Michèle Roméo

Heavy metal distribution in different fish species from the Mauritania coast.

Cadmium, copper, zinc and mercury concentrations were determined in pelagic and benthic fishes from the Mauritania coast. The pelagic fishes consisted of the round sardinella Sardinella aurita (Clupeidae), the chub mackerel Scomber japonicus (Scombridae) and the Atlantic horse mackerel Trachurus trachurus (Carangidae). Four benthic species were considered, three Serranidae: the painted comber Serranus scriba, the golden grouper Epinephelus costae, and the Niger hind Cephalopholis nigri and one Mullidae: the West African goatfish Pseudupeneus prayensis. Cadmium, copper, zinc and mercury concentrations are low in the edible muscles of the pelagic species. In the benthic fishes, metals could be determined also in the gills and liver. Cadmium and copper were present in relatively low levels in the muscle (< or = 0.06 microgram Cd g-1 dry wt., < or = 1.6 micrograms Cu g-1 dry wt.) and gills (< or = 0.23 microgram Cd g-1, < or = 3.1 micrograms Cu g-1) and higher levels are encountered in the livers (< or = 51 micrograms Cd g-1, < or = 49.1 micrograms Cu g-1). Zinc concentrations in the muscle are low (< or = 20 micrograms Zn g-1 dry wt.), the concentrations in the gills (< or = 120 micrograms Zn g-1) approach those of the livers for C. nigri and P. prayensis or are higher in the case of S. scriba and E. costae. Mercury concentrations in the gills and muscle of the pelagic species are very low, higher quantities are found in the livers of the benthic species. The metal concentrations in all the fishes analysed, which are restricted to approximately 40 specimens, are low except those found for cadmium in the livers of the benthic fishes.

Cadmium and copper display different responses towards oxidative stress in the kidney of the sea bass Dicentrarchus labrax

Copper and cadmium were i.p. injected into the fish Dicentrarchus labrax. Cu as Cd-treated fish showed an enlargement of the lysosomal membrane of the kidney (pronephros), Cu being more toxic than Cd. Following injection, metal uptake, measured in muscle, liver and kidney, was much higher with cadmium treatment than with copper, the kidney being the main accumulating organ of cadmium in which metal accumulation is correlated with increased zinc level, suggesting metallothionein induction. In vivo exposure to metal decreases the lysosomal membrane stability of pronephros with a half dose of 127 ng g(-1) Cu and 735 ng g(-1) Cd. Lipid peroxidation, expressed as malondialdehyde equivalents (MDA), and catalase activity were measured in kidney subcellular fractions. When added in vitro, Cu significantly raises the MDA level (365% at 200 µM), Cd having a lower effect (20% at 500 µM). Catalase activity is significantly reduced by Cd whereas Cu does not produce any significant effect at the tested concentrations. Results suggest that although both metals cause in vivo damage to pronephros lysosomal membrane, Cu activates the redox process generating oxyradicals but does not affect in vitro the protective catalase activity unlike Cd which appears to weakly participate in oxyradical generation but alters in vitro protective catalase activity.

Mussel transplantation and biomarkers as useful tools for assessing water quality in the NW Mediterranean.

Mussels, coming from an aquaculture farm located in a clean open bay, were transplanted to several stations of the bays of Nice and Cannes (NW Mediterranean) including a reference site for one month at three periods. Several biomarkers: activities of glutathione S-transferase (GST; exposure to organics), of catalase (exposure to oxidative stress) and of acetylcholinesterase (inhibited by some pesticides) and the lipid peroxidation (thiobarbituric acid reactive substances: TBARS) were measured in transplanted mussels. Cd, Cu and Zn concentrations were also measured as well as their condition index. The results demonstrated some seasonal variations in GST and catalase activities with higher levels in June compared to October. The condition index was also higher in June than in October. Principal component analyses performed with the whole set of data allowed to separate stations or groups of stations according to their responses. The mussels from the harbour of Nice were characterized by high TBARS levels and catalase activity in October 1999 whereas in the harbour of Cannes, animals presented very high copper concentrations and GST activities in June 2000. At the reference site, mussels generally presented low enzymatic activities (except AChE activity) and peroxidation levels and low heavy metal concentrations.

The use of acetylcholinesterase activity in Ruditapes decussatus and Mytilus galloprovincialis in the biomonitoring of Bizerta lagoon.

