Use of skin and blubber tissues of small cetaceans to assess the trace element content of internal organs.

In order to evaluate the use of biopsy samples as non-destructive tool for assessing trace element concentrations in small cetaceans, the concentrations of 14 trace elements were determined in skin, blubber, liver and kidneys of four species of small cetaceans (i.e. common dolphin Delphinus delphis, harbour porpoise Phocoena phocoena, bottlenose dolphin Tursiops truncatus and striped dolphin Stenella coeruleolba), stranded and/or by-caught along the NE Atlantic Ocean coast between 2001 and 2008. Only Cu, Fe, Hg, Mn, Ni and Zn were above the detection limit of the instruments and showed recoveries satisfactory enough to be interpreted. Among these trace elements, Hg was the only one showing a significant correlation between concentrations in and those in liver and kidneys. In consequence skin and blubber can only be used as non-invasive monitoring tissues to investigate Hg bioaccumulation in internal tissues for cetacean populations.

Bioaccumulation of persistent organic pollutants in female common dolphins (Delphinus delphis) and harbour porpoises (Phocoena phocoena) from western European seas: geographical trends, causal factors and effects on reproduction and mortality.

Concentrations of polychlorinated biphenyls (PCBs) in blubber of female common dolphins and harbour porpoises from the Atlantic coast of Europe were frequently above the threshold at which effects on reproduction could be expected, in 40% and 47% of cases respectively. This rose to 74% for porpoises from the southern North Sea. PCB concentrations were also high in southern North Sea fish. The average pregnancy rate recorded in porpoises (42%) in the study area was lower than in the western Atlantic but that in common dolphins (25%) was similar to that of the western Atlantic population. Porpoises that died from disease or parasitic infection had higher concentrations of persistent organic pollutants (POPs) than animals dying from other causes. Few of the common dolphins sampled had died from disease or parasitic infection. POP profiles in common dolphin blubber were related to individual feeding history while those in porpoises were more strongly related to condition.

Trace element levels in foetus-mother pairs of short-beaked common dolphins (Delphinus delphis) stranded along the French coasts.

Tissues of foetus-mother pairs of common dolphins (Delphinus delphis) stranded along the French coasts (Bay of Biscay and English Channel) were analysed for their Cd, Cu, Hg, Se and Zn contents. In the kidneys, foetal Cd levels were extremely low, and strong relationships between Cu and Zn suggested the involvement of metallothioneins since early foetal life. The results also indicated a limited maternal transfer of Hg during pregnancy since levels in the tissues of foetuses were below 1 microg g(-1) w.wt. However, hepatic Hg levels in foetuses increased with body length, and were also proportionate to maternal hepatic, renal and muscular Hg levels. Lastly, affinities between Hg and Se in tissues would participate in Hg neutralisation in both mothers--through tiemannite granules--and fetuses--through reduced glutathione--counteracting the toxic effects linked to the particularly high quantities of methyl-Hg to which marine mammals are naturally exposed.

Biological and ecological factors related to trace element levels in harbour porpoises (Phocoena phocoena) from European waters.

Selected trace elements (Cd, Cu, Hg, Se, Zn) were measured in the kidneys and the liver of 104 harbour porpoises (Phocoena phocoena) stranded along the coasts of France, Galicia (Spain), Ireland, Scotland (UK), and the Netherlands. Generally, relatively low concentrations of toxic elements were encountered in the tissues of European porpoises, except for two individuals, which displayed high hepatic Hg concentrations. Also, elevated Cd levels obtained in Scottish porpoises could be related to their feeding preferences and this result suggests an increase of the proportion of cephalopods in their diet with latitude. Moreover, significant geographical differences were seen in hepatic Zn concentrations; the elevated Zn concentrations displayed by porpoises from the Netherlands may relate their poor health status. Variation in metal concentrations within porpoises from the North Sea is likely to reflect a long-term segregation between animals from northern (Scotland) and southern areas (the Netherlands), making trace elements powerful ecological tracers.

Lead contamination of small cetaceans in European waters--the use of stable isotopes for identifying the sources of lead exposure.

