Variations of 210Po and 210Pb in various marine organisms from Western English Channel: contribution of 210Po to the radiation dose

Measurements of (210)Po were carried out in various marine matrices (mussels, oysters, seaweed, fish, and abalones) and in seawater at several points along the French coast, over a period of 2 years (2003-2005). These measurements contribute to a better knowledge of this element, since few recent data exist for the French coast. Marked seasonal variations have been revealed in some species and there are differences according to the way of life of these species. Activities in mussels (Mytilus edulis) and oysters (Crassostrea gigas) are similar and varying between 90 and 600 Bq kg(-1) (d.w.). Activities in macroalgae (Fucus serratus) are lowest, between 4 and 16 Bq kg(-1) (d.w.). In oyster, abalone (Haliotis tuberculata) and fish (Solea solea, Sparus sp.), the strongest activities are measured in the digestive glands, the gills and the gonads. (210)Po/(210)Pb ratios in all cases have values of more than one for all species. From a significant number of measurements, CFs were calculated for seaweed (between 4.6 x 10(3) and 5.0 x 10(3)) and for molluscs, with highest CFs (>10(5)) found for the digestive gland and gills of the oysters, the digestive gland of the abalones and the liver of fish. Finally, the activities measured have made it possible to estimate the internal dose from chronic exposure due to (210)Po received by the marine organisms (0.05 microGh(-1) for macroalgae, between 0.70 and 1.5 microGh(-1) for mussels and oyster), and the contribution of seafood to the dose received by humans (46-129 microSvy(-1)).     

Body burden of contaminants and biological effects in mussels: an integrated approach

An investigation of contaminants and biological effects in mussels from Halifax Harbour, Nova Scotia, Canada, focused on a 6 km section in the central most industrialized core of the harbour, where a site was previously identified as highly contaminated. The aim of the study was to compare the body burden of mussels in terms of polycyclic aromatic hydrocarbons (PAH), dichlorophenyltrichloroethane (DDT family), coprostanol and elements, relative to biological parameters such as condition indices, sex ratio, survival time in air, and to biochemical indicators of mussels' health analysed in gills, digestive gland and gonad tissues. These markers are total sugar and lipid content of gonads, mitochondrial electron transport activity in digestive gland and gonad tissues, lipid peroxidation in gill, digestive gland and gonad tissues, and heme oxidase activity in the digestive gland. At the north western end of the area, near a major sewage effluent, shorter survival time, higher oxidative stress and metabolism, gonad electron transport activity, levels of coporostanol, PAH, p,p'-dichlorophenyldichlo-roethylene (p,p'-DDE), Ag, Cu, Fe and P were observed. At the opposite south eastern end, longer survival time, higher lipid content, lowest condition indices and concentrations of coprostanol, PAH and p,p'-DDE, but higher concentrations of Sn and Cd, were detected. On-going improvements to sewage treatment in Halifax Harbour, including construction of sewage treatment plants that will discharge into deeper parts of the central harbour, should improve inter-tidal mussels' health in our study area.     

Stress-70 proteins in marine mussel Mytilus galloprovincialis as biomarkers of environmental pollution: a field study

In the present work we have investigated levels of stress-70 proteins in the gills of mussel Mytilus galloprovincialis collected seasonally from subtidal rocky shores at 6 different sites of the Rovinj coastal area (Northern Adriatic, Croatia). 1-D analysis (SDS-PAGE) using monoclonal mouse antibodies anti-HSP70 detected two bands of stress-70 proteins, 70 and 72 kDa constitutively present during the year. 2-D analysis (IEF+SDS-PAGE) proved that the antibodies used detected HSP70 (pI 5.7-5.9) and HSP72 (pI 5.5-5.6). The quantification of stress-70 proteins was possible using 200 ng of external HSP70 protein standard included on every blot. Maximal levels of HSP72 and HSP70 were observed in mussels in summer (September), and minimal levels in winter (December), and only HSP70 showed significant correlation with the sea temperature (r=+0.822, p<0.05). Acclimatization of mussels to a different lower salinity under experimental conditions proved that small changes in sea salinity (Delta=2 psu) could not cause significant stress-70 proteins induction. Results indicated that there are significant differences in HSP70 and HSP72 content in mussels from the control site (S-1) and mussels from other sampling sites with urban and industrial pollution. The usefulness of stress-70 proteins as biomarkers of environmental pollution is discussed.     

