Combined effects of high hydrostatic pressure and dispersed oil on the metabolism and the mortality of turbot hepatocytes (Scophthalmus maximus).

Since the DeepWater Horizon oil spill and the use at 1450 m depth of dispersant as a technical response, the need of relevant ecotoxicological data on deep-sea ecosystems becomes crucial. In this context, this study focused on the effect of high hydrostatic pressure (10.1 MPa) on turbot hepatocytes isolated from fish exposed either to chemically dispersed oil, mechanically dispersed oil or dispersant alone. Potential combined effects of oil/dispersant and hydrostatic pressure, were assessed on cell mortality (total cell death, necrosis and apoptosis), cell viability and on hepatocyte oxygen consumption (MO2). No change in cell mortality was observed in any of the experimental conditions, whereas, the results of cell viability showed a strong and significant increase in the two oil groups independently of the pressure exposure. Finally, oil exposure and hydrostatic pressure have additive effects on oxygen consumption at a cellular level. Presence of dispersant prevent any MO2 increase in our experimental conditions. These mechanistic effects leading to this increased energetic demand and its eventual inhibition by dispersant must be investigated in further experiments.

Deep-sea versus shallow conditions: a comparative ecobarotoxicological study.

In the context of new oil exploration/production areas, knowledge of the biological impact of dispersed oil in the deep-sea environment is essential. Hence, the aim of this study was to perform a comparison, at atmospheric pressure (0.1 MPa) and at a high hydrostatic pressure corresponding to 1000 m depth (10.1 MPa), of lethal concentrations (LC) on a model fish, Scophthalmus maximus, exposed to chemically dispersed oil. Fish were exposed concomitantly at 0.1 and 10.1 MPa using two exposure tanks connected to the same source tank thanks to a closed circuit. Acute toxicity was evaluated at 24 h through the determination of LC10 and LC50 (respectively, 10 and 50% of mortality) calculated from measured total petroleum hydrocarbon concentrations in the water. No statistical differences were observed between the LC10 at 0.1 MPa (46.1 mg L- 1) and the LC10 at 10.1 MPa (31.0 mg L- 1), whereas the LC50 of fish exposed to 0.1 MPa (90.8 mg L- 1) was significantly higher than the LC50 at 10.1 MPa (50.9 mg L- 1). These results clearly show an increase in oil toxicity under high hydrostatic pressure. This effect may be due to synergistic effects of pressure and oil contamination on fish energetic metabolism.

Akinetes and ancient DNA reveal toxic cyanobacterial recurrences and their potential for resurrection in a 6700-year-old core from a eutrophic lake.

In order to evaluate the recurrence of toxic cyanobacterial blooms and to determine the survival capabilities of the resistance cells through time, a sedimentary core spanning 6700 years was drilled in the eutrophic Lake Aydat. A multiproxy approach (density, magnetic susceptibility, XRF, pollen and non-pollen palynomorph analyses), was used initially to determine the sedimentation model and the land uses around the lake. Comparison with the akinete count revealed that Nostocales cyanobacteria have been present in Lake Aydat over a six thousand year period. This long-term cyanobacterial recurrence also highlights the past presence of both the anaC and mcyB genes, involved in anatoxin-a and microcystin biosynthesis, respectively, throughout the core. The first appearance of cyanobacteria seems to be linked to the natural damming of the river, while the large increase in akinete density around 1800 cal.yr BP can be correlated with the intensification of human activities (woodland clearance, crop planting, grazing, etc.) in the catchment area of the lake, and marks the beginning of a long period of eutrophication. This first investigation into the viability and germination potential of cyanobacteria over thousands of years reveals the ability of intact akinetes to undergo cell divisions even after 1800 years of sedimentation, which is 10 times longer than previously observed. This exceptional cellular resistance, coupled with the long-term eutrophic conditions of this lake, could partly explain the past and current recurrences of cyanobacterial proliferations.

