Cadmium and zinc bioaccumulation and metallothionein response in two freshwater bivalves (Corbicula fluminea and Dreissena polymorpha) transplanted along a polymetallic gradient

This study was designed to compare the metallothionein (MT) response capacity of two freshwater bivalves, Corbicula fluminea and Dreissena polymorpha, along an environmental gradient of polymetallic pollution. The bivalves were transplanted into 4 stations in southwestern France, with a cadmium and zinc pollution gradient. MT and metal concentrations were measured in the soft bodies of the clams and mussels over 2.5 months. In the zebra mussels, variations in MT concentrations were strictly correlated to progressive Cd and Zn bioaccumulation. In contrast, the faster response in the clams appeared positively correlated to Cd accumulation only, with the activation of efficient detoxification mechanisms which limited Cd bioaccumulation and reduced Zn concentrations over time. Nevertheless, despite stronger metal accumulation factors in D. polymorpha, C. fluminea revealed higher sensitivity of MT response along the pollution gradient.     

Comparative studies on the biokinetics of Cd, Cr, and Zn in the green mussel Perna viridis and the Manila clam Ruditapes philippinarum

A kinetic approach was employed to determine the rates of metal uptake (Cd, Cr and Zn) from the dissolved phase and the rate constants of metal depuration in the mussel Perna viridis and the clam Ruditapes philippinarum. The effects of ambient metal concentration, salinity, and body size on the metal influx rate were examined. A linear positive relationship was observed between the metal influx rate and the metal concentration in ambient seawater. There was some evidence that Zn uptake was regulated by the bivalves in response to an increase in ambient Zn concentration. The uptake rate constant was highest for Zn and lowest for Cr in both bivalves, and was higher in mussels than in clams. The metal influx rate decreased by 1.6-1.8 times for the three metals when the salinity was increased from 15 ppt to 30 ppt. However, the effect of salinity on Zn influx in mussels was not statistically significant. A negative relationship of Cd and Zn influx rates with tissue dry weight was also found in both bivalves. Cr uptake in mussels was not significantly correlated with body size, but its uptake in clams was significantly correlated with body size. Metal concentration in ambient seawater appeared to be the most determining factor on metal uptake from the dissolved phase in both bivalves. The efflux rate constants of the three metals were within the range of 0.01-0.03 d-1, and were comparable between the mussels and the clams. Using a simple bioenergetic-based kinetic model, it was shown that both dissolved uptake and food ingestion can contribute to metal accumulation in the bivalves. However, Zn accumulation in the clam R. philippinarum was dominated by uptake from food ingestion. Metal partitioning in ingested food was found to be critical in affecting the relative importance of metal uptake from the dissolved phase and food source, primarily because of the large variability of this parameter in natural environments.     

Selective uptake,distribution, and redistribution of 109Cd, 57Co, 65Zn, 63Ni,and 134Cs via xylem and phloem in the heavy metal hyperaccumulator Solanum nigrum L

The focus of this article was to explore the translocation of ¹⁰⁹Cd, ⁵⁷Co, ⁶⁵Zn, ⁶³Ni, and ¹³⁴Cs via xylem and phloem in the newly found hyperaccumulator Solanum nigrum L. Two experiments with the uptake via the roots and transport of ¹⁰⁹Cd, ⁵⁷Co, and ⁶⁵Zn labeled by roots, and the redistribution of ¹⁰⁹Cd, ⁶⁵Zn, ⁵⁷Co, ⁶³Ni, and ¹³⁴Cs using flap label in S. nigrum in a hydroponic culture with a standard nutrient solution were conducted. The results showed that ¹⁰⁹Cd added for 24 h to the nutrient medium of young plants was rapidly taken up, transferred to the shoot, and accumulated in the cotyledons and the oldest leaves but was not efficiently redistributed within the shoot afterward leading to a rather low content in the fruits. In contrast, ⁵⁷Co was more slowly taken up and released to the shoot, but afterward, this element was redistributed from older leaves to younger leaves and maturing fruits. ⁶⁵Zn was rapidly taken up and transferred to the shoot (mainly to the youngest leaves and not to the cotyledons). Afterward, this radionuclide was redistributed within the shoot to the youngest organs and finally accumulated in the maturing fruits. After flap labeling, all five heavy metals tested (¹⁰⁹Cd, ⁵⁷Co, ⁶⁵Zn, ⁶³Ni, ¹³⁴Cs) were exported from the labeled leaf and redistributed within the plant. The accumulation in the fruits was most pronounced for ⁶³Ni and ⁶⁵Zn, while a relatively high percentage of ⁵⁷Co was finally found in the roots. ¹³⁴Cs was roughly in the middle of them. The transport of ¹⁰⁹Cd differed from that previously reported for wheat or lupin and might be important for the potential of S. nigrum to hyperaccumulate cadmium.     

