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.     

Antioxidant responses to benzo[a]pyrene and Aroclor 1254 exposure in the green-lipped mussel, Perna viridis

In this study, the green-lipped mussel, Perna viridis (L.), was exposed to two concentrations of benzo[a]pyrene (B[a]P) (0.3 microg l(-1); 3 microg l(-1)) and two concentrations of Aroclor 1254 (0.5 microg l(-1); 5 microg l(-1)). In addition, a mixture of the contaminants was used (0.3 microg l(-1) B[a]P+0.5 microg l(-1) Aroclor 1254; 3 microg l(-1) B[a]P+5 microg l(-1) Aroclor 1254). All concentrations were nominal. A suite of enzymes [glutathione S transferase (GST), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR)], glutathione (GSH) level and lipid peroxidation (LPO) in the mussel gill and hepatopancreas were monitored over 18 days. CAT and GSH in gill tissue were positively correlated with concentration of Aroclor 1254. Activity of hepatic GST and SOD was significantly related to body burden of Aroclor 1254. LPO, GR and GPx in gill and hepatopancreas and hepatic GST were positively correlated with B[a]P concentration. The results indicate the importance of using biomarkers specific to the type of contaminant(s) that are likely to be present. Controlled laboratory experiments, such as this study, are useful in ascertaining biomarkers suitable for use with complex contaminant mixtures in the marine environment.     

Influences of dissolved and colloidal organic carbon on the uptake of Ag, Cd, and Cr by the marine mussel Perna viridis

The cross-flow ultrafiltration and radiotracer techniques were used to study the influences of natural dissolved organic carbon (DOC) and colloidal organic carbon (COC) on the bioavailability of Ag, Cd, and Cr to the green mussel Perna viridis. We examined the uptake of these metals by the mussels at different concentrations of DOC and COC from different origins (estuarine, coastal, and diatom decomposed). Using the DOC originating from the decomposed diatom (Thalassiosira pseudonana), we demonstrated that Cd and Cr uptake, quantified by the concentration factor (DCF), increased linearly with increasing DOC concentration. There was, however, no consistent influence of natural DOC concentration on the metal uptake when the DOC was obtained from different sources of seawater (coastal and estuarine). The influences of COC on metal bioavailability were metal-specific and dependent on the geochemical properties of colloids and colloid-metal complexation. Cd uptake rate was not influenced by the COC concentrations. Uptake of diatom-decomposed colloidal Cr was enhanced by 3.4x, whereas the uptake of diatom-decomposed colloidal Ag was decreased by 8.2x compared with the uptake of low molecular weight Cr and Ag (<1 kDa). The uptake of diatom-decomposed colloidal Cr and Ag was generally lower than the uptake of metals bound with the same type of colloids for 2 days. Further aging of the colloid-metal binding reduced metal bioavailability to the mussels. In the presence of different sizes of colloidal particles where there was no major binding of colloids with the metals, metal uptake by the mussels was not influenced by different COC concentrations. Overall, our study suggests that although metal dissociation from colloids may be an important step for the uptake of colloidal metals, other mechanisms such as pinocytosis and co-transport may also be involved in the uptake of these metals, especially in aquatic environments with high DOC and COC concentrations.     

Contamination by butyltin compounds and organochlorine residues in green mussel (Perna viridis, L.) from Thailand coastal waters

In order to elucidate marine pollution by butyltins and organochlorine residues in coastal waters of Thailand, green mussels (Perna viridis) were collected in 1994 and 1995 and analyzed. Butyltin compounds, such as tributyltin (TBT), dibutyltin (DBT) and monobutyltin (MBT), were detected in most mussel samples, ranging from 4 to 800 ng g(-1) wet wt (as total). The composition of butyltin derivatives in green mussel was in the order of TBT > DBT > MBT. The results indicated that butyltin contamination was widespread, particularly in high boating areas and in coastal aquaculture facilities. Concentrations of organochlorine residues were much lower than of butyltin compounds. Among organochlorine residues examined, dichlorodiphenyltrichloroethane compounds (DDTs) were found to be the highest, followed by polychlorinated biphenyls (PCBs) > chlordane compounds (CHLs) > hexachlorocyclohexanes (HCHs) > hexachlorobenzene (HCB). To our knowledge, this is a first report on the detection of butyltin residues in Thailand coastal waters.     

