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.     

Two heavy metal-binding proteins in the midgut gland of the crab Carcinus maenas

Two heavy metal-binding proteins occur naturally in the midgut glands of Carcinus maenas (L.) collected from the Firth of Clyde, Scotland. These proteins, of approximately 27,000 and 11,500 MW (molecular weight) have previously been described as Cd-binding proteins after their induction by high concentrations of cadmium in the laboratory. The 27,000 MW heavy metal-binding protein is bound to about 0.10 g-at of Cd, 0.70 g-at of Zn and 0.31 g-at of Cu per mole of protein; 7.7, 7.9 and 1.1%, respectively, of the soluble Cd, Zn and Cu in the midgut gland are associated with this 27,000 MW protein (6.7, 1.6 and 0.9% of the total midgut gland Cd, Zn and Cu). The 11,500 MW protein is bound to about 0.04 g-at of Cd, 0.37 g-at of Zn and 1.54 g-at of Cu per mole of protein; 29.3, 31.3 and 41.7%, respectively, of the soluble Cd, Zn and Cu in the midgut gland are associated with this protein (25.7, 6.3 and 34.4% of the total midgut gland Cd, Zn and Cu). Neither heavy metal-binding protein is bound to measurable amounts of lead.     

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.     

Metal contamination of soils and dusts in Seoul metropolitan city,Korea

To investigate the dispersion patterns and the characteristics of heavy metal contamination due to urbanisation and industrialisation, soils and dusts collected from the Seoul area were analysed for Cu, Pb, Zn and Cd. The metal concentrations in most soils and dusts are higher than the world averages. The pollution index (( Metal concentrations in soils/Permissible level for metal)÷(Number of metals)) of soils and dusts is > 1 in most of the Seoul area, a result that concurs with the industrialisation and urbanisation index of the Seoul area. The soils are contaminated with Cu, Zn, Cd and particularly Pb. This suggests that the contamination of the soils in the Seoul area are mainly caused by vehicular emissions. The pollution index of soil is the highest in the Kuro area where Cu and Zn contamination in soils are due to the indigenous brass and bronze factories. From the discriminant analysis, the Seoul area may be partitioned into control, traffic and industrialized areas by the metal concentrations in the order of Zn > Cu > Pb.     

Cadmium accumulation and toxicity in the kidney of the bay scallop Argopecten irradians

Specimes of bay scallops, Argopecten irradians, collected from shallow-water eel-grass beds near Beaufort, North Carolina (USA) in September 1979, were exposed to 0.7 ppm cadmium in flowing seawater for 5 d. This exposure resulted in a massive extrusion of the calcified concretions of most of the kidney epithelial cells, although marked morphological damage consisting of cytoplasmc degeneration was apparent in only a few focal areas of the kidney. In addition, unique cytoplasmic membrane-bound bodies were observed in epithelial cells of cadmium-treated but not control scallop kidneys. In some cells, these bodies appeared to fuse with the main concretion vacuole. Kidneys of cadmium-treated scallops accumulated cadmium to 200 ppm on a wet wt basis; of this 60% was associated with concretions (2 000 ppm dry wt) and 38% with the membranous pellet obtained after ultracentrifugation at 105 000 g for 1 h. Approximately 2% of total kidney cadmium was associated with the cytosolic fractions but, unlike zinc or manganese, which were bound to either high or low molecular weight species, a large component of the cadmium in this fraction was bound to a protein peak of approximately 21 000 daltons. Results of this study indicate that kidney concretions of A. irradians play a major role in the control of renal cadmium accumulation and excretion and hence the toxicity of cadmium to this organism.     

Barley lipid-transfer protein as heavy metal scavenger

A lipid-transfer protein was isolated from a domestic cultivar of brewers barley grain, Hordeum vulgare. The presence of Cu (II), Pb (II), Cd (II) and Zn (II) ions in its structure and its ability to bind Hg (II) and Ni (II) ions is known. We investigated its ability to bind other metal ions by differential pulse polarography. Here we demonstrate that the lipid-transfer protein has an affinity to bind Co (II) and Pb (II) and has no affinity towards Cd (II), Cu (II), Zn (II) and Cr (III). These results suggest a new possible role of barley lipid-transfer protein for phytoextraction.Selected article from the Regional Symposium on Chemistry and Environment, Krusevac, Serbia, June 2003, organised by Dr. Branimir Jovancicevic     

Lethal critical body residues as measures of Cd,Pb, and Zn bioavailability and toxicity in the earthworm<Emphasis Type="Italic">Eisenia fetida</Emphasis>

Background

Earthworm heavy metal concentrations (critical body residues, CBRs) may be the most relevant measures of heavy metal bioavailability in soils and may be linkable to toxic effects in order to better assess soil ecotoxicity. However, as earthworms possess physiological mechanisms to secrete and/or sequester absorbed metals as toxicologically inactive forms, total earthworm metal concentrations may not relate well with toxicity.

