Sub-lethal combined effects of illicit drug and decreased pH on marine mussels: A short-time exposure to crack cocaine in CO2 enrichment scenarios.

The increasing CO2-concentrations in the atmosphere promote ocean acidification. Seawater chemistry changes interact with contaminants, such as illicit drugs in the coastal zones. This work evaluates impacts of pH decrease and crack-cocaine exposure on the commercial mussel Perna perna through biomarker responses (lysosomal membrane stability, lipid peroxidation, and DNA strand breaks). The organisms were exposed to different crack-cocaine concentrations (0.5, 5.0, and 50 µg L-1) combined with different pH values (8.3, 8.0, 7.5, 7.0, 6.5, and 6.0) for 96 h. Crack-cocaine in the different acidification scenarios triggered cyto-genotoxicity, which affected the overall health of mussels exposed to cocaine environmentally relevant concentration. This study produced the first data on biomarker responses associated with CO2-induced acidification and illicit drugs (crack-cocaine) in marine organisms.

Experimental evidence of uncertain future of the keystone ragworm Hediste diversicolor (O.F. Müller, 1776) under climate change conditions.

It is currently assumed that climate change related factors pose severe challenges to biodiversity maintenance. This paper assesses the multi-stressor effects of elevated temperature (15 °C as control, 25 °C as elevated) and CO2 levels (pH 8.1 as control, 7.5 and 7.0 representing acidifying conditions) on the physiological (survival and regenerative capacity), behavioral (feeding and burrowing activities), and biochemical changes (metabolic capacity, oxidative status and biotransformation mechanisms) experienced by the keystone polychaete Hediste diversicolor. Temperature rise enlarged the adverse effect of marine acidification on the survival of H. diversicolor, delayed the beginning of the excavation activity, enhancing the negative effects that pH decrease had in the burrowing behavior of this polychaete. Additionally, regardless of the temperature, exposure of H. diversicolor to acidification results in a reduction in the feeding rate. It is the first time that this decreased feeding capacity is found related to seawater acidification in this species. The healing of the wound and the blastemal formation were retarded due to these two climatic factors which hinder the regenerative process of polychaetes. These vital physiological functions of H. diversicolor can be related to the oxidative stress induced by climate change conditions since free radicals overproduced will impair cells functioning, affecting species biochemical and physiological performance, including feeding and tissue regeneration. The present results also demonstrated that although polychaetes' metabolic capacity was enhanced under stress conditions, organisms were still able to increase or maintain their energy reserves. Our findings are of major environmental relevance considering that predicted climate change conditions will affect species vital and ecological and physiological capacities. These can be translated into shrinking not only at the individual and population level but also in microbial and endofaunal diversities, in the detritus processing in estuaries and biogeochemical cycles at the ecosystem level. Thus the conservation of H. diversicolor populations is vital for the normal functioning of estuarine mudflat ecosystems.

Sediment quality assessment in the Guadalquivir River (SW, Spain) using caged Asian clams: A biomarker field approach.

This study assesses the sediment quality of the Guadalquivir River watercourse between the Alcalá del Río dam and the city of Seville. The main objective of this work is to address sediment quality in the area using an integrative approach that links sediment contamination and toxicity using the Asiatic clam (Corbicula fluminea) under field conditions. This is the first study conducted in the area that use of a battery of biomarkers from exposure (GST, GPx) to adverse biological effect (DNA and histopathological damage) to identify the contamination adverse effects in a river area affected by a cocktail of different anthropogenic activities (urban, industrial, agricultural, etc.). The sediment quality characterized in the area shows a significant biological stress related to metal(loid)s at station located in Alcalá del Río in the river upper part of the studied area, being this stress toxic when approaching the city of Seville. The sediments located nearby this city showed toxicity by means of positive values in the biomarkers of effects measured in the caged clams and related to contaminants with an industrial and urban discharge origins. These results have shown the useful and strength of the biomarker approach used in this study that combines biomarker responses from exposure to effects and allows identifying the contamination adverse effects by means of using caging individuals of the Asian clam. It has been proved in the different experiments how once the exposure biomarkers reach a maximum value of their system the detoxification ability of the organisms is collapsed and then the biomarkers of effect are measured significantly in the different tissues. The use of field surveys using tolerant specie such as the Asian clam is recommendable to determine sediment quality under an integrative point of view as here reported.