Biomonitoring of heavy metals in the Western European Rivers Rhine and Meuse using the freshwater mussel Dreissena polymorpha

Since 1976, active and passive biological monitoring programs using the freshwater mussel Dreissena polymorpha have been carried out to study trends in the bioavailability of heavy metals in the rivers Rhine and Meuse. The Cd concentration in mussels exposed in the river Rhine has decreased from 74 mg kg(-1) in 1976 to 1.5 mg kg(-1) in 1988. In the river Meuse, however, the Cd concentration in Dreissena polymorpha has increased from 6 mg/kg (-1) to 22 mg kg(-1). Cu concentrations in mussels have not changed. Comparison of heavy metal concentrations in mussels from Lake Heerhugowaard, Lake Markermeer, Lake Maarsseveen and the river Dieze demonstrated that background concentrations for Cu, Zn, Cd and Pb in Dreissena polymorpha are about 12, 110, 1 and 0.5 mg kg(-1), respectively. Mussels from these four locations showed seasonal variation in heavy metal concentrations. This may be caused by both changes in the bioavailability of the metals and by the annual growth and reproductive cycle of the mussels. Therefore, one should take care that animals are collected at the same location and in the same section in long-term active biological monitoring programs.     

Trophic transfer of paralytic shellfish toxins from clams (Ruditapes philippinarum) to gastropods (Nassarius festivus)

A local strain of the dinoflagellate Alexandrium tamarense (ATCI01), which predominantly produces C2 toxin, was fed to the clams (Ruditapes philippinarum) under laboratory conditions. Concentrations of paralytic shellfish toxins (PSTs) in the dosed clams were determined by High Performance Liquid Chromatographic (HPLC) analyses, and the clams were homogenized and then fed to the gastropods (Nassarius festivus). In the toxin accumulation phase, which lasted for 42 days, concentrations of PSTs increased in the snails gradually, reaching a maximum of 1.10 nmole g(-1) at the end of the exposure period. The toxin content of the homogenized clams (food) was 13.18 nmole g(-1), which was about 12-fold higher than the PST content in the snails. Between day 43 and day 82, the snails were fed with non-toxic clams, and this period represented the depuration phase. Accumulation and depuration rates of PSTs in the snails, N. festivus, were determined by fitting the experimental data to user-defined parameters program using a one-compartment model. Two different modeling approaches were used to derive the accumulation and depuration rates. The first approach is to derive both values from the data for the toxin uptake. The second approach is to derive depuration rate from the depuration data and then to derive uptake rate, allowing for toxin depuration, from the data for toxin uptake. The first approach yielded more consistent results for the toxin concentration at the end of the uptake period, when compared with the experimental data. The toxin uptake and depuration rates were 1.64 (pmole of toxin into snail per day) per (nmole g(-1) of toxin in food) and 0.06+/-0.02 day(-1) (mean+/-SE), respectively. The toxin profiles of snails were similar to the clams, but different from the algae. Besides C toxins (C1 and C2), dcGTX2 and dcGTX3 were also detected in both clams and snails. The beta:alpha epimer ratio gradually decreased during trophic transfer and approached a ratio of 1:3 (26.4 mol%:73.6 mol% at day 42) in the snails, near the end of the accumulation period.     

