Asymmetric physiological response of a reef-building coral to pulsed versus continuous addition of inorganic nutrients.

Coral reefs, especially those located near-shore, are increasingly exposed to anthropogenic, eutrophic conditions that are often chronic. Yet, corals under unperturbed conditions may frequently receive natural and usually temporary nutrient supplementation through biological sources such as fishes. We compared physiological parameters indicative of long- and short-term coral health (day and night calcification, fragment surface area, productivity, energy reserves, and tissue stoichiometry) under continuous and temporary nutrient enrichment. The symbiotic coral Acropora intermedia was grown for 7 weeks under continuously elevated (press) levels of ammonium (14 µmol L-1) and phosphate (10 µmol L-1) as separate and combined treatments, to discern the individual and interactive nutrient effects. Another treatment exposed A. intermedia twice-daily to an ammonium and phosphate pulse of the same concentrations as the press treatments to simulate natural biotic supplementation. Press exposure to elevated ammonium or phosphate produced mixed effects on physiological responses, with little interaction between the nutrients in the combined treatment. Overall, corals under press exposure transitioned resources away from calcification. However, exposure to nutrient pulses often enhanced physiological responses. Our findings indicate that while continuous nutrient enrichment may pose a threat to coral health, episodic nutrient pulses that resemble natural nutrient supplementation may significantly benefit coral health and physiology.

Species sensitivity assessment of five Atlantic scleractinian coral species to 1-methylnaphthalene.

Coral reefs are keystone coastal ecosystems that are at risk of exposure to petroleum from a range of sources, and are one of the highest valued natural resources for protection in Net Environmental Benefit Analysis (NEBA) in oil spill response. Previous research evaluating dissolved hydrocarbon impacts to corals reflected no clear characterization of sensitivity, representing an important knowledge gap in oil spill preparedness related to the potential impact of oil spills to the coral animal and its photosymbiont zooxanthellae. This research addresses this gap, using a standardized toxicity protocol to evaluate effects of a dissolved reference hydrocarbon on scleractinian corals. The relative sensitivity of five Atlantic scleractinian coral species to hydrocarbon exposure was assessed with 48-h assays using the reference polycyclic aromatic hydrocarbon 1-methylnaphthalene, based on physical coral condition, mortality, and photosynthetic efficiency. The threatened staghorn coral Acropora cervicornis was found to be the most sensitive to 1-methylnaphthalene exposure. Overall, the acute and subacute endpoints indicated that the tested coral species were comparatively more resilient to hydrocarbon exposure than other marine species. These results provide a framework for the prediction of oil spill impacts and impact thresholds on the coral animal and related habitats, essential for informing oil spill response in coastal tropical environments.

Effect of Louisiana sweet crude oil on a Pacific coral, Pocillopora damicornis.

Recent oil spill responses such as the Deepwater Horizon event have underscored the need for crude oil ecotoxicological threshold data for shallow water corals to assist in natural resource damage assessments. We determined the toxicity of a mechanically agitated oil-seawater mixture (high-energy water-accommodated fraction, HEWAF) of a sweet crude oil on a branched stony coral, Pocillopora damicornis. We report the results of two experiments: a 96 h static renewal exposure experiment and a "pulse-chase" experiment of three short-term exposure durations followed by a recovery period in artificial seawater. Five endpoints were used to determine ecotoxicological values: 1) algal symbiont chlorophyll fluorescence, 2) a tissue regeneration assay and a visual health metric with three endpoints: 3) tissue integrity, 4) tissue color, and 5) polyp behavior. The sum of 50 entrained polycyclic aromatic hydrocarbons (tPAH50) was used as a proxy for oil exposure. For the 96 h exposure dose response experiment, dark-adapted maximum quantum yield (Fv/Fm) of the dinoflagellate symbionts was least affected by crude oil (EC50 = 913 µg/L tPAH50); light-adapted effective quantum yield (EQY) was more sensitive (EC50 =  428 µg/L tPAH50). In the health assessment, polyp behavior (EC50 = 27 µg/L tPAH50) was more sensitive than tissue integrity (EC50 = 806 µg/L tPAH50) or tissue color (EC50 = 926 µg/L tPAH50). Tissue regeneration proved to be a particularly sensitive measurement for toxicity effects (EC50 = 10 µg/L tPAH50). Short duration (6-24 h) exposures using 503 µg/L tPAH50 (average concentration) resulted in negative impacts to P. damicornis and its symbionts. Recovery of chlorophyll a fluorescence levels for 6-24 h oil exposures was observed in a few hours (Fv/Fm) to several days (EQY) following recovery in fresh seawater. The coral health assessments for tissue integrity and tissue color were not affected following short-term oil exposure durations, but the 96 h treatment duration resulted in significant decreases for both. A reduction in polyp behavior (extension) was observed for all treatment durations, with recovery observed for the short-term (6-24 h) exposures within 1-2 days following placement in fresh seawater. Wounded and intact fragments exposed to oil treatments were particularly sensitive, with significant delays observed in tissue regeneration. Estimating ecotoxicological values for P. damicornis exposed to crude oil HEWAFs provides a basis for natural resource damage assessments for oil spills in reef ecosystems. These data, when combined with ecotoxicological values for other coral reef species, will contribute to the development of species sensitivity models.

