8.15) in the surface water were observed. However, low DO (32-172 µmol kg-1), low pH (7.63-8.04) and low saturation state index of aragonite (ΩAr) (1.34-3.06) in the bottom water were observed. Relationships of Excess DIC with DO consumption, and pH and ΩAr with Excess DIC indicated that the hypoxia and acidification in the bottom water was due mainly to the remineralization of the marine-sourced organic matter. Nevertheless, both hypoxia and acidification were mitigated, i.e. the hypoxic area was smaller, the minimum DO concentration, pH and saturation state index of aragonite were higher in August of 2023 than under the general summer condition. The lower Changjiang discharge (∼60 % of the long-term monthly average) mitigated eutrophication of the East China Sea shelf and decreased the phytoplankton biomass in the surface water and subsequently the hypoxia and acidification in the bottom water. However, acidification of the bottom water on the East China Sea shelf was still severe even during the summer drought. Regulating the anthropogenic impact on the coastal marginal seas is still urgently needed to mitigate the acidification status.

Ocean acidification may alleviate the toxicity of zinc to the macroalga, Ulva lactuca.

We investigated the toxic effects of different zinc (Zn) concentrations (natural seawater, 25 µg/L, and 100 µg/L) under two CO2 concentrations (410 ppmv, and 1000 ppmv) on Ulva lactuca. A significant decrease in the relative growth rate of U. lactuca was observed with an increase in Zn concentration under the low CO2 treatment condition, and we observed a notable decrease at 100 µg/L Zn under the high CO2 treatment condition. Moreover, the net photosynthetic rate increased when thalli were cultured under 25 and 100 µg/L Zn under the high CO2 treatment condition. The concentrations of chlorophyll a and b were significantly increased under 100 µg/L Zn and the high CO2 treatment conditions. Malondialdehyde content decreased under high CO2 treatment conditions, compared with the low CO2 treatment conditions, regardless of the Zn concentration. These findings suggest that ocean acidification may alleviate the toxic effects of Zn pollution on U. lactuca.

Assessing the impact of simulated ocean acidification on the photodegradation of selected microplastics.

This study investigated the impact of ocean acidification on the photodegradation of three microplastics (MPs): polypropylene (PP), expanded polystyrene (EPS), and ethylene-vinyl acetate (EVA), under accelerated UV radiation at three pH levels (i.e., 8.1, 7.8, and 7.5), simulating marine conditions. The acidification system simulated current and projected future environmental conditions. As expected, an increase in partial pressure of CO2, total inorganic carbon, bicarbonate ion, and CO2 resulted in more acidic pH levels, with the reverse being true for the carbonate ion. Structural changes of MPs were evaluated, revealing that all weathered samples underwent higher degradation rate compared to the virgin samples. The oxidation state and crystallinity of PP and EVA MPs were higher in samples exposed to the lowest pH, whereas no significant increase in the degradation rate of EPS samples was observed. Saltwater acidification in this study contributed to enhance the photo-oxidation of MPs depending on their polymeric composition.

Interactions between ocean acidification and multiple environmental drivers on the biochemical traits of marine primary producers: A meta-analysis.

Ocean acidification (OA) interacts with multiple environmental drivers, such as temperature, nutrients, and ultraviolet radiation (UVR), posing a threat to marine primary producers. In this study, we conducted a quantitative meta-analysis of 1001 experimental assessments from 68 studies to examine the combined effects of OA and multiple environmental drivers (e.g., light, nutrient) on the biochemical compositions of marine primary producers. The results revealed significant positive effects of each environmental driver and their interactions with OA according to Hedge's d analysis. The results revealed significant positive effects of multiple environmental drivers and their interactions with OA. Additive effects dominated (71%), with smaller proportions of antagonistic (20%) and synergistic interactions (9%). The antagonistic interactions, although fewer, had a substantial impact, causing OA and other environmental drivers to interact antagonistically. Significant differences were observed among taxonomic groups: haptophytes and rhodophytes were negatively affected, while bacillariophytes were positively affected by OA. Our findings also indicated that the interactions between OA and multiple environmental drivers varied depending on specific type of the environmental driver, suggesting a modulating role of OA on the biochemical compositions of marine primary producers in response to global change. In summary, our study elucidates the complex interactions between OA and multiple environmental drivers on marine primary producers, highlighting the varied impacts on biochemical compositions and elemental stoichiometry.