Comparative bioaccumulation kinetics of trace elements in Mediterranean marine sponges

While marine organisms such as bivalves, seagrasses and macroalgae are commonly used as biomonitors for the environment pollution assessment, widely distributed sponges received little attention as potential helpful species for monitoring programmes. In this study, the trace element and radionuclide bioaccumulation and retention capacities of some marine sponges were estimated in a species-comparative study using radiotracers technique. Six Mediterranean species were exposed to background dissolved concentrations of ^110mAg, ²⁴¹Am, ¹⁰⁹Cd, ⁶⁰Co, ¹³⁴Cs, ⁵⁴Mn, ⁷⁵Se and ⁶⁵Zn allowing the assessment of the uptake and depuration kinetics for selected elements. Globally, massive demosponges Agelas oroides, Chondrosia reniformis and Ircinia variabilis displayed higher concentration factor (CF) than the erectile ones (Acanthella acuta, Cymbaxinella damicornis, Cymbaxinella verrucosa) at the end of exposure, suggesting that the morphology is a key factor in the metal bioaccumulation efficiency. Considering this observation, two exceptions were noted: (1) A. acuta reached the highest CF for ^110mAg and strongly retained the accumulated metal without significant Ag loss when placed in depuration conditions and (2) C. reniformis did not accumulate Se as much as A. oroides and I. variabilis. These results suggest that peculiar metal uptake properties in sponges could be driven by specific metabolites or contrasting biosilification processes between species, respectively. This study demonstrated that sponges could be considered as valuable candidate for biomonitoring metal contamination but also that there is a need to experimentally highlight metal-dependant characteristic among species.     

Uptake of 134Cs in the shoots of Amaranthus tricolor and Amaranthus cruentus

Amaranthus tricolor L. and Amaranthus cruentus L. were grown in pots containing 7.5 kg soils artificially contaminated with three levels of 134Cs activity: 5.55 x 10(5) Bq pot-1, 1.11 x 10(6) Bq pot(-)1, and 1.665 x 10(6) Bq pot(-1), respectively. Forty-nine days after sowing and growth, plants were harvested. The plants growing in soils with increasing 134Cs concentrations showed increasing concentration of this radionuclide in shoots. There were significant differences in uptake of 134Cs applied to soils between and within the plant species, depending on the initial 134Cs concentrations. The plant species showed different responses to the addition of (NH4)2SO4 to soils. Biomass production of both species was reduced in pots treated with (NH4)2SO4. (NH4)2SO4 application decreased the uptake of 134Cs by A. tricolor but increased the accumulation of 134Cs by A. cruentus, showing that chemicals with the highest efficiency to enhance the desorption of 134Cs might play an unexpected role in transferring the radionuclide to shoots.     

Accumulation and depuration of cyanobacterial toxin nodularin and biomarker responses in the mussel Mytilus edulis

Blue mussels (Mytilus edulis) were exposed to an extract made of natural cyanobacterial mixture containing toxic cyanobacterium Nodularia spumigena (70-110 microg nodularin l(-1), 24-h exposure followed by 144-h depuration period in clean water). Toxin concentration increased from initial 400 to 1100 mg kg(-1) after 24-h exposure, measured by liquid chromatography/mass spectrometry (LC/MS). Acetylcholinesterase activity (AChE), a biomarker of direct neurotoxic effects, showed inhibition after 12 and 24h exposure but returned to control level during the depuration period. Catalase (CAT) activity, an indicator of oxidative stress, showed significantly elevated levels in exposed mussels but only 72 h after the end of the exposure. No change in the activity of glutathione-S-transferase (GST) involved in conjugation reactions could be observed. A gradual yet incomplete elimination of nodularin (from 1100 to 600 mg kg(-1)) was observed during the depuration period, and the tissue levels were 30% lower in clean water after 24 h. The observed increase in oxidative stress indicated by elevated CAT activity is likely connected to detoxification reactions leading to the production of reactive oxygen species, including an apparent time lag in this specific enzymatic defence response. That no change in GST activity was observed suggests that this enzyme is not significantly involved in the detoxification process of nodularin-containing cyanobacterial extract in M. edulis.     