Objective

The objectives of this research were to: i) develop LD₅₀s (total earthworm metal concentration associated with 50% mortality) for Cd, Pb, and Zn; ii) evaluate the LD₅₀ for Zn in a lethal Zn-smelter soil; iii) evaluate the lethal mixture toxicity of Cd, Pb, and Zn using earthworm metal concentrations and the toxic unit (TU) approach; and iv) evaluate total and fractionated earthworm concentrations as indicators of sublethal exposure.

Methods

Earthworms (Eisenia fetida (Savigny)) were exposed to artificial soils spiked with Cd, Pb, Zn, and a Cd?Pb?Zn equitoxic mixture to estimate lethal CBRs and mixture toxicity. To evaluate the CBR developed for Zn, earthworms were also exposed to Zn-contaminated field soils receiving three different remediation treatments. Earthworm metal concentrations were measured using a procedure devised to isolate toxicologically active metal burdens via separation into cytosolic and pellet fractions.

Results and Discussion

Lethal CBRs inducing 50% mortality (LD₅₀, 95% CI) were calculated to be 5.72 (3.54–7.91), 3.33 (2.97–3.69), and 8.19 (4.78–11.6) mmol/kg for Cd, Pb, and Zn, respectively. Zn concentrations of dead earthworms exposed to a lethal remediated Zn-smelter soil were 3-fold above the LD₅₀ for Zn and comparable to earthworm concentrations in lethal Zn-spiked artificial soils, despite a 14-fold difference in total soil Zn concentration between lethal field and artificial soils. An evaluation of the acute mixture toxicity of Cd, Pb, and Zn in artificial soils using the Toxic Unit (TU) approach revealed an LD₅₀ (95% CI) of 0.99 (0.57–1.41) TU, indicating additive toxicity.

Conclusions

Total Cd, Pb, and Zn concentrations in earthworms were good indicators of lethal metal exposure, and enabled the calculation of LD₅₀s for lethality. The Zn-LD₅₀ developed in artificial soil was applicable to earthworms exposed to remediated Zn-smelter soil, despite a 14-fold difference in total soil Zn concentrations. Mixture toxicity evaluated using LD₅₀s from each single metal test indicated additive mixture toxicity among Cd, Pb, and Zn. Fractionation of earthworm tissues into cytosolic and pellet digesis yielded mixed results for detecting differences in exposure at the sublethal level.

Recommendation and Outlook

CBRs are useful in describing acute Cd, Pb, and Zn toxicity in earthworms, but linking sublethal exposure to total and/or fractionated residues may be more difficult. More research on detoxification, regulation, and tissue and subcellular partitioning of heavy metals in earthworms and other invertebrates is needed to establish the link between body residue and sublethal exposure and toxicity.

     

宝山堇菜多金属吸收特征和耐性策略

宝山堇菜Viola baoshanensis Shu,Liu et Lan是一种Cd超富集植物,但它对不同重金属的吸收和转运能力有待进一步研究。从湖南桂阳宝山多金属矿区中筛选4个宝山堇菜优势分布的小生境,分析这些生境中宝山堇菜及其根区土壤的重金属质量分数。化学分析结果显示,宝山堇菜优势分布土壤中Cd、Pb、Zn、Cu、Mn和Fe的平均质量分数（mg/kg）分别为471、15 044、8 273、1 776、4 702和69 054。宝山堇菜地上部Cd、Pb、Zn、Cu、Mn和Fe的平均质量分数（mg/kg）分别为387、1 077、1 037、99、379和1 812,其中Cd、Pb超过超富集植物标准,Zn、Cu的平均质量分数大约是Zn、Cu超富集植物标准的10%,Mn的平均质量分数低于Mn超富集植物标准的5%,Fe的平均质量分数高于1 000 mg/kg。上述结果表明,宝山堇菜可以超富集Cd和Pb,富集Zn、Cu和Fe以及低积累Mn。此外,宝山堇菜对不同重金属差别化吸收模式可能也代表了超富集植物适应重金属复合污染土壤的一种策略。     

