Description: Cold-water coral (CWC) reefs constitute one of the most complex deep-sea habitats harboring a vast diversity of associated species. Like other tropical or temperate framework builders, these systems are facing an uncertain future due to several threats, such as global warming and ocean acidification. In the case of Mediterranean CWC communities, the effect may be exacerbated due to the greater capacity of these waters to absorb atmospheric CO2 compared to the global ocean. Calcification in these organisms is an energy-demanding process, and it is expected that energy requirements will be greater as seawater pH and the availability of carbonate ions decrease. Therefore, studies assessing the effect of a pH decrease in skeletal growth, and metabolic balance are critical to fully understand the potential responses of these organisms under a changing scenario. In this context, the present work aims to investigate the medium- to long-term effect of a low pH scenario on calcification and the biochemical composition of two CWCs from the Mediterranean, Dendrophyllia cornigera and Desmophyllum dianthus. After 314 d of exposure to acidified conditions, a significant decrease of 70 % was observed in Desmophyllum dianthus skeletal growth rate, while Dendrophyllia cornigera showed no differences between treatments. Instead, only subtle differences between treatments were observed in the organic matter amount, lipid content, skeletal microdensity, or porosity in both species, although due to the high variability of the results, these differences were not statistically significant. Our results also confirmed a heterogeneous effect of low pH on the skeletal growth rate of the organisms depending on their initial weight, suggesting that those specimens with high calcification rates may be the most susceptible to the negative effects of acidification.

Description: Deep-water ecosystems are characterized by relatively low carbonate concentration values and, due to ocean acidification (OA), these habitats might be among the first to be exposed to undersaturated conditions in the forthcoming years. However, until now, very few studies have been conducted to test how cold-water coral (CWC) species react to such changes in the seawater chemistry. The present work aims to investigate the mid-term effect of decreased pH on calcification of the two branching CWC species most widely distributed in the Mediterranean, Lophelia pertusa and Madrepora oculata. No significant effects were observed in the skeletal growth rate, microdensity and porosity of both species after 6 months of exposure. However, while the calcification rate of M. oculata was similar for all colony fragments, a heterogeneous skeletal growth pattern was observed in L. pertusa, the younger nubbins showing higher growth rates than the older ones. A higher energy demand is expected in these young, fast-growing fragments and, therefore, a reduction in calcification might be noticed earlier during long-term exposure to acidified conditions.

Description: Knowledge on basic biological functions of organisms is essential to understand not only the role they play in the ecosystems but also to manage and protect their populations. The study of biological processes, such as growth, reproduction and physiology, which can be approached in situ or by collecting specimens and rearing them in aquaria, is particularly challenging for deep-sea organisms like cold-water corals. Field experimental work and monitoring of deep-sea populations is still a chimera. Only a handful of research institutes or companies has been able to install in situ marine observatories in the Mediterranean Sea or elsewhere, which facilitate a continuous monitoring of deep-sea ecosystems. Hence, today’s best way to obtain basic biological information on these organisms is (1) working with collected samples and analysing them post-mortem and / or (2) cultivating corals in aquaria in order to monitor biological processes and investigate coral behaviour and physiological responses under different experimental treatments. The first challenging aspect is the collection process, which implies the use of oceanographic research vessels in most occasions since these organisms inhabit areas between ca. 150 m to more than 1000 m depth, and specific sampling gears. The next challenge is the maintenance of the animals on board (in situations where cruises may take weeks) and their transport to home laboratories. Maintenance in the home laboratories is also extremely challenging since special conditions and set-ups are needed to conduct experimental studies to obtain information on the biological processes of these animals. The complexity of the natural environment from which the corals were collected cannot be exactly replicated within the laboratory setting; a fact which has led some researchers to question the validity of work and conclusions drawn from such undertakings. It is evident that aquaria experiments cannot perfectly reflect the real environmental and trophic conditions where these organisms occur, but: (1) in most cases we do not have the possibility to obtain equivalent in situ information and (2) even with limitations, they produce relevant information about the biological limits of the species, which is especially valuable when considering potential future climate change scenarios. This chapter includes many contributions from different authors and is envisioned as both to be a practical “handbook” for conducting cold-water coral aquaria work, whilst at the same time offering an overview on the cold-water coral research conducted in Mediterranean laboratories equipped with aquaria infrastructure. Experiences from Atlantic and Pacific laboratories with extensive experience with cold-water coral work have also contributed to this chapter, as their procedures are valuable to any researcher interested in conducting experimental work with cold-water corals in aquaria. It was impossible to include contributions from all laboratories in the world currently working experimentally with cold-water corals in the laboratory, but at the conclusion of the chapter we attempt, to our best of our knowledge, to supply a list of several laboratories with operational cold-water coral aquaria facilities.