Phototoxic effects of two common marine fuels on the settlement success of the coral Acropora tenuis.

Coral reefs are at risk of exposure to petroleum hydrocarbons from shipping spills and uncontrolled discharges during extraction. The toxicity of petroleum hydrocarbons can substantially increase in the presence of ultraviolet radiation (UVR), therefore spills in shallow coral reef environments may be particularly hazardous to reef species. Here we investigated the sensitivity of coral larvae (Acropora tenuis) to dissolved hydrocarbons from heavy fuel oil (HFO) and diesel in the absence and presence of UVR. Larval settlement success decreased with increasing concentrations of dissolved HFO, and co-exposure to UVR doubled the toxicity: 50% effect concentrations (EC50) decreased from 96 (-UVR) to 51 (+UVR) total petroleum aromatic hydrocarbons (TPAH). Toxic thresholds for HFO were similar to concentrations reported during marine spills: EC10s of 24 (-UVR) and 15 (+UVR) µg l-1. While less toxic, diesel also reduced settlement and exhibited phototoxicity: EC10s of 122 (+UVR) and 302 (-UVR) µg l-1. This study demonstrates that the presence of UVR increases the hazard posed by oil pollution to tropical, shallow-water coral reefs. Further research on the effects of oils in the presence of UVR is needed to improve the environmental relevance of risk assessments and ensure appropriate protection for shallow reef environments against oil pollution.

Comparative toxicity of five dispersants to coral larvae.

Oil spill responders require information on the absolute and relative toxicities of chemical dispersants to relevant receptor species to assess their use in spill response. However, little toxicity data are available for tropical marine species including reef-building corals. In this study, we experimentally assessed the sub-lethal toxicity of five dispersants to larvae of the coral Acropora millepora over three short exposure periods (2, 6 and 24 h) reflecting real-world spill response scenario durations. Inhibition of larval settlement increased rapidly between 2 and 6 h, and was highest at 24 h: EC50 Corexit EC9500A = 4.0 mg l-1; Ardrox 6120 = 4.0 mg l-1; Slickgone LTSW = 2.6 mg L-1; Slickgone NS = 11.1 mg L-1 and Finasol OSR52 = 3.4 mg L-1. Coral larvae were more sensitive to dispersants than most other coral life stages and marine taxa, but the toxic thresholds (EC10s) exceeded most realistic environmental dispersant concentrations. Estimating toxic threshold values for effects of dispersants on coral should benefit the decision-making of oil spill responders by contributing to the development of species sensitivity distributions (SSDs) for dispersant toxicity, and by informing net environmental benefit assessment (NEBA) for dispersant use.

Metabolomics reveals biotic and abiotic elicitor effects on the soft coral Sarcophyton ehrenbergi terpenoid content.