Molecular biomarkers of energy metabolism in mussels under anthropogenic pollution of Peter the Great Bay, the Sea of Japan

The functional state of the indicator species, the Gray mussel Crenomytilus grayanus (Bivalvia), has been analyzed in five areas of Peter the Great Bay exposed to anthropogenic pollution. The following indices of the state of mussels have been used: molecular biomarkers of energy metabolism—Na⁺,K⁺-ATPase, Mg²⁺-ATPase, and total ATPase activity—as well as the level of lipid peroxidation (LPO) and glutathione concentration in the hepatopancreas, gills, and gonads of mussels. The activity of ATPases, LPO level, and glutathione concentration significantly change in mussels from polluted areas relative to those in mussels from a conventionally unpolluted area (a bay in the Far Eastern State Marine Reserve). The molecular biomarkers used in the study provide reliable information on animal metabolism in impact areas. With consideration of the data obtained, it is concluded that the state of mussels in polluted areas is impaired.     

Effects of pyrene on mussels in different experimental conditions

The toxicity of the polyaromatic hydrocarbon pyrene to Mytilus edulis and Mytilus galloprovincialis was investigated in experiments conducted in the United Kingdom for M. edulis and in Turkey for M. galloprovincialis. Experimental conditions of temperature and salinity were chosen to be appropriate to the ambient conditions in which the mussels typically live. The effect of different feeding regimes on pyrene bioaccumulation and toxicity was also investigated. Feeding rate and neutral red retention biomarker techniques were used for toxicity assessment. An experiment with M. edulis demonstrated that mussels exposed to pyrene accumulated increasing amounts of this compound throughout a 15-day exposure period and that accumulation increased in relation to exposure concentration and with increasing concentration of unicellular algal food material. However, in these experiments, which were protected from UV light, there was no clear relationship between pyrene concentration in tissues and feeding rate. A clear concentration-response relationship was observed between tissue concentration and neutral red retention for days 1 and 7 of the experiment, but this relationship was lost by day 15, with evidence of the cells recovering. A similar experiment was conducted with M. galloprovincialis at a single (high) pyrene concentration in darkness. The feeding rate of the exposed mussels was always lower than the feeding rate of the control mussels, although the difference was insignificant at 7-day exposure. A decreasing trend in health status of the mussels was indicated by the neutral red retention assay results after 7 and 15 days of exposure to pyrene. In a third experiment (with M. galloprovincialis), an illuminated algal chemostat system containing pyrene was inserted in the exposure system as the food source for the mussels. Both biomarker results showed conclusively that toxic effects correlated with pyrene bioaccumulation, though there was no clear evidence for light-enhanced toxicity. These experiments showed that experimental conditions such as salinity, temperature and feeding regime and perhaps the species of mussel used may influence whether pyrene is perceived as being a toxic molecule.     

A biological and geochemical integrated approach to assess the environmental quality of a coastal lagoon (Ravenna, Italy)

The assessment of the environmental quality of coastal lagoons is rather difficult as their intrinsic high variability limits the power of analytical chemistry and ecotoxicological approaches. Aim of the present work was to integrate geochemical and biological data which were simultaneously collected from the Pialassa Baiona lagoon (Ravenna, Italy), and evaluate the usefulness of a biomonitoring strategy based on sentinel organisms and biomarkers. Geochemical analysis on suspended, surface, and subsurface sediments were performed at five different sites of the lagoon; moreover the heavy metal bioaccumulation and a battery of biomarkers was evaluated in Mytilus galloprovincialis transplanted in the same sites. Heavy metals appeared to be diffused throughout the lagoon reaching higher levels in the southernmost sites, and mainly concentrated in suspended sediments. Lysosome membrane stability, neutral lipid and lipofuscin accumulation, and metallothionein levels in mussels indicated the development of a stress syndrome in animals exposed to the polluted sites.     

Monitoring of the impact of Prestige oil spill on Mytilus galloprovincialis from Galician coast

