Effects of copper on lipid peroxidation,glutathione, metallothionein,and antioxidative enzymes in the freshwater mussel Anodonta anatina

Copper is an essential element to all animals. At elevated concentrations, it is toxic and can participate in the formation of reactive oxygen species, leading to cellular damage. In this study, the ecotoxicological relevance of copper was investigated with freshwater mussels, Anodonta anatina. When the mussels were exposed to copper at environmentally realistic concentrations, either via the water (0.3?µmol?L^?1 Cu) or fed with Cu-loaded algae (equivalent to 0.06?µmol?L^?1 Cu), the level of thiobarbituric acid-reactive substances rose and glutathione decreased. This was associated with the induction of metallothionein and, relative to total protein, of glutathione reductase and the antioxidative enzymes superoxide dismutase, catalase, and glutathione peroxidase. But, since the overall protein-synthetic capacity was hampered by the copper insult, the activities of the enzymes relative to tissue weight and copper concentrations were depressed. During depuration, most parameters started to normalize although not returning to control values within 12 days.     

Contribution to the ecotoxicological study of cadmium,copper and zinc in the mussel Mytilus edulis

A regulation of internal levels of some essential metals has been observed in various animals, whereas the bioaccumulation of several non-essential metals parallels their overloads in water. In the mussel Mytilus edulis L., we have attempted to determine if such a phenomenon exists by comparing the patterns of accumulation of copper and zinc vs cadmium. With this aim, mussels collected in the Bay of Bourgneuf (France) in November 1983 were exposed to these metals for 16 d. At external levels of zinc as high as 100 gl^-1, mussels were able to maintain a normal concentration in all groups of organs for 4 d. The ability of mussels to limit the bioaccumulation of copper and zinc varied from organ to organ, and decreased with higher levels of contamination and longer periods of exposure. In contrast, at the lowest experimental concentration and the lowest period of exposure, a significant increase of cadmium in mussel tissues was generally observed. Even at the highest sub-lethal doses, the levels of copper and zinc in mussel tissues were not much higher than the natural levels (contaminated:background ratios= 2.3 to 6.1), whereas the bioaccumulation of cadmium was less well restricted (contaminated:background ratios=136 to 192). The use of mussels as a bioindicator of pollution seems doubtful for essential metals, particularly as regards short-term pollution, since the levels of these trace elements in the organisms are largely independent of their concentration in the ambient seawater.     

Metallothioneins and lysosomes in metal toxicity and accumulation in marine mussels: the effect of cadmium in the presence and absence of phenanthrene

It has been confirmed that metallothioneins play an important role in the accumulation of cadmium (Cd) in the digestive gland cells of mussels (Mytilus galloprovincialis Lam.). The content of Cd in the tissue of mussels exposed for 9 d to the metal (estimated dosage of 180 g Cd mussel^-1 d^-1) was 66.2 ppm. This value is about the same as the metal content found in the digestive gland of Cd-exposed mussels kept in clean water for a recovery period of 28 d. At the end of the recovery period, however, the Cd bound to thionein had increased by approximately 250%. Our data demonstrate that the stability of lysosomes, a biological parameter adopted as a cellular stress index, is extremely low in mussels exposed to Cd for 9 d, but returns to control values in the digestive gland cells of mussels allowed to recover for 28 d in uncontaminated sea water. At this point most of the Cd present in the cytosol is bound to thionein. These data demonstrate the importance of metallothionein induction in the reduction of the cytotoxic effects exerted by high levels of Cd accumulation. The results of tests designed to clarify the reasons for the long biological half-life of Cd demonstrated that, in the digestive gland of mussels, the lysosomes are not able to eliminate Cd either bound to insoluble thionein polymers or to lipid peroxidation products such as lysosomal lipofuscin, both of which are apparently involved in the elimination of copper. The absence of these two mechanisms of metal sequestration and elimination via excretion of residual bodies (tertiary lysosomes) is in agreement with the persistence of cadmium in the digestive gland of mussels. Finally, the results also demonstrate that simultaneous exposure of mussels to Cd and phenanthrene, an established lysosomal membrane destabilizer, did not significantly alter the accumulation of Cd or the kinetics of the metal in mussels.     

