Modification of trace metal accumulation in the green mussel Perna viridis by exposure to Ag, Cu, and Zn

To examine the Cd, Hg, Ag, and Zn accumulation in the green mussel Perna viridis affected by previous exposure to Cu, Ag, or Zn, the dietary metal assimilation efficiency (AE) and the uptake rate from the dissolved phase were quantified. The mussel's filtration rate, metallothionein (MT) concentration, and metal tissue burden as well as the metal subcellular partitioning were also determined to illustrate the potential mechanisms underlying the influences caused by one metal pre-exposure on the bioaccumulation of the other metals. The green mussels were pre-exposed to Cu, Ag, or Zn for different periods (1-5 weeks) and the bioaccumulation of Cd, Hg, Ag, and Zn were concurrently determined. Pre-exposure to the three metals did not result in any significant increase in MT concentration in the green mussels. Ag concentration in the insoluble fraction increased with increasing Ag exposure period and Ag ambient concentration. Our data indicated that Cd assimilation were not influenced by the mussel's pre-exposure to the three metals (Cu, Ag, and Zn), but its dissolved uptake was depressed by Ag and Zn exposure. Although Hg assimilation from food was not affected by the metal pre-exposure, its influx rate from solution was generally inhibited by the exposure to Cu, Ag, and Zn. Ag bioaccumulation was affected the most obviously, in which its AE increased with increasing Ag tissue concentration, and its dissolved uptake decreased with increasing tissue concentrations of Ag and Cu. As an essential metal, Zn bioaccumulation remained relatively stable following the metal pre-exposure, suggesting the regulatory ability of Zn uptake in the mussels. Zn AE was not affected by metal pre-exposure, but its dissolved uptake was depressed by Ag and Zn pre-exposure. All these results indicated that the influences of one metal pre-exposure on the bioaccumulation of other metals were metal-specific due to the differential binding and toxicity of metals to the mussels. Such factors should be considered in using metal concentrations in mussel's soft tissues to evaluate the metal pollution in coastal waters.     

Distribution and relationships of selected trace metals in molluscs and associated sediments from the Gulf of Aden, Yemen

Concentrations of Cd, Pb, Zn, Cu, Ni, Co, Cr, Mn and Fe in the soft tissue of Turbo coronatus, Acanthopleura haddoni, Ostrea cucullata and Pitar sp., as well as in associated surface sediments (bulk and bioavailable metal concentrations) from the Gulf of Aden, Yemen, were determined by atomic absorption spectrophotometry method. Large differences between size-classes of molluscs in metal concentrations were recorded. Significant spatial differences in metal concentrations in both the soft tissue of the molluscs and associated sediments studied were mostly identified. Statistically significant correlations (p<0.01) between concentrations of selected metals were observed. A slope of the linear regression is significantly higher than unity for Fe (9.91) and Cd (3.45) in A. haddoni and for Ni (4.15) in T. coronatus, suggesting that the bioavailability of these metals is disproportionally increased with a degree of enrichment of the sediments in Fe, Cd and Ni, respectively. A slope constant approximating to unity (1.14) for Cu in A. haddoni relative to its concentration in sediment extract implies that bioavailability of this metal proportionally increased with growing concentrations of its labile forms in the associated sediment. The degree of contamination of Gulf of Aden waters by the metals studied is discussed and the potential ability of molluscs, especially A. haddoni and T. coronatus, as biomonitors of metallic pollutants is postulated.     

Interactions between copper and cadmium modify metal organ distribution in mature tilapia, Oreochromis mossambicus

Sexually mature female tilapia were exposed to sublethal concentrations of waterborne Cu and/or Cd over 6 days, and subsequent body concentrations of these metals were determined in several organs. The results show that the distribution of Cu and Cd was metal and organ specific. This is demonstrated, for example, by the observation that in tilapia, Cu exposure did not result in Cu accumulation in the liver, whereas in the intestinal wall, notably high concentrations of Cu and Cd were measured in metal exposed fish. In addition to single metal exposed fish, we also determined Cu and Cd body distribution in Cu?Cd co-exposed fish. The observed interactions in metal accumulation were most pronounced in the organs of fish exposed to low, environmentally realistic, metal concentrations.     

