Mussel watch: a review of Cu and other metals in various marine organisms in Taiwan, 1991-98

This study presents the distribution of Cu, Zn, Pb, Cd, Hg and As in various marine organisms collected along the western coast of Taiwan from 1991 to 1998, and also evaluates the time variation of Cu in oysters before (1980-85) and after (1986-98) the "green oyster" incident. The results show that relatively high geometric mean (GM) concentrations of Cu, Zn, Pb, Cd, As and Hg were generally found in Crassostrea gigas (Cu=229 microg/g, Zn=783 microg/g), Gomphina aeguialtera (Pb=30.3 microg/g), Tegillarca granosa (Cd=2.85 microg/g), Thais clavigera (As=96.9 microg/g) and Parapenaeopsis cornuta (Hg=1.35 microg/g), respectively. Especially, maximum Cu and Zn concentrations (GM=229 and 783 microg/g, respectively) in oysters (C. gigas) from different culture areas were much higher than those of the other organisms by about 1.13-458 and 2.40-63.7 times, respectively. Similarly, rock-shells (Thais clavigera) had a high capacity for accumulating Cu (GM=202 microg/g) and Zn (GM=326 microg/g) under the same physico-chemical conditions. The highest GM Cu and Zn concentrations of 1108 (range from 113 to 2806) and 1567 (range from 303 to 3593) microg/g were obtained in oysters from the Hsiangshan coastal area, one of the most important oyster culture areas in Taiwan. However, the highest GM Cd and As concentrations of 6.82 and 19.3 microg/g were found in oysters from the Machu Islands. Mean Cu concentrations in the oysters from the Erhjin Chi estuary declined from 2194+/-212 microg/g in 1986-90 to 545 microg/g (GM) in 1991-96. In the Hsiganshan area, GM Cu concentrations of 909 microg/g (1991-96) and 1351 microg/g (1997-98) in oysters were significantly higher than those of 201 microg/g (1980-85) and 682 microg/g (1986-90). The gradually increasing levels of Cu and Zn in the oysters from the Hsiangshan area have been observed year by year.     

Estimation of metal and organochlorine pesticide exposures and potential health threat by consumption of oysters in Taiwan

Pollutant concentrations detected in oysters from 12 different culture areas of Taiwan (especially for the Hsiangshan area and the Machu Islands) from 1991-98 were evaluated to investigate potential carcinogenic (inorganic As and organochlorine pesticides) and non-carcinogenic (Cu, Zn, Cd and inorganic As) risk to the public from ingestion of the oysters. The highest geometric mean (GM) Cu and Zn concentrations of 1108 (range 113-2806) and 1567 (range 303-3593) microg/g dry weight were obtained in oysters from the Hsiangshan coastal area. The maximum GM Cd and As concentrations of 6.82 and 19.3 microg/g dry weight were found in oysters from the Machu Islands area. The p,p'-DDE values range from not detectable in Penghu Islands' oysters to 164 ng/g dry weight in Machu Islands' oysters. The highest tDDT (sum of p,p'-DDE, p,p'-DDD and p,p'-DDT) concentrations of 337 and 340 ng/g dry weight were found in oysters from Kimmen and Machu Islands, respectively. A calculated target hazard quotient (THQ; daily intake/reference dose) of 11.4 (based on 139 g oysters/day) for Cu caused by consuming oysters from the Hsiangshan area is higher than that from other areas (range 0.124-5.95). The highest average Cu intake from Hsiangshan's oysters for individuals is 11.4 times (i.e. THQ=11.4) more than that of reference dose (40 microg/kg/day). However, the maximum THQ values for Cd and As caused by consuming oysters collected from the Machu Islands were 5.57 and 2.63 for Cd and As, respectively. Generally, the results of THQ showed that if only the maximally exposed individuals were considered, the value of 65.4% for oyster was higher than 1.0 in comparison with reference dose. All cancer risk estimates for inorganic As from consuming oysters were higher than 10(-6) (range from 128x10(-6) to 509x10(-6) for maximally exposed individuals and range from 17.1x10(-6) to 68.0x10(-6) for typically exposed individuals, respectively); that is the risk of the lower end of the range of acceptable risk. The highest risk estimate for inorganic As was 509x10(-6) for consumption of oysters by Machu Islands' residents. The lifetime cancer risks of 19.0x10(-6) for tDDT by consuming oysters from the Machu Islands was higher than those from the Penghu Islands (0.37x10(-6)). Therefore, the sum of lifetime cancer risks for tDDT and inorganic As had the highest risks (total risk=528x10(-6)) of consuming oysters from the Machu Islands. Furthermore, a 10(-6) upper limit on lifetime risk as the health protection standard would require maximum oyster consumption rates of approximately 0.26 g/day.     