The effects of six biotic and abiotic elicitors, i.e. MeJA (methyl jasmonate), SA (salicylic acid), ZnCl2, glutathione and ß-glucan BG (fungal elicitor), and wounding, on the secondary metabolite accumulation in the soft coral Sarcophyton ehrenbergi were assessed. Upon elicitation, metabolites were extracted and analysed by ultra-performance liquid chromatography-mass spectrometry (UPLC-MS). Except for MeJA, no differences in photosynthetic efficiency were observed after treatments, suggesting the absence of a remarkable stress on primary production. Chemometric analyses of UPLC-MS data showed clear segregation of SA and ZnCl2 elicited samples at 24 and 48 h post elicitation. Levels of acetylated diterpene and sterol viz., sarcophytonolide I and cholesteryl acetate, was increased in ZnCl2 and SA groups, respectively, suggesting an activation of specific acetyl transferases. Post elicitation, sarcophytonolide I level increased 132 and 17-folds at 48 h in 0.1 mM SA and 1 mM ZnCl2 groups, respectively. Interestingly, decrease in sarcophine, a major diterpene was observed only in response to ZnCl2, whereas no change was observed in sesquiterpene content following treatments. To the best of our knowledge, this study provides the first documentation for elicitation effects on a soft corals secondary metabolome and suggests that SA could be applied to increase diterpenoid levels in corals.

An evaluation of coral lophelia pertusa mucus as an analytical matrix for environmental monitoring: A preliminary proteomic study.

For the environmental monitoring of coral, mucus appears to be an appropriate biological matrix due to its array of functions in coral biology and the non-intrusive manner in which it can be collected. The aim of the present study was to evaluate the feasibility of using mucus of the stony coral Lophelia pertusa (L. pertusa) as an analytical matrix for discovery of biomarkers used for environmental monitoring. More specifically, to assess whether a mass-spectrometry-based proteomic approach can be applied to characterize the protein composition of coral mucus and changes related to petroleum discharges at the seafloor. Surface-enhanced laser desorption/ionization-time of flight mass spectrometry (SELDI-TOF MS) screening analyses of orange and white L. pertusa showed that the mucosal protein composition varies significantly with color phenotype, a pattern not reported prior to this study. Hence, to reduce variability from phenotype difference, L. pertusa white individuals only were selected to characterize in more detail the basal protein composition in mucus using liquid chromatography, mass spectrometry, mass spectrometry (LC-MS/MS). In total, 297 proteins were identified in L. pertusa mucus of unexposed coral individuals. Individuals exposed to drill cuttings in the range 2 to 12 mg/L showed modifications in coral mucus protein composition compared to unexposed corals. Although the results were somewhat inconsistent between individuals and require further validation in both the lab and the field, this study demonstrated preliminary encouraging results for discovery of protein markers in coral mucus that might provide more comprehensive insight into potential consequences attributed to anthropogenic stressors and may be used in future monitoring of coral health.

Acute ecotoxicology of natural oil and gas condensate to coral reef larvae.

Risks posed by oil spills to coral reefs are difficult to evaluate, partially due to the absence of studies that adequately assess toxicity to relevant coral reef species. Here we experimentally tested the acute toxicity of condensate, representing a fraction of light crude oil, to coral (Acropora tenuis) and sponge (Rhopaloeides odorabile) larvae. The metamorphosis of coral larvae was inhibited at total petroleum aromatic hydrocarbon (TPAH) concentrations of water accommodated fractions (WAF) as low as 103 µg l(-1), similar to concentrations detected in seawater following large spills. The sensitivity of coral larvae increased by 40% when co-exposed to UV light that they might encounter in shallow reefal systems. Condensate WAF was more toxic to coral larvae than predicted by summing the toxicity of its main components (benzene, toluene, p-xylene and napthalene). In contrast, the sensitivity of sponge larvae to condensate WAF (>10,000 µg l(-1) TPAH) was far less than coral in the presence and absence of UV, but similar to that of other marine invertebrates. While these results highlight the relative sensitivity of coral larvae to oil, further research is needed to better understand and predict the impacts and risks posed by hydrocarbons to tropical reef systems.

On the use of 31P NMR for the quantification of hydrosoluble phosphorus-containing compounds in coral host tissues and cultured zooxanthellae.

(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.

Nutrient enrichment coupled with sedimentation favors sea anemones over corals.