In November 2002, the Prestige oil tanker was wrecked in front of Galician coast (NW of Spain), spilling near 63,000 tons of heavy oil until February 2003. Contamination produced was very extensive (70% of Galician beaches were reached by the oil) but heterogeneous, alternating intensely affected zones with neighbour locations where the repercussion was minimal. The objective of this study was to monitor sea environment contamination caused by Prestige oil spill during an 11-month period (August 2003-June 2004, nine samplings) in two locations of Galician coast with different geographical properties (Lira and Ancoradoiro beaches), by means of chemical determination of total polycyclic aromatic hydrocarbons (TPAH) in seawater, and using as exposure biomarker TPAH content in mussel (Mytilus galloprovincialis) tissues, and as effect biomarker DNA damage in mussel gills, evaluated by the comet assay. In addition, recovery ability of the mussels was determined after a 7-day stay in the laboratory. TPAH contents in seawater were very high in the earliest samplings, but then they maintained below 200 ng L(-)(1), similar to reference seawater. However, TPAH levels in mussel tissues were more variable: they increased again from January 2004, probably due to the adverse meteorological conditions that turned over the sea bottom and dispersed the oil accumulated in sediments. In most samplings, these levels decreased during the recovery stage. DNA damage in oil-exposed mussels was significantly higher than in reference mussels, both before and after the recovery phase, but they did not differ to one another. Comet tail length was slightly reduced during the recovery stage, indicative of a certain DNA repair in exposed mussels. This study showed up the importance of monitoring sea contamination events during an extended time, not only in evaluating the presence of the contaminants in the environment but also in determining their bioaccumulation and their effect on the exposed organisms.     

The use of the shanny Lipophrys pholis for pollution monitoring: a new sentinel species for the northwestern European marine ecosystems

The contamination of aquatic ecosystems by organic contaminants such as polycyclic aromatic hydrocarbons (PAHs) is a matter of great concern. Mussels have been extensively used as sentinel species in a large number of monitoring programs. However, the use of bivalves as the sole species has some limitations, because they are not as responsive as fish to Aryl Hydrocarbon Receptor agonists. Hence, for many marine areas, there is the need to validate new sentinel fish species that can be used in the assessment of pollution by organic contaminants. The shanny Lipophrys pholis is an intertidal fish that combines many characteristics required in a sentinel species: is abundant and easy to catch, has a wide geographical distribution and restrict home range. After larvae recruitment to the intertidal rocky shores, they remain in the same area for the rest of the life-cycle, thus reflecting local pollutants exposure. In order to evaluate the species sensitivity to organic contaminants under field conditions, L. pholis were collected at six sites reflecting different degrees of anthropogenic contamination. The induction of two biomarkers extensively validated in the assessment of PAHs contamination ethoxyresorufin-O-deethylase activity (EROD) and Fluorescent Aromatic Compounds (FACs) was evaluated. In parallel, mussels were collected at the same locations and levels of 16 PAHs and selected heavy metals determined. Overall, the specimens collected in the urban areas showed a significant induction of EROD and FACs (up to a six-fold induction) if compared with the reference sites. Additionally, a positive correlation was observed between the biomarkers and PAHs levels in mussel tissues. Even though further validation is currently in progress, the available data indicate that L. pholis is responsive to organic contaminants such as PAHs, suggesting its future integration in monitoring programmes designed to evaluate the presence of these contaminants in European marine ecosystems.     

Genotoxic and biochemical responses in caged eel (Anguilla anguilla L.) after short-term exposure to harbour waters

European eel (Anguilla anguilla L.) were caged and exposed in situ for 8 and 48 h to the Aveiro offward fishing harbour water (HW) and to clean seawater under laboratory conditions (Control). Eel liver biotransformation (Phase I) was measured as ethoxyresorufin-O-deethylase (EROD) activity, cytochrome P450 (P450) and glutathione S-transferase (GST) activity (Phase II). Genotoxic responses were determined as blood, liver and kidney DNA strand breaks as well as erythrocytic nuclear abnormalities (ENAs). HW failed to significant increase liver EROD, GST activities and ENA frequency. Nevertheless, P450 content was significantly increased after 8 and 48 h exposure. Genotoxicity measured as DNA integrity decrease was found in blood after 8 and 48 h exposure to HW, whereas in liver and kidney, it was observed after 48 h exposure to HW. Blood, kidney and liver genotoxicity may be due to the presence of polycyclic aromatic hydrocarbons (PAHs) which are genotoxic compounds and the main HW organic contaminants.     

