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.     

Variability in dry weight and vertical distributions of decapod larvae in the Irish Sea and North Sea during the spring

Accumulation of waterborne cadmium in Littorina littorea, Mytilus edulis and Carcinus maenas (collected in 1988 and 1989 around the island of Funen, Denmark) was investigated in a matrix of salinities (10 to 30) and calcium concentrations (2.9 to 8.9 mM Ca⁺⁺). Cadmium accumulation rates in soft parts of L. littorina, soft parts and shells of M. edulis and whole bodies and exoskeletons of C. maenas decreased with increasing salinity. Changes in the calcium concentrations accounted for 72% of the salinity effect on cadmium accumulation rates in L. littorina, whereas calcium concentrations had little or no effect on cadmium accumulation in M. edulis. Cadmium accumulation in the whole body of C. maenas was affected equally by calcium concentrations and total salinity, whereas accumulation in the exoskeleton was mainly affected by changes in total salinity. Individual variability in cadmium accumulation in the organs of C. maenas was greater than the variation attributable either to changes in ambient calcium concentrations or total salinity. An appreciable amount of the inter-individual variability in the cadmium accumulation in all three species was correlated with wet:dry weight ratios of the tissues and size of the organisms.     

Effects of cadmium and low magnesium on cell division and calcification in the marine coccolithophorid Cricosphaera (Hymenomonas) carterae

Cadmium (Cd) added to normal media and to magnesium (Mg)-deficient media produced inhibitory effects on cell division and cell calcification in the marine coccolithophorid alga Cricosphaera (Hymenomonas) carterae. Compared with controls in normal media (with 25 mM Mg), cell growth decreased progressively with Cd at concentrations of 5–20 M. In Mg-deficient media (with 6 mM and 0.08 mM Mg) the inhibitory effects were more pronounced with complete arrest of cell division at 20 M Cd. The greatest Cd inhibition occurred in media with the lowest Mg concentration (0.08 mM). Cadmium (5–40 M) also decreased recalcification (coccolith formation) in cells previoasly decalcified with CO₂ with complete inhibition at 40 M Cd. Inhibition of recalcification in various Cd concentrations (5–40 M) was more pronounced in low-Mg medium (with 6 mM and 0.08 mM Mg) compared with normal medium (25 mM Mg). Partial or complete reversal of the inhibitory effects of Cd and low Mg media on cell division and calcification occurred following a wash and resuspension of the cells in normal control medium (25 mM Mg).     

Uptake of cadmium by the eelgrassZostera marina

The uptake of cadmium by the shoots ofZostera marina L. (eelgrass) was examined in the laboratory. Experiments were carried out in experimental chambers which allowed the separation of the leaves from the root-rhizome portions of intact shoots. Cadmium uptake by the root-rhizome portions over 24 h was directly related to substrate cadmium concentration (1.0, 5.0 and 10.0 g Cd ml^-1) and varied from 6.5 to 30.0 g Cd g^-1. Cadmium uptake by the root-rhizomes and the leaves in a substrate concentration of 1 g Cd ml^-1 was also related to exposure times (24, 48 and 72 h). Maximum uptake by the root-rhizomes and the leaves was observed after 72 h at a substrate concentration of 1 g Cd ml^-1 and was equivalent to 48 and 94 g Cd g^-1, respectively. Translocation of cadmium from the leaves to the root-rhizomes was observed after 24 h, and at the end of 72 h was equivalent to 27% of the total leaf uptake. No cadmium movement from the root-rhizomes to the leaves was detected.     

Cadmium accumulation,LC50 and oxygen consumption in the tropical marine amphipod Elasmopus rapax

Adult Elasmopus rapax, collected from the eastern coast of Venezuela in 1990, were exposed to seawater containing various CdCl₂ concentrations ranging from 0.25 to 5.5 mol l^-1. The 48-h and 96-h LC₅₀ values obtained were 4.0 and 1.6 mol Cd l^-1, respectively. In amphipods exposed to 1 mol Cd l^-1 for up to 240 h, the apparent rate of cadmium uptake was higher in dead animals (most of which had molted during the preceding 24 to 48 h) than in those which survived throughout the treatments without molting. Thus, whole-body cadmium content reached 1.74 mol g^-1 dry weight (dw) in the former and only 0.85 mol g^-1 dw in the latter; the higher body Cd-load may have caused the increased mortality observed in molters. On exposure to cadmium levels above 0.5 mol l^-1 the oxygen consumption rate of non-molters decreased from 2.2 to about 1.5 ml O₂ g^-1 dw h^-1 over the first 24 h, remaining unchanged thereafter. The results place E. rapax among the most sensitive marine organisms yet studied concerning cadmium toxicity, and emphasize the usefulness of the Amphipoda as bioindicators and research tools for bioassays.     