From Individual Calcifiers to Ecosystem Dynamics: Ocean Acidification Effects on Urchins and Abalone.

A central question in ecology is to what extent do trophic interactions govern the structure and function of communities? This question is becoming more pressing as trophic interactions shift with rapid climate change. Sea urchins and abalone are key invertebrates in the habitats where they reside. Sea urchins are critical members of exemplar trophic cascades in kelp forests due to their impact on kelp establishment and maintenance; yet their populations are controlled by predators, such as sea otters and sunflower sea stars. Abalone compete with urchins for macroalgal food resources and therefore can help regulate urchin populations in kelp forests. Given that both urchin tests and abalone shells used for predator defense are comprised of calcium carbonate, much research has been conducted on the impacts of ocean acidification (OA) on these calcified structures. A growing body of literature has shown that urchin tests are less calcified and break with less force under OA conditions. Less is known about abalone, but their shells also appear to respond negatively to OA. Using kelp forest communities as exemplar ecosystems, we discuss the morphological, biomechanical, and physiological responses to OA in urchins and abalone and consider how these individual level responses scale to trophic interactions and ultimately whole ecosystem processes. Although the impacts of OA on the calcified structures used for defense have been well studied, calcified mechanisms for food consumption, such as the Aristotle's lantern of urchins, are much less understood. Thus, examining both the feeding and defense sides of trophic interactions would greatly improve our understanding of OA responses across individual to ecosystem scales. More generally, measurements of morphological, biomechanical, and physiological responses to OA can be made in individuals to help predict higher level ecological responses, which would greatly contribute to broader predictions of whole ecosystem responses to OA.

Changes in gill neuroepithelial cells and morphology of threespine stickleback (Gasterosteus aculeatus) to hypoxia and simulated ocean acidification.

Coastal marine environments are characterized by daily, seasonal and long-term changes in both O2 and CO2, driven by local biotic and abiotic factors. The neuroepithelial cells (NECs) of fish are thought to be the putative chemoreceptors for sensing oxygen and CO2, and, thus, NECs play a key role in detecting these environmental changes. However, the role of NECs as chemosensors in marine fish remains largely understudied. In this study, the NECs of marine threespine sticklebacks (Gasterosteus aculeatus) were characterized using immunohistochemistry. We then determined if there were changes in NEC size and density, and in gill morphology in response to either mild (10 kPa) or moderate (6.8 kPa) hypoxia and two levels of elevated CO2 (1,500 and 3,000 µatm). We found that the NECs of stickleback contained synaptic vesicles and were innervated, and were 50-300% larger and 2 to 4 times more abundant than in other similar sized freshwater fishes. NEC size and density were largely unaffected by exposure to hypoxia, but there was a 50% decrease in interlamellar cell mass (ILCM) in response to mild and moderate hypoxia. NECs increased in size, but not abundance in response to elevated CO2. Moreover, fish exposed to moderate or elevated CO2 had 53-78% larger ILCMs compared to control fish. Our results demonstrated that adult marine sticklebacks have NECs that can respond to environmentally relevant pCO2 and likely hypoxia, which highlights the importance of NECs in marine fishes under the heterogeneity of environmental conditions in coastal areas.

Local differences in robustness to ocean acidification.

Ocean acidification (OA) caused by increased atmospheric carbon dioxide is affecting marine systems globally and is more extreme in coastal waters. A wealth of research to determine how species will be affected by OA, now and in the future, is emerging. Most studies are discrete and generally do not include the full life cycle of animals. Studies that include the potential for adaptation responses of animals from areas with different environmental conditions and the most vulnerable life stages are needed. Therefore, we conducted experiments with the widely distributed blue mussel, Mytilus edulis, from populations regularly exposed to different OA conditions. Mussels experienced experimental conditions prior to spawning, through embryonic and larval development, both highly vulnerable stages. Survivorship to metamorphosis of larvae from all populations was negatively affected by extreme OA conditions (pH 7.3, Ωar, 0.39, pCO2 2479.74), but, surprisingly, responses to mid OA (pH 7.6, Ωar 0.77, pCO21167.13) and low OA (pH 7.9, Ωar 1.53, pCO2 514.50) varied among populations. Two populations were robust and showed no effect of OA on survivorship in this range. One population displayed the expected negative effect on survivorship with increased OA. Unexpectedly, survivorship in the fourth population was highest under mid OA conditions. There were also significant differences in development time among populations that were unaffected by OA. These results suggest that adaptation to OA may already be present in some populations and emphasizes the importance of testing animals from different populations to see the potential for adaptation to OA.