Reduction of heavy metal content in Hiroshima Bay oysters (Crassostrea gigas) by purification

Cultured oysters from Hiroshima Bay were analysed to determine the content of nine trace elements, cadmium (Cd), lead (Pb), chromium (Cr), arsenic (As), manganese (Mn), nickel (Ni), copper (Cu), zinc (Zn), and iron (Fe) and compared with oysters treated for 48 h in both artificial sea water and a solution of 0.5% ethylenediaminetetraacetic acid (EDTA) in artificial sea water. It was found that the values of As and Fe in both groups of treated oysters were significantly lower than in the untreated oysters and that the Pb and Mn levels tended to be lower. It was also found that oysters treated in 0.5% EDTA solution in artificial sea water had lower levels of Cd and Cu than the oysters treated only in artificial sea water.     

Biomass-derived materials in the remediation of heavy-metal contaminated water: removal of Cadmium(II) and copper(II) from aqueous solutions

Manganese-coated activated carbon (MCAC) and activated carbon were used in batch experiments for the removal of cadmium(II) and copper(II). Results showed that uptake of Cd(II) and Cu(II) was unaffected by increases in pH (3.0 to 8.5) or concentration (1 to 20 mg/L). Increased ionic strength (from 0.001 to 1 M NaNO3), however, significantly affected the uptake of Cd(II); adsorption of Cu(II) was not affected. Freundlich adsorption isotherm results indicated that MCAC possessed higher sorption capacity than activated carbon. Second-order rate constants were found to be 0.0386 for activated carbon and 0.0633 g/mg x min for MCAC for Cd(II) and 0.0774 for AC and 0.1223 g/mg x min for MCAC for Cu(II). Column experiments showed that maximum sorption capacity of MCAC was 39.48 mg/g for Cu(II) and 12.21 mg/g for Cd(II).     

Bank voles as monitors of environmental contamination by heavy metals. A remote wilderness area in Poland imperiled

Concentrations of heavy metals (Cd, Pb, Cu, Zn, Fe) were determined in the tissues of bank voles collected in polluted forest sites in southern Poland and in Bia?owieza National Park. The highest cadmium concentrations were found in livers (0.9-12.8 microg g(-1)) and kidneys (3.2-29.6 microg g(-1)), whereas the lowest were in bones (0.2-0.9 microg g(-1)). The lead concentrations found in kidneys and bones ranged from 9.5 to 40.0 microg g(-1). The ranges of zinc, iron and copper concentrations in the tissues of animals were: 137-343 microg g(-1), 206-1017 microg(-1) and 12.0-73.8 microg(-1), respectively. Some of the differences between the levels found in various sites were statistically significant.     

Chronic effects of Cd on reproduction of Polypedilum nubifer (Chironomidae) through water and food

Chronic effects of Cd on reproduction of Polypedilum nubifer (Chironomidae) were investigated by Cd-exposure from the egg stage using a flow-through aquarium. No significant effect of Cd on reproduction was observed in the midge larvae which had been exposed to 10 or 20 microg Cd litre(-1). Emergence of the Cd-exposed larvae peaked several days before that of the control, although growth was impaired in 1st and/or 2nd instars. The percentage emergence success decreased to about 46% of the control at 40 microg Cd litre(-1). However, other reproductive processes (adult sex ratio, oviposition success and egg hatchability) were not impaired. The emergence success decreased to less than 3% of the control at 80 microg Cd litre(-1). The emergence success of midge larvae which had been fed Cd-contaminated food with 220 or 1800 microg Cd g(-1) decreased to nearly 60% of the control. However, other reproductive processes were not impaired. Emergence success of larvae exposed to food containing 22 microg Cd g(-1) was unaffected.     