Dispersed oil decreases the ability of a model fish (Dicentrarchus labrax) to cope with hydrostatic pressure.

Data on the biological impact of oil dispersion in deep-sea environment are scarce. Hence, the aim of this study was to evaluate the potential interest of a pressure challenge as a new experimental approach for the assessment of consequences of chemically dispersed oil, followed by a high hydrostatic pressure challenge. This work was conducted on a model fish: juvenile Dicentrarchus labrax. Seabass were exposed for 48 h to dispersant alone (nominal concentration (NC) = 4 mg L-1), mechanically dispersed oil (NC = 80 mg L-1), two chemically dispersed types of oil (NC = 50 and 80 mg L-1 with a dispersant/oil ratio of 1/20), or kept in clean seawater. Fish were then exposed for 30 min at a simulated depth of 1350 m, corresponding to pressure of 136 absolute atmospheres (ATA). The probability of fish exhibiting normal activity after the pressure challenge significantly increased from 0.40 to 0.55 when they were exposed to the dispersant but decreased to 0.26 and 0.11 in the case of chemical dispersion of oil (at 50 and 80 mg L-1, respectively). The chemical dispersion at 80 mg L-1 also induced an increase in probability of death after the pressure challenge (from 0.08 to 0.26). This study clearly demonstrates the ability of a pressure challenge test to give evidence of the effects of a contaminant on the capacity of fish to face hydrostatic pressure. It opens new perspectives on the analysis of the biological impact of chemical dispersion of oil at depth, especially on marine species performing vertical migrations.

Sensitivity of the deep-sea amphipod Eurythenes gryllus to chemically dispersed oil.

In the context of an oil spill accident and the following oil spill response, much attention is given to the use of dispersants. Dispersants are used to disperse an oil slick from the sea surface into the water column generating a cloud of dispersed oil droplets. The main consequence is an increasing of the sea water-oil interface which induces an increase of the oil biodegradation. Hence, the use of dispersants can be effective in preventing oiling of sensitive coastal environments. Also, in case of an oil blowout from the seabed, subsea injection of dispersants may offer some benefits compared to containment and recovery of the oil or in situ burning operation at the sea surface. However, biological effects of dispersed oil are poorly understood for deep-sea species. Most effects studies on dispersed oil and also other oil-related compounds have been focusing on more shallow water species. This is the first approach to assess the sensitivity of a macro-benthic deep-sea organism to dispersed oil. This paper describes a toxicity test which was performed on the macro-benthic deep-sea amphipod (Eurythenes gryllus) to determine the concentration causing lethality to 50% of test individuals (LC50) after an exposure to dispersed Brut Arabian Light (BAL) oil. The LC50 (24 h) was 101 and 24 mg L(-1) after 72 h and 12 mg L(-1) at 96 h. Based on EPA scale of toxicity categories to aquatic organisms, an LC50 (96 h) of 12 mg L(-1) indicates that the dispersed oil was slightly to moderately toxic to E. gryllus. As an attempt to compare our results to others, a literature study was performed. Due to limited amount of data available for dispersed oil and amphipods, information on other crustacean species and other oil-related compounds was also collected. Only one study on dispersed oil and amphipods was found, the LC50 value in this study was similar to the LC50 value of E. gryllus in our study. Since toxicity data are important input to risk assessment and net environmental benefit analyses, and since such data are generally lacking on deep-sea species, the data set produced in this study is of interest to the industry, stakeholders, environmental management, and ecotoxicologists. However, studies including more deep-sea species covering different functional groups are needed to evaluate the sensitivity of the deep-sea compartments to dispersed oil relative to other environmental compartments.

Growth and immune system performance to assess the effect of dispersed oil on juvenile sea bass (Dicentrarchus labrax).