Effects of CO2 enrichment on two microalgae species: A toxicity approach using consecutive generations.

As a result of the increasing pressure provoked by anthropogenic activities, the world climate is changing and oceans health is in danger. One of the most important factors affecting the marine environment is the well-known process called ocean acidification. Also, there are other natural or anthropogenic processes that produce an enrichment of CO2 in the marine environment (CO2 leakages from Carbon Capture and Storage technologies (CCS), organic matter diagenesis, volcanic vents, etc). Most of the studies related to acidification of the marine environment by enrichment of CO2 have been focused on short-term experiments. To evaluate the effects related to CO2 enrichment, laboratory-scale experiments were performed using the marine microalgae Tetraselmis chuii and Phaeodactylum tricornutum. Three different pH values (two treatments - pH 7.4 and 6.0 - and a control - pH 8.0) were tested on the selected species across four consecutive generations. Seawater was collected and exposed to different scenarios of CO2 enrichment by means of CO2 injection. The results showed different effects depending on the species and the generation used. Effects on T. chuii were shown on cell density, chlorophyll-a and metabolic activity, however, a slight adaptation across generations was found in this last parameter. P. tricornutum was more sensitive to acidification conditions through generations, with practically total growth inhibition in the fourth one. The conclusions obtained in this work are useful to address the potential ecological risk related to acidification by enrichment of CO2 on the marine ecosystem by using consecutive generations of microalgae.

Metal fractionation in marine sediments acidified by enrichment of CO2: A risk assessment.

Carbon-capture and storage is considered to be a potential mitigation option for climate change. However, accidental leaks of CO2 can occur, resulting in changes in ocean chemistry such as acidification and metal mobilization. Laboratory experiments were performed to provide data on the effects of CO2-related acidification on the chemical fractionation of metal(loid)s in marine-contaminated sediments using sequential extraction procedures. The results showed that sediments from Huelva estuary registered concentrations of arsenic, copper, lead, and zinc that surpass the probable biological effect level established by international protocols. Zinc had the greatest proportion in the most mobile fraction of the sediment. Metals in this fraction represent an environmental risk because they are weakly bound to sediment, and therefore more likely to migrate to the water column. Indeed, the concentration of this metal was lower in the most acidified scenarios when compared to control pH, indicating probable zinc mobilization from the sediment to the seawater.

Implications in studies of environmental risk assessments: Does culture medium influence the results of toxicity tests of marine bacteria?

Two marine bacterial populations (Roseobacter sp. and Pseudomonas litoralis) were exposed to different concentrations of zinc (300, 625, 1250, 2000, 2500 and 5000 mg L-1) and cadmium (75, 250, 340, 500 and 1000 mg L-1) using two culture media (full nutrient Marine Broth 2216 "MB" and 1:10 (vol/vol) dilution with seawater of Marine Broth 2216 "MBSW"), in order to assess population responses depending on the culture medium and also potential adverse effects associated with these two metals. Different responses were found depending on the culture medium (Bacterial abundance (cells·mL-1), growth rates (µ, hours-1), and production of Extracellular Polysaccharides Substances (EPS) (µg glucose·cells-1). Results showed negative effects in both strains after the exposure to Zn treatments. Both strains showed highest metal sensitivity at low concentrations using both culture media. However, different results were found when exposing the bacterial populations to Cd treatments depending on the culture medium. Highest toxicity was observed using MB at low levels of Cd concentrations, whereas MBSW showed toxicity to bacteria at higher concentrations of Cd. Results not only showed adverse effects on Roseobacter sp. and Pseudomonas litoralis associated with the concentration of Zn and Cd, but also confirm that depending on the culture medium results can differ. This work suggests MBSW as an adequate culture medium to study metal toxicity bioassays in order to predict realistic effects on marine bacterial populations.

