Claudio Leonzio

Heavy metals and glutathione metabolism in mussel tissues

Abstract Data are presented on the tissue glutathione content and on the activity of two of the main enzymes involved in GSH metabolism (γ-glutamyl-cysteine synthetase (GCS), GSH transferase (GST)) in the gills and digestive gland of mussels (Mytilus galloprovincialis Lam.) exposed for 1, 4 and 7 days to sublethal concentrations of copper and of mercury, both in the inorganic form of HgCl2 and in the organic form of CH3HgCl. The results indicate that Cu2+ (0.6 μM) and CH3Hg+ (0.2 μM) can affect, although to a different extent, glutathione metabolism in mussel tissues; common traits were a decrease in tissue glutathione content, stimulation of GST activity and inhibition of GSH synthesis rate; on the other hand, inorganic Hg2+ (0.2 μM) did not significantly affect glutathione content and metabolism in mussel tissues. Exposure to copper (0.6 μM) resulted in a decrease in total glutathione content in both gills and the digestive gland that was maximal after the first day, followed by a tendency to recover at longer exposure times. Such a decrease seemed to be mainly related to a stimulation of GST activity, which was transient in gills and persistent in the digestive gland. The tissue glutathione content was also decreased in mussel exposed to methylmercury (0.2 μM); however, such a decrease seemed to be mainly due to inhibition of the GSH synthesis rate and, to a lesser extent, to an increased GST activity. On the contrary, inorganic Hg2+ (0.2 μM) did not significantly affect glutathione metabolism, although mussel exposure to the same concentration of organic and inorganic Hg2+ resulted in similar total metal tissue load in mussel tissues. The role of glutathione as a potential defense against heavy metals is discussed in relation to different routes of metal toxicity and homeostasis in mussel tissues.

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.

The use of non-destructive biomarker in Mediterranean cetaceans: Preliminary data on MFO activity in skin biopsy

Abstract Cetaceans have been subjected to heavy anthropogenic pressure in the last century. In addition to hunting there is now the subtle threat of pollution which may be responsible for metabolic impairment and involved in stranding episodes and population decline. The need to study pollutants and their effect on cetacean populations is often in contrast with the need to protect these animals. In this paper a new method of collecting skin and hypodermic biopsies in the Fin whale (Balaenoptera physalus) and Striped dolphin (Stenella coeruleoalba) in order to analyse Mixed Function Oxidase (MFO) activity and organochlorine contents, is described. The results show marked differences between the two species. MFO activity in skin and organochlorines in blubber are higher in Striped dolphin than the Fin whale. The new method enables the main effects of exposure to lipophilic contaminants to be assessed in a nondestructive way in biomarker studies.

Trace elements in striped dolphins (Stenella coeruleoalba) from the western Mediterranean

Tissues obtained from Stenella coeruleoalba stranded along the Spanish and Italian Mediterranean coasts from 1987 to 1994 were analysed for cadmium (Cd), copper (Cu), mercury (Hg), selenium (Se) and zinc (Zn). The age, length and weight of the dolphins were recorded. Hg levels were also assayed in skin biopsies from dolphins of the same species in the waters off northeastern Spain and in the Tyrrhenian and Ligurian Seas. Levels of all elements differed in muscle of stranded dolphins from the two areas. Hg was higher in tissues from animals stranded on the Italian coasts and in skin biopsies obtained in the Tyrrhenian and Ligurian Seas, than in the respective Spanish samples. This is probably related to Hg pollution from the natural weathering of cinnabar ores in central Italy. Se and Cd levels had similar accumulation patterns to those of Hg. Accumulation of Hg and Se is explained by the existence of a detoxification pathway involving both elements, however the reason for the similar Cd trend is unclear. Geographical differences in the accumulation pattern of these elements may reflect the existence of two different populations of Stenella coeruleoalba in the western Mediterranean.

Effects of sublethal, environmentally relevant concentrations of hexavalent chromium in the gills of Mytilus galloprovincialis.