Lead concentrations and isotopic composition have been measured in bone and teeth of small cetaceans belonging to three species (Delphinus delphis, Phocoena phocoena and Stenella coeruleoalba), to evaluate the toxicological risk and to determine sources of lead in the European waters. Lead concentrations, far lower than threshold value inducing toxic effects in human, were higher in teeth than in bones, but highly correlated between the two tissues (r=0.92, p<0.001). Large variations of 206Pb/207Pb values in bone tissue showed that cetaceans must be submitted to various atmospheric influences. No geographical differences appeared which is consistent with studies on their distribution indicating seasonal movements between Brittany waters and the Bay of Biscay. The negative correlation between 206Pb/207Pb ratios and age of the individuals reflected the decrease in the production of alkyl lead in Europe, i.e., the increasing use of unleaded gasoline.

Total and organic Hg concentrations in cephalopods from the North Eastern Atlantic waters: influence of geographical origin and feeding ecology.

Total (T-Hg) and organic (O-Hg) mercury concentrations and tissue distribution were examined in 20 species of cephalopods (n=278) from the north eastern Atlantic waters, i.e. from the Bay of Biscay to the Faroe Islands. Concentrations of T-Hg in whole cephalopods showed elevated variations among species, i.e. from 40 to 3560ng g(-1) dwt, but a low variability within each species (mean CV%=39%). With the exception of oceanic squids, the digestive gland globally displayed higher T-Hg concentrations than the remaining tissues. In contrast, O-Hg concentrations determined in selected species were generally higher in the remaining tissues. Despite higher T-Hg concentrations, the digestive gland weakly contributed to the total body burden of both T-Hg and O-Hg (<25% and <15%, respectively). In fact, from 75% to 95% of the T-Hg and O-Hg were contained in the muscular remaining tissues. Therefore, O-Hg may have a strong affinity to proteins in cephalopods. Sex and size only significantly influenced the bioaccumulation of Hg for the Loliginidae family. T-Hg and O-Hg concentrations were also influenced by geographical origin: Celtic Sea>Bay of Biscay>Faroe Islands, corresponding to the seawater Hg concentrations in these areas. In the Faroe Islands and the Celtic Sea, benthic cephalopods contained significant higher Hg concentrations compared to pelagic ones. This suggests that diet is not the main pathway of Hg uptake in cephalopods as pelagic species were expected to be more exposed to O-Hg through fish consumption than benthic ones.

New insights from age determination on toxic element accumulation in striped and bottlenose dolphins from Atlantic and Mediterranean waters.

This study aimed at comparing toxic element (Hg, Cd) bioaccumulation in relation to age for bottlenose (Tursiops truncatus) and striped dolphins (Stenella coeruleoalba) from Mediterranean and Atlantic waters. Metal concentrations were also measured in selected prey to infer metal exposure through the diet. As expected, Mediterranean prey exhibited the highest Hg levels, probably as a consequence of the Hg enrichment of the Mediterranean Sea. Comparing the predators from each area and taking age into account, Mediterranean bottlenose dolphins displayed higher Hg levels than Atlantic dolphins (p = 0.032), whereas Mediterranean striped dolphins did not (p = 0.691). The consumption of Myctophid fish, which showed the highest Hg levels (105+/-80 ngg(-1) w.wt.) among Atlantic prey, may explain the high Hg levels in the liver of the Atlantic striped dolphins and suggested a preferential offshore feeding behaviour in this area. Concerning Cd, no clear differences were found between geographical areas.

Trace element (Cd, Cu, Hg, Se, Zn) accumulation and tissue distribution in loggerhead turtles (Caretta caretta) from the Western Mediterranean Sea (southern Italy).

