Laboratory simulation system, using Carcinus maenas as the model organism, for assessing the impact of CO2 leakage from sub-seabed injection and storage.

The capture and storage of CO2 in sub-seabed geological formations has been proposed as one of the potential options to decrease atmospheric CO2 concentrations in order to mitigate the abrupt and irreversible consequences of climate change. However, it is possible that CO2 leakages could occur during the injection and sequestration procedure, with significant repercussions for the marine environment. We investigate the effects of acidification derived from possible CO2 leakage events on the European green crab, Carcinus maenas. To this end, a lab-scale experiment involving direct release of CO2 was conducted at pH values between 7.7 and 6.15. Female crabs were exposed for 10 days to sediment collected from two different coastal areas, one with relatively uncontaminated sediment (RSP) and the other with known contaminated sediment (MZ and ML), under the pre-established seawater pH conditions. Survival rate, histopathological damage and metal (Fe, Mn, Cu, Zn, Cr, Cd and Pb) and As accumulation in gills and hepatopancreas tissue were employed as endpoints. In addition, the obtained results were compared with the results of the physico-chemical characterization of the sediments, which included the determination of the metals Fe, Mn, Cu, Zn, Cr, Pb and Cd, the metalloid As, certain polycyclic aromatic hydrocarbons (PAHs) and polychlorinated biphenyls (PCBs), as well as nonchemical sediment properties (grain size, organic carbon and total organic matter). Significant associations were observed between pH and the histological damage. Concentrations of Fe, Mn, Cr, Pb, Cd and PAHs in sediment, presented significant negative correlations with the damage to gills and hepatopancreas, and positive correlations with metal accumulation in both tissues. The results obtained in this study reveal the importance of sediment properties in the biological effects caused by possible CO2 leakage. However, a clear pattern was not observed between metal accumulation in tissues and pH reduction. Animals' avoidance behavior and degree of tolerance to acidification are confounding factors for assessing metal bioaccumulation. Further research is required to find a suitable assay that would allow us to predict the risk to environmental health of possible negative side effects of metal mobility derived from CO2 leakage during its injection and storage in sub-seabed formations.

Predicting the impacts of CO2 leakage from subseabed storage: effects of metal accumulation and toxicity on the model benthic organism Ruditapes philippinarum.

The urgent need to minimize the potential harm deriving from global climate change and ocean acidification has led governmental decision-makers and scientists to explore and study new strategies for reducing the levels of anthropogenic CO2. One of the mitigation measures proposed for reducing the concentration of atmospheric CO2 is the capture and storage of this gas in subseabed geological formations; this proposal is generating considerable international interest. The main risk associated with this option is the leakage of retained CO2, which could cause serious environmental perturbations, particularly acidification, in marine ecosystems. The study reported is aimed at quantifying the effects of acidification derived from CO2 leakage on marine organisms. To this end, a lab-scale experiment involving direct release of CO2 through marine sediment was conducted using Ruditapes philippinarum as a model benthic organism. For 10 days bivalves were exposed to 3 sediment samples with different physicochemical characteristics and at pre-established pH conditions (8.0-6.1). End points measured were: survival, burrowing activity, histopathological lesions, and metal accumulation (Fe, Al, Mn, Cu, and Zn) in whole body. Correlations analyses indicated highly significant associations (P < 0.01) between pH and the biological effects measured in R philippinarum, except for metal concentrations in tissues. Further research to understand and predict the biological and economic implications for coastal ecosystems deriving from acidification by CO2 leakages is urgently needed.

An early approach for the evaluation of repair processes in fish after exposure to sediment contaminated by an oil spill.

A chronic bioassay was carried out under laboratory conditions using juvenile Solea senegalensis to determine the toxicity of contaminants from an oil spill(Prestige). Also, the repair processes in fish affected by contaminants due to oil exposure were evaluated. Over 30 days individuals were exposed to clean sediment (control) and to sediment contaminated by a mixture of polyaromatic hydrocarbons (PAHs) and other substances. The physicochemical parameters of the tanks (salinity, temperature, pH and dissolved oxygen) were controlled during the exposure period. Clean sediment from the Bay of Cadiz (Spain) was used as negative control and was mixed with fuel oil to prepare the dilution (0.5% w:w dry-weight). After the exposure period, fish were labeled and transferred to "clean tanks" (tanks without sediment) in order to study the recovery and the repair processes in the exposed organisms. A biomarker of exposure (ethoxyresorufin-O-deethylase activity - EROD activity) and a biomarker of effect (histopathology) were analyzed during the exposure and recovery period. After 10, 20 and 30 days of exposure, individuals showed significant induction (P < 0.05) of the EROD activity and also presented diverse histopathological damages. The analysis of both the biomarkers of exposure and effect, after the 5th and 10th day of recovery in the "clean tank", enabled a first evaluation of the repair process of the induced damages due to the fuel oil exposure. After the recovery phase, control individuals showed a more significant decrease (P < 0.05) of the alteration of the measured biomarkers than in the oil-exposed fish. While in the oil-exposed fish the EROD activity showed some recovery, the histopathological damages did hardly improve. According to our results, tissue repair processes probably need longer recovery periods to observe significant improvement of the affected organs. This will be further investigated in the future.

