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Concentrations of dissolved nitrogen in seawater can affect the resilience of the cnidarian-dinoflagellate symbiosis to climate change-induced bleaching. However, it is not yet known how the assimilation and translocation of the various nitrogen forms change during heat stress, nor how the symbiosis responds to nutrient depletion, which may occur due to increasing water stratification. Here, the tropical scleractinian coral Stylophora pistillata, in symbiosis with dinoflagellates of the genus Symbiodinium, was grown at different temperatures (26°C, 30°C and 34°C), before being placed in nutrient-replete or -depleted seawater for 24 h. The corals were then incubated with 13C-labelled sodium bicarbonate and different 15N-labelled nitrogen forms (ammonium, urea and dissolved free amino acids) to determine their assimilation rates. We found that nutrient depletion inhibited the assimilation of all nitrogen sources studied and that heat stress reduced the assimilation of ammonium and dissolved free amino acids. However, the host assimilated over 3-fold more urea at 30°C relative to 26°C. Overall, both moderate heat stress (30°C) and nutrient depletion individually decreased the total nitrogen assimilated by the symbiont by 66%, and combined, they decreased assimilation by 79%. This led to the symbiotic algae becoming nitrogen starved, with the C:N ratio increasing by over 3-fold at 34°C, potentially exacerbating the impacts of coral bleaching., Competing Interests: Competing interests The authors declare no competing or financial interests., (© 2024. Published by The Company of Biologists Ltd.)

Dissolved inorganic phosphorus (DIP) is a limiting nutrient in the physiology of scleractinian corals. Anthropogenic addition of dissolved inorganic nitrogen (DIN) to coastal reefs increases the seawater DIN:DIP ratio and further increases P limitation, which is detrimental to coral health. The effects of imbalanced DIN:DIP ratios on coral physiology require further investigation in coral species other than the most studied branching corals. Here we investigated the nutrient uptake rates, elemental tissue composition and physiology of a foliose stony coral, Turbinaria reniformis , and a soft coral, Sarcophyton glaucum , exposed to four different DIN: DIP ratios (0.5:0.2, 0.5:1, 3:0.2, 3:1). The results show that T. reniformis had high uptake rates of DIN and DIP, proportional to the seawater nutrient concentrations. DIN enrichment alone led to an increase in tissue N content, shifting the tissue N:P ratio towards P limitation. However, S. glaucum had 5 times lower uptake rates and only took up DIN when the seawater was simultaneously enriched with DIP. This double uptake of N and P did not alter tissue stoichiometry. This study allows us to better understand the susceptibility of corals to changes in the DIN:DIP ratio and predict how coral species will respond under eutrophic conditions in the reef.