Interactions between copper and cadmium modify metal organ distribution in mature tilapia, Oreochromis mossambicus

Sexually mature female tilapia were exposed to sublethal concentrations of waterborne Cu and/or Cd over 6 days, and subsequent body concentrations of these metals were determined in several organs. The results show that the distribution of Cu and Cd was metal and organ specific. This is demonstrated, for example, by the observation that in tilapia, Cu exposure did not result in Cu accumulation in the liver, whereas in the intestinal wall, notably high concentrations of Cu and Cd were measured in metal exposed fish.In addition to single metal exposed fish, we also determined Cu and Cd body distribution in Cu---Cd co-exposed fish. The observed interactions in metal accumulation were most pronounced in the organs of fish exposed to low, environmentally realistic, metal concentrations.     

Metal bioaccumulation and metallothionein concentrations in larvae of Crassostrea gigas

Larval stages of bivalve molluscs are highly sensitive to pollutants. Oysters from a hatchery from Normandy (English Channel) were induced to spawn, and fertilized eggs were exposed to copper or cadmium for 24 h. Metal accumulation (from 0.125 to 5 microg Cu L(-1) and from 25 to 200 microg Cd L(-1)) and MT concentrations were measured in larvae. Compared to controls, larvae accumulated copper and cadmium with an increase in MT concentrations particularly with cadmium (i.e. 130.96 ng Cu (mg protein)(-1) and 12.69 microg MT (mg protein)(-1) at 1 microg Cu L(-1) versus 23.19 ng Cu (mg protein)(-1) and 8.92 microg MT (mg protein)(-1) in control larvae; 334.3 ng Cd (mg protein)(-1) and 11.70 microg MT (mg protein)(-1) at 200 microg Cd L(-1) versus 0.87 ng Cd (mg protein)(-1) and 4.60 microg MT (mg protein)(-1) in control larvae). Larvae were also obtained from oysters of a clean area (Arcachon Bay) and a polluted zone (Bidassoa estuary) and exposed to copper in the laboratory, their MT concentration was measured as well as biomarkers of oxidative stress. Biomarker responses and sensitivity to copper for the larvae from Arcachon oysters were higher than for those from Bidassoa.     

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.     

Heavy metal accumulation and metallothionein concentration in the frog Rana ridibunda after exposure to chromium or a mixture of chromium and cadmium

The accumulation of two heavy metals (chromium (Cr) and cadmium (Cd)) in the liver, kidney and gut of Rana ridibunda exposed to Cr or to a mixture of Cr and Cd was investigated. The concentration of metallothioneins (MTs) in the same tissues was also studied. Both metals accumulated mainly in the kidney. Cr accumulation in the liver and gut was not affected by the presence of Cd. Furthermore, Cr concentration in the kidney was doubled when Cd was present. MT concentration did not increase after Cr treatment but it increased two- to six-fold over control values in mixture-exposed frogs, the highest value being observed in the gut. MTs in the gut could act as a barrier preventing ingested heavy metals from entering the blood stream. MT concentration correlated positively with Cd concentrations in both the liver and the gut of mixture-exposed animals.     

Interactions of metals affect their distribution in tissues of Phragmites australis

Culms of Phragmites australis were grown in vermiculite in a greenhouse. Some plants were exposed to 1000 microg/g Cu or Pb or Zn, or combinations of two or three of those metals. When plants reached senescence, they were harvested and analyzed for metal concentrations in upper leaves, lower leaves, stems and roots. While all metals accumulated in highest concentrations in the roots, Zn accumulated in aboveground tissues far more than the other metals. Furthermore, the concentration of any one metal in the different tissues was affected by the presence of other metals. The amount of copper in upper leaves increased when Zn was also present with the Cu. The amount of Cu in roots was increased in the presence of Pb and/or Zn. The amount of Zn in lower leaves was reduced when Cu was also present along with Zn. Thus, when combinations of metals were present, the distribution of metals was altered. The most important interactions appeared to be of Cu and Zn, which may reflect competition for binding sites on metal-binding proteins.     