Will temperature and salinity changes exacerbate the effects of seawater acidification on the marine microalga Phaeodactylum tricornutum?

To evaluate the effects related to the combination of potential future changes in pH, temperature and salinity on microalgae, a laboratory experiment was performed using the marine diatom Phaeodactylum tricornutum. Populations of this species were exposed during 48h to a three-factor experimental design (3×2×2) with two artificial pH values (6, 7.4), two levels of temperature (23°C, 28°C), two levels of salinity (34psu, 40psu) and a control (pH8, Temp 23°C, Sal 34psu). The effects on growth, cell viability, metabolic activity, and inherent cell properties (size, complexity and autofluorescence) of P. tricornutum were studied using flow cytometry. The results showed adverse effects on cultures exposed to pH6 and high temperature and salinity, being the inherent cell properties the most sensitive response. Also, linked effects of these parameters resulted on cell viability and cell size decrease and an increase of cell autofluorescence. The conclusions obtained from this work are useful to address the potential effects of climate change (in terms of changes on pH, salinity and temperature) in microalgae.

Effects of a hypothetical escape of CO2 gas from subterranean storage sites on water flea Daphnia magna.

The impacts of a hypothetical CO2 gas leak from freshwater sediments on the survival and reproduction of freshwater flea Daphnia magna were analyzed. Another objective was to assess the performance of standard toxicity testing protocols for CO2-induced acidification research in freshwaters. Four pH levels (7.5, 7.0, 6.5, and 6.0) and two sediments with different contamination level were tested. The results revealed that the D. magna are susceptible to a gradual but relatively rapid CO2 enrichment of the water column causing a change from circumneutral to acidic conditions. Standard 48-h immobilization test with D. magna tended to underestimate the toxicity at CO2-induced acidity condition. Dissolved aluminum may be implicated in the toxicity to the parental daphnids exposed. Metal outflux from sediments and behavior in elutriate have been discussed.

Simulating CO2 leakage from sub-seabed storage to determine metal toxicity on marine bacteria.

CO2 storage in sub-seabed marine geological formations has been proposed as an adequate strategy to mitigate high CO2 concentration from the atmosphere. The lack of knowledge about the potential risks of this technology on marine bacteria population in presence of metals has lead us to perform laboratory-scale experiments in order to evaluate its consequences. Thus, the effects of Zn and Cd were studied under acid conditions on Roseobacter sp. and Pseudomonas litoralis. Bacterial abundance (cellsmL-1), growth rates (µ, h-1), relative inhibitory effects of CO2 (RICO2), and production of Extracellular Polysaccharides Substances (EPS) (µgGlucosecells-1) were evaluated. A decreasing exopolysaccharides (EPS) production was found under low pH. Bacterial abundance as well as growth rates showed negative effects. Data obtained in this work are useful to determine the potential effects associated with enrichment of CO2 and metals on the marine ecosystem.

The influence of ph and waterborne metals on egg fertilization of the blue mussel (Mytilus edulis), the oyster (Crassostrea gigas) and the sea urchin (Paracentrotus lividus).

This study evaluated the combined effect of pH and metals on the egg fertilization process of two estuarine species, the blue mussel (Mytilus edulis), the oyster (Crassostrea gigas) and a marine species, the sea urchin (Paracentrotus lividus). The success of egg fertilization was examined after exposure of gametes to sediment extracts of various degrees of contamination at pH 6.0, 6.5, 7.0, 7.5 and 8.0. At the pH levels from 6.5 to 8.0, the egg fertilization of the different species demonstrated different sensitivity to metal and/or acidic exposure. In all species, the results revealed that egg fertilization was almost completely inhibited at pH 6.0. The egg fertilization of the blue mussel M. edulis was the least sensitive to the exposure while that of the sea urchin P. lividus demonstrated a concentration-dependent response to the pH levels from 6.5 to 8.0. The results of this study revealed that acidity increased the concentration of several metal ions (Cr, Ni, Cu, Zn, Cd, and Pb) but reduced its availability to the organisms, probably related to the reactivity of the ions with most non-metals or to the competition among metals and other waterborne constituents.