Chronic bioassay in benthic fish for the assessment of the quality of sediments in different areas of the coast of Spain impacted by acute and chronic oil spills.

More and more, the coastal regions of the world suffer from the contamination of petroleum hydrocarbon [principally polycyclic aromatic hydrocarbons (PAHs)]. This contamination can be acute, as it happened in the Coast of Galicia (NW, Spain) by the oil spill from a tanker, or chronic by the existence of high maritime traffic and a lot of industries as it is the case of the Bay of Algeciras (BA) (SW, Spain). It is of a great concern due to the toxicity, especially in sediments and ecosystem associated to it. The objective of this study is to assess, through chronic bioassay, sediment toxicity in samples collected in different littoral areas of Spain and to compare the damage caused in benthic fish, Solea senegalensis, according to that which suffers acute spill (Coast of Galicia) or chronic spill (the BA) by means of histopathology methods and enzymatic activities studies. Organisms were exposed to different sediments from Galician Coast and the BA during 42 days and every sample was analyzed by triplicate in glass aquaria. At the end of the bioassay, histopathological diseases were analyzed in the gills, target organ. Likewise, stress parameters as ethoxyresorufin-O-deethylase (EROD) and the glutathione S-transferases (GSTs) activities were determined in the liver. Results showed a significant positive correlation between the biomarkers of exposure (EROD and GST), biomarkers of effect (histopathology), and PAHs concentrations in the sediments.

Determining sediment quality for regulatory proposes using fish chronic bioassays.

Sediment quality assessments for regulatory purposes (i.e. dredged material disposal) are characterized by linking chemical and acute ecotoxicological data. The design of chronic bioassays that incorporate more sensible endpoints than acute tests is discussed to address sediment quality for environmental quality assessment and regulatory proposes. The chronic tests use juveniles of commercial species of fish Sparus aurata and Solea senegalensis, to assess sediment toxicity in samples collected along different littoral areas in the North and the South of Spain. The organisms were exposed during 60 days and sublethal endpoints were selected including biomarkers of exposure to metals (metallothioneins - MTs) and to organic contaminants (ethoxyresorufin-O-deethylase activity - EROD activity) and biomarkers of effect (histopathology in different tissues, gill and liver). A Multivariate Analysis Approach was conducted in order to associate these biological responses with sediment metal concentration from the ports and with chemical residues in biological tissues exposed to sediments under laboratory conditions.

Kinetic of biomarker responses in juveniles of the fish Sparus aurata exposed to contaminated sediments.

Sediments in the National Park of the Atlantic Islands (Galicia, Spain) were affected by the spill of the tanker Prestige (November, 2002) and still present high levels of Polycyclic aromatic hydrocarbons. The adverse effects associated with the contaminants in sediments were tested using a chronic bioassay, exposing juveniles of the fish Sparus aurata (seabream). A toxicokinetic approach is proposed to evaluate sediment quality by linking chemical and ecotoxicological data along the time. Sediment samples were physicochemically characterized and the concentration of contaminants (Polycyclic aromatic hydrocarbons - PAHs - and metals) was measured. Fishes were exposed to contaminated sediments, and samples from different tissues were collected every 15 days throughout the 60 days that lasted the experiment. A biomarker of exposure (ethoxyresorufin O-deethylase activity - EROD activity) and a biomarker of effect (histopathology) were analyzed during the exposure period. Results show a relationship between the biomarkers and the concentrations in sediments of polycyclic aromatic hydrocarbons-PAHs. Besides, the toxicokinetic approach links biomarkers response providing information about the relationship between the detoxification process and the damages observed in the different tissues. The frequency of the histological damage is highest when the EROD activity slightly decreases in accordance with the mechanism of detoxification of this enzymatic system against PAHs and other organic contaminants.

Ecotoxicity of sediments contaminated by the oil spill associated with the tanker "Prestige" using juveniles of the fish Sparus aurata.

In November 2002, the oil spill from the tanker Prestige in the Galician Coast caused an ecological catastrophe in Spain. The adverse effects associated with the contaminants bound to sediments were tested using juveniles of the fish Sparus aurata (seabream). The approach evaluates sediment quality by using an integrated assessment including chemical and ecotoxicological data. Sediment samples were physicochemically characterized, and the concentration of contaminants (polycyclic aromatic hydrocarbons-(PAHs) and metals) was measured. Different biomarkers of exposure (metallothioneins and ethoxyresorufin O-deethylase activity (EROD)) and biomarkers of effect (histopathology) were analyzed along the time. A multivariate analysis approach was used to correlate concentration of contaminants and sublethal effects measured in individuals of fish. Results show that increasing concentrations of PAHs in sediments were related to increased EROD activities and histopathological lesions. This is the first evidence showing adverse effects associated with petroleum contamination of PAHs in sediments after this spill, and it demonstrates the utility of the sublethal toxicity tests for monitoring the impact of petroleum spills.