Lysosomal responses and metallothionein induction in the blue mussel Mytilus edulis from the south-west coast of Iceland

It has recently been emphasized that high levels of inorganic and organic micropollutants (particularly organometals, POPs and PAHs) may be present in coastal waters at high latitudes, stressing the need to evaluate the effects of contaminants on marine organisms from sub-arctic zones. With this aim, specimens of the blue mussel Mytilus edulis were sampled in polluted and reference areas along the south-west coast of Iceland in July 2004. Samples were collected from the intertidal zone at three sites in Reykjavik harbour which are differently exposed to contaminants, and at three reference coastal sites, two located along the Reykjanes Peninsula and the third one on the northern part of Hvalfjordur fiord. Lipofuscin content, neutral lipid accumulation and lysosomal enlargement were evaluated in digestive cells from cryostat sections of the mussel hepatopancreas, and quantified by automated image analysis. Metallothionein induction was also determined in the same tissue. Results indicate that mussels from the inner part of Reykjavik harbour, which is the most sheltered and most influenced by extensive shipping traffic, were the worst affected, with the highest values in neutral lipids, lipofuscin and lysosomal swelling. At the other two harbour sites, mussels exhibited lower values, similar to those observed in organisms collected in Hvalfjordur fiord and in bay of Osar. Mussels from Kuagerdi had the lowest values.     

Cellular alterations in Mytilus galloprovincialis (LMK) and Tapes philippinarum (Adams and Reeve, 1850) as biomarkers of environmental stress: field studies in the Lagoon of Venice (Italy)

A long-term biomonitoring study was carried out in the Lagoon of Venice (North-East Italy) with the aim of evaluating variations in biological responses to environmental stress in estuarine bivalves. Two different species, the mussel Mytilus galloprovincialis and the clam Tapes philippinarum, both widespread in the Lagoon, were studied in several sites. Two cellular biomarkers: lysosomal membrane stability in digestive cells and thickness of digestive epithelia, were evaluated in native organisms (on a seasonal basis), and in organisms which have been transferred from a reference site to several differently influenced ones. Results indicate that, to some extent, both test and organisms were able to highlight site-specific differences, but the effects of pollution were generally more easily detected by reduction in lysosomal stability than by reduction in digestive tubule epithelium. Further findings show that the inherent variability of a number of natural parameters, particularly in the reference sites, produced less effective results when biological responses in the reference organisms were compared with the polluted ones. The assessment of the two conditions was most valuable when they referred to the 25-75% range of values comprised within the seasonal medians, recorded respectively in control and polluted sites. Impaired from steady states were most effectively distinguished when the control values were medians from two reference locations. Lately, the overall results indicate that both biomarkers are more suitably deployed through the translocation approach, revealing it to be more sensitive than traditional biomonitoring, at least in the sense that it may overcome problems related to the adaptation of native organisms to sub lethal chronic pollution levels.     

Metal and metallothionein level in the heat-treated cytosol of gills of transplanted mussels Mytilus galloprovincialis Lmk

A stock of mussels Mytilus galloprovincialis was transplanted over 1 year to four sites in a semi-enclosed bay in Croatia which is under the influence of various sources of pollution. The positive correlation of metal (Cd, Zn, Cu, Mn, Fe) and metallothionein (MT) tissue contents based on the analysis of heat-treated cytosol of gills with shell mass, as an indicator of mussel age, indicated to accumulation of metals, as well as the increase of MTs with mussel age. The principal component analysis (PCA) revealed that 74% of total variance of obtained results could be explained through two principal components. The first principal component was highly correlated with MT, Cd and Zn indices (metal or MT content/shell mass), and the second one with Cu, Mn, and Fe indices, as well as the gill index. High correlation of MTs with Zn and Cd is consistent with their affinity for binding to MTs.     

Factors affecting 210Po and 210Pb activity concentrations in mussels and implications for environmental bio-monitoring programmes

The activity of ²¹⁰Po and ²¹⁰Pb was determined in mussels of the same size (3.5-4.0 cm shell length) sampled monthly over a 17-month period at the Atlantic coast of Portugal. Average radionuclide concentration values in mussels were 759 ± 277 Bq kg⁻¹ for ²¹⁰Po (range 460-1470 Bq kg⁻¹ dry weight), and 45 ± 19 Bq kg⁻¹ for ²¹⁰Pb (range 23-96 Bq kg⁻¹ dry weight). Environmental parameters and mussel biometric parameters were monitored during the same period. Although there was no seasonal variation of radionuclide concentrations in sea water during the study period, the concentration of radionuclide activity in mussels varied seasonally displaying peaks of high concentrations in winter and low concentrations in summer. Analysis of radionuclide data in relation to the physiological Condition Index of mussels revealed that ²¹⁰Po and ²¹⁰Pb activities in the mussel (average activity per individual) remained nearly constant during the investigation period, while mussel body weight fluctuated due to fat storage/expenditure in the soft tissues. Similar variation of radionuclide concentrations was observed in mussels transplanted from the sea coast into the Tejo Estuary. However, under estuarine environmental conditions and with higher food availability throughout the year, transplanted mussel Condition Index was higher than in coastal mussels and average radionuclide concentrations were 210 ± 75 Bq kg⁻¹ (dry weight) for ²¹⁰Po and 10 ± 4 Bq kg⁻¹ (dry weight) for ²¹⁰Pb, therefore lower than in coastal mussels with similar shell length. It is concluded that the apparent seasonal fluctuation and inter-site difference of radionuclide concentrations were mostly caused by mussel body weight fluctuation and not by radionuclide body burden fluctuation. This interpretation can be extended to the apparent seasonal fluctuation in concentrations of lipophilic and lipophobic contaminants in mussels, and provides an explanation for occasional high concentrations of ²¹⁰Po and man-made contaminants measured in mussels far from pollution sources.     