The common mussel Mytilus edulis as an indicator of pollution by zinc,cadmium, lead and copper. I. Effects of environmental variables on uptake of metals

The net uptake of zinc, cadmium, lead and copper by the common mussel Mytilus edulis (L.) exposed to different conditions was investigated with a view to using this species as an indicator of contamination of the marine environment by these metals. The variables studied were season, position of the mussel in the water column, water salinity, water temperature, and the simultaneous presence of all four metals. Each of these 5 variables affected the net uptake of some or all of the metals studied under some conditions. Seasonal variation in concentrations of zinc, cadmium and copper was found in samples collected at three separate locations. The relationship of seasonal variation to tissue weight and absorption route of the metals is discussed. Near to freshwater inputs of trace metals, the concentrations of zinc, cadmium and lead in mussels were found to vary according to the depth at which the mussels were collected; in summer when freshwater run-off is less, this effect was absent. Low salinities did not affect the net uptake of zinc by mussels, but increased the net uptake of cadmium and decreased that of lead. Low temperatures had no effect on the net uptake of zinc or lead; the net uptake of cadmium was unaffected by low temperatures at high salinities but was decreased by low temperatures at low salinities. The presence of the other metals had no effect on the individual net uptake of either zinc, cadmium or lead. A sampling programme was devised to eliminate the effects of these environmental variables and to allow the use of M. edulis as an indicator of zinc, cadmium and lead in marine and estuarine environments. In contrast to the other metals, the net uptake of copper by the mussel was extremely erratic, and was affected by salinity and temperature changes and by the presence of the other metals and changes in their relative concentrations. The effects of other metals on the net uptake of copper cannot be easily eliminated or allowed for; it is, therefore, suggested that the mussel should not be used as an indicator of copper in the marine environment.     

Metal interaction during accumulation by the mussel Mytilus edulis planulatus

Mytilus edulis planulatus (Lamarck) were collected from Howden, South-east Tasmania in autumn 1981. Interaction effects of cadmium, copper and zinc during accumulation by mussels exposed for ten days to all three metals simultaneously were examined in a series of experiments in which each metal was tested at three concentrations. In general, interaction effects were most evident at the highest concentrations tested (20 g l^-1 Cd; 20 g l^-1 Cu; 200 g l^-1 Zn) and led to a reduction in the accumulation of cadmium and an increase in that of copper and zinc. More specifically, high levels of zinc caused a decrease in cadmium uptake and an increase in copper accumulation. The presence of copper resulted in depressed cadmium accumulation while zinc accumulation increased. Cadmium tended to enhance zinc accumulation, but copper accumulation was only affected to any great extent when zinc was also present.     

Effects of copper on the latency of lysosomal hexosaminidase in the digestive cells of Mytilus edulis

Mytilus edulis collected from Tomales Bay, California, USA, during mid-winter 1979 were exposed to increased concentrations of dissolved copper under controlled laboratory conditions. A dose-dependent reduction in the latency of lysosomal hexosaminidase activity in digestive cells was induced after a 30 d exposure to copper. The half-time of the hexosaminidase staining reaction in sections of digestive gland from control mussels was 15.5 min; for mussels exposed to 25, 50, and 75 g Cu l^-1 it was 11.8, 8.5 and 5.5 min, respectively. In addition, the dyecoupled reaction product was seen earlier in sections from individuals exposed to 50 and 75 g Cu l^-1 (after 30 s) and 25 g Cu l^-1 (1 min) than in sections from control individuals (2.5 min). Copper accumulations were demonstrated histochemically to have the same distribution as the hexosaminidase reaction product, indicating that copper is sequestered in lysosomes. Copper concentrations in digestive gland tissue, were related to the concentrations of copper in the water to which the mussels were exposed.     