Phosphorus (P) is an essential but limiting nutrient for coral growth due to low concentrations of dissolved inorganic concentrations (DIP) in reef waters. P limitation is often exacerbated when concentrations of dissolved inorganic nitrogen (DIN) increase in the reef. To increase their access to phosphorus, corals can use organic P dissolved in seawater (DOP). They possess phosphatase enzymes that transform DOP into DIP, which can then be taken up by coral symbionts. Although the concentration of DOP in reef waters is much higher than DIP, the dependence of corals on this P source is still poorly understood, especially with different concentrations of DIN in seawater. As efforts to predict the future of corals increase, improved knowledge of the P requirements of corals living under different DIN concentrations may be key to predicting coral health. In this study, we investigated P content and phosphatase activities (PAs) in Stylophora pistillata maintained under nutrient starvation, long-term nitrogen enrichment (nitrate or ammonium at 2 μM) and short-term (few hours) nitrogen pulses. Results show that under nutrient depletion and ammonium-enriched conditions, a significant increase in PAs was observed compared to control conditions, with no change in the N:P ratio of the coral tissue. On the contrary, under nitrate enrichment, there was no increase in PAs compared to control conditions, but an increase in the N:P ratio of the coral tissue. These results suggest that under nitrate enrichment, corals were unable to increase their ability to rely on DOP and replenish their cellular P content. An increase in cellular N:P ratio is detrimental to coral health as it increases the susceptibility of coral bleaching under thermal stress. These results provide an overall view of the P requirements of corals exposed to different nutrient conditions and improve our understanding of the effects of nitrogen enrichment on corals., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Coral reefs depend on the highly optimized mutualistic relationship between corals and Symbiodiniaceae dinoflagellates. Both partners exchange nutrients obtained through heterotrophy of the host and autotrophy of the symbionts. While heterotrophy helps corals withstand the harmful effects of seawater warming, the exchange of heterotrophic nutrients between the two partners is poorly understood. Here, we used compound-specific δ 15 N and δ 13 C of amino acids ( δ 15 N AA and δ 13 C AA ) and a 15 N pulse-chase experiment with Artemia salina nauplii in two coral-dinoflagellate associations to trace the assimilation and allocation of heterotrophic nutrients within the partners. We observed that changes in the trophic position (TP Glx-Phe ), δ 15 N AA , and δ 13 C AA with heterotrophy were holobiont-dependent. Furthermore, while TP Glx-Phe and δ 15 N of all AAs significantly increased with heterotrophy in the symbionts and host of Stylophora pistillata, only the δ 15 N AA of the symbionts changed in Turbinaria reniformis. Together with the pulse-chase experiment, the results suggested a direct transfer of heterotrophically acquired AAs to the symbionts of S. pistillata and a transfer of ammonium to the symbionts of T. reniformis. Overall, we demonstrated that heterotrophy underpinned the nutrition of Symbiodinaceae and possibly influenced their stress tolerance under changing environmental conditions. IMPORTANCE Coral reefs rely upon the highly optimized nutritional symbiosis between corals and Symbiodiniaceae dinoflagellates. Heterotrophic feeding on plankton is key to the resistance of corals to environmental stress. Yet, a detailed understanding of heterotrophic nutrient assimilation and utilization within the symbiosis is lacking. Here, we used the advanced tools of compound-specific isotope analysis of amino acids and 15 N-labeling of plankton to show that heterotrophy underpinned the nutrition of Symbiodinaceae. Symbionts received either heterotrophically acquired amino acids or recycled ammonium due to their association with the coral host. This study brought new insight into the nutrient exchanges in coral-Symbiodiniaceae associations and allowed a better understanding of the mechanisms involved in coral resistance to environmental stress.

Climate change-related increase in seawater temperature has become a leading cause of coral bleaching and mortality. However, corals from the northern Red Sea show high thermal tolerance and no recorded massive bleaching event. This specific region is frequently subjected to intense dust storms, coming from the surrounding arid deserts, which are expected to increase in frequency and intensity in the future. The aerial dust deposition supplies essential bioelements to the water column. Here, we investigated the effect of dust deposition on the physiology of a Red Sea coral, Stylophora pistillata. We measured the modifications in coral and Symbiodiniaceae metallome (cellular metal content), as well as the changes in photosynthesis and oxidative stress status of colonies exposed during few weeks to dust deposition. Our results show that 1 mg L -1 of dust supplied nanomolar amounts of nitrate and other essential bioelements, such as iron, manganese, zinc and copper, rapidly assimilated by the symbionts. At 25°C, metal bioaccumulation enhanced the chlorophyll concentration and photosynthesis of dust-exposed corals compared to control corals. These results suggest that primary production was limited by metal availability in seawater. A 5°C increase in seawater temperature enhanced iron assimilation in both control and dust-enriched corals. Temperature rise increased the photosynthesis of control corals only, dust-exposed ones having already reached maximal photosynthesis rates at 25°C. Finally, we observed a combined effect of temperature and bioelement concentration on the assimilation of molybdenum, cadmium, manganese and copper, which were in higher concentrations in symbionts of dust-exposed corals maintained at 30°C. All together these observations highlight the importance of dust deposition in the supply of essential bioelements, such as iron, to corals and its role in sustaining coral productivity in Red Sea reefs., (© 2022 John Wiley & Sons Ltd.)