Fine sediments, which account for the majority of total fluvial sediment flux, have been suggested to degrade coral reefs on a global scale. Furthermore, sediment impacts can be exacerbated by extreme rainfall events associated with global climate change and anthropogenic nutrient enrichment. We report the findings from a series of mesocosm experiments exploring the effects of short-term sedimentation and nutrient enrichment on the interactions between the hard coral Acropora muricata, the sea anemone Mesactinia ganesis, and the green macroalga Codium edule. Mesocosms were manipulated to simulate either unimpacted reefs or reefs exposed to elevated levels of fine sediments for 10 or 14 days to simulate the effects of heavy rainfall. The first and second experiments were aimed to examine the effects of inorganic and organic sediments, respectively. The third experiment was designed to examine the interactive effects of nutrient enrichment and elevated sediment loads. Neither inorganic nor organic sediment loadings significantly affected the physiological performance of the coral, but, importantly, did reduce its ability to compete with other organisms. Photosynthetic efficiencies of both the green macroalga and the sea anemone increased in response to both sediment loadings when they were simultaneously exposed to nutrient enrichment. While organic sediment loading increased the nitrogen content of the green macroalga in the first experiment, inorganic sediment loading increased its phosphorus content in the second experiment. The coral mortality due to sea anemones attack was significantly greater upon exposure to enriched levels of organic sediments and nutrients. Our findings suggest that the combined effects of short-term sedimentation and nutrient enrichment could cause replacement of corals by sea anemones on certain coral reefs.

Context-dependent effects of nutrient loading on the coral-algal mutualism.

Human-mediated increases in nutrient availability alter patterns of primary production, impact species diversity, and threaten ecosystem function. Nutrients can also alter community structure by disrupting the relationships between nutrient-sharing mutualists that form the foundation of communities. Given their oligotrophic nature and the dependence of reef-building corals on symbiotic relationships, coral reefs may be particularly vulnerable to excess nutrients. However, individual studies suggest complex, even contradictory, relationships among nutrient availability, coral physiology, and coral growth. Here, we used meta-analysis to establish general patterns of the impact of nitrogen (N) and phosphorus (P) on coral growth and photobiology. Overall, we found that over a wide range of concentrations, N reduced coral calcification 11%, on average, but enhanced metrics of coral photobiology, such as photosynthetic rate. In contrast, P enrichment increased average calcification rates by 9%, likely through direct impacts on the calcification process, but minimally impacted coral photobiology. There were few synergistic impacts of combined N and P on corals, as the nutrients impact corals via different pathways. Additionally, the response of corals to increasing nutrient availability was context dependent, varying with coral taxa and morphology, enrichment source, and nutrient identity. For example, naturally occurring enrichment from fish excretion increased coral growth, while human-mediated enrichment tended to decrease coral growth. Understanding the nuances of the relationship between nutrients and corals may allow for more targeted remediation strategies and suggest how other global change drivers such as overfishing and climate change will shape how nutrient availability impacts corals.

Toxicity of Deepwater Horizon source oil and the chemical dispersant, Corexit® 9500, to coral larvae.

Acute catastrophic events can cause significant damage to marine environments in a short time period and may have devastating long-term impacts. In April 2010 the BP-operated Deepwater Horizon (DWH) offshore oil rig exploded, releasing an estimated 760 million liters of crude oil into the Gulf of Mexico. This study examines the potential effects of oil spill exposure on coral larvae of the Florida Keys. Larvae of the brooding coral, Porites astreoides, and the broadcast spawning coral, Montastraea faveolata, were exposed to multiple concentrations of BP Horizon source oil (crude, weathered and WAF), oil in combination with the dispersant Corexit® 9500 (CEWAF), and dispersant alone, and analyzed for behavior, settlement, and survival. Settlement and survival of P. astreoides and M. faveolata larvae decreased with increasing concentrations of WAF, CEWAF and Corexit® 9500, however the degree of the response varied by species and solution. P. astreoides larvae experienced decreased settlement and survival following exposure to 0.62 ppm source oil, while M. faveolata larvae were negatively impacted by 0.65, 1.34 and 1.5 ppm, suggesting that P. astreoides larvae may be more tolerant to WAF exposure than M. faveolata larvae. Exposure to medium and high concentrations of CEWAF (4.28/18.56 and 30.99/35.76 ppm) and dispersant Corexit® 9500 (50 and 100 ppm), significantly decreased larval settlement and survival for both species. Furthermore, exposure to Corexit® 9500 resulted in settlement failure and complete larval mortality after exposure to 50 and 100 ppm for M. faveolata and 100 ppm for P. astreoides. These results indicate that exposure of coral larvae to oil spill related contaminants, particularly the dispersant Corexit® 9500, has the potential to negatively impact coral settlement and survival, thereby affecting the resilience and recovery of coral reefs following exposure to oil and dispersants.

Physiological and biochemical performances of menthol-induced aposymbiotic corals.