Trace Metal Studies on the Starfish Asterias rubens L. From the Western Baltic Sea

In 1984, 115 samples of the starfish Asterias rubens L. collected in the south-eastern part of Cadet Trench (Mecklenburg Bay/Western Baltic Sea) were analyzed for their contents of a few major (calcium, magnesium) and trace elements (cadmium, copper, iron, mercury, manganese, nickel, lead, selenium and zinc). Distinct differences were found between starfish from different stations, and these are attributed to the composition of the sediments acting as a substrate for their prey (mussels, snails). Except for cadmium, the concentrations of the elements studied all correlated negatively with the diameter and weight of the starfish.

Parallel analyses of starfish arms and the central discs of the same animals showed that iron, zinc, copper and mercury levels were 16 to 30% higher, and selenium, manganese, magnesium, calcium and lead were 4 to 9% higher in the arms. Cadmium concentrations were 20% higher in the central discs than in the arms.

Stock estimations (about 52,000 tons fresh weight) show that starfish play a significant role in the benthic ecosystem of the western Baltic Sea. They can consume up to 200,000 tons of mussels and clams (Mytilus edulis, Macoma baltica) a year and may therefore represent a notable factor in the trace metal balance of the region.     

Trace Metal Studies on the Starfish Asterias rubens L. From the Western Baltic Sea

In 1984, 115 samples of the starfish Asterias rubens L. collected in the south-eastern part of Cadet Trench (Mecklenburg Bay/Western Baltic Sea) were analyzed for their contents of a few major (calcium, magnesium) and trace elements (cadmium, copper, iron, mercury, manganese, nickel, lead, selenium and zinc). Distinct differences were found between starfish from different stations, and these are attributed to the composition of the sediments acting as a substrate for their prey (mussels, snails). Except for cadmium, the concentrations of the elements studied all correlated negatively with the diameter and weight of the starfish.

Parallel analyses of starfish arms and the central discs of the same animals showed that iron, zinc, copper and mercury levels were 16 to 30% higher, and selenium, manganese, magnesium, calcium and lead were 4 to 9% higher in the arms. Cadmium concentrations were 20% higher in the central discs than in the arms.

Stock estimations (about 52,000 tons fresh weight) show that starfish play a significant role in the benthic ecosystem of the western Baltic Sea. They can consume up to 200,000 tons of mussels and clams (Mytilus edulis, Macoma baltica) a year and may therefore represent a notable factor in the trace metal balance of the region.     

Marine Organisms as Indicators of Heavy Metal Pollution-Experience from 16 Years of Monitoring at a Lead Zinc Mine in Greenland

In monitoring the impact of a lead-zinc mine in Greenland, species of fish, prawns, seaweed and mussels have been analysed for cadmium, copper, lead and zinc for several years. These metals have been released to the marine environment in significant amounts from the mining operation.     

Marine Organisms as Indicators of Heavy Metal Pollution-Experience from 16 Years of Monitoring at a Lead Zinc Mine in Greenland

In monitoring the impact of a lead-zinc mine in Greenland, species of fish, prawns, seaweed and mussels have been analysed for cadmium, copper, lead and zinc for several years. These metals have been released to the marine environment in significant amounts from the mining operation.     

Schwermetallbelastung von Dreissena polymorpha in Donau und Drau und ihre Bedeutung als Bioindikator

Goal and Scope

This study was undertaken to investigate the differences in heavy metal burden between the organisms and environmental compartments and to evaluate the role of Dreissena polymorpha as a bioindicator organism.

Methods

The concentrations of zinc, copper, cadmium and lead in whole soft body and selected tissues of D. polymorpha at two river habitats in Austria were examined using atomic absorption spectroscopy (AAS). Concentrations in organisms were compared to those in sediment and water.