Evaluation of cadmium, copper, zinc, and iron concentrations and tissue distributions in the benthic crab, Dorippe granulata (De Haan, 1841) from Tolo Harbour, Hong Kong

The distributions of copper, zinc, iron, and cadmium among the tissues of Dorippe granulata were determined. The highest copper concentrations were found in the haemolymph (c. 53 microg ml(-1)) while the highest iron concentrations occurred in the gills (c. 720 microg g(-1) dry weight) and the highest zinc concentrations in the exoskeleton (c. 200 microg g(-1) dry weight). By comparison, concentrations of the non-essential metal, cadmium, were low in all tissues (mean = 10 microg g(-1) dry weight). The highest value was recorded from the midgut gland of a female crab (18.5 microg Cd g(-1) dry weight). Concentrations of copper, zinc, and iron were positively correlated with tissue-hydration levels. Such a relationship was not found for cadmium. The findings are discussed with regard to trace-metal levels found in temperate and tropical brachyurans from clean and polluted localities.     

The induction of metallothionein in tissues of the Norway lobster Nephrops norvegicus following exposure to cadmium, copper and zinc: the relationships between metallothionein and the metals

Nephrops norvegicus were exposed simultaneously to cadmium, copper and zinc over an 18-day period. Exposure concentrations were control, 1, 5 and 25 microg litre(-1) for cadmium and copper and 8, 40 and 200 microg litre(-1) for zinc. Concentrations of cadmium, copper, zinc and metallothionein were measured in homogenates of both the gill and the hepatopancreas. Quantification of metallothionein was carried out by differential pulse polarography. Cadmium concentrations increased significantly in the gill and hepatopancreas of both male and female animals in response to increases in exposure concentration. In contrast, the concentration of copper and zinc increased significantly in the gills of males, but not in females. In the hepatopancreas, neither copper nor zinc resulted in significant changes in concentrations of these metals. Metallothionein concentrations in the gill and hepatopancreas were increased significantly in relation to metal exposure in both males and females. Concentrations of cadmium and metallothionein in both the gill and hepatopancreas of males and females were positively correlated. Copper in the hepatopancreas also showed positive relationships with MT concentrations in males, but not in females. This study suggested that cadmium MTs in the gill and hepatopancreas of Nephrops norvegicus could be used as a sensitive tool to detect cadmium contamination in the lobsters, although this was not true for copper and zinc.     

Copper regulation and homeostasis of Daphnia magna and Pseudokirchneriella subcapitata: influence of acclimation

This study aimed to evaluate (1) the capacity of the green alga Pseudokirchneriella subcapitata and the waterflea Daphnia magna to regulate copper when exposed to environmentally realistic copper concentrations and (2) the influence of multi-generation acclimation to these copper concentrations on copper bioaccumulation and homeostasis. Based on bioconcentration factors, active copper regulation was observed in algae up to 5 microg Cu L(-1) and in daphnids up to 35 mug Cu L(-1). Constant body copper concentrations (13+/-4 microg Cu g DW(-1)) were observed in algae exposed to 1 through 5 microg Cu L(-1) and in daphnids exposed to 1 through 12 microg Cu L(-1). At higher exposure concentrations, there was an increase in internal body copper concentration, while no increase was observed in bioconcentration factors, suggesting the presence of a storage mechanism. At copper concentrations of 100 microg Cu L(-1) (P. subcapitata) and 150 microg Cu L(-1) (D. magna), the significant increases observed in body copper concentrations and in bioconcentration factors may be related to a failure of this regulation mechanism. For both organisms, internal body copper concentrations lower than 13 microg Cu g DW(-1) may result in copper deficiency. For P. subcapitata acclimated to 0.5 and 100 microg Cu L(-1), body copper concentrations ranged (mean+/-standard deviation) between 5+/-2 microg Cu g DW(-1) and 1300+/-197 microg Cu g DW(-1), respectively. For D. magna, this value ranged between 9+/-2 microg Cu g DW(-1) and 175+/-17 microg Cu g DW(-1) for daphnids acclimated to 0.5 and 150 microg Cu L(-1). Multi-generation acclimation to copper concentrations >or =12 microg Cu L(-1) resulted in a decrease (up to 40%) in body copper concentrations for both organisms compared to the body copper concentration of the first generation. It can be concluded that there is an indication that P. subcapitata and D. magna can regulate their whole body copper concentration to maintain copper homeostasis within their optimal copper range and acclimation enhances these mechanisms.     