Sub-cellular partitioning of Cd, Cu and Zn in tissues of indigenous unionid bivalves living along a metal exposure gradient and links to metal-induced effects

We studied organ and sub-cellular distributions of several trace metals in a freshwater bivalve that has been proposed for use as a metal biomonitor. Specimens of Pyganodon grandis were collected from nine lakes located along a Cd, Cu and Zn concentration gradient (Rouyn-Noranda area, Quebec). Gills and digestive gland were isolated, homogenized and six sub-cellular fractions were separated by differential centrifugation and analysed for their Cd, Cu and Zn content. Metallothionein was quantified independently. Gill tissues contained abundant calcium concretions that accounted for over 60% of the total gill burden of each metal. Cadmium and Zn concentrations in this granule fraction reflected ambient metal concentrations. Metal concentrations in the digestive gland also responded to the metal contamination gradient, but to a lesser extent than the gills, reflecting the lower abundance of granules in the digestive gland. Metals (Cd, Cu) in this organ were present largely in the "heat-stable proteins" fraction, and metal concentrations in this fraction were strongly correlated with those of both metallothionein and, to a lesser extent, the "lysosomes+microsomes" and "mitochondria" fractions. In both organs, Cd concentrations in the "heat-denaturable protein" fraction remained low and constant, suggesting reasonably effective metal detoxification. Some evidence for oxidative stress was noted in the gills but not in the digestive gland. Overall, we conclude that in nature metals in P. grandis are bound differently in the gills and in the digestive gland and that metal detoxification in the former organ may be less effective than in the latter.     

Responses of wild small mammals to a pollution gradient: Host factors influence metal and metallothionein levels

We investigated how host factors (species, age, gender) modulated Cd, Pb, Zn, and Cu concentrations, metallothionein levels (MTs) and their relationships in 7 sympatric small mammal species along a pollution gradient. Cd concentrations in liver and kidneys increased with age in all species. Age effect on other metals and MTs differs among species. Gender did not influence metal and MT levels except in the bank vole. Three patterns linking internal metal concentrations and MTs were observed along the gradient: a low metal accumulation with a (i) high (wood mouse) or (ii) low (bank vole) level of MTs accompanied by a slight or no increase of MTs with Cd accumulation; (iii) an elevated metal accumulation with a sharp increase of MTs (common and pygmy shrews). In risk assessment and biomonitoring perspectives, we conclude that measurements of MTs and metals might be associated because they cannot be interpreted properly when considered separately.     

To what extent are hepatic concentrations of heavy metals in Anguilla anguilla at a site in a contaminated estuary related to body size and age and reflected in the metallothionein concentrations?

We explored how hepatic [metal]s in Anguilla anguilla at a contaminated estuarine site are influenced by body size, age and season, and the extent that [Cu], [Cd] and [Zn]s are reflected in [metallothionein (MT)]s. Although each [metal] and [MT] increased significantly with length, weight and age, those biotic variables explained <10% of the variation in each [metal] and only 11-16% for [MT]. Seasonal changes in [Cu] and [Cd] were paralleled by [MT]. The variation in [MT] explained by Cu (42%) was greater than by Zn (16%) and Cd (13%), and seasonally lay between 43 and 69% for those metals collectively. Thus, hepatic [MT] in eels is closely correlated with hepatic [heavy metal]s. However, the great variability among [MT]s for eels of similar sizes and ages, which reflects marked variability in hepatic [heavy metal]s, means that this variable reflects imprecisely the contamination level at a particular site.     

Trade-offs between elimination and detoxification in rainbow trout and common bivalve molluscs exposed to metal stressors

The purpose of this paper was to examine trade-offs between elimination and detoxification in rainbow trout and three common bivalve molluscs (clam, oyster, and scallop) exposed to cadmium (Cd), copper (Cu), and zinc (Zn) based on recent reported experimental data. We incorporated metal influx threshold with subcellular partitioning to estimate rate constants of detoxification (k_d) and elimination (k₂). We found that the relationships between k₂ and k_d were negative for rainbow trout and positive for bivalve molluscs. However, the relationships between k_d and % metal in metabolically detoxified pool were found positive for rainbow trout and negative for bivalve molluscs. Our results also indicated that rainbow trout had higher accumulation (∼60-90%) in metabolically active pool when exposed to essential metals of Cu and Zn and had only 10-50% accumulation in response to non-essential metal of Cd. Based on a cluster analysis, this study indicated that similarity of physiological regulations among study species was found between Cd and Zn. Our study suggested that detoxification can be predicted by an elimination-detoxification scheme with the known elimination rate constant. We concluded that quantification of trade-offs between subcellular partitioning and detoxification provides valuable insights into the ecotoxicology of aquatic organisms and enhances our understanding of the subcellular biology of trace metals.     