Crassostrea virginica as an indicator of lead pollution

After treatment with 1.0 and 3.3 g Pb kg^-1 for 20 weeks in flowing seawater at ambient salinity and temperature, Crassostrea virginica accumulated as much as 6.57 and 11.42 g g^-1 dry weight, respectively, and no mortalities were recorded. Lead uptake was curvilinear; however, the general trend was an increase in lead concentration over time. Dry weight of the oyster had no significant relation with tissue lead concentration (g g^-1); however, a significant positive relationship existed between weight and lead content (g). A decrease in lead concentration in the tissues occurred from mid-August to mid-September despite continuous addition of lead to the seawater. Addition of lead to the seawater was terminated after 20 weeks (October), and lead loss was studied for the following 12 weeks. Lead loss was studied well after spawning in an attempt to eliminate any influence this might have on lead concentration in the tissues. After 4 weeks, approximately 54% of the accumulated lead was lost under conditions of a natural temperature decline. The biological half-life of lead in C. virginica tissues was calculated to be 5.5 weeks. In the ensuing 8 weeks, little or no lead loss was observed. Larvae from each treatment developed to the straight-hinged stage within 48 h with no apparent abnormalities. Lead appeared to have no adverse effect on larvae when parents had been treated with 1.0 and 3.3 g kg^-1 seawater for 10 weeks.     

Relationship between cadmium concentrations in seawater and those in the mussel Mytilus edulis

Various forms of regression analysis are presented which show that there is a significant equilibrium relationship between total recoverable cadmium in seawater and its concentration in the mussel Mytilus edulis (P<0.001). The calculations show that the concentration of cadmium in seawater should not exceed 0.20 g l^-1 if the mussel is not to reach a cadmium concentration of 2 mg kg^-1 wet weight: a value frequently used as a food standard for human consumption. Further, when a cadmium value of 2 mg kg^-1 wet weight is reached, the concentration factor by mussels for cadmium from seawater is 9 950.     

Influence of temperature and salinity on the uptake,distribution and depuration of mercury,cadmium and lead by the black-lip oyster Saccostrea echinata

Saccostrea echinata (Quoy and Gaimard) were exposed to 10 g 1^-1 of either mercury, cadmium or lead at 30 °C, 36S; 30 °C, 20S; 20°C, 36S and 20°C, 20S for 30 d and were then transferred to clean seawater for a further 30 d to depurate. Specimens were removed at regular intervals during the exposure and depuration periods, dissected into gills, mantle, visceral mass and adductor, and analysed for the appropriate metal by atomic absorption spectroscopy. Mercury was concentrated more than the other metals in all tissues under all conditions. Cadmium uptake was greater than lead in all tissue in the high-temperature experiments, whereas both metals were concentrated to similar extents at low temperature. The gill tissue generally accumulated the greatest amount of all 3 metals, whilst the adductor concentrated the least amount. At both salinities, mercury and cadmium accumulation by all tissues was significantly greater at the higher temperature whereas lead uptake was only marginally increased. The accumulation rates of mercury at high temperature were significantly greater in all tissues at low compared with high salinity, whereas at low temperature, differences were not significant. Accumulation rates of cadmium and lead in the majority of tissues examined were significantly greater in lowsalinity water at both temperatures. In general, lead was lost the most rapidly from oyster tissues, followed by mercury and then cadmium. The residence times for mercury and cadmium differed significantly between tissues, with the gills showing the highest turnover rate. In contrast, residence times for lead were similar between tissues. Losses of all 3 metals from oyster tissues were not obviously influenced by temperature and only mercury losses differed significantly between salinities.     