The unique mutualism between corals and their photosynthetic zooxanthellae (Symbiodinium spp.) is the driving force behind functional assemblages of coral reefs. However, the respective roles of hosts and Symbiodinium in this endosymbiotic association, particularly in response to environmental challenges (e.g., high sea surface temperatures), remain unsettled. One of the key obstacles is to produce and maintain aposymbiotic coral hosts for experimental purposes. In this study, a simple and gentle protocol to generate aposymbiotic coral hosts (Isopora palifera and Stylophora pistillata) was developed using repeated incubation in menthol/artificial seawater (ASW) medium under light and in ASW in darkness, which depleted more than 99% of Symbiodinium from the host within 4∼8 days. As indicated by the respiration rate, energy metabolism (by malate dehydrogenase activity), and nitrogen metabolism (by glutamate dehydrogenase activity and profiles of free amino acids), the physiological and biochemical performances of the menthol-induced aposymbiotic corals were comparable to their symbiotic counterparts without nutrient supplementation (e.g., for Stylophora) or with a nutrient supplement containing glycerol, vitamins, and a host mimic of free amino acid mixture (e.g., for Isopora). Differences in biochemical responses to menthol-induced bleaching between Stylophora and Isopora were attributed to the former digesting Symbiodinium rather than expelling the algae live as found in the latter species. Our studies showed that menthol could successfully bleach corals and provided aposymbiotic corals for further exploration of coral-alga symbioses.

P-glycoprotein (multi-xenobiotic resistance) and heat shock protein gene expression in the reef coral Montastraea franksi in response to environmental toxicants.

The deleterious impacts of marine pollutants on reef corals and their symbiotic algae are an important element of global coral reef decline. In the current study we examined the impacts of toxicants on the reef coral Montastraea franksi by analysing the expression of three stress-related genes belonging to the coral host, using Taqman real-time quantitative reverse transcription-PCR. Gene expression profiles of P-glycoprotein (or multi-xenobiotic resistance protein) (P-gp); heat shock protein 70 (Hsp70) and heat shock protein 90 (Hsp90) were examined following 4 and 8h exposures to the heavy metal copper (3, 10, 30 and 100 microgL(-1)) or the third generation oil dispersant Corexit9527 (1, 5, 10 and 50 ppm). Additionally, the expression of P-gp was examined following exposure to 0.5 and 5 microM concentrations of the chemotherapeutic drug vinblastine, a classic substrate of P-gp. The expression of P-gp increased significantly in corals treated with vinblastine and also increased following exposure to copper and Corexit9527. Hsp70, and to a lesser extent Hsp90 expression increased following exposure to copper and Corexit9527 indicating a general cellular stress response. Densities of symbiotic algae in the tissues of the corals did not change significantly during the experiments, nor was any loss or paling of coral tissues observed. These findings provide important insight into how corals defend themselves against pollution and complement ongoing initiatives developing molecular biomarkers of stress in reef-building corals.

Heat stress stimulates nitric oxide production in Symbiodinium microadriaticum: a possible linkage between nitric oxide and the coral bleaching phenomenon.

Nitric oxide (NO) is a gas displaying multiple physiological functions in plants, animals and bacteria. The enzymes nitrate reductase and NO synthase have been suggested to be involved in the production of NO in plants and algae, but the implication of those enzymes in NO production under physiological conditions remains obscure. Symbiodinium microadriaticum, commonly referred to as zooxanthellae, is a marine microalga commonly found in symbiotic association with a cnidarian host including reef-building corals. Here we demonstrate NO production in zooxanthellae upon supplementation of either sodium nitrite or L-arginine as a substrate. The nitrite-dependent NO production was detected electrochemically and confirmed by the application of 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (cPTIO), a specific NO scavenger. Cells stained with the diaminofluorescein, DAF-2 DA, an NO fluorescent probe, showed an increase in fluorescence intensity upon supplementation of both sodium nitrite and L-arginine. Microscopic observations of DAF-stained cells verified that NO was produced inside the cells. NO production in S. microadriaticum was found to increase upon exposure of cells to an acute heat stress which also caused a decline in the photosynthetic efficiency of PSII (F(v)/F(m)). This study provides substantial evidence to confirm that zooxanthellae can synthesize NO even when they are not in a symbiotic association with a coral host. The increase in NO production at high temperatures suggests that heat stress stimulates the microalgal NO production in a temperature-dependent manner. The implications of these findings are discussed in the light of the coral bleaching phenomenon which is associated with elevated sea surface temperature due to global warming.