The distribution and intracellular compartmentation of metals in the endogeic earthworm Aporrectodea caliginosa sampled from an unpolluted and a metal-contaminated site

The tissue distribution of Cd, Cu, Pb, Zn and Ca in the endogeic earthworm Aporrectodea caliginosa living in a non-polluted and a heavy metal polluted soil was investigated. The tissues of animals from the contaminated soil contained greater concentrations of Cd, Pb and Zn than the corresponding tissues of animals from the unpolluted soil. The greatest concentrations of Cd, Pb, Zn, and Ca were primarily accumulated within the posterior alimentary canal (PAC), a tissue fraction which contained the greatest proportion of the whole-worm burdens of the respective metals. Cu was distributed fairly evenly in the tissue fractions investigated. The pattern of accumulation for the 'heavy' metals is broadly similar to that for epigeic earthworms; in contrast, a different pattern of tissue accumulation was found for Ca. In animals from the uncontaminated site, the major elemental constituents of the chloragosomes were P, Ca, Zn and S. A significant positive correlation exists between P and Ca within the chloragosomal matrix. These intracellular vesicles are major foci for Pb and Zn accumulation within the PAC, with 'excess' metals associated with P ligands within the chloragosome matrix. The incorporation of Pb and Zn appears to involve the cationic displacement of Ca. Such compartmentation appears to prevent dissemination of large concentrations of these metals into other earthworm tissues, and may thus represent a detoxification strategy based on accumulative immobilization. No intracellular localization of Cd was identified in the study, although the Cd concentration in the metalliferous soils examined was not exceptionally high. The observations are discussed in the context of a contribution to enhanced understanding of metal ecotoxicology in earthworms by providing baseline data on a little investigated ecophysiological group of earthworms. Comparisons of metal distribution and mechanisms of metal sequestration are made with other ecophysiological groups of earthworms, and the significance of the findings to biomonitoring and toxicity-testing programmes is considered.     

Heavy metal concentrations in Mullus barbatus and Pagellus erythrinus in relation to body size,gender, and seasonality

Marine environments have been subjected to an increase in heavy metal pollution. Investigations were conducted in the bioaccumulation of heavy metals for both a benthic (Mullus barbatus) and a benthopelagic fish species (Pagellus erythrinus). The aim of this study was to examine the concentration levels of four metals in the body tissue of two fish species, in Pagasitikos Gulf in Greece, and to determine if metal concentration levels were affected by season, size, and species. Fish samples were collected monthly from September 2009 to August 2010. Chromium (Cr), Copper (Cu), Zinc (Zn), and Cadmium (Cd) concentrations were measured in muscle, gills, vertebral column, and in the “remaining fish sample.” Statistical analysis pinpointed substantial differences in metal concentration levels between some size classes. Significant differences were observed between two fish species’ tissues concerning Cu, Zn, and Cd concentrations. Cu and Zn concentrations varied amongst red mullet tissues as did Zn and Cd concentrations in common pandora. Ample variations were found seasonally in metal concentration levels; however, nonsignificant statistical differences were found among sexes.     

Trace metals in seagrass, algae and molluscs from an uncontaminated area in the Mediterranean

The concentrations of Cd, Cr, Cu, Pb and Zn were measured in specimens of four marine organisms--the seagrass Posidonia oceanica (L.) Delile, the brown algae Padina pavonica (L.) Thivy, and the two gastropod molluscs Monodonta turbinata Born and Patella caerulea L.--selected as possible cosmopolitan biomonitors of trace metals in the Mediterranean area. The organisms were collected at five coastal sites in Favignana Island (Sicily, Italy), an area virtually uninfluenced by anthropogenic activities. In order to gain a more complete picture of both the environmental conditions of the experimental area and the bioaccumulation patterns of the selected organisms, soluble and total metal concentrations were determined in coastal water samples collected at the same stations. The picture of bioavailable metal loads in the different sites of the selected area provided by the four species was rather univocal. An overall trend of increased metal concentrations at the station in which the local harbour is located was clear. On the other hand, the metal concentrations recorded at the 'clean' stations generally fall in the range of the lowest values available in the literature and may be considered as useful background levels to which to refer for intraspecific comparison within the Mediterranean area. Implications in biomonitoring of the observed accumulation patterns, especially in the different tissues of Posidonia oceanica, are discussed.     