Accumulation of cadmium from contaminated water and sediment by the shrimp Callianassa australiensis

The burrowing marine shrimp Callianassa australiensis (Dana) was collected from an uncontaminated area in Western Port, Victoria, Australia in 1977. The shrimp were exposed to cadmium-contaminated water and sediment for 8 wk. The concentrations ranged from 0.5 to 63 g Cd 1^-1 for water and 0.5 to 63 g Cd g^-1 for sediment. The shrimp accumulated cadmium from water at a rate commensurate with increases in the concentration of cadmium in water and the duration of the experiment. Although the cadmium concentration in the sediments was 10³ times higher than that in water, it hat no effect on cadmium uptake by the shrimp. The concentration factors decreased with increasing concentration of cadmium in water but increased as the duration of exposure increased. The shrimp dry weight decreased with increasing concentration of cadmium in water and duration of exposure. As was the case with cadmium uptake by the shrimp, these two factors acted interactively on the shrimp dry weight, but the third factor, concentration of cadmium in sediment, had no effect.     

Cadmium bioaccumulation in organs of the scallopMizuhopecten yessoensis

Cadmium and Zinc content was determined in organs of the Japanese scallopMizuhopecten yessoensis, collected in 1984 from the Sea of Japan and aged between 1 and 8 yr. Under conditions of background Cd, Cd concentration in hepatopancreas and kidney increased linearly with age from 39 to 400µg g^?1 dry wt in hepatopancreas, and from 100 to 640µg g^?1 dry wt in kidney. Such a pattern was not determined for Zn. Cd concentration in muscle, mantle and gill did not exceed 6µg g^?1 dry wt in the oldest scallops. In subcellular fractions of the hepatopancreas, cytosolic Cd accounted for 71.7% in 1-yr-old scallops and 98.8% in 8-yr-olds. A similar ratio was established for gills, although gill Cd content was an order of magnitude lower. Analysis of Cd distribution in cytoplasmic proteins (of different molecular weight) in the hepatopancreas, showed that the amount of Cd bound to metallothionein-like proteins increases with scallop growth. A considerable amount of Cd also was detected in high molecular weight proteins. Thus, Cd accumulation, with age, in organs of the Japanese scallop is due to metal binding by cytoplasmic proteins.     

Uptake and loss of radioactive cadmium by the sea-skater Halobates robustus (Heteroptera: Gerridae)

Sea-skaters, Halobates robustus Barber, caught around the Galápagos Islands, were exposed over a period of 3 days to radioactive cadmium dissolved in seawater. Although ^115mCd (metastable ¹¹⁵Cd) was accumulated by the insects, they did not concentrate it above the seawater level. The experiments clearly showed that cadmium is taken up into the body, not merely adsorbed onto surfaces. The uptake of Cd followed a biphasic time course. When sea-skaters labelled in this way were transferred back into unlabelled seawater, the rate of loss of Cd was likewise biphasic. During uptake and loss, the biological half-life times for the first compartments were almost identical (0.64 and 0.79 h), indicating that the same compartment might be involved. On the other hand, the biological half-life times for the second compartments were different (1.7 days during uptake, 19.5 days during loss), indicating that the site or chemical form of bound Cd in the sea-skater had changed. For an adult female, weighing about 8.5 mg wet weight, the drinking rate of seawater was estimated at 2.5 l day^-1, equivalent to 29% body weight.     

Studies on the uptake of cadmium by the crab Carcinus maenas in the laboratory. I. Accumulation from seawater and a food source

The crab Carcinus maenas (L.) was exposed to radioactively labelled cadmium dissolved in seawater at concentrations of 0.1, 1 and 10 ppm, the latter concentration being toxic to the crabs (50% mortality after 12.3 days). Net accumulation of cadmium from solution was proportional to the level and time period of cadmium exposure. Total absorbed cadmium levels reached 0.0043 and 0.0412 mg Cd g^-1 dry weight after 40 days exposure to 0.1 and 1 ppm Cd, respectively, and 0.1115 mg Cd g^-1 dry weight after 12.3 days average exposure to 10 ppm Cd. The highest tissue concentration was found in the midgut gland, reaching 0.786 mg Cd g^-1 dry weight after 12.3 days average exposure to 10 ppm Cd. The midgut gland only contained about 10% of the total cadmium absorbed from solution, while the exoskeleton contained the bulk of obsorbed cadmium (59 to 80%) probably passively adsorbed onto the surface. When cadmium was absorbed by the crabs from a food source, the midgut gland contained 16.9% of the total absorbed cadmium whereas the exoskeleton now contained only 22.2%. Ten percent of the cadmium available in the food source (Artemia salina) was accumulated by the crabs. When placed in cadmium-free seawater, crabs that had accumulated cadmium from solution lost 69% of the absorbed cadmium in 10 days, mostly from the exoskeleton which lost 78% of its original absorbed cadmium concentration.     