The variations of acetylcholinesterase activity, considered to be a biochemical biomarker of organophosphorus and carbamate compounds, was followed in Mediterranean clams (Ruditapes decussatus) and mussels (Mytilus galloprovincialis) as a function of season and sampling sites in the lagoon of Bizerta (Tunisia). Results show that acetylcholinesterase activity in clams collected from station A was the lowest and was so throughout the year. This low activity may be due to the input of non-treated waste waters in the lagoon. Heavy metal contamination has been reported in the sediments of this area, suggesting that acetylcholinesterase activity in clams may be inhibited by this type of pollution. Clams harvested from station F, a site adjacent to an agricultural area, were characterized by a decrease in acetylcholinesterase activity, especially after rain periods. Clams from station J presented the highest activity throughout the greater part of the year. This phenomenon may imply that this location is less affected by pollutants inhibiting AChE activity than the others. In the mussels from station C, located in the channel from the lagoon to the Mediterranean and submitted to pollutants (urban wastes from Bizerta and hydrocarbons from the maritime traffic), acetylcholinesterase activities were lower than in those from station J. In conclusion, the variations in acetylcholinesterase activity observed between stations in both species may be the result of pollution and of the environmental conditions.

Evaluation of biomarkers in caged fishes and mussels to assess the quality of waters in a bay of the NW Mediterranean Sea

Specimens of sea bass (Dicentrarchus labrax) were placed in cages for 1 month in spring and autumn at different locations in the Bay of Cannes (NW Mediterranean). Biochemical markers evaluated were: ethoxyresorufin O-deethylase (EROD) and glutathione S-transferase (GST) activities in fish livers and acetylcholinesterase (AChE) activity in fish muscle. EROD and GST activities were higher in front of the outlet for the wastewater plant of Cannes and in the harbour than outside the marina. High EROD and GST activities may be induced by petrol hydrocarbons and/or polychlorinated biphenyls (PCBs). AChE was low in the muscles of the fish caged in the harbour compared with samples from the other cages. Low AChE activity could suggest the presence of organophosphorus and carbamate compounds in seawater from the harbour. Mussels (Mytilus galloprovincialis) were caged off Cannes for the same periods as the fishes. Heavy metal, metallothionein (MT) concentrations and lysosomal membrane stability were evaluated in the digestive gland of the mussels. Results show low heavy metal and MT concentrations, implying low metal concentrations in the surrounding waters. High lysosomal membrane stability revealed a good physiological status of these animals after caging. The whole set of data indicates that seawater in the Bay of Cannes appeared to be unpolluted as regards pollutants which may induce the measured biomarkers, except in restricted areas.

Biomarkers in Ruditapes decussatus: a potential bioindicator species

The clam Ruditapes decussatus is distributed worldwide and due to its ecological and economical interest has been proposed as a bioindicator in areas where mussels are not available. The accumulation of several anthropogenic compounds in their tissues suggests that they possess mechanisms that allow them to cope with the toxic effects of these contaminants. Besides pollutant uptake, the use of biomarkers is pointed out in this paper since it is a promising approach to monitor the effect of these contaminants in the marine environment. Biomarkers complement the information of the direct chemical characterization of different types of contaminants. Therefore, the aim of this paper is to review the role of several biomarkers: (metallothioneins (MT), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidases (GPx) (total and selenium-dependent), lipid peroxidation (measured as MDA, one of the final products of lipid peroxidation), glutathione S-transferase (GST) and acetylcholinesterase (AChE), measured in different tissues of the clam R. decussatus, in laboratory conditions and under various environmental stresses, in two ecosystems (Ria Formosa lagoon- Portugal) and Bizerta lagoon (Tunisia) in a perspective of a multibiomarker approach to assess environmental changes. Experiment and field studies are in good agreement since MT levels, especially in the gills, the first target tissue of these contaminants, can be used as biomarker of exposure to Cd. GPx and MDA may also be determined in this respect. AChE activity is inhibited by pesticide and, to a less extent, by metal exposure in the gills and whole soft body of clams. However, the induction of GST isoforms experimentally demonstrated is not observed in the field because only global GST activity was determined. The whole set of results opens new research perspectives for the use of this species to assess the effect of mixtures of pollutants in the aquatic environment.