Comparative study of the cadmium and mercury kinetics between the short-lived gastropod Viviparus georgianus (Lea) and pelecypod Elliptio complanata (Lightfoot), under laboratory conditions

A laboratory study of the cadmium and mercury accumulation and elimination kinetics was conducted on the pelecypod Elliptio complanata (Lightfoot) and the short-lived gastropod Viviparus georgianus (Lea), according to age-classes. Preliminary results (metal concentration vs time of exposure) have demonstrated that uptake of Cd and Hg, in the two molluscs studied, follow a biphasic pattern, whereby a steady state is reached after approximately 16 days' exposure and then accumulation increases again for the rest of exposure period. The elimination of the two metals is also characterized in a biphasic way: fast excretion for the first four days followed by a slower depuration for the rest of exposure time. A two-compartment bioaccumulation model has been used to described the different kinetic parameters: (1) the rate constant for depuration; (2) the rate constant for uptake; (3) the theoretical bioconcentration factor extrapolated to steady-state conditions; and (4) the biological half-life of the metals.     

Identification of multixenobiotic defence mechanism (MXR) background activities in the freshwater bivalve Dreissena polymorpha as reference values for its use as biomarker in contaminated ecosystems

The biological defence mechanism called MXR or MXD for multixenobiotic resistance or defence protects cells against the entry and the accumulation of xenobiotics. As the defence is modulated by man made chemicals, MXR is used as a biomarker of organisms' exposure to environmental contamination. However, to reliably assess and evidence MXR induction, the use of a reference level is required. In this context, we focused on MXR background level in a freshwater bivalve, the zebra mussel Dreissena polymorpha, in order to propose its use as a reference during MXR evaluation. We monitored the MXR transport activity in mussels collected either in a natural population or in a caged population and then transplanted to clean water in the laboratory. The results showed that MXR activity was decreased to its baseline level after an eight to nine day depuration period (13.1+/-3.1; 7+/-2.6; 13.7+/-3.9 pmol RB min(-1)org(-1) after three experiments of laboratory depurations). Moreover, significant MXR induction was measured in depurated zebra mussels exposed to contaminated sites (39.6+/-3.7; 59.2+/-20.3 pmol RB min(-1)org(-1) after two experiments of field exposure), showing that the laboratory depuration did not affect the induction potential. The MXR responses (decrease as well as increase) occurred in few days and were highly significant, highlighting its reactivity in zebra mussels. Finally, this paper confirms the usefulness of MXR as a tool in biomonitoring studies and provides a protocol for field experiments that enables to establish and use the background level of MXR activity as a reference.     

The fate of trace metals during the metamorphosis of chironomids (diptera, chironomidae)

The fate of the trace metals zinc, cadmium and copper during the metamorphosis of chironomids was studied under field and experimental conditions. Field observations demonstrated a significant decrease in the trace metal body burden of larvae and imagines of Stictochironomus histrio Fabricius. No such difference were noted between successive developmental stages of Chironomus anthracinus Zett. The exuviae of both species contained small amounts of trace metals. Experiments, in which larvae of S. histrio and C. riparius Meigen were individually exposed, showed that larvae of both species accumulated substantial amounts of zinc and cadmium. For these metals, a transfer to pupae and imagines occurred. However, pupae had a lower body burden than larvae, and imagines a lower body burden than the pupae. Copper was also accumulated in the larvae, but it was excreted almost completely before the pupal stage. Elimination pathways of the trace metals during metamorphosis and the ecotoxicological consequences of trace metal transfer are discussed.     