Simulating CO2 leakages from CCS to determine Zn toxicity using the marine microalgae Pleurochrysis roscoffensis.

Due to the current climate change and ocean acidification, a new technology for CO2 mitigation has been proposed, the Carbon dioxide Capture and Storage (CCS). However, there is an ecological risk associated with potential CO2 leakages from the sub-seabed storages sites. To evaluate the effects related to CO2 leakages, laboratory-scales experiments were performed using the marine microalgae Pleurochrysis roscoffensis. Five Zn concentrations were tested at different pHs to study Zn toxicity under acidified conditions. Seawater was collected and submitted to acidification by means of CO2 injection and by HCl addition. Results showed differences between both acidification techniques: while microalgae growth was enhanced by CO2 supply, reaching the optimal growth at pH 6.5 and full inhibition at pH 5.5, HCl acidification growth was inhibited at pH 6.5. Although small concentrations of Zn were positive for P. roscoffensis growth, Zn toxicity increased at lower pHs, and more severely on samples acidified with HCl. The conclusions obtained in this work are useful to address the potential effects on the marine ecosystem related to changes in metal bioavailability during CO2 leakages scenarios.

Dredged material characterization and management frameworks: A case study at the port Vilagarcia (NW, Spain).

The potential impact of dredged sediment has been assessed at sixteen areas of the high-traffic port of Vilagarcia (Northwest Spanish Atlantic coast). The assessment has been done by three weight-of-evidence tools, which integrated data on sediment characteristics and toxicity responses of Ampelisca brevicornis, Vibrio fischeri and eggs and embryos of Paracentrotus lividus. Two of the tools also represented management options regarding the disposal of dredged material. The comparison of the logic in these tools revealed essential differences in the type and the necessity of bioassays and threshold values for chemical concentrations. However, despite this difference, assessment results and the derived management options coincided in most of the sediments. The potential toxicity of sediments was relatively low especially for eggs and embryos possibly due to different contaminant availability in solid and liquid phases. The importance of a battery of toxicity tests in the dredged material quality assessment has been emphasized to avoid an underestimation of sediment toxicity for solid phase organisms, if only liquid phase responses are considered. The potential false implications, which may result from the application of the third tool, were highlighted. The strengths and weaknesses of the tools were discussed from the dredged material management perspective.

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.

Evaluation of the threat of marine CO2 leakage-associated acidification on the toxicity of sediment metals to juvenile bivalves.

The effects of the acidification associated with CO2 leakage from sub-seabed geological storage was studied by the evaluation of the short-term effects of CO2-induced acidification on juveniles of the bivalve Ruditapes philippinarum. Laboratory scale experiments were performed using a CO2-bubbling system designed to conduct ecotoxicological assays. The organisms were exposed for 10 days to elutriates of sediments collected in different littoral areas that were subjected to various pH treatments (pH 7.1, 6.6, 6.1). The acute pH-associated effects on the bivalves were observed, and the dissolved metals in the elutriates were measured. The median toxic effect pH was calculated, which ranged from 6.33 to 6.45. The amount of dissolved Zn in the sediment elutriates increased in parallel with the pH reductions and was correlated with the proton concentrations. The pH, the pCO2 and the dissolved metal concentrations (Zn and Fe) were linked with the mortality of the exposed bivalves.

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.

Simulation of the potential effects of CO2 leakage from carbon capture and storage activities on the mobilization and speciation of metals.

One of the main risks associated with carbon capture and storage (CCS) activities is the leakage of the stored CO2, which can result in several effects on the ecosystem. Laboratory-scale experiments were performed to provide data on the possible effects of CO2 leakage from CCS on the mobility of metals previously trapped in sediments. Metal-contaminated sediments were collected and submitted to acidification by means of CO2 injection using different pH treatments. The test lasted 10 days, and samples were collected at the beginning and at the end of the experiment for metal analysis. The results revealed increases in the mobility of metals such as Co, Cu, Fe, Pb and Zn due to pH decreases. Geochemical modeling demonstrated that acidification influenced the speciation of the metals, increasing the concentrations of their free forms. These data suggest the possible sediment contamination consequences of accidental CO2 leakage during CCS activities.