Reef ecosystems are under increasing pressure from global and local stressors. Rising seawater temperature and high ultraviolet radiation (UVR) levels are the main drivers of the disruption of the coral-dinoflagellate symbiosis (bleaching). Bleaching can also be exacerbated by nitrate contamination in coastal reefs. However, the underlying physiological mechanisms are still poorly understood. Here, we assessed the physiological and oxidative state of the scleractinian coral Pocillopora damicornis, maintained eight weeks in a crossed-factorial design including two temperatures (26 °C or 30 °C), and two nitrate (0.5 and 3 μM-enriched), and UVR (no UVR and 25/1.5 Wm -2 UVA/B) levels. Nitrate enrichment, and high temperature, significantly impaired coral photosynthesis. However, UVR alleviated the nitrate and temperature-induced decrease in photosynthesis, by increasing the coral's antioxidant capacity. The present study contributes to our understanding of the combined effects of abiotic stressors on coral bleaching susceptibility. Such information is urgently needed to refine reef management strategies., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Artificial Light at Night (ALAN), which is the alteration of natural light levels as the result of anthropogenic light sources, has been acknowledged as an important factor that alters the functioning of marine ecosystems. Using LEDs light to mimic ALAN, we studied the effect on the physiology (symbiont and chlorophyll contents, photosynthesis, respiration, pigment profile, skeletal growth, and oxidative stress responses) of two scleractinian coral species originating from the Red Sea. ALAN induced the photoinhibition of symbiont photosynthesis, as well as an overproduction of reactive oxygen species (ROS) and an increase in oxidative damage to lipids in both coral species. The extent of the deleterious effects of ALAN on the symbiotic association and coral physiology was aligned with the severity of the oxidative stress condition experienced by the corals. The coral species Sylophora pistillata, which experienced a more severe oxidative stress condition than the other species tested, Turbinaria reniformis, also showed a more pronounced bleaching (loss of symbionts and chlorophyll content), enhanced photoinhibition and decreased photosynthetic rates. Findings of the present study further our knowledge on the biochemical mechanisms underpinning the deleterious impacts of ALAN on scleractinian corals, ultimately shedding light on the emerging threat of ALAN on coral reef ecology. Further, considering that global warming and light pollution will increase in the next few decades, future studies should be taken to elucidate the potential synergetic effects of ALAN and global climate change stressors., Competing Interests: Declaration of competing interest The authors declare no competing financial interests., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Mass coral bleaching represents one of the greatest threats to coral reefs and has mainly been attributed to seawater warming. However, reduced water quality can also interact with warming to increase coral bleaching, but this interaction depends on nutrient ratios and forms. In particular, nitrate (NO 3 - ) enrichment reduces thermal tolerance while ammonium (NH 4 + ) enrichment tends to benefit coral health. The biochemical mechanisms underpinning the different bleaching responses of corals exposed to DIN enrichment still need to be investigated. Here, we demonstrated that the coral Stylophora pistillata underwent a severe oxidative stress condition and reduced aerobic scope when exposed to NO 3 - enrichment combined with thermal stress. Such condition resulted in increased bleaching intensity compared to a low-nitrogen condition. On the contrary, NH 4 + enrichment was able to amend the deleterious effects of thermal stress by favoring the oxidative status and energy metabolism of the coral holobiont. Overall, our results demonstrate that the opposite effects of nitrate and ammonium enrichment on coral bleaching are related to the effects on corals' energy/redox status. As nitrate loading in coastal waters is predicted to significantly increase in the future due to agriculture and land-based pollution, there is the need for urgent management actions to prevent increases in nitrate levels in seawater. In addition, the maintenance of important fish stocks, which provide corals with recycled nitrogen such as ammonium, should be favoured.