Description: We evaluated the effects of low pH on Corallium rubrum from aquaria experiments. Several colonies of C. rubrum were long-term maintained for 314 days in aquaria at two different pH levels (8.10 and 7.81, pHT). Calcification rate, spicule morphology, major biochemical constituents (protein, carbohydrates and lipids) and fatty acids composition were measured periodically. Exposure to lower pH conditions caused a significant decrease in the skeletal growth rate in comparison to the control treatment. Similarly, the spicule morphology clearly differed between both treatments at the end of the experiment, with aberrant shapes being observed only under the acidified conditions. On the other hand, while total organic matter was significantly higher under low pH conditions, no significant differences were detected between treatments regarding total carbohydrate, lipid, protein and fatty acid composition. However, the lower variability found among samples maintained in acidified conditions relative to controls, suggests a possible effect of pH decrease on the metabolism of the colonies. Our results show, for the first time, evidence of detrimental ocean acidification effects on this valuable and endangered coral species.

Description: In recent years, some of the ramifications of the ocean acidification problematic derived from the anthropogenic rising of atmospheric CO2 have been widely studied. In particular, the potential effects of a lowering pH on tropical coral reefs have received special attention. However, only a few studies have focused on testing the effects of ocean acidification in corals from the Mediterranean Sea, despite the fact that this basin is especially sensitive to increasing atmospheric CO2. In this context, we investigated the response to ocean acidification of the two zooxanthellate coral species capable of constituting the main framework of the community, the endemic Cladocora caespitosa and the non-native Oculina patagonica. To this end, we examined the response of both species to pCO2 concentrations expected by the end of the century, 800 ppm, vs the present levels. Calcification rate measurements after 92 days of exposure to low pH conditions showed the same negative response in both species, a decrease of 32-35% compared to corals reared under control conditions. In addition, we detected in both species a correlation between the calcification rate of colonies in control conditions and the degree of impairment of the same colonies at low pH. Independent of species, faster growing colonies were more affected by decreased pH. After this period of decreased pH, we conducted a recovery experiment, in which corals reared in the acidic treatment were brought back to control conditions. In this case, normal calcification rates were reached in both species. Overall, our results suggest that O. patagonica and C. caespitosa will both be affected detrimentally by progressive ocean acidification in the near future. They do not display differences in response between native and non-native species but do manifest differential responses depending on calcification rate, pointing to a role of the coral genetics in determining the response of corals to ocean acidification.

Description: Ocean acidification (OA) could become a serious threat for the Antarctic marine ecosystem over coming years, as the solubility of atmospheric CO2 and CaCO3 minerals increases at lower temperatures. We evaluated the effect of OA on the stress response of the limpet Nacella concinna by measuring gene expression levels. The experiment was performed with the two ecotypes (Littoral and Sublittoral) of the species during 54 days (IPCC, 2019 scenario RCP8.5; control, 375 ppm; low-pH treatment, 923 ppm). Exposure to low-pH treatment during 15 days triggered the down-regulation of two heat-shock protein genes (HSP70A, HSP70B) only in sublittoral individuals. Little variation in the relative expression values of all genes in both ecotypes was observed probably, due to a historical exposure to the substantial daily natural pH fluctuations recorded in the study area during the experiment. This study provides relevant baseline data for future OA experiments on coastal species in Antarctica.

