Cristina Fossi

Heavy metals and selenium in stranded dolphins of the Northern Tyrrhenian (NW Mediterranean)

Heavy metal (Hg, Cd, Pb and Zn) and selenium levels were determined in striped (Stenella coeruleoalba) and bottle-nosed dolphins (Tursiops truncatus) stranded along the coast of Tuscany and Latium, Italy in the period 1987-1989. Lead and zinc concentrations were quite low and there was modest accumulation of cadmium in the kidney of both species. Mercury levels were very high, especially in the liver where they reached peaks of 4400 ppm (dry weight) in the striped dolphin and 13,150 ppm (dry weight) in the bottle-nosed dolphin. Selenium levels were also high and were significantly correlated with mercury levels in some organs and tissues. The toxicological significance of the selenium-mercury interaction is discussed.

Selection of reference genes for quantitative RT-PCR studies in striped dolphin (Stenella coeruleoalba) skin biopsies

BackgroundOdontocete cetaceans occupy the top position of the marine food-web and are particularly sensitive to the bioaccumulation of lipophilic contaminants. The effects of environmental pollution on these species are highly debated and various ecotoxicological studies have addressed the impact of xenobiotic compounds on marine mammals, raising conservational concerns. Despite its sensitivity, quantitative real-time PCR (qRT-PCR) has never been used to quantify gene induction caused by exposure of cetaceans to contaminants. A limitation for the application of qRT-PCR is the need for appropriate reference genes which allow the correct quantification of gene expression. A systematic evaluation of potential reference genes in cetacean skin biopsies is presented, in order to validate future qRT-PCR studies aiming at using the expression of selected genes as non-lethal biomarkers.ResultsTen commonly used housekeeping genes (HKGs) were partially sequenced in the striped dolphin (Stenella coeruleoalba) and, for each gene, PCR primer pairs were specifically designed and tested in qRT-PCR assays. The expression of these potential control genes was examined in 30 striped dolphin skin biopsy samples, obtained from specimens sampled in the north-western Mediterranean Sea. The stability of selected control genes was determined using three different specific VBA applets (geNorm, NormFinder and BestKeeper) which produce highly comparable results. Glyceraldehyde-3P-dehydrogenase (GAPDH) and tyrosine 3-monooxygenase (YWHAZ) always rank as the two most stably expressed HKGs according to the analysis with geNorm and Normfinder, and are defined as optimal control genes by BestKepeer. Ribosomal protein L4 (RPL4) and S18 (RPS18) also exhibit a remarkable stability of their expression levels. On the other hand, transferrin receptor (TFRC), phosphoglycerate kinase 1 (PGK1), hypoxanthine ribosyltransferase (HPRT1) and β-2-microglobin (B2M) show variable expression among the studied samples and appear as less suitable reference genes for data normalization.ConclusionIn this work, we have provided essential background information for the selection of control genes in qRT-PCR studies of cetacean skin biopsies, as a molecular technique to investigate ecotoxicological hazard in marine mammals. Of 10 HKGs tested, those encoding for YWHAZ and GAPDH appear as the most reliable control genes for the normalization of qRT-PCR data in the analysis of striped dolphin skin biopsies. Potentially useful reference genes are also those encoding for ribosomal proteins L4 and S18.

Persistent organochlorine residues in fish and water birds from the Biobio River, Chile

Concentrations of polychlorinated biphenyls (PCBs), DDT and its metabolites, HCH isomers and hexachlorobenzene (HCB) were determined in fish and birds from different locations in the Biobio river basin (central Chile). Samples collected near the mouth of the river contained high concentrations of PCBs, reflecting the massive use of these xenobiotics in the urban and industrial areas of Concepcion and Talcauano. Samples collected in the central part of the basin contained very high concentrations of lindane that coincide with the widespread use of lindane-based pesticides (purified γ-HCH) in this area. DDT was distributed homogeneously throughout the basin, except at Laguna Icalma, the source of the river in the Andes. Most PCB residues in fish and birds consisted of congeners between penta- and hepta-chlorobiphenyls. In fish, the predominant congeners were the pentachlorobiphenyl 23′44′5 (IUPAC number 118) and the hexachlorobiphenyl 22′344′55′ (PCB-153); in birds 22′44′55′ (PCB-180) prevailed.

Chlorinated hydrocarbons in the tissues of South Polar Skuas (Catharacta maccormicki) and Adélie Penguins (Pygoscelis adeliea) from Ross Sea, Antarctica

High rates of egg infertility and embryo death in a colony of South Polar Skuas breeding in the Antarctic were similar to those in polluted North Atlantic populations of the Great Skua. Such loss could not be linked to factors such as organochlorine pollutants, as levels of DDE and PCBs in the contents of skua eggs from the population were only a small fraction of those in polluted skua populations from the Northern Hemisphere. Average eggshell thickness for skuas nesting on Ross Island has shown no significant change since the introduction of DDT. Concentrations of DDE and PCBs in South Polar Skuas were 13 and 22 times higher, respectively, than those in the eggs of sympatric Adélie Penguins, and this probably reflects the greater exposure of skuas to pollution when they migrate north of the Antarctic Convergence in winter. Residues in liver tissue showed a similar trend, and a higher rate of mixed function oxidase induction in skua liver compared to that of penguins is consistent with the trends seen in pollutant levels. The same PCB congener predominated in both skua and penguin samples. Comparisons with historical residue data suggest that global levels of DDT residues are declining.