Results and Conclusion

Zebra mussels of the river Drau showed generally higher heavy metal concentrations as compared to mussels of the river Danube and contained elevated zinc and cadmium levels as compared to metal concentrations found in soft tissues of zebra mussels from uncontaminated sites in Germany and The Netherlands. The essential metals zinc and copper were mainly accumulated in gills, foot and byssal gland tissue of the mussel, in contrast to the non-essential metals cadmium and lead which were found predominantly in the midgut gland. The heavy metal concentrations in both, sediments and mussel tissue, were higher than in water samples. There was no correlation between the concentrations in water and in the organisms except for zinc. In contrast, correlations were found between concentrations in sediments and mussel soft tissue.

Recommendation and Perspective

Further investigations should include the examination of sediments and consider the mechanism of food uptake to assess the role of D. polymorpha as a bioindicator organism.     

Combined effects of copper and cadmium on Chlorella pyrenoidosa H.Chick: subcellular accumulation,distribution, and growth inhibition

Disposal of waste into aquatic ecosystems may cause microalgae to be exposed to various metals, e.g. copper and cadmium. The effects caused by combinations of metals may be more serious. Evaluations of subcellular fate, bioaccumulation, and biological effects of metals on aquatic organisms are generally derived from experiments with individual metals. The present study aims to evaluate the effects of exposure of Chlorella pyrenoidosa to copper and cadmium in combination on subcellular accumulation, distribution, and growth. The algae were exposed for 72 h to copper at concentrations of 13 – 25 µmol L^?1, cadmium at about 6 µmol L^?1, and combinations thereof. The levels of copper and cadmium in subcellular organelles, heat-denaturated protein, metal-rich granules, and heat-stable protein were determined by atomic absorption spectrometry. Exposure of C. pyrenoidosa to copper and cadmium in combination inhibited growth more strongly than copper and cadmium individually. Highest accumulation was observed in metal-rich granules and heat-stable proteins. Administration of both metals in combination affected their subcellular distribution: copper was mainly distributed into the metal-rich granules (70%–80%) and heat-stable proteins (9%–24%), cadmium in the metal-rich granules (88%–98%).     

Metallothionein induction inMytilus edulis exposed to cadmium

The exposure of mussels,Mytilus edulis, collected from Whitsand Bay, southwest England, in August 1988, to sublethal concentrations of cadmium (400µg l^–1) for 65 d resulted in the induction of metallothionein (MT) synthesis in the soft tissues. In cadmium-exposed mussels, metallothionein concentrations, measured by differential pulse polarography, increased by a factor of three, from 2 to 3 mg g^–1 to a maximum of 9 mg g^–1 after 30 d. No significant changes could be detected in controls. Cadmium accumulated in the soft tissues of mussels correlated significantly with metallothionein concentrations and can be described by the relationship: MT (mg g^–1)=0.045 Cd (µg g^–1)+3.03 (r=0.803,P<0.001). Gel chromatography of heat-treated cytosolic extracts showed that the accumulated cadmium is bound principally to the newly formed metallothioneins. Copper and zinc were also analysed in the whole soft-tissues and in subcellular fractions of cadmium-exposed mussels. Although copper concentrations were not affected by cadmium-exposure, zinc levels were significantly reduced. The results demonstrate that the induction of metallothioneins inM. edulis is a quantifiable biological response to sublethal levels of cadmium exposure.     

Effect of calcium and magnesium on copper,cadmium, and some essential elements of Indian major carps Labeo rohita (Hamilton) and Catla catla (Hamilton) treated with copper and cadmium