Labile dissolved organic carbon (DOC) is a major pollutant in coastal marine environments affected by anthropogenic impacts, and may significantly contribute to coral bleaching and subsequent mortality on coastal reefs. DOC can cause bleaching indirectly through the rapid proliferation of copiotrophic and pathogenic bacteria. Here we demonstrate that labile DOC compounds can also impair the coral-dinoflagellate symbiosis by directly affecting coral physiology on both the host and algal symbiont level. In a controlled aquarium experiment, we monitored over several weeks key physiological parameters of the tropical coral Stylophora pistillata exposed to ambient and elevated labile DOC levels (0.1 and 1.0 mM) in combination with low and high nitrogen (i.e. ammonium) conditions (0.2 and 4.0 μM). At the symbiont level, DOC exposure under low ammonium availability decreased the photosynthetic efficiency accompanied by ∼75 % Chl a and ∼50 % symbiont cell reduction. The photosynthetic functioning of the symbionts recovered once the DOC enrichment ceased indicating a reversible shift between autotrophic and heterotrophic metabolism. At the host level, the assimilation of exogenous DOC sustained the tissue carbon reserves, but induced a depletion of the nitrogen reserves, indicated by ∼35 % decreased protein levels. This suggests an imbalanced exogenous carbon to nitrogen supply with nitrogen potentially limiting host metabolism on the long-term. We also demonstrate that increased ammonium availability delayed DOC-induced bleaching likely by keeping symbionts in a photosynthetically competent state, which is crucial for symbiosis maintenance and coral survival. Overall, the present study provides further insights into how coastal pollution can de-stabilize the coral-algal symbiosis and cause coral bleaching. Therefore, reducing coastal pollution and sustaining ecological integrity are critical to strengthen the resilience of coral reefs facing climate change., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Soft corals often constitute one of the major benthic groups of coral reefs. Although they have been documented to outcompete reef-building corals following environmental disturbances, their physiological performance and thus their functional importance in reefs are still poorly understood. In particular, the acclimatization to depth of soft corals harboring dinoflagellate symbionts and the metabolic interactions between these two partners have received little attention. We performed stable isotope tracer experiments on two soft coral species (Litophyton sp. and Rhytisma fulvum fulvum) from shallow and upper mesophotic Red Sea coral reefs to quantify the acquisition and allocation of autotrophic carbon within the symbiotic association. Carbon acquisition and respiration measurements distinguish Litophyton sp. as mainly autotrophic and Rhytisma fulvum fulvum as rather heterotrophic species. In both species, carbon acquisition was constant at the two investigated depths. This is a major difference from scleractinian corals, whose carbon acquisition decreases with depth. In addition, carbon acquisition and photosynthate translocation to the host decreased with an increase in symbiont density, suggesting that nutrient provision to octocoral symbionts can quickly become a limiting factor of their productivity. These findings improve our understanding of the biology of soft corals at the organism-scale and further highlight the need to investigate how their nutrition will be affected under changing environmental conditions.

Dinitrogen (N 2 ) fixing bacteria (diazotrophs) are an important source of new nitrogen in oligotrophic environments and represent stable members of the microbiome in tropical corals, while information on corals from temperate oligotrophic regions is lacking. Therefore, this study provides new insights into the diversity and activity of diazotrophs associated with the temperate coral Oculina patagonica from the Mediterranean Sea by combining metabarcoding sequencing of amplicons of both the 16S rRNA and nifH genes and 15 N 2 stable isotope tracer analysis to assess diazotroph-derived nitrogen (DDN) assimilation by the coral. Results show that the diazotrophic community of O. patagonica is dominated by autotrophic bacteria (i.e. Cyanobacteria and Chlorobia). The majority of DDN was assimilated into the tissue and skeletal matrix, and DDN assimilation significantly increased in bleached corals. Thus, diazotrophs may constitute an additional nitrogen source for the coral host, when nutrient exchange with Symbiodinium is disrupted (e.g. bleaching) and external food supply is limited (e.g. oligotrophic summer season). Furthermore, we hypothesize that DDN can facilitate the fast proliferation of endolithic algae, which provide an alternative carbon source for bleached O. patagonica. Overall, O. patagonica could serve as a good model for investigating the importance of diazotrophs in coral recovery from bleaching., (© 2018 Society for Applied Microbiology and John Wiley & Sons Ltd.)

