Thomas Milinkovitch

Liver antioxidant and plasma immune responses in juvenile golden grey mullet (Liza aurata) exposed to dispersed crude oil

Dispersants are often used after oil spills. To evaluate the environmental cost of this operation in nearshore habitats, the experimental approach conducted in this study exposed juvenile golden grey mullets (Liza aurata) for 48 h to chemically dispersed oil (simulating, in vivo, dispersant application), to dispersant alone in seawater (as an internal control of chemically dispersed oil), to mechanically dispersed oil (simulating, in vivo, natural dispersion), to the water-soluble fraction of oil (simulating, in vivo, an oil slick confinement response technique) and to seawater alone (control condition). Biomarkers such as fluorescence of biliary polycyclic aromatic hydrocarbon (PAH) metabolites, total glutathione liver content, EROD (7-ethoxy-resorufin-O-deethylase) activity, liver antioxidant enzyme activities, liver lipid peroxidation and an innate immune parameter (haemolytic activity of the alternative complement pathway) were measured to assess the toxicity of dispersant application. Significant responses of PAH metabolites and total glutathione content of liver to chemically dispersed oil were found, when compared to water-soluble fraction of oil. As was suggested in other studies, these results highlight that priority must be given to oil slick confinement instead of dispersant application. However, since the same patterns of biomarker responses were observed for both chemically and mechanically dispersed oil, the results also suggest that dispersant application is no more toxic than the natural dispersion occurring in nearshore areas (due to, e.g. waves). The results of this study must, nevertheless, be interpreted cautiously since other components of nearshore habitats must be considered to establish a framework for dispersant use in nearshore areas.

Effects of dispersed oil exposure on the bioaccumulation of polycyclic aromatic hydrocarbons and the mortality of juvenile Liza ramada

Dispersing an oil slick is considered to be an effective response to offshore oil spills. However, in nearshore areas, dispersant application is a controversial countermeasure: environmental benefits are counteracted by the toxicity of dispersant use. In our study, the actual toxicity of the dispersant response technique in the nearshore areas was evaluated through an experimental approach using juvenile Liza ramada. Fish were contaminated via the water column (i) by chemically dispersed oil, simulating dispersant application, (ii) by dispersant, as an internal control of chemical dispersion, (iii) by mechanically dispersed oil, simulating only the effect of natural mixing processes, without dispersant application, and (iv) by the water soluble fraction of oil, simulating the toxicity of an oil slick before recovery. Bioconcentrations of polycyclic aromatic hydrocarbons (PAH) and mortality were evaluated, and related to both total petroleum hydrocarbon (TPH) and polycyclic aromatic hydrocarbon (PAH) concentrations in seawater. Fish exposed to chemically dispersed oil showed both a higher bioconcentration of PAH and a higher mortality than fish exposed to either the water soluble fraction of oil or the mechanically dispersed oil. These results suggest that (i) dispersion is a more toxic response technique than containment and recovery of the oil slick; (ii) in turbulent mixing areas, dispersant application increases the environmental risk for aquatic organisms living in the water column. Even if the experimental aspects of this study compel us to be cautious with our conclusions, responders could consider these results to establish a framework for dispersant use in nearshore areas.

Toxicity of dispersant application: Biomarkers responses in gills of juvenile golden grey mullet (Liza aurata)

Dispersant use in nearshore areas is likely to increase the exposure of aquatic organisms to petroleum. To measure the toxicity of this controversial response technique, golden grey mullets (Liza aurata) were exposed to mechanically dispersed oil, chemically dispersed oil, dispersant alone in seawater, water-soluble fraction of oil and to seawater as a control treatment. Several biomarkers were assessed in the gills (enzymatic antioxidant activities, glutathione content, lipid peroxidation) and in the gallbladder (polycylic aromatic hydrocarbons metabolites). The significant differences between chemically dispersed oil and water soluble fraction of oil highlight the environmental risk to disperse an oil slick when containment and recovery can be conducted. The lack of significance between chemically and mechanically dispersed oil suggests that dispersant application is no more toxic than the natural dispersion of the oil slick. The results of this study are of interest in order to establish dispersant use policies in nearshore areas.

Toxicological effects of crude oil and oil dispersant: biomarkers in the heart of the juvenile golden grey mullet (Liza aurata).