Accumulation of cadmium by Artemia salina

The relative importance of accumulation of cadmium by Artemia salina (L.) directly from solution and from ingested food has been studied at 3 cadmium concentrations (0.1, 1 and 10 ppm) under controlled experimental conditions. At each cadmium concentration, A. salina were exposed to cadmium in 4 ways; in solution; in solution in the presence of latex food particles; in solution with cadmium-rich Dunaliella tertiolecta as a food source; and to cadmium-rich D. tertiolecta alone. Net accumulation of cadmium by A. salina continued throughout 5 days exposure under all 4 conditions. When the brine shrimp were removed to cadmium-free conditions, their accumulated cadmium concentrations declined and levelled off to a stabilised plateau after 10 days. From consideration of these stabilised levels it was shown that at least 30% of cadmium accumulation directly from solution occurs via uptake through the alimentary tract. The ratio of cadmium accumulated from solution to cadmium accumulated from food was found to be 1:4.9, 1:6.7 and 1:1.1 at 0.1, 1 and 10 ppm Cd exposure, respectively. At lower cadmium exposures uptake from food is the major route for cadmium accumulation, but at higher exposures the cadmium-saturated food source displaced cadmium-rich water from the gut and therefore actually inhibited cadmium accumulation. This study, therefore, concludes that the food chain will be the major source of cadmium as long as the previous trophic level has the ability to accumulate the metal to such an extent as to make it more available to the consumer than by direct uptake from seawater.     

Seasonal variations in biochemical constituents during the reproductive cycle of the female dog cockle Glycymeris glycymeris

In an attempt to describe the biochemical events associated with the main stages of the annual and reproductive cycles of the female dog cockle Glycymeris glycymeris L., we studied seasonal variations in the various stages of oocyte development of the ovaries, and the glycogen, total protein and total lipid content of five body tissues – adductor muscle, foot, tunic coat, visceral mass and mantle. From November 1991 to November 1994, microscopic examination of the ovaries and measurement of the tissue concentrations of glycogen, total proteins and total lipids in these five body tissues were made monthly on ten female dog cockles originating from the sea area around Douarnenez (south Brittany, France). Morphological studies revealed that in the population investigated the annual cycle is characterised by three major periods: a first period of vitellogenesis extending from February/March to April/May and preceding a spawning in spring; a second period of vitellogenesis extending from May/June to September/October and leading to either no spawning, a single autumnal spawning event, or to two spawning events in summer and autumn; and a third period extending from October/November to February/March and characterised by a high level of oocyte lysis. In the muscular body tissues of the dog cockle, i.e. the adductor muscles, the foot and the tunic coat (the muscular envelope containing the visceral mass), the concentrations of glycogen, total proteins and total lipids underwent very similar variations during the annual cycle. During each stage of vitellogenesis, a typical glycogen–protein–lipid sequence was observed in the muscular tissues that was characterised firstly by a peak of glycogen concentration 2 to 3 mo before spawning, followed by a peak in total proteins 1 mo before spawning, and finally by a peak in lipid content just before spawning. A similar glycogen–protein–lipid sequence was also recorded in the first half of the winter period. However, these events were followed by general atresia affecting all oocytes in the gonad. Maximum energetic value of biochemical constituents in females coincided with peaks in lipid content in the visceral mass and mantle. These biochemical events occurred principally immediately before and at the end of oocyte lysis (December/January). A drop in the total energetic value, affecting mainly the visceral mass and the mantle, was recorded each year during the period January to March, coinciding with the period of shell growth in this species. Our data clearly indicate that in female G. glycymeris all muscular tissues contribute to the storage of glycogen and proteins, and suggest that glycogen may be the source of energy triggering vitellogenesis. Biochemical and microscopic observations revealed that oocyte development takes place during the first half of winter, but that these oocytes undergo atresia in December/January. The metabolites produced from oocyte lysis could contribute to somatic growth, which occurs in late winter. Received: 3 March 1997 / Accepted: 23 July 1997     