Coral reefs are of major ecological and socio-economic interest. They are threatened by global warming and natural pressures such as solar ultraviolet radiation. While great efforts have been made to understand the physiological response of corals to these stresses, the signalling pathways involved in the immediate cellular response exhibited by corals remain largely unknown. Here, we demonstrate that c-Jun N-terminal kinase (JNK) activation is involved in the early response of corals to thermal and UV stress. Furthermore, we found that JNK activity is required to repress stress-induced reactive oxygen species (ROS) accumulation in both the coral Stylophora pistillata and human skin cells. We also show that inhibiting JNK activation under stress conditions leads to ROS accumulation, subsequent coral bleaching and cell death. Taken together, our results suggest that an ancestral response, involving the JNK pathway, is remarkably conserved from corals to human, protecting cells from the adverse environmental effects.

Tropical corals are associated with a diverse community of dinitrogen (N 2 )-fixing prokaryotes (diazotrophs) providing the coral an additional source of bioavailable nitrogen (N) in oligotrophic waters. The overall activity of these diazotrophs changes depending on the current environmental conditions, but to what extent it affects the assimilation of diazotroph-derived N (DDN) by corals is still unknown. Here, in a series of 15 N 2 tracer experiments, we directly quantified DDN assimilation by scleractinian corals from the Red Sea exposed to different environmental conditions. We show that DDN assimilation strongly varied with the corals' metabolic status or with phosphate availability in the water. The very autotrophic shallow-water (~5 m) corals showed low or no DDN assimilation, which significantly increased under elevated phosphate availability (3 µM). Corals that depended more on heterotrophy (i.e., bleached and deep-water [~45 m] corals) assimilated significantly more DDN, which contributed up to 15% of the corals' N demand (compared to 1% in shallow corals). Furthermore, we demonstrate that a substantial part of the DDN assimilated by deep corals was likely obtained from heterotrophic feeding on fixed N compounds and/or diazotrophic cells in the mucus. Conversely, in shallow corals, the net release of mucus, rich in organic carbon compounds, likely enhanced diazotroph abundance and activity and thereby the release of fixed N to the pelagic and benthic reef community. Overall, our results suggest that DDN assimilation by corals varies according to the environmental conditions and is likely linked to the capacity of the coral to acquire nutrients from seawater., Importance: Tropical corals are associated with specialized bacteria (i.e., diazotrophs) able to transform dinitrogen (N 2 ) gas into a bioavailable form of nitrogen, but how much of this diazotroph-derived nitrogen (DDN) is assimilated by corals under different environmental conditions is still unknown. Here, we used 15 N 2 labeling to trace the fate of DDN within the coral symbiosis. We show that DDN is assimilated by both the animal host and the endosymbiotic algae. In addition, the amount of assimilated DDN was significantly greater in mesophotic, bleached, or phosphorus-enriched corals than in surface corals, which almost did not take up this nitrogen form. DDN can thus be of particular importance for the nutrient budget of corals whenever they are limited by the availability of other forms of dissolved nutrients., (Copyright © 2017 Bednarz et al.)

During the 20(th) century, seawater temperatures have significantly increased, leading to profound alterations in biogeochemical cycles and ecosystem processes. Elevated temperatures have also caused massive bleaching (symbiont/pigment loss) of autotrophic symbioses, such as in coral-dinoflagellate association. As symbionts provide most nutrients to the host, their expulsion during bleaching induces host starvation. However, with the exception of carbon, the nutritional impact of bleaching on corals is still unknown, due to the poorly understood requirements in inorganic nutrients during stress. We therefore assessed the uptake rates of nitrogen and phosphate by five coral species maintained under normal and thermal stress conditions. Our results showed that nitrogen acquisition rates were significantly reduced during thermal stress, while phosphorus uptake rates were significantly increased in most species, suggesting a key role of this nutrient. Additional experiments showed that during thermal stress, phosphorus was required to maintain symbiont density and photosynthetic rates, as well as to enhance the translocation and retention of carbon within the host tissue. These findings shed new light on the interactions existing between corals and inorganic nutrients during thermal stress, and highlight the importance of phosphorus for symbiont health.