Dispersant use is a controversial oil spill response technique in coastal areas. Using an experimental approach, this study evaluated the toxicity of dispersant use upon juveniles of golden grey mullet (Liza aurata). Fish were exposed for 48 h to either dispersant only, chemically dispersed oil, mechanically dispersed oil, the water soluble fraction of oil or to control conditions. Following exposure and a depuration period, biomarkers were assessed in fish hearts, namely the total glutathione content and the activity of four enzymes (glutathione S-transferase, superoxide dismutase, catalase and glutathione peroxides). Comparing biomarker responses between the different treatments, this study revealed that 48 h exposure to dispersed oil (whether mechanically or chemically dispersed) resulted in a toxicity that was still detectable after a 14 days depuration period. Comparing biomarkers responses after an exposure to chemically and mechanically dispersed oil, this study suggests that chemical dispersion of the oil slick would not be more toxic than its natural dispersion under certain turbulent meteorological conditions (e.g. waves). Furthermore, the results indicated that the heart could be a target organ of interest in further studies investigating the toxicity of hydrocarbons. This study, which has been integrated into the DISCOBIOL project (Dispersant et techniques de lutte en milieu côtier: effets biologiques et apport à la réglementation), presents information of interest when attempting to provide a framework for dispersant applications in coastal areas.

Effect of dispersed crude oil exposure upon the aerobic metabolic scope in juvenile golden grey mullet (Liza aurata)

This study evaluated the toxicity of dispersant application which is, in nearshore area, a controversial response technique to oil spill. Through an experimental approach with juveniles of Liza aurata, the toxicity of five exposure conditions was evaluated: (i) a chemically dispersed oil simulating dispersant application; (ii) a single dispersant as an internal control of chemically dispersed oil; (iii) a mechanically dispersed oil simulating natural dispersion of oil; (iv) a water soluble fraction of oil simulating an undispersed and untreated oil slick and (v) uncontaminated seawater as a control exposure condition. The relative concentration of PAHs (polycyclic aromatic hydrocarbons) biliary metabolites showed that the incorporation of these toxic compounds was increased if the oil was dispersed, whether mechanically or chemically. However, toxicity was not observed at the organism level since the aerobic metabolic scope and the critical swimming speed of exposed fish were not impaired.

Trace metal concentrations in post-hatching cuttlefish Sepia officinalis and consequences of dissolved zinc exposure

In this study, we investigated the changes of 13 trace metal and metalloid concentrations (i.e. Ag, As, Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb, Se, V, Zn) and their subcellular fractionation in juvenile cuttlefish Sepia officinalis reared in controlled conditions between hatching and 2 months post-hatching. In parallel, metallothionein concentrations were determined. Our results highlighted contrasting changes of studied metals. Indeed, As and Fe concentrations measured in hatchlings suggested a maternal transfer of these elements in cuttlefish. The non-essential elements Ag and Cd presented the highest accumulation during our study, correlated with the digestive gland maturation. During the 6 first weeks of study, soluble fractions of most of essential trace metals (i.e. Co, Cr, Cu, Fe, Se, Zn) slowly increased consistently with the progressive needs of cuttlefish metabolism during this period. In order to determine for the first time in a cephalopod how metal concentrations and their subcellular distributions are impacted when the animals are trace metal-exposed, we studied previously described parameters in juveniles exposed to dissolved Zn at environmental (i.e. 50 μg l(-1)) and sublethal (i.e. 200 μg l(-1)) levels. Moreover, oxidative stress (i.e. glutathione S-transferase (GST), superoxide dismutase (SOD) and catalase activities, and lipid peroxidation (LPO)) was assessed in digestive gland and gills after 1 and 2 months exposures. Our results highlighted no or low ability of this stage of life to regulate dissolved Zn accumulation during the studied period, consistently with high sensitivity of this organism. Notably, Zn exposures caused a concentration-dependent Mn depletion in juvenile cuttlefish, and an increase of soluble fraction of Ag, Cd, Cu without accumulation modifications, suggesting substitution of these elements (i.e. Mn, Ag, Cd, Cu) by Zn. In parallel, metallothionein concentrations decreased in individuals most exposed to Zn. Finally, no perturbations in oxidative stress management were detected in gills, whereas modifications of GST, SOD and catalase activity levels were recorded in digestive gland, resulting in an increase of LPO content after a 6-week exposure to low Zn concentration. Altogether, these perturbations are consistent with previously described high sensitivity of juvenile cuttlefish towards Zn. Our results underlined the need to study deeply contamination impact on this animal at this stage of life.