Flux of radioactive cadmium through the sea-skater Halobates (Heteroptera: Gerridae)

Uptake of ¹⁰⁹Cd in freshly caught sea-skaters Halobates nereis and H. flaviventris was measured under the influence of abiotic parameters as salinity and stable Cd concentration. After 6 d of accumulation a concentration factor of 8 over the seawater level was reached with no indication for an equilibration. The calculated biological half-life of ¹⁰⁹Cd amounted to 9.3 d. Cadmium uptake from food (wingless Drosophila sp. flies reared on cadmium enriched agar) can be considerable. A large pond-skater Limnogonus fossarum, measured for comparative purposes, accumulated cadmium, but at a much lower rate. The results are discussed with respect to observed natural cadmium concentrations in open ocean sea-skater populations.     

Elemental composition of the hydrothermal vent clam Calyptogena magnifica from the East Pacific Rise

Specimens of the hydrothermal vent clam Calyptogena magnifica were collected at a depth of 2 600 m from 21° N on the East Pacific Rise in spring, 1982. The elemental composition was determined for the total soft tissues, individual organs, and the shell. The soft parts contained high concentrations of a number of trace metals. Iron, copper, and zinc, at 760, 148 and 2 152 g g^-1 dry weight, respectively, were present at the highest concentrations. Silver and antimony exhibited the greatest enrichments when their concentrations in C. magnifica were compared with those measured in a shallow-water marine mussel, Mytilus edulis. In contrast to the soft parts, the shell was not a site of metal accumulation or deposition. Within the soft parts; gills, kidney, pericardium, and mantle were involved in the concentration of various metals; the degree of concentration varied depending on the particular metal under consideration. C. magnifica possesses metal body burdens which are ordinarily considered to be very high and potentially toxic in other species.     

Lead uptake from sea water and food,and lead loss in the common mussel Mytilus edulis

Common mussels, Mytilus edulis (shell length 19 to 21 mm, average dry weight 30 mg) were maintained for 6 weeks in sea water containing different concentrations of lead (0.005 to 5 mg · l^-1). The lead concentration in the mussels' whole soft parts was analysed at different times during the experiment. A constant rate of lead uptake, linearly dependent on the lead concentration of the medium, was observed. Thus, the temporal change of the concentration factor is also linear (regression coefficient 149.9 daily). Rate of lead loss, measured after transferring the mussels into natural sea water, is linearly dependent on the original lead concentration in the soft parts. Rates of uptake and loss in large mussels (shell length 45 to 55 mm, average dry weight 750 mg) are less than those in small mussels (shell length 19 to 21 mm, average dry weight 30 mg). During a much more extended experimental period, adjustment to a steady state is expected to occur; rates of lead uptake and loss are then non-linear. Lead uptake by individual organs (kidney, gills, adductor muscle, digestive gland, foot, mantle with gonads) of large M. edulis (shell length 45 to 55 mm, average dry weight 750 mg) was analysed in 2 test series. In the test series medium, the mussels were kept in a seawater medium containing 0.01 mg. Pb.l^-1. In the test series food, the mussels were kept in natural sea water but fed with the green algae Dunaliella marina containing lead (approximately 600 g.g^-1 dry weight). The lead quantity given per mussel per day was about 2 g in both test series. Within 35 days, the mussels of test series medium took up 29% of the total amount of lead given, those of test series food took up 23.5%. In all organs, lead concentration increased, but rates of uptake differed; the kidney displayed by far the highest rate of uptake. With these physiological properties M. edulis is an ideal indicator organism for lead pollution in the marine environment. A biologic calibration curve, the relationship between lead concentration in the mussels' whole soft parts at equilibrium and lead concentration in sea water, is presented.This paper forms part of a doctoral thesis in biology at Hamburg University     