60Co accumulation from sediment and planktonic algae by midge larvae (Chironomus luridus)

This paper reports the results of several experiments carried out to evaluate uptake and retention by a limicolous midge larva of 60Co retained in sediment, either adsorbed on mineral particles or bound to planktonic algae. In order to determine their relative contributions in radionuclide accumulation, the different vectors (water, algae and sediment) were first labelled individually and then simultaneously. 60Co accumulation from water and from algae results in a maximum concentration factor of 30 and in a mean trophic transfer factor of 4.5 x 10(-3). The level of contamination of midge larvae from sediment is markedly influenced by the presence of endogenous organic matter. Thus the radionuclide transfer factor is about twice as high for larvae placed in labelled raw sediment than for larvae placed in labelled incinerated sediment, in the presence as in the absence of contaminated planktonic algae. Irrespective of the contamination conditions, 60Co depuration from midge larvae is a very rapid phenomenon that corresponds, in all cases, to a radionuclide half-life of only a few days.     

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.     

Are native naiads more tolerant to pollution than exotic freshwater bivalve species? An hypothesis tested using physiological responses of three species transplanted to mercury contaminated sites in the Ebro River (NE, Spain)

In the lower Ebro River exist the paradoxical convergence of relatively well preserved river dynamics with the historical presence of a chloralkali plant with a long history of mercury discharges and the recent invasion of foreign bivalves species. Here we performed a comparative study on two alien bivalves, the Zebra mussel and the Asian clam (Dreissena polymorpha and Corbicula fluminea), and one protected species of naiads (Psilunio littoralis), which is the most common species of the freshwater mussel assemblages in this river. Individuals of the three species were transplanted to three sites that included a clean unpolluted upstream site, a contaminated location next to the mercury source and a downstream one. The study focused on digestive gland antioxidant and oxidative stress responses such as antioxidant enzymes, glutathione S transferase, glutathione levels, metallothionein proteins, DNA strand breaks and lipid peroxidation levels. Results evidenced interspecies differences on accumulation levels of mercury, antioxidant defensive systems and oxidative tissue damage. The naiad species, despite of accumulating more mercury showed the greatest antioxidant defensive potential, which was characterized by having high constitutive activities of glutathione S transferase and inducible activities and levels of key antioxidant enzymes and glutathione. Exposed individuals of C. fluminea had moderate levels of metal accumulation, the highest activities of antioxidant enzymes but also high levels of lipid peroxidation. D. polymorpha mussels showed the lowest levels of mercury but the lowest antioxidant responses and consequently the highest levels of oxidative injuries in the DNA and of mortality. Our results support the hypothesis that naiad species might be more tolerant to pollution than exotic species.     

Evaluation of transport and storage of 60Co, 134Cs, 137Cs and 65Zn by river sediments in the lower Susquehanna River

The Peach Bottom Atomic Power Station (PBAPS) has contributed measurable quantities of radioactivity to Conowingo Reservoir, an impoundment of the lower Susquehanna River. As part of an ongoing radiological assessment program, concentrations of plant-related radionuclides in sediments have been monitored in spring and fall since 1980. Mass balance estimates derived from grab samples of surface sediments (< 10 cm) indicate that less than 20% of reactor released (60)Co, (65)Zn, (134)Cs and (137)Cs is present in these sediments. Significant seasonal variations in radionuclide trapping efficiency by the reservoir are not apparent. Deep core samples (c. 200 cm) confirm that some, but not all, of this surface sediment radionuclide inventory remains within the reservoir-trapped in discrete locations by subsequent sediment accumulation. The remaining radionuclide mass, in dissolved or particle-associated form, appears to be transported downstream, through Conowingo Dam, to upper Chesapeake Bay. The detection of PBAPS-derived radionuclides in the sediments of upper Chesapeake Bay, primarily the Susquehanna Flats, confirms the transport of these radionuclides from the lower Susquehanna River.     