Depuration of copper and zinc by green oysters and blue mussels of Taiwan

This paper describes depuration processes of copper and zinc in green oysters (Crassostrea gigas) and in blue mussels (Mytilus smarangdium) collected from an environment with heavy copper contamination, and then transferred to natural clean seawater. Results show that the total loss of copper content per oyster is an exponential function of exposure time for the first 6 days with a depuration rate of 351 microg g(-1) day(-1) and then levels off. During this exponential decrease period approximately 67% of the copper accumulated in green oysters was depurated. However, when the copper contents in the oysters decreased from 2225 +/- 111 microg g(-1) to 344 +/- 18.7 microg g(-1) the depuration rates decreased from 245 microg g(-1) day(-1) to 0.08 microg g(-1) day(-1). This means that green oysters had a 16-fold higher copper depuration rate (351 microg g(-1) day(-1)) than normal oysters (21.5 microg g(-1) day(-1)) for the first 6 days. However, the depuration of accumulated copper and zinc by the mussels was a fast process in natural clean seawater. About 91% of the accumulated copper was lost during the first 6-day period; copper contents declined from 20.2 +/- 3.41 microg g(-1) to 1.80 +/- 0.21 microg g(-1). Only 36% of the accumulated zinc was lost during a depuration period of 6 days. Calculations show that the biological half-lives of copper in green and normal oysters were 11.6 and 25.1 days, respectively. The biological half-lives of zinc in green and normal oysters were 16.7 and 30.1 days, respectively. In spite of the relatively low initial copper content in blue mussels being 20.2 +/- 3.41 microg g(-1), the biological half-life is only 6.40 days. From these results it is important to emphasise that the fastest turnover rate is for copper in blue mussels. However, zinc is more retentive in blue mussels than copper.     

Heavy metal concentrations in green-lipped mussels collected from Tolo Harbour and markets in Hong Kong and Shenzhen

Green-lipped mussels, Perna viridis, were collected from Kat O, Yim Tin Tsai, Ma Liu Shui and Tap Mun around Tolo Harbour and six local markets in Hong Kong (Aberdeen, Shau Kei Wan, Kowloon City, Mongkok, Yuen Long) and Shenzhen (Dongmun) between July 1994 and February 1995 and analysed for cadmium (Cd), chromium (Cr), copper (Cu), nickel (Ni), lead (Pb) and zinc (Zn). The metal concentrations of mussels collected from the study sites were Cd (0.45-1.44 microg/g), Cr (0.82-4.89 microg/g), Cu (6.02-23.99 microg/g), Ni (3.25-6.87 microg/g), Pb (2.02-4.36 microg/g) and Zn (90-135 microg/g), while those from the markets were Cd (0.27-1.44 microg/g), Cr (1.09-3.30 microg/g), Cu (9.05-17.8 microg/g), Ni (2.44-5.25 microg/g), Pb (1.17-5 microg/g) and Zn (51-103 microg/g). The metal concentrations were below the maximum permissible levels set by the Hong Kong Government. In addition, seasonal variation of metal accumulation in mussels was investigated in Yim Tin Tsai and Ma Liu Shui and a reduction in the total heavy metal concentrations during winter was noted. The non-carcinogenic hazard index of mussels collected from Tolo Harbour and from Hong Kong markets was between 0.46 and 1.36 compared with those from Shenzhen markets (0.85-1.46), which indicated a low but possible risk in consuming the mussels.     