(31)P Nuclear Magnetic Resonance (NMR) was assessed to investigate the phosphorus-containing compounds present in the tissues of the scleractinian coral Stylophora pistillata as well as of cultured zooxanthellae (CZ). Results showed that phosphorus-containing compounds observed in CZ were mainly phosphate and phosphate esters. Phosphate accounted for 19 ± 2% of the total phosphorus compounds observed in CZ maintained under low P-levels (0.02 μM). Adding 5 mM of dissolved inorganic phosphorus (KH2PO4) to the CZ culture medium led to a 3.1-fold increase in intracellular phosphate, while adding 5 mM of dissolved organic phosphorus led to a reduction in the concentration of phosphorus compounds, including a 2.5-fold intracellular phosphate decrease. In sharp contrast to zooxanthellae, the host mainly contained phosphonates, and to a lesser extent, phosphate esters and phosphate. Two-months of host starvation decreased the phosphate content by 2.4 fold, while bleaching of fed corals did not modify this content. Based on (31)P NMR analyses, this study highlights the importance of phosphonates in the composition of coral host tissues, and illustrates the impact of phosphorus availability on the phosphorus composition of host tissues and CZ, both through feeding of the host and inorganic phosphorus enrichment of the CZ.

Anthropogenic nutrient enrichment affects the biogeochemical cycles and nutrient stoichiometry of coastal ecosystems and is often associated with coral reef decline. However, the mechanisms by which dissolved inorganic nutrients, and especially nitrogen forms (ammonium versus nitrate) can disturb the association between corals and their symbiotic algae are subject to controversial debate. Here, we investigated the coral response to varying N : P ratios, with nitrate or ammonium as a nitrogen source. We showed significant differences in the carbon acquisition by the symbionts and its allocation within the symbiosis according to nutrient abundance, type and stoichiometry. In particular, under low phosphate concentration (0.05 µM), a 3 µM nitrate enrichment induced a significant decrease in carbon fixation rate and low values of carbon translocation, compared with control conditions (N : P = 0.5 : 0.05), while these processes were significantly enhanced when nitrate was replaced by ammonium. A combined enrichment in ammonium and phosphorus (N : P = 3 : 1) induced a shift in nutrient allocation to the symbionts, at the detriment of the host. Altogether, these results shed light into the effect of nutrient enrichment on reef corals. More broadly, they improve our understanding of the consequences of nutrient loading on reef ecosystems, which is urgently required to refine risk management strategies., (© 2015 The Author(s).)

Reef-building corals form symbioses with dinoflagellates from the diverse genus Symbiodinium. This symbiotic association has developed adaptations to acquire and share nutrients, which are essential for its survival and growth in nutrient-poor tropical waters. The host is thus able to prey on a wide range of organic food sources (heterotrophic nutrition) whereas the symbionts acquire most of the inorganic nutrients (autotrophic nutrition). However, nutrient fluxes between the two partners remain unclear, especially concerning heterotrophically acquired carbon and nitrogen. We combined physiological measurements and pulse-chase isotopic labeling of heterotrophic carbon and nitrogen, as well as autotrophic carbon to track nutrient fluxes in two coral species, Stylophora pistillata and Turbinaria reniformis, in symbiosis with Symbiodinium clades A, and C,D respectively. We showed a rapid acquisition, exchange and a long-term retention of heterotrophic nutrients within the symbiosis, whereas autotrophic nutrients were rapidly used to meet immediate metabolic needs. In addition, there was a higher retention of heterotrophic nitrogen compared with carbon, in agreement with the idea that tropical corals are nitrogen-limited. Finally, a coupling between auto- and heterotrophy was observed in the species S. pistillata, with a higher acquisition and retention of heterotrophic nutrients under low irradiance to compensate for a 50% reduction in autotrophic nutrient acquisition and translocation. Conversely, T. reniformis conserved an equivalent heterotrophic nutrient acquisition at both light levels because this coral species did not significantly reduce its rates of gross photosynthesis and autotrophic carbon acquisition between the two irradiances. These experiments advance the current understanding of the nutrient exchanges between the two partners of a symbiotic association, providing evidence of the complexity of the host-symbiont relationship., (© 2015. Published by The Company of Biologists Ltd.)