Cadmium (Cd), copper (Cu), mercury (Hg), selenium (Se) and zinc (Zn) were determined in the liver, kidney and muscle of 29 loggerhead turtles, Caretta caretta, from the South Tyrrhenian Sea (Western Mediterranean). No significant differences (p>0.05) were detected between males and females. Trace element concentrations were not influenced by the size of the specimen except Se in the liver, which was negatively correlated with the curved carapace length (p<0.001). Muscles generally displayed the lowest trace element burdens, with the exception of Zn which contained concentrations as high as 176 microgg-1dwt. Kidneys displayed the highest Cd and Se mean concentrations (57.2+/-34.6 and 15.5+/-9.1 microgg-1dwt, respectively), while liver exhibited the highest Cu and Hg levels (37.3+/-8.7 and 1.1+/-1.7 microgg-1dwt, respectively). Whichever tissue is considered, the toxic elements had elevated coefficients of variation (i.e. from 60% to 177%) compared to those of the essential ones (i.e. from 14% to 65%), which is a consequence of homeostatic processes for Cu, Se and Zn. Globally, the concentrations of Hg remained low in all the considered tissues, possibly the result of low trophic level in sea turtles. In contrast, the diet of loggerhead turtles would result in a significant exposure to Cd. Highly significant correlations between Cd and Cu and Zn in the liver and kidney suggest that efficient detoxification processes involving MT occur which prevent Cd toxicity in loggerhead turtles.

Distribution of trace elements in the tissues of benthic and pelagic fish from the Kerguelen Islands.

New information on the concentrations of Cd, Cu, Hg and Zn in the liver, kidney and muscles of eight marine benthic and pelagic sub-Antarctic fish species are presented to determine the importance of these metals in the marine systems of the Kerguelen Islands. Compared to the reported metal concentrations in other Antarctic fish species, the present results are globally within the same range of concentrations, although Cd displayed a very high interspecific variability in liver and kidney. Indeed, the highest Cd concentrations in liver, ranging from 10.0 to 52.1 microg x g(-1) dry wt. but also the lowest Cd concentrations in muscles (<0.030 microg x g(-1) dry wt.) have been displayed by the pelagic Myctophidae Gymnoscopelus piabilis. Metal concentrations differences might be related to diet and feeding habits of benthic and pelagic fish species. However, Cd and Hg concentrations in the edible muscle are lower than the French limit values (

Trace elements in two odontocete species (Kogia breviceps and Globicephala macrorhynchus) stranded in New Caledonia (South Pacific).

Liver, muscle and blubber tissues of two short-finned pilot whales (Globicephala macrorhynchus) and two pygmy sperm whales(Kogia breviceps) stranded on the coast of New Caledonia have been analysed for 12 trace elements (Al, Cd, Co, Cr, Cu. Fe, organic and total Hg, Mn, Ni, Se, V, and Zn). Liver was shown to be the most important accumulating organ for Cd, Cu, Fe, Hg, Se, and Zn in both species, G. macrorhynchus having the highest Cd, Hg, Se and Zn levels. In this species, concentrations of total Hg are particularly elevated, reaching up to 1452 microg g(-1) dry wt. Only a very low percentage of the total Hg was organic. In both species,the levels of Hg are directly related to Se in liver. Thus, a molar ratio of Hg:Se close to 1.0 was found for all specimens, except for the youngest K. breviceps. Our results suggest that G. macrorhynchus have a physiology promoting the accumulation of high levels of naturally occurring toxic elements. Furthermore, concentrations of Ni, Cr and Co are close to or below the detection limit in the liver and muscles of all specimens. This suggests that mining activity in New Caledonia, which typically elevates the levels of these contaminants in the marine environment, does not seem to be a significant source of contamination for these pelagic marine mammals.

Cadmium detoxification processes in the digestive gland of cephalopods in relation to accumulated cadmium concentrations.

The high concentrations of cadmium recorded in the digestive gland of cephalopods from various temperate and subpolar waters suggest that these molluscs have developed efficient cadmium detoxification mechanisms. The subcellular distribution of cadmium in the digestive gland cells was investigated in seven cephalopod species from the Bay of Biscay (France) and the Faroe Islands. In most species, cadmium was mainly found in the cytosolic fraction of the digestive gland cells, reaching up to 86% of the total cadmium for the squid Loligo vulgaris from the Bay of Biscay. But species with the highest total level of cadmium showed a higher percentage of cadmium associated to insoluble compounds. The quantification of metallothioneins (MTs) by the polarographic method was performed in order to evaluate the involvement of these proteins in the detoxification of the high amounts of bioaccumulated cadmium. Metallothionein levels in cephalopods ranged form 742 +/- 270 to 3478 +/- 1572 microg/g wet weight. No relationship could be established between total cadmium, cytosolic cadmium and MT levels suggesting the occurrence of other Cd-binding ligands. Although these proteins have not been characterised, as cadmium in the digestive gland of cephalopods is mainly associated with soluble ligands, a high potential transfer to predators can be predicted.