Hexavalent chromium Cr(VI) is an important contaminant released from both domestic and industrial effluents, and represents the predominant chemical form of the metal in aquatic ecosystems. In the marine bivalve Mytilus galloprovincialis exposure to non-toxic, environmentally relevant concentrations of Cr(VI) was shown to modulate functional parameters and gene expression in both the digestive gland and hemocytes. In this work, the effects of exposure to Cr(VI) (0.1-1-10 μg L(-1) animal(-1) for 96 h) in mussel gills were investigated. Gill morphology and immunolocalization of GSH-transferase (GST), of components involved in cholinergic (AChE and ChAT), adrenergic (TH) and serotoninergic (5-HT(3) receptor) systems, regulating gill motility, were evaluated. Total glutathione content, activities of GSH-related enzymes (glutathione reductase - GSR, GST), of catalase, and of key glycolytic enzymes (phosphofructokinase - PFK and pyruvate kinase - PK) were determined. Moreover, mRNA expression of selected Mytilus genes (GST-π, metallothionein isoforms MT10 and MT20, HSP70 and 5-HT receptor) was assessed by RT-q-PCR. Cr(VI) exposure induced progressive changes in gill morphology and in immunoreactivity to components involved in neurotransmission that were particularly evident at the highest concentration tested, and associated with large metal accumulation. Cr(VI) increased the activities of GST and GSR, and total glutathione content to a different extent at different metal concentrations, this suggesting Cr(VI) detoxication/reduction at the site of metal entry. Cr(VI) exposure also increased the activity of glycolytic enzymes, indicating modulation of carbohydrate metabolism. Significant changes in transcription of different genes were observed. In particular, the mRNA level for the 5-HTR was increased, whereas both decreases and increases were observed for GST-π, MT10, MT20 and HSP70 mRNAs, showing sex- and concentration-related differences. The results demonstrate that Cr(VI) significantly affected functional and molecular parameters in mussel gills, and indicate that this tissue represents the major target of exposure to environmentally relevant concentrations of the metal.

Use of nondestructive biomarkers and residue analysis to assess the health status of endangered species of pinnipeds in the south-west Atlantic

Total porphyrins, EROD, PROD, BROD, organochlorines, PAHs and total Hg were determined in specific organs and tissues of pinnipeds collected from the south-west Atlantic. The results confirm the existence of metabolic imbalance between high toxifying and low detoxifying potential in pinnipeds, as reported in other marine mammals. The non destructive biomarker approach based on the analysis of metabolic responses (porphyrins) and residues (DDTs, PCBs, PAHs and Hg) in non-invasive samples (excreta and fur) appear to be a useful tool for the hazard assessment of endangered species.

Heavy metals in organisms from the Northern Tyrrhenian sea

The concentrations of zinc, manganese, copper, cadmium, lead and mercury have been determined in soft tissues of four marine organisms (Mytilus galloprovincialis Lmk., Nephrops norvegicus L., Mullus barbatus L., Engraulis encrasicolus L.) collected seasonally from the winter of 1976 to the spring of 1980 in various areas of the Tyrrhenian Sea (western Mediterranean). In all four species levels of all metals, except mercury in all areas are alike while mercury levels vary and, furthermore, are higher than in specimens from other areas of the Mediterranean and also from other seas. The possible natural origin of the mercury from cinnabar (HgS) ore as well as the suitability of the four species as bioindicators is discussed.

Serum esterase inhibition in birds: A nondestructive biomarker to assess organophosphorus and carbamate contamination

With the aim of proposing a nondestructive biomarker for monitoring the toxicological risk to birds of exposure to the organophosphorus insecticide azamethiphos and the carbamate insecticide methomyl, laboratory studies were performed on serum “B” esterases in Japanese quail (Coturnix coturnix japonica). The birds received two single dose treatments of each compound (azamethiphos and methomyl), i.e., 50 mg/kg and 250 mg/kg respectively. In the first treatment, serum butyrylcholinesterase (BChE) and carboxylesterase (CbE) were drastically inhibited in the azamethiphos-treated group, 24 h after the dose. No inhibition was detected for BChE and CbE activities in the methomyl-treated group, 24 h after the dose. In the second treatment, the birds died or were sacrified 3 h after the dose. Serum BChE and brain acetylcholinesterase (AChE) were strongly inhibited after treatment with both insecticides. Serum CbE, hepatic microsomal CbE and 7-ethoxyresorufin dealkylation activities were also inhibited. A statistically significant correlation between serum BChE and brain AChE was found at lethal and sublethal doses of these xenobiotics. The experimental results indicate that the nondestructive biomarker BChE can give an early qualitative and semi-quantitative warning of the toxic effects of organophosphate and carbamate insecticides in birds.

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.