垃圾填埋场渗滤液灌溉地土壤和植物中重金属积累

分加在广州市李坑垃圾填埋场有渗滤液回灌的地表和未经渗滤液回灌的地表随机采集土样,深度达１ｍ,同时分别采集长于灌溉地和非灌溉地上的三个优势植物种（牛筋草,孔雀稗和狗牙根）为代表,调查渗滤液灌溉后封和植物的重金属（Ｃｕ,Ｚｎ,Ｐｂ,Ｃｄ）积累效应。结果表明,渗滤液灌溉地的不同深度土壤和地表植物体内都有不同程度重金属积累。经ｔ－检验,Ｃｕ,Ｐｂ和Ｃｄ在所调查的各土层内的积累效应α＝０．０５时都达到显著水     

Geographical and seasonal variation of trace metal bioavailabilities in the Gulf of Gdansk,Baltic Sea using mussels (<Emphasis Type="Italic">Mytilus trossulus</Emphasis>) and barnacles (<Emphasis Type="Italic">Balanus improvisus</Emphasis>) as biomonitors

The barnacle Balanus improvisus and the mussel Mytilus trossulus have been used as biomonitors of the trace metals Cu, Zn, Cd, Fe, Pb, Mn and Ni at five sublittoral sites in the Gulf of Gdansk (Baltic Sea) between February 2000 and September 2001. The study has established a benchmark against which future biomonitoring programmes will be able to establish changes in local metal pollution, particularly if metal loadings in the river Vistula (draining into the Gulf) alter in the future. The study highlighted differences in trace metal bioavailabilities to both barnacles and mussels, geographically and over time. Accumulated metal concentrations of Cu, Zn, Fe, Pb and Ni, but not Cd or Mn, were correlated in the barnacles and mussels, suggesting that the bioavailabilities of the former metals to the two biomonitors were similar. The barnacles showed greater discriminatory power than the mussels as trace metal biomonitors. Concentrations of trace metals in surficial sediments (<63 m) did not correlate significantly with accumulated metal concentrations in either barnacles or mussels, indicating that sediment metal concentrations are not necessarily good proxy measures of ambient trace metal bioavailabilities to the local coastal filter feeders.Communicated by J.P. Thorpe, Port Erin     

Interactions between metals accumulated in the narrow-clawed crayfish Astacus leptodactylus (Eschscholtz, 1823) in Dikilitaş Lake,Turkey

The accumulations of Al, Cd, Cr, Cu, Fe, Ni, Pb and Zn in the exoskeleton, gills, hepatopancreas and abdominal muscles of crayfish Astacus leptodactylus (Eschscholtz, 1823) were determined. The strongest correlation observed was between Cr and Ni in the gills (r?=?0.904); moderate to strong correlations between Al, Cr, Fe, Ni and Cu were also observed in gill tissue. Disregarding the gills, the strongest correlation was found between Cu and Zn in the hepatopancreas (r?=?0.808); the correlation between these two metals might have been a result of metallothionein activity. The accumulation of Pb was found to correlate with that of Cd in the exoskeleton, Cd and Zn in the gills, Zn and Cu in the hepatopancreas and Cu in the abdominal muscle. None of these correlations were present in lakewater and sediment samples, suggesting that the crayfish metabolism may be responsible for the co-accumulation of metal–metal pairs. As all correlations in non-gill tissues are observed between divalent metals, a shared transporter such as divalent metal transporter 1 might be involved in the accumulation of these metals.     

Temporal variation of metals in the seagrass Posidonia australis and its potential as a sentinel accumulator near a lead smelter

Temporal variation in the concentrations of Cd, Cu, Mn, Ni, Pb and Zn in leaves of the seagrass Posidonia australis was studied at three sites near a lead smelter on the shore of Spencer Gulf, a large hypersaline marine embayment in South Australia, on four occasions from October 1980 to September 1981. Concentrations of Cd, Mn, Pb and Zn of up to 541, 537, 379 and 4241 g g^-1, respectively, were found in leaves collected from the site nearest to the smelter. A substantial temporal variation in the concentrations of these metals in samples from all sites resulted from the combined effect of leaf age and collection strategy. Concentrations of Cd, Cu and Zn in the leaf epibiota were lower than those in the leaves, but the reverse was true for Mn and Ni. The use of seagrass leaves as sentinel accumulators for Cd, Pb and Zn must be based on collections made at the same time of year, or otherwise account for the effect of leaf age on concentrations of the metals in the samples.     