Short and long term toxicity of crude oil and oil dispersants to two representative coral species.

Oil dispersants, the tool of choice for treating oil spills in tropical marine environments, is potentially harmful to marine life, including reef corals. In a previous study, we found that dispersed oil and oil dispersants are harmful to soft and hard coral species at early life stages. In this broader study, we employed a "nubbin assay" on more than 10 000 coral fragments to evaluate the short- and long-term impacts of dispersed oil fractions (DOFs) from six commercial dispersants, the dispersants and water-soluble-fractions (WSFs) of Egyptian crude oil, on two Indo Pacific branching coral species, Stylophora pistillata and Pocillopora damicornis. Survivorship and growth of nubbins were recorded for up to 50 days following a single, short (24 h) exposure to toxicants in various concentrations. Manufacturer-recommended dispersant concentrations proved to be highly toxic and resulted in mortality for all nubbins. The dispersed oil and the dispersants were significantly more toxic than crude oil WSFs. As corals are particularly susceptible to oil detergents and dispersed oil, the results of these assays rules out the use of any oil dispersant in coral reefs and in their vicinity. The ecotoxicological impacts of the various dispersants on the corals could be rated on a scale from the least to the most harmful agent, as follows: Slickgone > Petrotech > Inipol = Biorieco > Emulgal > Dispolen.

The ecotoxicology of vegetable versus mineral based lubricating oils: 3. Coral fertilization and adult corals.

Biodegradable vegetable-derived lubricants (VDL) might be less toxic to marine organisms than mineral-derived oils (MDL) due to the absence of high molecular weight aromatics, but this remains largely untested. In this laboratory study, adult corals and coral gametes were exposed to various concentrations of a two-stroke VDL-1A and a corresponding MDL to determine which lubricant type was more toxic to each life stage. In the fertilization experiment, gametes from the scleractinian coral Acropora microphthalma were exposed to water-accommodated fractions (WAF) of VDL-1A and MDL for four hours. The MDL and VDL-1A WAFs inhibited normal fertilization of the corals at 200 microg l(-1) total hydrocarbon content (THC) and 150 microg l(-1) THC respectively. Disturbance of a stable coral-dinoflagellate symbiosis is regarded as a valid measure of sub-lethal stress in adult corals. The state of the symbiosis in branchlets of adult colonies of Acropora formosa was monitored using indicators such as dinoflagellate expulsion and dark-adapted photosystem II yields of dinoflagellate (using pulse amplitude modulation fluorescence). An effect on symbiosis was measurable following 48 h exposure to the lubricants at concentrations of 190 microg l(-1) and 37 microg l(-1) THC for the MDL and VDL-1A respectively. GC/MS revealed that the main constituent of the VDL-1A WAF was the compound coumarin, added by the manufacturer to improve odour. The fragrance containing coumarin was removed from the lubricant formulation and the toxicity towards adult corals re-examined. The coumarin-free VDL-2 exhibited significantly less toxicity towards the adult corals than all of the other oil types tested, with the only measurable effect being a slight but significant drop in photosynthetic efficiency at 280 microg l(-1).

The use of coral nubbins in coral reef ecotoxicology testing.

While there is an urgent demand to establish reliable ecotoxicological assays for reef corals, there has not been yet an available source material that can supply the high number of colony replicates needed for reliable tests. In past experiments, the major obstacle to obtaining as many fragments as possible had been the damage inflicted to donor colonies by pruning. In this paper, we present the application of coral nubbins, a novel source material for coral ecotoxicology assays. Nubbins from the branching Red Sea coral Stylophora pistillata (n>450) were used for evaluating the impacts of water soluble fractions from a crude oil, an oil dispersant and dispersed oil. Coral nubbins (minute coral fragments in the size of one to several polyps) harvested from a single colony are genetically identical to each other, may be obtained in any quantity needed and whenever research activities demand their use. Several dozens of nubbins can be obtained from a single small branch in branching coral species, a procedure that has minimal impact on donor genotypes. Nubbins production is a low cost procedure and requires limited maintenance space. Results of short and long-term acute ecotoxicological tests are revealed and discussed here, indicating the advantageous use of nubbins as ubiquitous coral material for toxicology assays and physiological studies.