Multivariate analysis of elements content of Larch Bolete (Suillus grevillei) mushroom

Larch Bolete (Suillus grevillei) mushrooms collected at spatially distant sites in Poland showed on relative abundance of K, P, Mg, Rb, Al, Ca, Fe (at mg/g dw), Ag, Ba, Cd, Cu, Mn, Na and Zn (at microg/g dw) and less of Co, Cr, Ni, Pb and Sr (<1 microg/g dw). Amongst these elements Rb and K were highly bioconcentrated by Larch Bolete (BCF around 1000 and 300 for the caps), and weaker were Cu, Mg, Na, P and Zn (BCF between 4.1+/-2.5 and 62+/-21). Ag and Cd were bioconcentrated mainly in caps with BCF value of 25+/-16 and 13+/-10. Al, Ba, Ca, Co, Cr, Fe, Mn, Ni, Pb and Sr were excluded (BCF<1) by this species. Intake of meal prepared of 300 g of fresh fruiting bodies of Larch Bolete picked-up at the Western Beskid or Pogórze Wielickie sites in the south of Poland will result in exceeding of oral reference dose set for cadmium. In the case of silver, copper or zinc content a meal of 300 g of fresh or 30 g dried Larch Bolete will not result in exceeding reference doses for these elements. Extent of dose that customer would be exposed after eating of three 300 g fresh the Larch Bolete meals during a week will led to intake of lead and zinc within margin of 0.9-5.1% and 9-16% of PTWI for these elements, respectively. In the case of cadmium and specimens from the Western Beskid or Pogórze Wielickie sites in the south of Poland intake will slightly (around 10%) exceed PTWI value, and for other sites will not exceed (between 2% and 26% of PTWI).     

Development and application of a sediment toxicity test using the benthic cladoceran Chydorus sphaericus

This study reports on the development and application of a whole sediment toxicity test using a benthic cladoceran Chydorus sphaericus, as an alternative for the use of pelagic daphnids. A C. sphaericus laboratory culture was started and its performance under control conditions was optimised. The test was firstly validated by determining dose-response relationships for aqueous cadmium and copper and ammonia, showing a sensitivity of C. sphaericus (96 h LC(50) values of 594 microg Cd/L, 191 microg Cu/L and 46 mg ammonia/L at pH 8) similar to that of daphnids. Next, sediment was introduced into the test system and a series of contaminated sediments from polluted locations were tested. A significant negative correlation between survival and toxicant concentrations was observed. It is concluded that the test developed in the present study using the benthic cladoceran C. sphaericus is suitable for routine laboratory sediment toxicity testing.     

The effect of humic acid on uptake/adsorption of copper by a marine bacterium and two marine ciliates

The effect of humic acid (HA) on Cu uptake by a bacterium and two bacterivorus ciliates was investigated. The presence of HA resulted in a statistically significant (p<0.001) decrease in Cu associated with bacteria that were exposed to 67 microg Cu L(-1). Complexation of Cu appears to lower the availability of Cu with respect to bacterial cell surface binding and uptake. For ciliates, 10 mg HA L(-1) significantly reduced uptake of Cu by Uronema, but did not reduce uptake of Cu by Pleuronema. Uronema exposed to 67 microg Cu L(-1) accumulated 54% less Cu when 10 mg HA L(-1) was present (0.50 pg ciliate(-1) vs 0.23 pg ciliate(-1)). Uronema feeding on V. natriegens, took up less than half as much Cu as unfed Uronema when exposed to Cu without HA (0.41 pg Cu fed ciliate(-1) vs 0.86 pg Cu unfed ciliate(-1), but only 40% less when exposed to Cu and HA (0.31 pg Cu fed ciliate(-1) vs 0.51 pg Cu unfed ciliate(-1)). The lower % reduction attributable to fed ciliates in the presence of HA suggests that some of the Cu associated with HA is available through trophic processes.     