Scleractinian corals are essential constituents of tropical reef ecological diversity. They live in close association with diazotrophs [dinitrogen (N2)-fixing microbes], which can fix high rates of N2. Whether corals benefit from this extrinsic nitrogen source is still under debate. Until now, N2 fixation rates have been indirectly estimated using the acetylene reduction assay, which does not permit assessment of the amount of nitrogen incorporated into the different compartments of the coral holobiont. In the present study, the (15)N2 technique was applied for the first time on three Red Sea coral species. Significant (15)N enrichment was measured in particles released by corals to the surrounding seawater. N2 fixation rates were species specific and as high as 1.6-2 ng N day(-1) l(-1). However, no significant enrichment was measured in the symbiotic dinoflagellates or the coral host tissues, suggesting that corals do not benefit from diazotrophic N2 fixation., (© 2014. Published by The Company of Biologists Ltd.)

Corals live in symbiosis with dinoflagellates of the genus Symbiodinum. These dinoflagellates translocate a large part of the photosynthetically fixed carbon to the host, which in turn uses it for its own needs. Assessing the carbon budget in coral tissue is a central question in reef studies that still vexes ecophysiologists. The amount of carbon fixed by the symbiotic association can be determined by measuring the rate of photosynthesis, but the amount of carbon translocated by the symbionts to the host and the fate of this carbon are more difficult to assess. In the present study, we propose a novel approach to calculate the budget of autotrophic carbon in the tissue of scleractinian corals, based on a new model and measurements made with the stable isotope (13)C. Colonies of the scleractinian coral Stylophora pistillata were incubated in H(13)CO (-)(3)-enriched seawater, after which the fate of (13)C was followed in the symbionts, the coral tissue and the released particulate organic carbon (i.e. mucus). Results obtained showed that after 15 min, ca. 60% of the carbon fixed was already translocated to the host, and after 48 h, this value reached 78%. However, ca. 48% of the photosynthetically fixed carbon was respired by the symbiotic association, and 28% was released as dissolved organic carbon. This is different from other coral species, where <1% of the total organic carbon released is from newly fixed carbon. Only 23% of the initially fixed carbon was retained in the symbionts and coral tissue after 48 h. Results show that our (13)C-based model could successfully trace the carbon flow from the symbionts to the host, and the photosynthetically acquired carbon lost from the symbiotic association.

Temperate symbiotic corals, such as the Mediterranean species Cladocora caespitosa, live in seasonally changing environments, where irradiance can be ten times higher in summer than winter. These corals shift from autotrophy in summer to heterotrophy in winter in response to light limitation of the symbiont's photosynthesis. In this study, we determined the autotrophic carbon budget under different conditions of irradiance (20 and 120 µmol photons m(-2) s(-1)) and feeding (fed three times a week with Artemia salina nauplii, and unfed). Corals were incubated in H(13)CO(3) (-)-enriched seawater, and the fate of (13)C was followed in the symbionts and the host tissue. The total amount of carbon fixed by photosynthesis and translocated was significantly higher at high than low irradiance (ca. 13 versus 2.5-4.5 µg cm(-2) h(-1)), because the rates of photosynthesis and carbon fixation were also higher. However, the percent of carbon translocation was similar under the two irradiances, and reached more than 70% of the total fixed carbon. Host feeding induced a decrease in the percentage of carbon translocated under low irradiance (from 70 to 53%), and also a decrease in the rates of carbon translocation per symbiont cell under both irradiances. The fate of autotrophic and heterotrophic carbon differed according to irradiance. At low irradiance, autotrophic carbon was mostly respired by the host and the symbionts, and heterotrophic feeding led to an increase in host biomass. Under high irradiance, autotrophic carbon was both respired and released as particulate and dissolved organic carbon, and heterotrophic feeding led to an increase in host biomass and symbiont concentration. Overall, the maintenance of high symbiont concentration and high percentage of carbon translocation under low irradiance allow this coral species to optimize its autotrophic carbon acquisition, when irradiance conditions are not favourable to photosynthesis.