Lysosomal membrane stability,phagocytosis and tolerance to emersion in the mussel Perna viridis (Bivalvia: Mytilidae) following exposure to acute,sublethal, copper

The mytilid mussel Perna viridis is distributed throughout the Indo-Pacific region and is potentially a suitable candidate for biological effects (biomarker) monitoring in the subtropics. A suite of cytological and physiological responses to acute (48-72 h) copper exposures of 50-200 microgl(-1) were assessed in order to determine the suitability of P. viridis for marine pollution monitoring. Copper elicited significant destabilisation of the haemocyte lysosomal membranes and also impaired phagocytosis. Survival during emersion following exposure to copper was not related to the experimental copper exposures suggesting that higher metal concentrations may be required to interfere with anaerobic enzymes responsible for suppression of metabolism. Based on this preliminary study, cytological biomarkers evaluated in the haemocytes extracted from P. viridis should prove an effective non-destructive means of assessing metal pollution throughout the mussels subtropical range.     

Cu accumulation in the earthworm Dendrobaena veneta in a heavy metal (Cu, Pb, Zn) contaminated site compared to Cu accumulation in laboratory experiments

Earthworms (Dendrobaena veneta) were exposed to heavy metal contaminated soil from a field site under both laboratory and field conditions. In the laboratory study, D. veneta were analysed for Cu after 2 weeks' exposure. The tissue Cu concentration (Cu(W)) increased proportionally with the total extractable soil Cu content (Cu(T)) in soils that contained less than 150 mg Cu kg(-1). In D. veneta exposed to soils containing more than Cu(T) = 150 mg kg(-1), however, we observed no further increase in Cu(W). In a 64-m(2) field plot at a contaminated site, 81 soil samples were taken and analysed for Cu, Pb, Zn and pH. In June 1994, 1000 D. veneta were homogeneously distributed in the plot and sampled after 1, 2 and 5 weeks to analyse for Cu. Additionally, 2000 D. veneta were introduced in September 1994 and sampled after 1, 2 and 4 weeks. At locations where earthworms were removed, the soil Cu content was estimated by disjunctive kriging. Cu concentration factors (ratio of Cu(W) to Cu(T)) in the field experiment were in excellent agreement with those of the laboratory experiment. This indicates that, with a sufficiently high sampling density, it may be feasible to predict earthworm Cu accumulation in the field using a concentration factor determined in the laboratory.     

Sublethal effect of silver and chromium in the green mussel Perna viridis with reference to alterations in oxygen uptake, filtration rate and membrane bound ATPase system as biomarkers

Perna viridis is an ideal animal for studying the impairment caused by the effects of heavy metals that are often detected in coastal areas. Preliminary bioassay tests revealed that the lethal (LC(100)), median lethal (LC(50)) and sublethal (LC(0)) concentration of silver and chromium to P. viridis were 6.5, 4.0, 2.0 mg l(-1) and 4.5, 2.5, 1.0 mg l(-1), respectively. Toxic effect of silver and chromium was evaluated in the green mussel P. viridis, with reference to oxygen consumption, filtration rate and ATPase system in laboratory experiments. These parameters were selected as the end point of sublethal stress. Oxygen consumption and filtration rates were calculated as a measure of decline in the dissolved oxygen level and algal concentration (feed) in the aquaria water, respectively. Silver and chromium affects both oxygen consumption and filtration rate significantly (P<0.01) at 96 h when compared to control. The activity of ATPases system in the gills, hepatopancreas, ovary and muscle of mussels were inhibited by silver and chromium indicating that metals exerted significant toxic effect. The inhibition of Na(+)K(+) ATPase, Ca(2+) ATPase and Mg(2+) ATPase in the mussels were significant (P<0.05) for silver and highly significant (P<0.01) for chromium, which indicates that chromium was more toxic to mussels when compared to silver. The assessment of oxygen consumption, filtration and ATPases system can thus be used as a valid biomarker in aquatic ecotoxicology studies.     