Honeybee biomarkers as promising tools to monitor environmental quality

The aim of this study was to distinguish the impacts of two different anthropogenic conditions using the honeybee Apis mellifera as a bioindicator associated with a battery of biomarkers previously validated in the laboratory. Both the urban (RAV, Ravine des Cabris) and semi-natural (CIL, Cilaos) sites in La Reunion Island were compared in order to assess the impacts of two types of local pollution using the discriminating potential of biomarkers. Hives were placed at the CIL and RAV sites and honeybees were collected from each hive every three months over one year. Honeybee responses were evaluated with respect to several biochemical biomarkers: glutathione-S-transferase (GST), acetylcholinesterase (AChE), alkaline phosphatase (ALP) and metallothioneins (MT). The results showed a significant difference between the localities in terms of GST, AChE and ALP activities, as regarding midgut MT tissue levels. Compared to the CIL site, ALP and MT tissue levels were higher at the RAV site, although AChE activity was lower. GST displayed more contrasted effects. These results strongly suggest that the honeybees based in the more anthropized area were subjected to sublethal stress involving both oxidative stress and detoxification processes with the occurrence of neurotoxic pollutants, amongst which metals were good candidates. A classification tree enabled defining a decision procedure to distinguish the sampling locations and enabled excellent classification accuracy (89%) for the data set. This field study constitutes a strong support in favour of the in situ assessment of environmental quality using honeybee biomarkers and validates the possibility of performing further ecotoxicological studies using honeybee biomarkers.     

Changes in mussel Mytilus galloprovincialis protein profile as a reaction of water pollution

In the present report the changes in mussels Mytilus galloprovincialis protein profile, as a reaction of water pollution, is presented. The antioxidant capacity was determined by the efficacy to scavenge the derived radical in mussel samples using the Trolox Equivalent Antioxidant Capacity (TEAC) assay. The highest percentage of inhibition was estimated in protein mussels from the dirty area (DM) in comparison with the clean samples (CM). The amounts of both zinc and copper determined by atomic spectroscopy were significantly higher in the protein mussels from the dirty than from the clean areas (P<0.05). Fourier Transform Infrared (FT-IR) spectroscopy and fluorescence revealed specific qualitative changes in secondary and tertiary structures of mussel proteins from the dirty area by the shift in the amides I and II positions and fluorescence intensity. Estimated temperature and enthalpy of denaturation in the protein mussels well correlated with fluorescence and spectroscopic measurements and showed the changes influenced by water pollution.     

Evidence of genetic damage in grass gobies and mussels from the Venice lagoon

The presence of genetic damage has been investigated in two native species of the Venice lagoon: the common mussel Mytilus galloprovincialis and the grass goby Zosterisessor ophiocephalus. Two sampling campaigns were performed in summer 1998 and 1999. Aromatic-like DNA adducts were analysed in selected tissues of gobies and mussels by using the 32P-postlabelling assay. In 1999, micronuclei and other nuclear abnormalities were additionally scored on gill cells and haemocytes of individual mussels whereas inorganic (As, Cd, Cr, Hg, Ni, Pb, Sn) as well as organic contaminants (polycyclic aromatic hydrocarbons, polychlorinated biphenyls and other chlorinated compounds) were measured in the total mussel pulp. Compared to the lagoon inlet area, gobies and mussels from the industrial district (Marghera) showed significant DNA adduct levels and increased frequencies of cytogenetic alterations (evidence of genetic damage was absent or inconsistent in other sites). The substantial levels of aromatic and chlorinated contaminants detected in mussels from Marghera also support the exposure of native organisms to genotoxic agents.     

The accumulation and retention of 95mTc by the Norway lobster, Nephrops norvegicus L.