Several untested aspects of the regulation of inorganic nutrient uptake were examined using nutrient depletion experiments with the symbiotic coral Stylophora pistillata. The total inhibition of phosphate uptake in artificial seawater lacking sodium indicates the involvement of a sodium/phosphate symporter for the uptake of phosphate across host membranes. Addition of ammonium or nitrate (up to 6.0 μmol l(-1)) did not enhance saturated phosphate uptake rates, thus indicating that corals, or their symbiotic algae, were not, or not sufficiently, nitrogen limited to modify their phosphate needs. Conversely, the saturated uptake rate of ammonium increased by 2.5-fold in the presence of 3.0 μmol l(-1) of phosphate, thus indicating that the corals or their symbionts were lacking intracellular phosphate to take advantage of the inorganic nitrogen compounds dissolved in their surrounding medium. Overall, these results highlight some greater limitation in phosphate rather than in nitrogen. Finally, the rate of phosphate uptake decreased with particulate feeding of the host (organic phosphate source). Indeed, corals that were fed 1 and 3 days before the uptake experiment took up phosphate 42 and 19% slower, respectively, than corals that were fed 21 days before. This result provides additional evidence of phosphate limitation in S. pistillata. This study therefore brings new insights into the relationships between nutrients and symbiotic corals, and may provide a rapid and effective tool to investigate which nutrient is the most limiting for coral metabolism.

The effects of ocean acidification and elevated seawater temperature on coral calcification and photosynthesis have been extensively investigated over the last two decades, whereas they are still unknown on nutrient uptake, despite their importance for coral energetics. We therefore studied the separate and combined impacts of increases in temperature and pCO(2) on phosphate, ammonium, and nitrate uptake rates by the scleractinian coral S. pistillata. Three experiments were performed, during 10 days i) at three pH(T) conditions (8.1, 7.8, and 7.5) and normal temperature (26°C), ii) at three temperature conditions (26°, 29°C, and 33°C) and normal pH(T) (8.1), and iii) at three pH(T) conditions (8.1, 7.8, and 7.5) and elevated temperature (33°C). After 10 days of incubation, corals had not bleached, as protein, chlorophyll, and zooxanthellae contents were the same in all treatments. However, photosynthetic rates significantly decreased at 33°C, and were further reduced for the pH(T) 7.5. The photosynthetic efficiency of PSII was only decreased by elevated temperature. Nutrient uptake rates were not affected by a change in pH alone. Conversely, elevated temperature (33°C) alone induced an increase in phosphate uptake but a severe decrease in nitrate and ammonium uptake rates, even leading to a release of nitrogen into seawater. Combination of high temperature (33°C) and low pH(T) (7.5) resulted in a significant decrease in phosphate and nitrate uptake rates compared to control corals (26°C, pH(T) = 8.1). These results indicate that both inorganic nitrogen and phosphorus metabolism may be negatively affected by the cumulative effects of ocean warming and acidification.

This study was designed to assess the importance of dissolved free amino acids (DFAA) as a nitrogen source for the scleractinian coral Stylophora pistillata. For this purpose, experiments were performed using (15)N-enriched DFAAs, and %(15)N enrichment was measured both in animal tissue and zooxanthellae at different DFAA concentrations, incubation time and light levels. As previously observed for urea, which is another source of organic nitrogen, DFAA uptake exhibited a biphasic mode consisting of an active carrier-mediated transport for concentrations below 3 micromol l(-1) and a linear uptake for higher concentrations. The value of the carrier affinity (K(m)=1.23 micromol l(-1) DFAA) indicated good adaptation of the corals to the low levels of DFAA concentrations measured in most oligotrophic waters. DFAA uptake was also correlated with light. The DFAA contribution to the nitrogen requirements for tissue growth was compared to the contribution of ammonia, nitrate and urea, for which uptake was also measured in S. pistillata. Inorganic sources (NH(4)(+) and NO(3)(-)) contributed 75% of the daily nitrogen needs against 24% for organic sources. Taken altogether, dissolved organic and inorganic nitrogen can supply almost 100% of the nitrogen needs for tissue growth.