Mussel-based monitoring of trace metal and organic contaminants along the east coast of China using Perna viridis and Mytilus edulis

Green lipped mussels, Perna viridis, and blue mussels, Mytilus edulis, were collected from seven locations along the east coast of China in September and October 2001. The mussel tissues were analyzed for metals (Ag, As, Cd, Cr, Ni, Pb, Se, Zn, Cu, Fe and Hg), and trace organic contaminants including organochlorine compounds (OCs), polycyclic aromatic hydrocarbons (PAHs; based on 24 individual PAHs), polychlorinated biphenyls (PCBs) and petroleum hydrocarbons (PHCs). Maximum concentrations of Ag, As, Cd, Cr, Ni, Pb, Se, Zn, Cu, Fe and Hg in the mussel tissues were 0.14, 26.76, 5.31, 15.72, 4.78, 2.93, 7.40, 231.0, 54.17, 1002 and 317.3 microg/g dry weight respectively. Levels of DDTs, PAHs, PCBs and PHCs in the mussel samples were 14-640, 456-3495, 1-13 ng/g and 621-2863 microg/g dry weight, respectively. Results of this study indicated that contaminant levels were, in general, higher or at least comparable to those reported in other local or regional studies. Mussel samples collected in Chongming Dao in Shanghai and Jiao Zhou Wan in Qingdao had significantly higher levels of metal and trace organic contaminants among the seven sampling stations. Examination of the contaminant profiles suggests that PHCs originated from petrogenic sources, while both petrogenic and pyrolytic sources were important for PAHs. The generally high levels of metals and organochlorine compounds are probably the result of increasingly intense industrial activities along the east coast of China. An assessment of potential risks to human health due to consumption of the mussels was undertaken for the metals, PCBs, DDTs and chlordanes, and the results indicated that all metals, except Ni, could pose a health risk to heavy seafood consumers, while Hg appears to be of concern even for low level consumers.     

Monitoring of organochlorine pesticides residues in green mussels (Perna viridis) from the coastal area of Thailand

The utilization of organochlorine pesticides for pest control chemical has been of great interest on residue contamination from biological organisms in the environment. Green mussel (Perna viridis) samples were monitored as bioindicators for assessment of the water quality in coastal waters along the Gulf of Thailand. Thirty-six samples were collected from 12 stations during 1997-1999 and analysed for 26 organochlorine pesticide compounds. This paper focuses on the contamination of organochlorine pesticide residues in green mussel (P. viridis) during 1997-1999. The limit of detection of all organochlorine pesticides compounds was at the range of 0.1-8.3 ng g(-1) wet weight and recovery 75-95%. The concentration of organochlorine pesticides residues in green mussel was lower than the maximum residue limit for aquatic animals as recommended by the Ministry of Public Health of Thailand. The trend of organochlorine pesticide residue contamination in this area decreased from 1989 to 1999.     

Measurements of Cd, Cu, Pb and Zn in the lower reaches of major Eurasian arctic rivers using trace metal clean techniques

Concentrations of dissolved and particulate Cd, Cu, Pb and Zn were determined in samples collected in summer 1998 from the lower reaches of six major Eurasian arctic rivers: the Onega, Severnaya Dvina, Mezen, Pechora, Ob and Yenisey. These data comprise some of the earliest measurements of trace metals in Eurasian arctic rivers above the estuaries using recognized clean techniques. Significant (α = 0.05) differences were observed among mean concentrations of particulate metals in the individual rivers (F ≤ 0.006), with highest levels overall observed in the Severnaya Dvina and Yenisey. No significant differences were observed among mean concentrations of dissolved metals in the individual rivers (F = 0.10-0.84). Contributions from anthropogenic sources are suggested by comparison of trace metal ratios in the samples to crustal abundances. These results establish a baseline for assessing future responses of Eurasian arctic river systems to climate-related environmental changes and shifting patterns of pollutant discharge.     

Tissue-specific heavy metal (Cd, Pb, Cu, Zn) deposition in a natural population of the zebra mussel Dreissena polymorpha Pallas.