Laboratory experiments were carried out to study bioaccumulation and determine a concentration factor (CF) for technetium (95mTc) in the homarid crustacean Nephrops norvegicus L. The steady state CF for accumulation from seawater was estimated to be about 2,000 and the biological half-time was about 50 days. The highest tissue Tc concentrations were found in the green gland and the digestive gland. Depuration following accumulation from water was slow with a half-time of about 165 days. Tc accumulation from labelled food followed a biphasic model with one compartment containing about 94 percent of the ingested activity and with a half-time of about 1 day and the second compartment containing about 6 percent of the ingested activity with a half-time of about 56 days. Most retained activity was found in the digestive gland.     

Effects of natural and chemical stressors on Enchytraeus albidus: Can oxidative stress parameters be used as fast screening tools for the assessment of different stress impacts in soils?

Enchytraeids are important organisms of the soil biocenosis. They improve the soil pore structure and the degradation of organic matter. These organisms are used in standardized testing, using survival and reproduction (6 weeks) as endpoints. The use of biomarkers, linked to ecologically relevant alterations at higher levels of biological organization, is a promising tool for Environmental Risk Assessment. Here, enchytraeids were exposed for different time periods (two days and three weeks) to different soils (OECD artificial soil, different compositions in its organic matter, clay or pH value, and LUFA 2.2 natural soil) and different chemicals (Phenmedipham and copper). The main question addressed in the present study was if the effects of chemicals and different soil properties are preceded by alterations at the sub-cellular level, and if these endpoints may be used reliantly as faster screening tools for the assessment of different stress conditions in soils. The parameters measured in E. albidus whole body were: lipid peroxidation (LPO), total glutathione (TG), as well as the enzymatic activities of superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR), glutathione peroxidase (GPx) and glutathione S-transferase (GST). The results showed that biomarker responses in E. albidus were significantly affected by the soil type (GST, CAT, GPx, GR and LPO) and the duration of exposure in OECD artificial soil (GST, GPx, GR, CAT and LPO) but not in LUFA 2.2 natural soil. For the abiotic factors studied, after 2 days, low pH decreased significantly the TG levels and the activities of CAT and GR,and low OM also significantly decreased CAT and GR activities. After 3 weeks, differences in soil properties caused a decrease in GR and GPx activities, whereas increased GST activity was observed due to low organic matter and pH. Copper significantly increased the activities of CAT, GPx and GR, and decreased the activity of GST after 2 days as well as inscreasing LPO levels after 3 weeks. Phenmedipham increased LPO levels, associated with increased levels of TG as well as increased activities of CAT and GPx and decreased GST activity after 3 weeks exposure. This study shows that both abiotic and chemical stresses could be followed through biomarker analysis and that some of these determinations are potential endpoints in a quick soil contamination assessment procedure.     

Histopathology and Changes of Reproductive Parameters in Peled,Coregonus peled,from the Lower Ob Basin

Russian Journal of Ecology - Histopathological changes in the gills, liver, kidneys and gonads and heavy metal accumulation have been studied in peled (Coregonus peled) from the Lower Ob basin. It...     

Bio-accumulation of some trace metals in the short-neck clam Paphia malabarica from Mandovi estuary, Goa

Monthly collections of the clam Paphia malabarica were made for period of 1 year from May 2002 to April 2003 from Verem, Mandovi estuary, Goa. Clams were categorised into two groups, 25-35 mm (small size) and 35-40 mm (big size). Significant difference was not observed in the accumulation of metals in the whole soft tissue between the two size groups. Irrespective of size, annual mean, metal content recorded as microg/g dry weight in the whole soft tissue was 3.8, 30.3, 13.5, 36.6 and 105.7, respectively, for Cd, Pb, Cu, Zn and Fe. Of the two toxic elements Cd and Pb, Cd content was almost uniform throughout the year except for a rise in September for the small size and October for the big size. Pb, on the other hand, was low from the beginning of the monsoon and exhibited distinct accumulation from December onwards up to April, May and, to some extent, June. The pattern was similar in both the size groups, the values being higher for the bigger size group. The three essential elements Cu, Fe and Zn exhibit trends similar to one another with peaks in September and December-January in both the size groups. Cadmium accumulation was highest in the mantle and adductor muscle, Lead, in foot, Copper, in digestive gland and gonad, and Zn and Fe, in gills. Correlation coefficient between different metal couplings as tested statistically revealed significant coupling for Zn-Fe (r 0.92) in the bigger size group, the same was observed between Cu-Fe (r 0.62) and Cd-Zn (r 0.94). Seasonal difference in Pb accumulation was highly significant.