Carbon export from submarine groundwater discharge in a semi-enclosed bay: Impact for the buffering capacity against coastal ocean acidification.

Submarine groundwater discharge (SGD) serves as an important pathway for the transport of dissolved carbon from land to ocean, significantly affecting the coastal biogeochemical cycles. However, the impact of SGD-derived dissolved carbon on the coastal carbon budget remains poorly understood. This study first quantified SGD and associated dissolved organic carbon (DOC), dissolved inorganic carbon (DIC) and total alkalinity (TA) fluxes in Daya Bay using mass balance models based on radium isotopes (223Ra, 224Ra, 226Ra and 228Ra). We then constructed carbon mass balance models to evaluate the impact of SGD-derived carbon on the buffering capacity against coastal ocean acidification. The estimated SGD fluxes ranged from 0.80 × 107 to 2.64 × 107 m3d-1. The DIC, DOC and TA fluxes from SGD were 17.90-36.44 mmol m-2d-1, 0.93-2.13 mmol m-2d-1, and 21.19-28.47 mmol m-2d-1, respectively. Based on carbon mass balances, the DIC flux from SGD was 19-39 times the riverine input, accounting for 27.16 % ∼ 37.64 % of the total carbon source. These results suggest that SGD is a major contributor to DIC, significantly affecting the coastal carbon budget. Furthermore, the average TA:DIC ratio of groundwater discharging into Daya Bay was approximately 1.13. High TA exports enhance the buffering capacity of the coastal ocean and contribute bicarbonate to the ocean, playing a significant role in the ocean carbon sequestration process. This study demonstrates the importance of SGD-derived dissolved carbon in the assessment of coastal carbon budgets.

Ocean acidification alters microeukaryotic and bacterial food web interactions in a eutrophic subtropical mesocosm.

Ocean acidification (OA) is known to influence biological and ecological processes, mainly focusing on its impacts on single species, but little has been documented on how OA may alter plankton community interactions. Here, we conducted a mesocosm experiment with ambient (∼410 ppmv) and high (1000 ppmv) CO2 concentrations in a subtropical eutrophic region of the East China Sea and examined the community dynamics of microeukaryotes, bacterioplankton and microeukaryote-attached bacteria in the enclosed coastal seawater. The OA treatment with elevated CO2 affected taxa as the phytoplankton bloom stages progressed, with a 72.89% decrease in relative abundance of the protist Cercozoa on day 10 and a 322% increase in relative abundance of Stramenopile dominated by diatoms, accompanied by a 29.54% decrease in relative abundance of attached Alphaproteobacteria on day 28. Our study revealed that protozoans with different prey preferences had differing sensitivity to high CO2, and attached bacteria were more significantly affected by high CO2 compared to bacterioplankton. Our findings indicate that high CO2 changed the co-occurrence network complexity and stability of microeukaryotes more than those of bacteria. Furthermore, high CO2 was found to alter the proportions of potential interactions between phytoplankton and their predators, as well as microeukaryotes and their attached bacteria in the networks. The changes in the relative abundances and interactions of microeukaryotes between their predators in response to high CO2 revealed in our study suggest that high CO2 may have profound impacts on marine food webs.

Preliminary investigation on the potential involvement of an ABC-like gene in Halomicronema metazoicum (Cyanobacteria) tolerance to low seawater pH in an ocean acidification scenario.

Decreasing ocean surface pH, called ocean acidification (OA), is among the major risks for marine ecosystems due to human-driven atmospheric pCO2 increase. Understanding the molecular mechanisms of adaptation enabling marine species to tolerate a lowered seawater pH could support predictions of consequences of future OA scenarios for marine life. This study examined whether the ATP-binding cassette (ABC)-like gene slr2019 confers tolerance to the marine cyanobacterium Halomicronema metazoicum to low seawater pH conditions (7.7, 7.2, 6.5) in short- and long-term exposures (7 and 30 d). Photosynthetic pigment content indicated that the species can tolerate all three lowered-pH conditions. At day 7, slr2019 was up-regulated at pH 7.7 while no changes were observed at lower pH. After 30-d exposure, a significant decrease in slr2019 transcript levels was observed in all low-pH treatments. These first results indicate an effect of low pH on the examined transporter expression in H. metazoicum.