Impact of humic substances on the aqueous solubility, uptake and bioaccumulation of platinum, palladium and rhodium in exposure studies with Dreissena polymorpha

Zebra mussels (Dreissena polymorpha) were exposed to different types of water containing PGE salts (PtCl4, PdSO4, RhCl3) to investigate the influence of humic substances on the aqueous solubility, uptake and bioaccumulation of noble metals. The results showed a time dependent decrease of the aqueous PGE concentrations in tank water for all groups. This could mainly be related to non-biological processes. The aqueous solubility of Pd and Rh was higher in humic water compared with non-chlorinated tap water, whereas Pt showed opposing results. Highest metal uptake rates and highest bioaccumulation plateaus were found for Pd, followed by Pt and Rh. Pd uptake and bioaccumulation was significantly hampered by humic substances, whose presence appear to increase Pt uptake and bioaccumulation. No clear trend emerged for Rh. Differences in effects of humic matter among the PGE may be explained by formation of metal complexes with different fractions of humic substances.     

The effects of cadmium or zinc multigenerational exposure on metal tolerance of Spodoptera exigua (Lepidoptera: Noctuidae)

The effects of ten generational zinc or cadmium pre-exposure on metal tolerance among beet armyworm Spodoptera exigua individuals were compared. These effects were assessed in animals from the 11th generation, reared on a diet either uncontaminated or contaminated with metal (cadmium or zinc). The survival rate of larvae and the degree of metal accumulation (in larvae, pupae and moths; among larval organs: gut and fat body) were analysed. Catalase, superoxide dismutase and glutathione transferase activity in larval organs of individuals subjected to different metal treatments were also measured. Animals transferred from control rearing to metals (cadmium or zinc) in the 11th generation, as well as those from multigenerational zinc treatment, but not from multigenerational cadmium treatment, had a significantly lower survival rate than control animals. Insects from the groups with the high metal treatment had high bioaccumulation factors (above 3.7 and 2.3 following cadmium and zinc, respectively). Cadmium (but not zinc) pre-exposure had a significant effect on metal accumulation in larvae. Multigenerational metal pre-exposure seemed to have mainly a negative effect on glutathione transferase activity in the gut of larvae from the 11th generation, in the case of the individuals exposed to metal other than that used in pre-exposure treatment or kept in control conditions. However, in the case of zinc pre-exposure, such effect was only apparent when zinc was replaced by cadmium. The long-term effect of cadmium on catalase activity in larvae was found.     

Effect of ectomycorrhizae and ammonium on 134Cs and 85Sr uptake into Picea abies seedlings

Microorganisms play an important role in the fixation of radionuclides in forest soils. In particular, fungi have the capacity to absorb and translocate radionuclides. The role of the ectomycorrhizal fungus Hebeloma crustuliniforme in the uptake of radiocaesium (134Cs) and radiostrontium (85Sr) into seedlings of Norway spruce (Picea abies) was investigated in a pouch test system. Inoculated and non-inoculated seedlings; seedlings inoculated during 8 and 15 weeks; seedlings exposed during 2 and 3 weeks to the radioactive solution; and seedlings grown under low and high ammonium conditions prior to the application of the radionuclides were compared. The final 134Cs and 85Sr activity was determined in fine-roots, main-roots, stems and needles. The results showed that ectomycorrhizae reduced the uptake of 134Cs and 85Sr. The degree of ectomycorrhization was of crucial importance and seemed to be governed by the period during which ectomycorrhizae were allowed to develop and by the ammonium concentration in the nutrient solution. The radionuclide uptake increased with increasing exposure time. Both radionuclides were predominantly accumulated in fine-roots. However, needles proved to describe best the result of net root uptake and translocation to the shoot. The uptake-and and translocation-rates of 85Sr were smaller than those of 134Cs. It is assumed that the translocation is coupled with the intensity of water fluxes through the xylem and that 85Sr is more readily adsorbed into mycelium or plant tissue relative to 134Cs. The effect of high ammonium growth conditions was overcome by the effect of ectomycorrhization, except in needles with a very large biomass which behaved as a strong sink and led to a high accumulation of 134Cs.     

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.