The zebra mussel Dreissena polymorpha was tested as an indicator of heavy metal exposure in urban waters of Vienna, Austria. Mussels, two sediment fractions, suspended matter, and filtrate were collected over one annual cycle at five sampling stations. Dreissena was dissected into five body parts: viscera, gill, foot, byssus, and shell, to determine tissue-specific metal accumulation. Cadmium, lead, copper, and zinc were measured by AAS. There was no clear relationship between (relatively low) metal concentrations in ambient compartments and in zebra mussel bodies. Therefore, D. polymorpha must be regarded as a poor suitable indicator tool under near-background contamination situations. Tissue-specific metal accumulation showed that cadmium was mainly stored in soft body parts. Lead, copper, and zinc showed significantly highest concentrations in the byssal threads. Metal concentrations and distribution patterns within the mussel's body must be interpreted as a result of (unknown) internal metal treatment/regulation. Excretion of lead, copper, and zinc via the byssus complex probably is an effective strategy for preventing toxic injury in D. polymorpha.     

Transcriptional response of stress genes to metal exposure in zebra mussel larvae and adults

Development of stress markers for the invader freshwater zebra mussel (Dreissena polymorpha) is of great interest for both conservation and biomonitoring purposes. Gene expression profiles of several putative or already established gene expression stress markers (Metallothionein, Superoxide dismutase, Catalase, Glutathione S transferase, Glutathione peroxidase, Cytochrome c oxidase, the multixenobiotic resistance P-gp1, and heat shock proteins HSP70 and HSP90) were analyzed by quantitative Real-Time PCR in adults and pediveliger larvae after exposure to metals (Hg, Cu, Cd). A defined pattern of coordinated responses to metal exposure and, presumably, to oxidative stress was observed in gills and digestive gland from adults. A similar, albeit partial response was observed in larvae, indicating an early development of stress-related gene responses in zebra mussel. The tools developed in this study may be useful both for future control strategies and for the use of zebra mussel as sentinel species in water courses with stable populations.     

Genotypic variations in the accumulation of Cd, Cu, Pb and Zn exhibited by six commonly grown vegetables

Metal contaminants in garden and allotment soils could possibly affect human health through a variety of pathways. This study focused on the potential pathway of consumption of vegetables grown on contaminated soil. Five cultivars each of six common vegetables were grown in a control and in a soil spiked with Cd, Cu, Pb and Zn. Highly significant differences in metal content were evident between cultivars of a number of vegetables for several of the contaminants. Carrot and pea cultivars exhibited significant differences in accumulated concentrations of Cd and Cu with carrot cultivars also exhibiting significant differences in Zn. Distinctive differences were also identified when comparing one vegetable to another, legumes (Leguminosae) tending to be low accumulators, root vegetables (Umbelliferae and Liliaceae) tending to be moderate accumulators and leafy vegetables (Compositae and Chenopodiaceae) being high accumulators.     

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.     

Total and subcellular distribution of trace elements (Cd, Cu and Zn) in the liver and kidney of green turtles (Chelonia mydas) from the Mediterranean Sea

This study investigated the subcellular distribution of Cd, Cu and Zn in liver and kidney of green turtles (Chelonia mydas) stranded along the Italian coast of the South Adriatic Sea (Eastern Mediterranean). Cd and Zn mean concentrations did not differ significantly between liver (4.26microgg(-1) and 34.53microgg(-1), respectively) and kidney (5.06microgg(-1) and 26.39microgg(-1), respectively), whereas the levels of Cu were significantly higher in liver (32.75microgg(-1)) than in kidney (8.20microgg(-1)) (p<0.009). Most of Cd, Cu and Zn was present in hepatic and renal cytosol, and their concentrations increased with total levels in both organs, indicating that cytosol has a crucial role in metal accumulation. Cd and Cu in hepatic and renal cytosol were present mostly in metallothionein fractions (MTs), whereas Zn was fractionated into MTs and high-molecular-weight-substances (HMWS). The comparison with the results of other investigations on individuals of the same species collected in different marine areas shows good agreement relatively to essential metals. For Cd our data are comparable with those encountered in specimens from the Mediterranean Sea (Cyprus) confirming the homogeneity of the area comprising the south-eastern basin of the Mediterranean Sea from an ecological point of view.     