Description: Ocean acidification (OA) and warming related to the anthropogenic increase in atmospheric CO2 have been shown to have detrimental effects on several marine organisms, especially those with calcium carbonate structures such as corals. In this study, we evaluate the response of two Mediterranean shallow-water azooxanthellate corals to the projected pH and seawater temperature (ST) scenarios for the end of this century. The colonial coral Astroides calycularis and the solitary Leptopsammia pruvoti were grown in aquaria over a year under two fixed pH conditions, control (8.05 pHT units) and low (7.72 pHT units), and simulating two annual ST cycles, natural and high (+3 °C). The organic matter (OM), lipid and protein content of the tissue and the skeletal microdensity of A. calycularis were not affected by the stress conditions (low pH, high ST), but the species exhibited a mean 25 % decrease in calcification rate at high-ST conditions at the end of the warm period and a mean 10 % increase in skeletal porosity under the acidified treatment after a full year cycle. Conversely, an absence of effects on calcification and skeletal microdensity of L. pruvoti exposed to low-pH and high-ST treatments contrasted with a significant decrease in the OM, lipid and protein content of the tissue at high-ST conditions and a 13 % mean increase in the skeletal porosity under low-pH conditions following a full year of exposure. This species-specific response suggests that different internal self-regulation strategies for energy reallocation may allow certain shallow-water azooxanthellate corals to cope more successfully than others with global environmental changes.

Description: Deep-water ecosystems are characterized by relatively low carbonate concentration values and, due to ocean acidification (OA), these habitats might be among the first to be exposed to undersaturated conditions in the forthcoming years. However, until now, very few studies have been conducted to test how cold-water coral (CWC) species react to such changes in the seawater chemistry. The present work aims to investigate the mid-term effect of decreased pH on calcification of the two branching CWC species most widely distributed in the Mediterranean, Lophelia pertusa and Madrepora oculata. No significant effects were observed in the skeletal growth rate, microdensity and porosity of both species after 6 months of exposure. However, while the calcification rate of M. oculata was similar for all colony fragments, a heterogeneous skeletal growth pattern was observed in L. pertusa, the younger nubbins showing higher growth rates than the older ones. A higher energy demand is expected in these young, fast-growing fragments and, therefore, a reduction in calcification might be noticed earlier during long-term exposure to acidified conditions.

Description: We investigated the effects of an increase in dissolved CO2 on the microbial communities of the Mediterranean Sea during two mesocosm experiments in two contrasting seasons: winter, at the peak of the annual phytoplankton bloom, and summer, under low nutrient conditions. The experiments included treatments with acidification and nutrient addition, and combinations of the two. We followed the effects of ocean acidification (OA) on the abundance of the main groups of microorganisms (diatoms, dinoflagellates, nanoeukaryotes, picoeukaryotes, cyanobacteria, and heterotrophic bacteria) and on bacterial activity, leucine incorporation, and extracellular enzyme activity. Our results showed a clear stimulation effect of OA on the abundance of small phytoplankton (pico- and nanoeukaryotes), independently of the season and nutrient availability. A large number of the measured variables showed significant positive effects of acidification in summer compared with winter, when the effects were sometimes negative. Effects of OA were more conspicuous when nutrient concentrations were low. Our results therefore suggest that microbial communities in oligotrophic waters are considerably affected by OA, whereas microbes in more productive waters are less affected. The overall enhancing effect of acidification on eukaryotic pico- and nanophytoplankton, in comparison with the non-significant or even negative response to nutrient-rich conditions of larger groups and autotrophic prokaryotes, suggests a shift towards medium-sized producers in a future acidified ocean.