Cadmium-containing granules in kidney tissue of the Atlantic white-sided dolphin (Lagenorhyncus acutus) off the Faroe Islands.

Top predators from the northern sub-polar and polar areas exhibit high cadmium concentrations in their tissues. In the aim to reveal possible adverse effects, samples of five Atlantic white-sided dolphins Lagenorhyncus acutus have been collected on the occasion of the drive fishery in the Faroe Islands, for ultrastructural investigations and energy dispersive X-ray microanalyses. Cadmium concentrations were less than the limit of detection in both immature individuals and ranged from 22.7 to 31.1 microg x g(-1) wet weight in the mature individuals. Two individuals with the highest cadmium concentrations exhibited electron dense mineral concretions in the basal membranes of the proximal tubules. They are spherocrystals made up of numerous strata mineral deposit of calcium and phosphorus together with cadmium. Cadmium has been detected with a molar ratio of Ca:Cd of 10:1 in the middle of these concretions. To our knowledge, this is the first report of such granules in a wild vertebrate. The role of these granules in the detoxification of the metal and the possible pathological effects are considered.

Cephalopods as a vector for the transfer of cadmium to top marine predators in the north-east Atlantic Ocean.

Three hundred and fifty individuals of 12 species of cephalopods which differed in their feeding habitats were sampled from the French Atlantic coast to the sub-Arctic region (Bay of Biscay, English Channel, west Irish coast and Faroe Islands) and analysed for their cadmium contents. Comparison of the Cd levels of the cephalopods showed that those from the sub-Arctic area contained very high Cd concentrations compared to those from lower latitudes such as along the French Atlantic coast. High Cd levels in cephalopods from the sub-Arctic zone correspond closely to the reported high Cd concentrations in the tissues of top vertebrate predators from the same area. Comparison of the weekly Cd intakes for the Faroe Island pilot whales with the 'Provisional Tolerable Weekly Intake' for humans recommended by the World Health Organisation, showed that top vertebrate predators are often subjected to Cd doses far in excess of those recommended for humans. Our limited survey results suggest that cephalopods constitute an important source of Cd for cephalopod predators, and that this bioaccumulation effect is most evident at high latitudes.

Mercury in pilot whales: possible limits to the detoxification process.

The ability of pilot whales (Globicephala melas) to concentrate cadmium and mercury is well established. The levels of these metals were generally higher than those encountered in other species of marine mammals. The biological data have not revealed a major toxic problem in the population, and this suggests a remarkable tolerance of this species to heavy metals. Cellular distribution of mercury was carried out in liver samples. The presence of metallothionein-like proteins in the soluble fraction has been demonstrated, but 95% of mercury was mainly bound to the insoluble fraction, showing that these proteins had no role in this metal detoxification. The molar ratio between mercury and selenium suggests that the major mechanism of detoxification is through the formation of a complex Hg-Se which leads to the demethylation of mercury. The site of this process is the liver in which mercury mainly appeared as inorganic, whereas in the muscle the percentage of organic to total mercury was much higher. Nevertheless, this detoxification is limited in lactating females and in all the individuals of one school. This could be the result of changes in the diet and could constitute a toxicological risk for the species.

Organomercury determination in biological reference materials: application to a study on mercury speciation in marine mammals off the Faröe Islands.

The potential use of graphite furnace atomic absorption spectrometry (GF-AAS) for the organic mercury determination in marine biological tissues was evaluated. Following its isolation by acid extraction in toluene, organic mercury was recovered in aqueous thiosulfate and measured by GF-AAS. The detection limit was 0.01 microgram Hg/g (as methyl mercury). Analyses were conducted on three reference standard materials certified for their methyl mercury content, DOLT-1, DORM-1, and TORT-1, provided by the National Research Council of Canada. The method resulted in very good recovery and reproducibility, indicating that GF-AAS can provide results comparable to those obtained by using more expensive and time consuming analytical techniques. The method was applied to the analysis of liver tissues of pilot whale specimens (Globicephala melas) from the drive fishery of the Faröe Islands (northeast Atlantic). The results provided useful information on the proportion of different mercury forms in the liver of these marine mammals.