Toxic and essential trace elements in human milk from Greek lactating women: association with dietary habits and other factors

The aim of this study was to determine the concentration of some essential and toxic metals in the colostrum and transitory human milk in conjunction with various factors that may influence their concentrations i.e. diet, supplementation, place of residence, smoking, as well as socioeconomic and somatometric characteristics. Zinc, iron, copper, manganese, cadmium and lead were measured by AAS in 180 colostrum samples from healthy lactating women collected on third day postpartum. A second milk sample was collected in 95 (53%) subjects 14 days later. Dietary habits were assessed by a 7-day food frequency questionnaire and various characteristics and socio-economic factors were also recorded. The mean (+/-standard deviation) values of colostrum samples were: Zn 4905 +/- 1725 microg l(-1), Fe 544 +/- 348 microg l(-1), Mn 4.79 +/- 3.23 microg l(-1), Cu 381 +/- 132 microg l(-1), Cd 0.190 +/- 0.150 microg l(-1), Pb 0.48 +/- 0.60 microg l(-1). All metals with the exception of copper were found in lower concentrations in transitory samples. Cadmium and lead weekly intakes were found to be below the Maximum Tolerable Weekly Intakes as they have been established for infants by WHO or NRC. Our results revealed: higher Pb concentration in the samples from urban areas; effect of smoking on Cu level; dietary habits seem to play a role in metal levels in human milk as the logistic regression models revealed.     

Rapid behavioural and morphological responses of hydropsychid larvae (Trichoptera, Hydropsychidae) to sublethal cadmium exposure

Larvae of Hydropsyche contubernalis and H. siltalai were exposed to sublethal cadmium concentrations of 0, 0.012, 0.16 and 10 mg/litre(-1) for 72 h. Linear logit models revealed a significant increase in the frequency and degree of damage of the anal papillae of both species with increasing Cd concentration. Hydropsyche contubernalis showed a stronger and earlier anal papillae response under Cd exposure than H. siltalai, whereas in the latter species darkening of the ventral sides of the abdomen was also observed. The lowest Cd level altered the competition behaviour of Hydropsyche contubernalis larvae after only 24 h exposure, yet no visual signs of morphological damage were detected. The exposure intruder larvae spent significantly less time trying to enter the nets of resident larvae than did unexposed intruders. In addition, both the exposed intruders and exposed residents pursued different behavioural tactics during the encounters compared to their unexposed counterparts.     

Green oysters caused by copper pollution on the Taiwan coast

The first case of green oysters (Crassostrea gigas) broke out along the Charting mariculture area of south-western Taiwan in January 1986. The green color was found to be due to high copper content in the oyster tissue. Since then, a long-term survey around this area shows that total dissolved copper ranges from 4.99 to 23.6 microg/liter and particulate copper ranges from 1.09 to 5.51 microg/liter in sea-water. The green oysters collected from the Erhjin Chi estuary on 26 January, 1989 gave the highest copper content, 4401+/-79 ppm dry wt. Other green oyster cases were occasionally observed in the Hsiangsan and Anpin mariculture areas. Meanwhile, an experiment of copper accumulation in oysters was conducted at three stations (south-western Taiwan) for up to 90 days. Multiple regression analysis indicates that the food pathway may predominate in copper accumulation by green oysters. This bioaccumulation experiment shows that the total uptake of copper per oyster is an exponential function of exposure time for the first 2 weeks with an accumulation rate of 214 ppm Cu/day and then levels off. The average values of concentration factors for oysters (about 5 x 10(5)) were very close to steady-state values under the natural conditions at each station.     