全氟辛烷磺酸盐(PFOS)胁迫对翡翠贻贝抗氧化酶的影响

为了解全氟辛烷磺酸盐(PFOS)对海洋贝类抗氧化防御系统的毒性效应及致毒机理,在实验室条件下研究了PFOS对翡翠贻贝的96h急性毒性,同时探讨PFOS胁迫和净水恢复过程中翡翠贻贝外套膜和内脏团组织中抗氧化指标(SOD活性、GSH和MDA含量)的变化。结果显示,PFOS对翡翠贻贝的96h半致死浓度(LC50)为68.3mg·L-1,安全浓度为6.83mg·L-1。在PFOS胁迫阶段,1mg·L-1浓度组外套膜SOD活性显著性升高(p<0.05),内脏团SOD酶活性显著降低(p<0.05);而PFOS浓度高于1mg·L-1时,外套膜SOD活性显著性降低(p<0.05),内脏团SOD活性显著升高(p<0.05)。PFOS胁迫对翡翠贻贝外套膜和内脏团中GSH含量均有显著的诱导作用(p<0.05),PFOS胁迫15d后各浓度组GSH含量均受到显著的抑制(p<0.05)。翡翠贻贝外套膜MDA含量受PFOS胁迫后显著升高(p<0.05),内脏团MDA含量的变化呈先降低、后升高的规律。净水释放阶段,翡翠贻贝两组织中SOD活性在释放7d后均恢复至对照组水平,GSH含量和MDA含量呈显著升高的趋势(p<0.05)。研究结果表明,PFOS暴露能够引起翡翠贻贝外套膜和内脏团氧化胁迫,但这种损伤的效果不明显,释放短时间后即可自我恢复。     

Concentrations of Mn,Fe, Cu,Zn and Cd in the mesopelagic decapod Systellaspis debilis from the East Atlantic Ocean

Specimens of the oceanic decapod Systellaspis debilis were collected from six sites in the East Atlantic Ocean between 1970 and 1984, and were analysed for Mn, Fe, Cu, Zn and Cd. The data confirm that there are small but significant differences in mean metal concentrations from some sites which showed no obvious pattern in relation to geographic location of the samples. As a result, ranges of site means are quoted as baseline levels for each metal (g g^-1 dry wt): 2.3 to 2.9 g Mn g^-1, 31.2 to 77.8 g Fe g^-1, 25.9 to 83.4 g Cu g^-1, 41.9 to 92.9 gZn g^-1, 11.1 to 31.8 g Cd g^-1. The concentration of cadmium in S. debilis from all sites was raised relative to cadmium concentrations reported for coastal decapods, perhaps as a result of dietary enrichment. Metal accumulation may provide useful information for understanding the complex feeding behaviour of many oceanic animals.     

Quantitative determination of metallothionein-like proteins in mussels. Methodological approach and field evaluation

Electrochemical quantitation of metallothionin-like proteins (MLP) in mussels was based on the determination of their constituent cysteinyl residues according to Brdika's catalytic reaction. Calibration was performed by an internal MLP standard isolated from the digestive gland of Mytilus galloprovincialis for which protein concentration had been estimated by Bradford's spectrophotometric method. For that purpose three metal-binding proteins [MLP-I, MLP-II and Cu-BP (binding protein)] were separated by DEAE-Sephadex A-25 chromatography from the digestive gland of mussels previously exposed to Cd. The most negatively changed MLP-II fraction was characterized by the fact that it contained the largest amount of both total metal and sulphydryl (-SH) content per mass of protein, although this was approximately two times lower than the-SH level of commercially available MT from rabbit liver. Exposure of mussels to a relatively low level of cadmium (0.2 g Cd l^-1) added into the seawater either by itself or as a mixture with other metals (2 g Cu l^-1 and 1.6 g Pb l^-1) resulted in a measurable level of MLP induction within 14 d in comparison to the control specimens. The effect of the metal mixture on MLP synthesis appears to be less than additive, suggesting a competitive interaction between metal ions for uptake and binding sites as well as differing potentials for MLP induction. Variations in the MLP content observed in the digestive gland of mussels seasonally collected from the same location are in the range 2.1±0.4 mg g^-1 on a wet weight basis. The methodological and conceptual aspects of the application of MLP induction in the Mytilus sp. as a biomarker in seawater trace metal monitoring are critically evaluated.