Distribution of mercury in the organs and tissues of five toothed-whale species of the Mediterranean

Mercury levels were determined in the tissues and organs (lung, liver, kidney, skin, muscle, bone) of five toothed-whales stranded along the Corsican coast between November 1993 and February 1996. The species taken into consideration were the bottlenose dolphin Tursiops truncatus, the common dolphin Delphinus delphis, the striped dolphin Stenella coeruleoalba, the pilot whale Globicephala melas and the Rissos dolphin Grampus griseus. The variation in mercury levels between the different tissues and organs (lung, liver, kidney, skin, muscle, bone) of the cetacean species are discussed as regards storage, biotransformation and elimination. In all cases, the liver appears to be the preferential organ for mercury accumulation (with concentrations as high as 4250 microg Hg/g dw and 3298 microg Hg/g in the livers of Tursiops truncatus and Grampus griseus, respectively). The kidney and lung are the next organs in terms of mercury uptake followed by the muscle, bone and skin. The stomach contents of Grampus griseus and D. delphis were determined and consisted of cephalopods for Grampus griseus, and of sardines Sardina pilchardus and mackerels Trachurus sp. for D. delphis. Cephalopods had higher mercury concentrations (25.4 microg Hg/g dw) than fish (ca 1 microg Hg/g). These contents represent only one meal and mercury levels found in livers may integrate mercury uptake having occurred during the whole life span of animals.

Effect of cadmium in the clam Ruditapes decussatus assessed by proteomic analysis.

Cadmium, an environmental stressor due to its toxicity, persistence and accumulation in biota, is widespread in the aquatic environment. Cadmium accumulation kinetics have revealed that Ruditapes decussatus has a high affinity to this metal. Proteomics is an effective tool to evaluate the toxic effects of contaminants. The aim of this study was to investigate the Cd effects in the gill and digestive gland of the sentinel species R. decussatus. Protein expression profiles (PEPs) in the clam tissues exposed to Cd (40 microg l(-1), 21 days) were compared to unexposed ones. Cd induces major changes in tissue-specific protein expression profiles in gill and digestive gland. This tissue dependent response results mainly from differences in Cd accumulation, protein inhibition and/or autophagy. An overall decrease of protein spots was detected in both treated tissues, being higher in gill. Some of the spots more drastically altered after pollutants exposure were excised and nine were identified by micro liquid chromatography tandem mass spectrometry (LC-MS/MS). Proteins identified by homology search in databases included: three proteins (8-fold) up-regulated, one down-regulated, four suppressed and one induced. Cd induces major changes in proteins involved in cytoskeletal structure maintenance (muscle-type actin, adductor muscle actin and beta-tubulin), cell maintenance (Rab GDP) and metabolism (ALDH and MCAD, both identified by de novo sequencing) suggesting potential energetic change. They provide a valuable knowledge of Cd effects at biochemical and molecular levels in the gill and digestive gland of R. decussatus.

Metal distribution in different tissues and in subcellular fractions of the Mediterranean clam Ruditapes decussatus treated with cadmium, copper, or zinc

Kinetics of accumulation of Cd (500 μg/l), Cu (150 μg/l) and Zn (1000 μg/l) were measured in organs (gills, digestive gland and remainder) of the clam Ruditapes decussatus. A significant accumulation was observed in the organs of animals treated with Cd or Cu, whereas no difference was obvious between controls and animals exposed to Zn. A slight increase was noted only in the gills of Zn-exposed animals. After subcellular fractionation, a major part of Cu and Cd found in the gills and in the digestive gland was associated with the cytosol, whereas zinc was mainly bound to the particulate fraction. Metallothionein (MT) concentrations were higher in the gills of Cu-treated animals than in those of controls or cadmium-exposed clams. As regards the digestive gland, no difference was found in MT concentrations of controls or of animals treated with Cu or Cd.

Size dependent bioaccumulation and ecotoxicity of gold nanoparticles in an endobenthic invertebrate: the Tellinid clam Scrobicularia plana.

Gold nanoparticles (AuNPs) have important technological applications resulting in an increased potential for release to the environment, and a greater possibility of toxicological effects. The marine bivalve Scrobicularia plana was exposed to AuNPs of size 5, 15 and 40 nm during a 16 d laboratory exposure at 100 μg Au L(-1). After exposure to AuNPs forming aggregates (>700 nm), the clams accumulated Au in their soft tissues. Biochemical (biomarkers) and behavioral (burrowing and feeding) responses were investigated. Au NPs were responsible of metallothionein induction (5, 40 nm), increased activities of catalase (15, 40 nm) and superoxide dismutase (40 nm) and of glutathione S-transferase by the three sizes of AuNPs indicating defense against oxidative stress. Exposure to AuNPs impaired burrowing behavior. However, it must be underlined that these effects were observed at a dose much higher than expected in the environment.