Bioaccumulation of cadmium by the freshwater isopod Asellus aquaticus (L.) from aqueous and dietary sources

Experiments were conducted to determine the kinetics and relative importance of aqueous and dietary uptake of cadmium by the freshwater isopod Asellus aquaticus (L.). Test animals were exposed during 30 days to aqueous Cd in a continuous flow system (exposure levels: 0.2 - 10 microg litre(-1)) and kept on a diet of previously contaminated Elodea sp. (range of Cd concentrations: 2-350 microg g(-1), dry weight). Preceding semi-static experiments on dosage-control of the dietary factor revealed a rapid uptake of Cd by Elodea, with relatively high concentration factors (CF), which ranged from 4.8 to 5.5 (dry weight log (CF) after 16 days). For Asellus uptake from water appeared to be the predominant route. Highly significant bioconcentration of cadmium from water was observed in the animals, even at exposure levels below 1.0 microg litre(-1). In the various treatments, direct uptake from water accounted for 50-98% of the body burdens after 30 days exposure. The experimental results were described with a first order one-compartment bioaccumulation model. Model parameter estimates (mean +/- standard error) were obtained for rate constant of uptake (560 +/- 110 day(-1)), rate constant of elimination (0.032 +/- 0.017 day(-1)) and assimilation efficiency of Cd uptake from food (1.1 +/- 0.7%). The (dry weight) bioconcentration factor (BCF) and bioaccumulation factor (BAF) for extrapolated steady state conditions were estimated at 18 000 (BCF) and 0.08 (BAF). Experiments conducted at two different pH levels (5.9 versus 7.6) revealed no significant effects of pH on the uptake of aqueous Cd by the isopods. The results are discussed in relation to their potential significance to the field situation.     

Multi-generation cadmium acclimation and tolerance in Daphnia magna Straus

The cladoceran Daphnia magna was acclimated for seven generations to cadmium concentrations ranging from 0 (control) to 250 microg/l Cd (corresponding to a free ion activity of 4.60 nM Cd2+). Acute and chronic cadmium tolerance as well as cadmium accumulation were monitored as a function of acclimation time. After two to three generations of acclimation to concentrations ranging from 0.23 to 1.11 nM Cd2+ increases in acute tolerance were maximal (factor 7.2) and significant. Acclimation for seven generations to the same acclimation concentrations did result in an increased chronic cadmium tolerance (21 days EC50 values increased). Organisms acclimated to 1.93 nM Cd2+ were equally or more sensitive than non-acclimated daphnids in acute and chronic toxicity tests. Cadmium contents in D. magna increased significantly as a function of the acclimation concentration. Maximum body burdens of 236+/-30 microg Cd/g dry weight were measured in organisms exposed to 4.60 nM Cd2+, but detoxification mechanisms were only successful up to 82+/-20 microg Cd/g dry weight as this concentration did not cause major decreases in survival and reproduction in chronic toxicity tests. As the potential positive effect of acclimation on cadmium tolerance disappeared with successive acclimation generations and increasing acclimation concentrations, it is concluded that multi-generation acclimation studies are important for the evaluation of the long-term effects of environmental toxicants.     

Molecular cloning and expression of novel metallothionein (MT) gene in the polychaete Perinereis nuntia exposed to metals

To report a novel metallothionein (MT) gene and evaluate its potency as a biomarker, we clone this MT gene and measured the expression levels in the metal-exposed polychaete Perinereis nuntia. Accumulated metal contents and metallothionein-like proteins (MTLPs), which have been recognized as potential biomarkers, were compared with the relative mRNA expressions of the MT gene of P. nuntia (Pn-MT). In addition, the metal-binding affinity was estimated by recombinant Pn-MT protein. Pn-MT having high cysteine residues with three metal response elements in the promoter region closely clusters with those of other invertebrates. The accumulation patterns of metals were dependent on the exposure times in lead (Pb), cadmium (Cd), and copper (Cu) exposure. Particularly, both MTLP levels and relative mRNA expressions of MT were increased with accumulated metal contents and exposure time in P. nuntia exposed to Pb and Cd. There was no significant modulation of the Pn-MT gene in polychaetes exposed to Zn and As. However, the metal-binding ability of the recombinant Pn-MT protein provides a clear evidence for a high affinity of MT to several metal elements. These results suggest that Pn-MT would play an important role in the detoxification and/or sequestration of specific metals (e.g., Pb and Cd) in P. nuntia and have potential as a molecular biomarker in the monitoring of the marine environment using a polychaete.