The potential impact of chemically and mechanically dispersed oil was assessed in a model fish of European coastal waters, the sea bass Dicentrarchus labrax. Juvenile sea bass were exposed for 48h to dispersed oil (mechanically and chemically) or dispersants alone. The impact of these exposure conditions was assessed using growth and immunity. The increase observed in polycyclic aromatic hydrocarbon metabolites in bile indicated oil contamination in the fish exposed to chemical and mechanical dispersion of oil without any significant difference between these two groups. After 28 days of exposure, no significant differences were observed in specific growth rate,apparent food conversion efficiency and daily feeding). Following the oil exposure, fish immunity was assessed by a challenge with Viral Nervous Necrosis Virus (VNNV). Fish mortality was observed over a 42 day period. After 12 days post-infection, cumulative mortality was significantly different between the control group (16% p≤0.05) and the group exposed to chemical dispersion of oil (30% p≤0.05). However, at the end of the experiment, no significant difference was recorded in cumulative mortality or in VNNV antibodies secreted in fish in responses to the treatments. These data suggested that in our experimental condition, following the oil exposure, sea bass growth was not affected whereas an impact on immunity was observed during the first days. However, this effect on the immune system did not persist over time.

Innate immunity and antioxidant systems in different tissues of sea bass (Dicentrarchus labrax) exposed to crude oil dispersed mechanically or chemically with Corexit 9500.

The aim of the study was to evaluate effects of chemically dispersed oil by the dispersant Corexit 9500 on innate immunity and redox defenses in a marine model fish. Sea bass (Dicentrarchus labrax) were exposed 48h to four experimental conditions: a control group (C), a group only exposed to the dispersant (D; 3.6mg/L) and two groups exposed to 80mg/L oil mechanically or chemically dispersed (MD; CD). Alternative pathway of complement activity and lysozyme concentration was measured in plasma in order to evaluate the general fish health status. Total glutathione, glutathione peroxidase (GPX) and superoxide dismutase (SOD) were analyzed in gills, liver, brain, intestine and muscle. The chemical dispersion induced a significant reduction of lysozyme concentration when compared to the controls, and the hemolytic activity of the alternative complement pathway was increased in mechanical and chemical dispersion. The analysis of SOD, GPX and total glutathione showed that antioxidant defenses were activated in liver and reduced in intestine and brain. Dispersant was also responsible for an SOD activity inhibition in these two last tissues, demonstrating a direct effect of this dispersant on reactive oxygen species homeostasis that can be interpreted as a signal of tissue toxicity. This result should raise concern about the use of dispersants and show that they can lead to adverse effects on marine species.

Effect of dispersed crude oil on cardiac function in seabass Dicentrarchus labrax.

In this study, the impact of dispersed oil was assessed in Dicentrarchus labrax, a fish frequently used as an oil contamination indicator species. Fish were exposed for 48h to (mechanically and chemically) dispersed oil and dispersant alone. The impact of these exposure conditions was assessed on cardiac function by measuring (i) the contraction strength, the contraction and the relaxation speeds (ii) the cardiac energy metabolism using respirometry on permeabilized cardiac fibers. Compared to control, the increase of polycyclic aromatic metabolites observed in the bile indicated oil contamination in our fish. Following 48h of oil exposure at realistic oil concentrations, alterations of cardiac performances were observed. A decrease in contraction strength, contraction and relaxation speeds was observed in the presence of oil without effect of dispersant on these three parameters. Looking at cardiac energy metabolism, dispersant alone decreases all the activity of the respiratory chain and increases the proton leak. From these results, it appears that the observed decrease in cardiac performance in fish exposed to oil was not linked to a decrease in energy availability.

Acute toxicity of chemically and mechanically dispersed crude oil to juvenile sea bass (Dicentrarchus labrax): Absence of synergistic effects between oil and dispersants.