The effect of calcium and magnesium either singly or in combination on accumulation of cadmium and copper in Labeo rohita (rohu) and Catla catla (catla) was investigated in this study under laboratory conditions. The investigation showed that copper accumulation in rohu exposed to 0.25 mg L^?1 of copper for 14 days reduced from 31.0 ± 0.4 mg kg^?1 at no calcium/magnesium treatment to 3.5 ± 0.2 and 2.2 ± 0.1 mg kg^?1, respectively, at 75 mg L^?1 calcium or magnesium treatment. The copper level in catla exposed to 0.20 mg L^?1 of copper for 14 days reduced from 5.7 ± 0.1 mg kg^?1 at no calcium/magnesium treatment to 3.4 ± 0.2 and 3.3 ± 0.1 mg kg^?1, respectively, at 300 mg L^?1 calcium or magnesium treatment. The cadmium accumulation in rohu exposed to 0.20 mg L^?1 of cadmium for 14 days reduced from 1.7 ± 0.1 mg kg^?1 at no magnesium treatment to 1.4 ± 0.1 mg kg^?1 at 120 mg L^?1 magnesium treatment. The cadmium accumulation in catla exposed to 0.20 mg L^?1 of cadmium for 14 days reduced from 0.8 ± 0.2 mg kg^?1 at no magnesium treatment to 0.6 ± 0.2 mg kg^?1 at 80 mg L^?1 magnesium treatment. Copper and cadmium treatments also reduced some essential microelements of rohu and catla. Both the fishes restored these elements at different levels of calcium and magnesium.     

Uptake and effects of copper and cadmium in the gonad of the scallop Placopecten magellanicus: concurrent metal exposure

Sea scallops Placopecten magellanicus in early gametogenesis from the southern shelf of Hudson Canyon, New Jersey, USA, were exposed in late winter 1984 to sublethal levels of Cu and Cd in a flowing seawater system at the NMFS Milford Laboratory. Exposure was to copper (10 and 20 gl^-1: low-Cu and high-Cu groups) or to a combination of copper and equimolar cadmium (10g Cu+17.7 g Cdl^-1: low-Cu/Cd group) for eight weeks, with sampling at 2-wk intervals. Copper had a strongly inhibitory effect on gamete production and maturation, which in some respects was partially moderated in the presence of cadmium in the female gonad only. Total gamete weight per scallop doubled in control individuals but dropped by 60% in both high-metal exposure groups over the 8-wk exposure period, with a smaller, temporary decrease in the low-Cu group. Cadmium did not add to the inhibition by copper of gamete development in the low-Cu/Cd group, but there was no partial recovery at 8 wk, as was seen in female scallops exposed to low-Cu alone. Gonadal RNA, higher in the females, decreased proportionately more in that sex than in the males of the metal exposure groups. Conversely, DNA levels were higher in the male than in the female gonad, and decreased sharply in all metal-exposed males. Gonadal protein concentration also dropped in all metal-exposed scallops with time and degree of metal exposure. Copper uptake in the gonad increased with time and metal exposure concentration, and cadmium increased in the low-Cu/Cd group with time. In contrast, manganese decreased significantly in the gonads of Cu-exposed scallops, especially in the high-Cu group. In the low-Cu/Cd group, manganese concentrations stabilized after an initial sharp drop at 2 wk in the low-Cu/Cd group, then rose at 6 and 8 wk. We ascribe this phenomenon to the induction by cadmium of metal-binding proteins (Fowler and Megginson 1986). In both sexes, gonadal magnesium concentrations did not change with increasing tissue burdens of copper and cadmium, but instead rose initially in proportion to the degree of metal exposure, homeostasis being maintained thereafter.     

Effects of copper and cadmium on osmoregulation and oxygen consumption in two species of estuarine crabs

Green crabs (Carcinus maenas) and rock crabs (Cancer irroratus) were exposed to various concentrations of copper as cupric chloride (CuCl₂ · 2 H₂O), and cadmium as cadmium chloride (CdCl₂ · 21/2 H₂O) for 48 h. The exposures were conducted at 5 different salinities. At the end of each exposure period, tests of blood-serum osmolality and gill-tissue oxygen consumption were performed. Copper-exposed crabs exhibited loss of osmoregulatory function with increasing copper concentration until normally hyperosmotic serum became isosmotic with the surrounding medium. Cadmium elevated greencrab serum above its normal hyperosmotic state. Copper had no effect on gill-tissue oxygen consumption; however, cadmium reduced the rate of oxygen consumption in both species tested.     

Mytilus edulis planulatus: an “integrator” of cadmium pollution?