Diverse inorganic carbon uptake strategies in Antarctic seaweeds: Revealing species-specific responses and implications for Ocean Acidification.

Seaweeds are important components of coastal benthic ecosystems along the Western Antarctic Peninsula (WAP), providing refuge, food, and habitat for numerous associated species. Despite their crucial role, the WAP is among the regions most affected by global climate change, potentially impacting the ecology and physiology of seaweeds. Elevated atmospheric CO2 concentrations have led to increased dissolved inorganic carbon (Ci) with consequent declines in oceanic pH and alterations in seawater carbonate chemistry, known as Ocean Acidification (OA). Seaweeds possess diverse strategies for Ci uptake, including CO2 concentrating mechanisms (CCMs), which may distinctly respond to changes in Ci concentrations. Conversely, some seaweeds do not operate CCMs (non-CCM species) and rely solely on CO2. Nevertheless, our understanding of the status and functionality of Ci uptake strategies in Antarctic seaweeds remains limited. Here, we investigated the Ci uptake strategies of seaweeds along a depth gradient in the WAP. Carbon isotope signatures (δ13C) and pH drift assays were used as indicators of the presence or absence of CCMs. Our results reveal variability in CCM occurrence among algal phyla and depths ranging from 0 to 20 m. However, this response was species specific. Among red seaweeds, the majority relied solely on CO2 as an exogenous Ci source, with a high percentage of non-CCM species. Green seaweeds exhibited depth-dependent variations in CCM status, with the proportion of non-CCM species increasing at greater depths. Conversely, brown seaweeds exhibited a higher prevalence of CCM species, even in deep waters, indicating the use of CO2 and HCO3-. Our results are similar to those observed in temperate and tropical regions, indicating that the potential impacts of OA on Antarctic seaweeds will be species specific. Additionally, OA may potentially increase the abundance of non-CCM species relative to those with CCMs.

Predictable patterns within the kelp forest can indirectly create temporary refugia from ocean acidification.

Kelps are recognized for providing many ecosystem services in coastal areas and considered in ocean acidification (OA) mitigation. However, assessing OA modification requires an understanding of the multiple parameters involved in carbonate chemistry, especially in highly dynamic systems. We studied the effects of sugar kelp (Saccharina latissima) on an experimental farm at the north end of Hood Canal, Washington-a low retentive coastal system. In this field mesocosm study, two oyster species (Magallana gigas, Ostrea lurida) were exposed at locations in the mid, edge, and outside the kelp array. The Hood Head Sugar Kelp Farm Model outputs were used to identify dominating factors in spatial and temporal kelp dynamics, while wavelet spectrum analyses helped in understanding predictability patterns. This was linked to the measured biological responses (dissolution, growth, isotopes) of the exposed organisms. Positioned in an area of high (sub)-diel tidal fluxes with low retention potential, there were no measurable alterations of the seawater pH at the study site, demonstrating that the kelp array could not induce a direct mitigating effect against OA. However, beneficial responses in calcifiers were still observed, which are linked to two causes: increased pH predictability and improved provisioning through kelp-derived particulate organic resource utilization and as such, kelp improved habitat suitability and indirectly created refugia against OA. This study can serve as an analogue for many coastal bay habitats where prevailing physical forcing drives chemical changes. Future macrophyte studies that investigate OA mitigating effects should focus also on the importance of predictability patterns, which can additionally improve the conditions for marine calcifiers and ecosystem services vulnerable to or compromised by OA, including aquaculture sustainability.

Effects of water flow and ocean acidification on oxygen and pH gradients in coral boundary layer.

Reef-building corals live in highly hydrodynamic environments, where water flow largely controls the complex chemical microenvironments surrounding them-the concentration boundary layer (CBL). The CBL may be key to alleviate ocean acidification (OA) effects on coral colonies by partially isolating them. However, OA effects on coral CBL remain poorly understood, particularly under different flow velocities. Here, we investigated these effects on the reef-building corals Acropora cytherea, Pocillopora verrucosa, and Porites cylindrica. We preconditioned corals to a control (pH 8.0) and OA (pH 7.8) treatment for four months and tested how low flow (2 cm s-1) and moderate flow (6 cm s-1) affected O2 and H+ CBL traits (thickness, surface concentrations, and flux) inside a unidirectional-flow chamber. We found that CBL traits differed between species and flow velocities. Under OA, traits remained generally stable across flows, except surface pH. In all species, the H+ CBL was thin and led to lower surface pH. Still, low flow thickened H+ CBLs and increased light elevation of surface pH. In general, our findings reveal a weak to null OA modulation of the CBL. Moreover, the OA-buffering capacity by the H+ CBL may be limited in coral species, though low flow could enhance CBL sheltering.