Studying the effect of CO2-induced acidification on sediment toxicity using acute amphipod toxicity test.

Carbon capture and storage is increasingly being considered one of the most efficient approaches to mitigate the increase of CO2 in the atmosphere associated with anthropogenic emissions. However, the environmental effects of potential CO2 leaks remain largely unknown. The amphipod Ampelisca brevicornis was exposed to environmental sediments collected in different areas of the Gulf of Cádiz and subjected to several pH treatments to study the effects of CO2-induced acidification on sediment toxicity. After 10 days of exposure, the results obtained indicated that high lethal effects were associated with the lowest pH treatments, except for the Ría of Huelva sediment test. The mobility of metals from sediment to the overlying seawater was correlated to a pH decrease. The data obtained revealed that CO2-related acidification would lead to lethal effects on amphipods as well as the mobility of metals, which could increase sediment toxicity.

Simulation of CO2 leakages during injection and storage in sub-seabed geological formations: metal mobilization and biota effects.

To assess the potential effects on metal mobilization due to leakages of CO2 during its injection and storage in marine systems, an experimental set-up was devised and operated, using the polychaete Hediste diversicolor as the model organism. The objective was to study the effects of such leakage in the expected scenarios of pH values between 8.0 and 6.0. Polychaetes were exposed for 10 days to seawater with sediment samples collected in two different coastal areas, one with relatively uncontaminated sediment as reference (RSP) and the other with known contaminated sediment (ML), under pre-determined pH conditions. Survival and metal accumulation (Al, Fe, Mn, Cu, Zn, As and Hg) in the whole body of H. diversicolor were employed as endpoints. Mortality was significant at the lowest pH level in the sediment with highest metal concentrations. In general, metal concentrations in tissues of individuals exposed to the contaminated sediment were influenced by pH. These results indicate that ocean acidification due to CO2 leakages would provoke increased metal mobilization, causing adverse side effects in sediment toxicity.

Effects on the mobility of metals from acidification caused by possible CO2 leakage from sub-seabed geological formations.

Carbon dioxide capture and storage (CCS) in submarine geological formations has been proposed as a mitigation measure for the prevention of global warming. However, leakage of CO2 to overlying sediments may occur over time, leading to various effects on ecosystems. Laboratory-scale experiments were performed, involving direct release of carbon dioxide into sediment, inside non-pressurized chambers, in order to provide data on the possible effects of CO2 leakage from geological storage sites on the fate of several metals. Marine sediments from three sites with different levels of contamination were sampled and submitted to acidification by means of CO2 injection. The experiment lasted 10 days and sediment samples were collected at the beginning and end of the experiment and pore water was extracted for metal analysis. The results revealed that mobility of metals from sediment to pore water depends on the site, metal and length of time exposed. Mobilization of the metals Al, Fe, Zn, Co, Pb and Cu increases with acidification, and this response generally increases with time of exposure to CO2 injection. The geochemical model applied suggests that acidification also influences the speciation of metals, transforming metals and metalloids, like As, into species much more toxic to biota. The data obtained from this study will be useful for calculating the potential risk of CCS activities to the marine environment.

Bioaccumulation and effects of metals bound to sediments collected from Gulf of Cádiz (SW Spain) using the polychaete Arenicola marina.

A short-term whole-sediment test using the polychaete Arenicola marina was conducted under laboratory conditions to assess the bioavailability of metals bound to sediments collected from 12 sites of the Gulf of Cádiz. To achieve this objective, the rate of increase of metal bioaccumulation and the induction of a typical biomarker, metallothioneinlike proteins (MTLPs) were determined. Results of the multivariate analysis showed associated metal-rich sediments, increased rate of Cu and Zn accumulations, but lower toxicity with an increased MTLP induction, whereas sedimentary Ni and Co concentrations were related to higher toxicity to lugworms, although it might be caused by other contaminants present in these sediments. The linear kinetic approach was shown to be valid in certain circumstances, but more validation studies of this parameter are required before it can be recommended for use in evaluating metal bioavailability in sediments.