Mussels, Mytilus edulis planulatus, were collected from Port Phillip Bay, Victoria, Australia, in February 1984, to test the assumption that they are integrators of cadmium pollution. Groups of mussels were subjected to the same average dose of cadmium (21 g l^-1), administered according to different dosing regimes over 4 wk (Regimes 1, 2, 3, 5); other groups received twice the average dose (42 g l^-1) in half the time (Regime 4). During each regime, the mussels were exposed to the different cadmium concentrations for one week at each concentration. Nominally, the regimes were: (1) 36 g (Wk 1) to 26 g (Wk 2) to 16 g (Wk 3) to 6 g (Wk 4) Cd l^-1; (2) 6 to 16 to 26 to 36 g Cd l^-1; (3) 21 to 21 to 21 to 21 g Cd l^-1; (4) 42 to 42 g Cd l^-1; (5) 42 to 42 g Cd l^-1 to background (<0.5 g) to background. Differences in cadmium accumulation by mussels from Regimes 1 and 4 were not statistically significant, nor were differences in accumulation between mussels from Regimes 2 and 3. However, mussels from Regimes 1 and 4 had accumulated significantly more cadmium than had mussels from Regimes 2 and 3. Accumulation by mussels from Regime 5 was not significantly different from that by mussels from any of the other regimes. These results suggest that, at least for cadmium, the assumption that mussels are integrators of pollution should be treated with caution. They also have implications with regard to the quantitative biological monitoring of pollution. For example, even in a carefully controlled monitoring program, using mussels of standard size and condition, significant differences in cadmium content between mussels need not indicate exposure to different levels of contamination. Rather, these differences could reflect differences in the regime by which the contamination was received.     

Biochemical constituents and bioaccumulation as biomarkers in the green mussel Perna viridis with reference to silver and chromium toxicity

The sublethal effect of silver and chromium on some biochemical constituents was studied in the green mussel Perna viridis. The results revealed an overall reduction in total protein, total DNA, total RNA, glycogen, protein bound sugars and total lipid in the gill, hepatopancreas, and ovary of the mussels exposed to metals individually and in combination when compared with control. It was apparent that mussels exposed to mixtures of metals exhibited highly significant (P?P?P?相似文献   

The common mussel Mytilus edulis as an indicator of pollution by zinc,cadmium, lead and copper. II. Relationship of metals in the mussel to those discharged by industry

Concentrations of zinc, cadmium, lead and copper were determined in whole soft parts of the common mussel Mytilus edulis (L.) sampled in Port Phillip Bay and Western Port Bay, both in Victoria, Australia. The mussels were sampled according to procedures suggested by previous studies in order to eliminate the effects of natural environmental variables. Results of the analytical studies were compared to data on the quantities of trace metals known to be discharged by industry into the catchments of each Bay. This allowed an evaluation of the indicator ability of the mussel without the need for multiple analyses of water samples. The results suggest that the mussel is capable of acting as an efficient time-integrated indicator of zinc, cadmium and lead over a wide variety of environmental conditions. Although additional variables may remain to be investigated, the mussel is recommended as an alternative to the analysis of water and sediments. With further development this organism may also be useful for monitoring industrial effluent characteristics at the discharge site, allowing rapid, reliable and inexpensive control of water quality. In contrast, the mussel is of doubtful use as an indicator of copper; other indicator types such as macroalgae may be better-suited to the study of this element.     

Copper-induced differential mortality in the mussel Mytilus edulis

Copper toxicity experiments were performed with 3 000 mussels (Mytilus edulis L.) from a population that is a mixture of individuals originating in the Kattegat and in the Baltic. Subsequently, the electrophoretic patterns of the enzymes phosphoglucose isomerase (PGI) and phosphogluco mutase (PGM) were analysed in the dead and surviving mussels in order to detect possible differential mortality. Significant differences in allele frequencies between dead and surviving mussels were found for PGI. The results are discussed in relation to genotypic differences in the PGI and in relation to the composite structure of the investigated population. It is concluded that the differential mortality is due to genotypic differences in the PGI locus and/or to different susceptibility to copper by mussels originating in the two geographic areas.