Transcriptomic responses in the nervous system and correlated behavioural changes of a cephalopod exposed to ocean acidification.

BACKGROUND: The nervous system is central to coordinating behavioural responses to environmental change, likely including ocean acidification (OA). However, a clear understanding of neurobiological responses to OA is lacking, especially for marine invertebrates. RESULTS: We evaluated the transcriptomic response of the central nervous system (CNS) and eyes of the two-toned pygmy squid (Idiosepius pygmaeus) to OA conditions, using a de novo transcriptome assembly created with long read PacBio ISO-sequencing data. We then correlated patterns of gene expression with CO2 treatment levels and OA-affected behaviours in the same individuals. OA induced transcriptomic responses within the nervous system related to various different types of neurotransmission, neuroplasticity, immune function and oxidative stress. These molecular changes may contribute to OA-induced behavioural changes, as suggested by correlations among gene expression profiles, CO2 treatment and OA-affected behaviours. CONCLUSIONS: This study provides the first molecular insights into the neurobiological effects of OA on a cephalopod and correlates molecular changes with whole animal behavioural responses, helping to bridge the gaps in our knowledge between environmental change and animal responses.

Rubble persistence under ocean acidification threatened by accelerated bioerosion and lower-density coral skeletons.

As the balance between erosional and constructive processes on coral reefs tilts in favor of framework loss under human-induced local and global change, many reef habitats worldwide degrade and flatten. The resultant generation of coral rubble and the beds they form can have lasting effects on reef communities and structural complexity, threatening the continuity of reef ecological functions and the services they provide. To comprehensively capture changing framework processes and predict their evolution in the context of climate change, heavily colonized rubble fragments were exposed to ocean acidification (OA) conditions for 55 days. Controlled diurnal pH oscillations were incorporated in the treatments to account for the known impact of diel carbonate chemistry fluctuations on calcification and dissolution response to OA. Scenarios included contemporary pH (8.05 ± 0.025 diel fluctuation), elevated OA (7.90 ± 0.025), and high OA (7.70 ± 0.025). We used a multifaceted approach, combining chemical flux analyses, mass alteration measurements, and computed tomography scanning images to measure total and chemical bioerosion, as well as chemically driven secondary calcification. Rates of net carbonate loss measured in the contemporary conditions (1.36 kg m-2 year-1) were high compared to literature and increased in OA scenarios (elevated: 1.84 kg m-2 year-1 and high: 1.59 kg m-2 year-1). The acceleration of these rates was driven by enhanced chemical dissolution and reduced secondary calcification. Further analysis revealed that the extent of these changes was contingent on the density of the coral skeleton, in which the micro- and macroborer communities reside. Findings indicated that increased mechanical bioerosion rates occurred in rubble with lower skeletal density, which is of note considering that corals form lower-density skeletons under OA. These direct and indirect effects of OA on chemical and mechanical framework-altering processes will influence the permanence of this crucial habitat, carrying implications for biodiversity and reef ecosystem function.

Out of shape: Ocean acidification simplifies coral reef architecture and reshuffles fish assemblages.

Climate change stressors are progressively simplifying biogenic habitats in the terrestrial and marine realms, and consequently altering the structure of associated species communities. Here, we used a volcanic CO2 seep in Papua New Guinea to test in situ if altered reef architecture due to ocean acidification reshuffles associated fish assemblages. We observed replacement of branching corals by massive corals at the seep, with simplified coral architectural complexity driving abundance declines between 60% and 86% for an assemblage of damselfishes associated with branching corals. An experimental test of habitat preference for a focal species indicated that acidification does not directly affect habitat selection behaviour, with changes in habitat structural complexity consequently appearing to be the stronger driver of assemblage reshuffling. Habitat health affected anti-predator behaviour, with P. moluccensis becoming less bold on dead branching corals relative to live branching corals, irrespective of ocean acidification. We conclude that coral reef fish assemblages are likely to be more sensitive to changes in habitat structure induced by increasing pCO2 than any direct effects on behaviour, indicating that changes in coral architecture and live cover may act as important mediators of reef fish community structures in a future ocean.