Metal stoichiometry in predicting Cd and Cu toxicity to a freshwater green alga Chlamydomonas reinhardtii

In this study, we quantified the accumulation and toxicity of cadmium and copper in a freshwater green alga, Chlamydomonas reinhardtii, under different phosphate conditions. The accumulated Cd and Cu concentrations increased significantly with increasing ambient P concentrations and free metal ion concentrations. The metal:P ratio remained independent of the ambient P concentration. For the three pulse-amplitude-modulated parameters, the median inhibition concentrations were 1.5-1.6x and 2.0x higher, but the medium inhibition cellular quota was 2.2x and 1.2x lower for cells maintained at 0.1 microM P than for cells maintained at 10 microM P for Cd and Cu, respectively. Furthermore, the difference in metal toxicity decreased (for Cd) or disappeared (for Cu) when the toxicity was expressed by the metal:P ratio in the cells, indicating that the stoichiometry of metals and P can be better used to predict the toxicity of metals. It is necessary to consider the stoichiometry of metals in predicting metal toxicity in phytoplankton.     

An integrated approach using bioaccumulation and biomarker measurements in female shore crab, Carcinus maenas

The shore crab Carcinus maenas, was used as a biomonitoring organism to measure the potential impact that the mining spill in the Guadalquivir Estuary (SW, Spain), in 1998 may have exerted on local biota. Female intermoult C. maenas were exposed to dissolved cadmium, copper and zinc at concentrations found in local waters following the spill (3 microgl-1 Cd, 15 microgl-1 Cu, 700 microgl-1 Zn). Groups of shore crabs were exposed for 21 days to these metals, both singularly and in combination. Residues of metals were measured in gill, midgut gland and muscle tissues, together with metallothioneins in the midgut gland, at the end of the exposure period. Haemolymph samples were taken every seven days, and vitellogenin/vitellin analysed to determine the flux of these compounds. Biomarker results suggested that they would be good indicators of the effects of heavy metal environmental contamination, particularly in the case of Cd and Zn. Variation in results was obtained between responses measured in crabs exposed to single metals and those exposed to combinations of metals.     

Heavy metal concentrations in molluscs from the Atlantic coast of southern Spain

Trace metals were determined in the two most abundant species of bivalve molluscs along the Atlantic coast of southern Spain (Donax trunculus and Chamelea gallina) and in the sediments where they live. The results show that the area near the mouth of the Huelva estuary is where the highest metal concentrations are found in sediments and in the two bivalve species. This is not surprising, considering that the Huelva estuary is the mouth of the Tinto and Odiel rivers, which have one of the highest levels of metal pollution of all the rivers of Europe. The two species of bivalves have different amounts of metals in their tissues. The concentrations of Cr, Cu, Pb, Zn, As and Hg in D. trunculus were significantly higher (p < 0.05) than in C. gallina; however, C. gallina contained more Ni and Cd. In both species the most abundant elements were Cu and Zn, while Hg showed the lowest values. There is a significant correlation (p < 0.05) for concentrations of Cu, Pb, Zn and Hg in D. trunculus and C. gallina relative to their concentrations in surface sediments.     

Metal accumulation in the polychaete Hediste japonica with emphasis on interaction between heavy metals and petroleum hydrocarbons

The accumulation of cadmium (Cd) and copper (Cu) in the polychaete Hediste japonica exposed to the mixture of Cd (or Cu) and petroleum hydrocarbons (PHCs) was investigated and compared with that exposed to single Cd (or Cu). The increased bioavailability of Cd or Cu with exposure concentrations resulted in an increase in the accumulation and net accumulation rate of Cd or Cu during single metal exposure. The net accumulation rate of Cd increased, but the net accumulation rate of Cu decreased with exposure time during single metal exposure, suggesting that H. japonica could actively regulate Cu burden in their body by inhibition of absolute uptake or promotion of excretion. The interactions between Cd (or Cu) and PHCs had complicated influences on the net accumulation rate of Cd and Cu in H. japonica under the condition of the binary mixture, which are dependent on their concentration combinations and exposure time.