Use of zebra mussel (Dreissena polymorpha) to assess trace metal contamination in the largest Italian subalpine lakes.

Trace metal (Cd, Co, Cr, Cu, Hg, Ni, Pb and Zn) contamination was evaluated in zebra mussels from the lakes Maggiore, Lugano, Como, Iseo and Garda, which are located in the most highly populated and industrialised area in Italy. Zebra mussels from Lake Maggiore contained the highest concentrations (3.44, 1.51, 4.97, 0.158, 5.87, 346 microg g(-1) for Cd, Co, Cr, Hg, Pb, Zn, respectively) of all metals analysed except Cu and Ni. The lowest levels of most metals were in animals from Garda and Lugano (0.78 and 0.60 microg g(-1) for Cd, 2.87 and 2.03 microg g(-1) for Cr, 0.065 and 0.049 microg g(-1) for Hg, 12.1 and 11.9 microg g(-1) for Ni, 1.96 and 2.46 microg g(-1) for Pb, 158 and 163 microg g(-1) for Zn). The most contaminated sites and possible local sources of metals were identified for each lake, and the lakes classified into quality classes concerning metal pollution.     

Prosopis juliflora--a green solution to decontaminate heavy metal (Cu and Cd) contaminated soils

Soil and plant samples (root and shoot) of Prosopis juliflora were collected in the vicinity of metal based foundry units in Coimbatore and assessed for their heavy metal content (Cu and Cd) to ascertain the use of P. juliflora as a green solution to decontaminate soils contaminated with Cu and Cd. The results showed that Cu and Cd content was much higher in plant components compared to their extractable level in the soil. Furthermore, there exist a strong correlation between the distance of the sources of industrial units and accumulation of heavy metals in plants. Accumulation of Cd in roots is comparatively higher than that of shoots. However, in case of Cu no such clear trend is seen. Considering the accumulation efficiency and tolerance of P. juliflora to Cd and Cu, this plant can be explored further for the decontamination of metal polluted soils. On the other hand, in view of heavy metal accumulate the practice of providing foliage and pods as fodder for live stock should be avoided.     

Linking biosensor responses to Cd, Cu and Zn partitioning in soils

Soils bind heavy metals according to fundamental physico-chemical parameters. Bioassays, using bacterial biosensors, were performed in pore waters extracted from 19 contrasting soils individually amended with Cd, Cu and Zn concentrations related to the EU Sewage Sludge Directive. The biosensors were responsive to pore waters extracted from Zn amended soils but less so to those of Cu and showed no toxicity to pore water Cd at these environmentally relevant amended concentrations. Across the range of soils, the solid-solution heavy metal partitioning coefficient (K(d)) decreased (p<0.01) with increasing amendments of Cu and Zn; Cu exhibited the highest K(d) values. Gompertz functions of Cu and Zn, K(d) values against luminescence explained the relationship between heavy metals and biosensors. Consequently, biosensors provide a link between biologically defined hazard assessments of metals and standard soil-metal physico-chemical parameters for determining critical metal loadings in soils.     

浮游球衣菌对Pb2+、Cu2+、Zn2+、Cd2+的吸附性能研究

研究了浮游球衣菌（Sphaerotilus natans）在不同吸附条件下对溶液中Pb^2＋、Cu^2＋、Zn^2＋、Cd^2＋的吸附规律。结果表明,Sphaerotilus natans对这4种重金属离子均有一定的吸附作用,并在20min内达到吸附平衡,pH对吸附过程影响较大,pH为5.5时Sphaerotilus natans对这4种金属离子的吸附效果最好,Sphaerotilus natans对它们的吸附选择性为Pb^2＋〉Cu^2＋〉Zn^2＋〉Cd^2＋,Pb^2＋、Cu^2＋能部分置换出已被菌体吸附的Zn^2＋、Cd^2＋。HCI和EDTA溶液可有效地将金属离子从菌体上解吸下来,解吸后的菌体可重复使用。