The goal of the present experiment was to assess the relative acute toxicities of mechanically and chemically dispersed oil (crude Arabian Light) in controlled conditions. Juvenile sea bass (Dicentrarchus labrax) were exposed to 4 commercial formulations of dispersants (Corexit EC9500A, Dasic Slickgone NS, Finasol OSR 52, Inipol IP 90), to mechanically dispersed oil, and to the corresponding chemical dispersions. Acute toxicity was evaluated at 24 h, 48 h, 72 h, and 96 h through the determination of 10%, 50%, and 90% lethal concentrations calculated from measured total petroleum hydrocarbon (TPH) concentrations; Kaplan-Meyer mortality analyses were based on nominal concentrations. Animals were exposed to the dissolved fraction of the oil and to the oil droplets (ranging from 14.0 µm to 42.3 µm for the chemical dispersions). Kaplan-Meyer analyses demonstrated an increased mortality in the case of chemical dispersions. This difference can be attributed mainly to differences in TPH, because the chemical lethal concentrations were not reduced compared with mechanical lethal concentrations (except after 24 h of exposure). The ratios of lethal concentrations of mechanical dispersions to the different chemical dispersions were calculated to allow direct comparisons of the relative toxicities of the dispersions. The results ranged from 0.27 to 3.59, with a mean ratio close to 1 (0.92). These results demonstrate an absence of synergistic effect between oil and chemical dispersants in an operational context.

Impact of dispersed fuel oil on cardiac mitochondrial function in polar cod Boreogadus saida.

In this study, impact of dispersed oil on cardiac mitochondrial function was assessed in a key species of Arctic marine ecosystem, the polar cod Boreogadus saida. Mature polar cod were exposed during 48 h to dispersed oil (mechanically and chemically) and dispersants alone. The increase observed in ethoxyresorufin-O-deethylase activity and polycyclic aromatic hydrocarbon metabolites in bile indicated no difference in contamination level between fish exposed to chemical or mechanical dispersion of oil. Oil induced alterations of O2 consumption of permeabilised cardiac fibres showing inhibitions of complexes I and IV of the respiratory chain. Oil did not induce any modification of mitochondrial proton leak. Dispersants did not induce alteration of mitochondrial activity and did not increase oil toxicity. These data suggest that oil exposure may limit the fitness of polar cod and consequently could lead to major disruption in the energy flow of polar ecosystem.

Effects of oil exposure and dispersant use upon environmental adaptation performance and fitness in the European sea bass, Dicentrarchus labrax.

The worldwide increasing recourse to chemical dispersants to deal with oil spills in marine coastal ecosystems is a controversial issue. Yet, there exists no adequate methodology that can provide reliable predictions of how oil and dispersant-treated oil can affect relevant organism or population-level performance. The primary objective of the present study was to examine and compare the effects of exposure to untreated oil (weathered Arabian light crude oil), chemically dispersed oil (Finasol, TOTAL-Fluides) or dispersant alone, upon the ability of fish for environmental adaptation. To reach that goal, we implemented high-throughput, non-lethal challenge tests to estimate individual hypoxia and heat tolerance as surrogate measures of their capacity to face natural contingencies. Experimental populations were then transferred into semi-natural tidal ponds and correlates of individuals' fitness (growth and survival) were monitored over a period of 6 months. In accordance with our stated objectives, the contamination conditions tested corresponded to those observed under an oil slick drifting in shallow waters. Our results revealed that the response of control fish to both challenges was variable among individuals and temporally stable (repeatable) over a 2-month period. Exposure to chemical dispersant did not affect the repeatability of fish performance. However, exposure to oil or to a mixture of oil plus dispersant affected the repeatability of individuals' responses to the experimental challenge tests. At population level, no difference between contamination treatments was observed in the distribution of individual responses to the hypoxia and temperature challenge tests. Moreover, no correlation between hypoxia tolerance and heat tolerance was noticed. During the field experiment, hypoxia tolerance and heat tolerance were found to be determinants of survivorship. Moreover, experimental groups exposed to oil or to dispersant-treated oil displayed significantly lower survival than control or dispersant-exposed groups. Finally, from the four experimental populations tested, the one exposed to chemically dispersed oil presented the lowest growth rate.