Lead, mercury, cadmium and selenium in two species of gull feeding on inland dumps, and in marine areas

The influence of the foraging area on the intake of trace elements was evaluated in tissues of two species of gull: the black-headed gull (Larus ridibundus) and the herring gull (Larus argentatus). Mercury levels were higher in birds from the coastal environment than in those caught on inland dumps; birds found at one dump site showed high levels of lead. Interspecific differences in the levels of mercury and cadmium are discussed.

Trace elements and chlorinated hydrocarbons in eggs of Pelecanus crispus, a world endangered bird species nesting at Lake Mikri Prespa, North-western Greece

Eggs of the Dalmatian pelican, Pelecanus crispus, collected from 1984 to 1986 at Lake Mikri Prespa, north-western Greece, contained residues of trace elements and PCBs at low concentrations, and of DDE at rather high concentrations. DDE is negatively related to eggshell thickness. Eggshell thickness was reduced by 12-20% in comparison with the pre-1947 (before DDT use) eggshell thickness. However, this decrease did not affect the reproductive success of this species. The main fish species eaten by the Dalmatian pelican at Lake Mikri Prespa were analyzed for pollutants. All the fish contained low concentrations of residues. The contribution of the diet while the birds are on the wintering grounds is unknown. Recommendations for a monitoring programme of pollutants in eggs of the Dalmatian pelican, a world-endangered species, are proposed.

Mixed function oxidase activity and cytochrome P-450 forms in black-headed gulls feeding in different areas

Abstract The aim of the present study was to evaluate the mixed function oxidase (MFO) system and the forms of supposed cytochrome P-450 in the liver of black-headed gulls (Larus ridibundus) feeding in a lagoon and on a garbage dump. Tissue levels of polychlorinated biphenyls (PCBs) were also determined. Ethoxyresorufin 10-deethylase (EROD) and aldrin epoxidase activities were higher in the dump gulls than in the lagoon gulls. A marked thickening of the proteins in the cytochrome P-450 area, particularly of P-450c and P-450d, was also found in the dump gulls. No relationship was found between MFO activity and tissue chlorinated hydrocarbon levels.

Heavy metals and selenium variation in a migratory bird wintering in a mercury-polluted lagoon

Recent papers have demonstrated that water birds collected in the Mediterranean area accumulate remarkable amounts of mercury, usually greater than in the same species obtained from other areas. The critical exposure to the pollutant could negatively influence the survival of some species of birds, particularly those high in the food chain. Nevertheless animals belonging to various phyla have the ability to adapt themselves to environmental-pollutant damage. Of the various defense mechanisms employed by birds and mammals, the retention of selenium as an antagonist to mercury toxicity is the best known. In the present paper the authors report the results of a long term monitoring of mercury, cadmium, lead and selenium and their interactions in tissues of the Black-necked grebe (Podiceps nigricollis) wintering in the lagoon of Marano (NE Italy), an environment moderately polluted by mercury.

An assessment of pollutants in eggs of audouin's gull (Larus audouinii), a rare species of the Mediterranean sea

Audouins gull (Larus audouinii) is a very rare species endemic to the Mediterranean basin. A sub-population of an estimated 130 pairs is breeding in the Tuscan Archipelago, an area heavily polluted by mercury and chlorinated hydrocarbons. Here we present the data from 5 years spent monitoring the contaminant levels in the eggs of this species. Mercury, selenium, polychlorobiphenyls (PCBs) and DDE concentrations are two to five times higher than in similar species, such as the yellow-legged herring gull, nesting in the same area. The annual trends in these contaminant levels are discussed. In view of the endangered status of Audouins gull, there is a need to observe closely the developing trend of contamination in this species.

Increase of organochlorines and MFO activity in water birds wintering in an Italian lagoon

The Black-necked grebe (Podiceps nigricollis) is a migratory and dispersive birds species. Its breeding areas are located in central and eastern Europe; its wintering quarters are in western Europe, around the Mediterranean, Adriatic, Black and Caspian Seas. Recently Focardi et al. (1984) have reported that specimens of Black-necked grebes accumulate polychlorinated biphenyls and mercury in their tissues during their wintering in the Northern Adriatic lagoon of Marano. Residues of some chlorinated hydrocarbons and the levels of the mixed function oxidases (one of the most efficient hepatic detoxication systems) have been evaluated in specimens of Black-necked grebes collected in the lagoon of Marano during the wintering period (October and December 1984; April 1985) and are the subject of this report.