In situ evaluation of oxidative stress and immunological parameters as ecotoxicological biomarkers in a novel sentinel species (Mimachlamys varia)

Although the variegated scallop Mimachlamys varia seems to be a suitable sentinel species for contaminant monitoring, no study has identified biomarkers in this species. In order to fill this gap, this study conducted an in situ biomarker approach. M. varia were collected in contaminated and uncontaminated areas and responsiveness of oxidative stress and immunological biomarkers was evaluated in the digestive gland. In parallel, 14 trace element concentrations were evaluated in the same organ. Superoxide dismutase activity and malondialdehyde content responded efficiently to in situ contamination when a certain degree of contamination was reached. Laccase-type phenoloxidase showed a high sensitivity but saturation of the response was highlighted for the highest contaminations. Additionally, correlations were found between biomarkers and trace element concentrations. Taken together, results showed that biomarker approach conducted in the digestive gland of M. varia represents a sensitive analytical tool to highlight ecotoxicological issues in coastal marine ecosystems.

Biomarker modulation associated with marine diesel contamination in the Iceland scallop (Chlamys islandica)

The decrease of ice cover in the Arctic will lead to an increase of ship traffic in the upcoming decades. Consequently, oil pollution is expected. In this context, the goals of this study were to evaluate the biological impact of marine diesel contamination and, on this basis, to determine analytical tools of interest (biomarkers) for future biomonitoring of diesel spills. Using a 7-day contamination protocol, this study investigated biochemical modulations in the digestive gland of the Iceland scallop (Chlamys islandica). Incorporation of contaminants was verified assessing haemolymph metabolites. Results showed a response of glutathione-S-transferase to contamination suggesting detoxification processes and the suitability of such a tool for diesel spill biomonitoring. The lack of modulation of superoxide dismutase activity and lipid peroxidation suggests no oxidative stress and the unsuitability of these molecular tools for biomonitoring.

First evidence of marine diesel effects on biomarker responses in the Icelandic scallops, Chlamys islandica

The decrease of ice cover in polar areas is expected to lead to an increase in ship traffic. In this context, the risk associated with exposure of the aquatic environment to oil-related chemicals from spills and/or accidental fuel discharges from ships will increase in the near future, potentially causing negative impacts on sensitive Arctic species. This study investigated for the first time the biomarker responses of the Icelandic scallops (Chlamys islandica) to marine diesel exposure. Antioxidant response, neurotoxic effect, gonad maturation, and lipid content were assessed in male and female scallops, Chlamys islandica after a 7-day exposure to marine diesel. At the end of exposure, results showed an inhibition of acetylcholinesterase activity in Icelandic scallops exposed to high concentration of diesel. At the end of exposure, results showed an increase in naphthalene residues in hemolymph, together with an inhibition of AChE activity in Icelandic scallops exposed to high concentration of diesel. In addition, alteration of the escape capacity was observed suggesting effects on behavior responses. Overall, this study contributes to the better knowledge of physiological effects of marine diesel in Arctic marine organisms.

Influence of temperature on age-related lipid peroxidation in a short-lived vertebrate ( Nothobranchius furzeri )

In ectotherms, temperature modulates oxidative stress, a key driver of aging. However, the effects of temperature on oxidative stress have not been investigated at several life stages of an ectotherm. In order to improve understanding of aging processes, we conducted a cross-sectional study in short-lived ectotherm vertebrates, the killifish Nothobranchius furzeri. Malondialdehyde contents were assessed at three stages of life, in the liver and muscles of fish acclimatized to optimal or sub-optimal temperatures during all their life cycle. In accordance with the “free-radical theory,” our results highlighted an increase in lipid peroxidation in senescent organisms. In the liver, this lipid peroxidation increase was more intense in senescent fish acclimatized to sub-optimal temperature than in fish acclimatized to their optimal temperature.