Concentration and distribution of heavy metals in tissues of two cephalopods,Eledone cirrhosa and Sepia officinalis,from the French coast of the English Channel

The concentrations of 11 heavy metals (Ag, Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb, V and Zn) were measured in the tissues (digestive gland, branchial hearts, gills, digestive tract, kidney, genital tract, muscle, skin, shell) of the two cephalopods Eledone cirrhosa (d'Orb.) and Sepia officinalis (L.) collected from the French coast of the English Channel in October 1987. The tissues of both species displayed a similar pattern of heavy-metal accumulation: the digestive gland, branchial hearts and kidney were the major sites of concentration for all 11 metals; the digestive gland accumulated silver, cadmium, cobalt, copper, iron, lead and zinc, the branchial hearts high concentrations of copper, nickel and vanadium, and the kidney high concentrations of manganese, nickel and lead. The digestive gland, which constituted 6 to 10% of the whole-animal tissue, contained >80% of the total body burden of Ag, Cd and Co and from 40 to 80% of the total body burden of the other metals. The ratios between heavy metal concentrations in the digestive gland and those in the muscle separated the elements into three groups, those with a ratio 10 (Cr, Mn, Ni, Pb, V, Zn), those with a ratio >10 to <50 (Co, Cu, Fe), and those with a ratio 50 (Ag, Cd). The digestive gland of cephalopods (carnivorous molluscs whose age can be easily calculated with great accuracy) would seem to constitute a good potential indicator of heavy metal concentrations in the marine environment.     

Trace metal contents ofTaraxacum officinale (dandelion) as a convenient environmental indicator

Some vascular plants are known to concentrate trace metals and are regarded to be suitable indicators of atmospheric metal deposition. Among plant species used for biogeochemical studies dandelion (Taraxacum officinale.) is convenient for monitoring air/soil pollution. The plant commonly occurs in different ecosystems with relatively parallel stages of ontogenesis over a broad area of geographical regions. Its leaves and roots are easily accessible for sampling. Leaf to root ratios of metal concentrations in dandelion may indicate the source of metals during the growing season. Trace metals in leaves and roots of dandelion from 132 sites in Poland showed higher concentrations in the plants from the SW region compared to those from the NE region of the country. However, the differences were only statistically significant ( = 0.05) for Cd, Ni, and Pb. Geometric means of metal concentrations (mg kg^–1, air dried weight) in dandelion leaves of the SW and NE regions were: Cd 0.85, 0.52; Cr 0.99, 0.81; Cu 11.2, 11.1; Fe 184.4, 100.0; Mn 59.7, 51.4; Ni 2.1, 1.5; Pb 4.4, 3.0; and Zn 49.6, 41.3, respectively. Markedly higher concentrations of Cd, Pb, and Zn were found in the leaves of the dandelion over roots in the SW region. These metals are the most serious aerial pollutants in that part of the country.     

A comparison of metals in sediments and water in the River Nahr-Ibrahim,Lebanon: 1996 and 1999

Rivers whose basins are underlain by carbonate rocks exhibit high pH, lower desorption of metals and possess high buffering capacity against acidic inputs to the river. The catchment of River Nahr-Ibrahim, Lebanon, is largely underlain by limestone. Compared to neighbouring countries, Lebanon is relatively fortunate since precipitation is high. However, recently a warming in temperature and a drop in precipitation has occurred, thus causing low water levels in rivers. The objective of this study is to investigate the variation of the total metal content (Fe, Mn, Zn, Cu, Pb and Cd) in bed sediments and water of River Nahr-Ibrahim between 1996 and 1999 (two years); and relate these variations to the effect of changes in human activities and/or due to the variations of precipitation rate, temperature and pH of water. Bed load sediments and stream water were collected simultaneously from five sampling sites. Water pH and temperature were determined in situ. Sediment samples were dried at room temperature and sieved; the sediment size <75 m size was retained. Water was analysed for major constituents and trace metals. Metals were extracted from sediments with aqua regia. Metal concentration in water and sediments were determined using ICP-MS technique. Data revealed a drop in metal concentrations (Zn, Cu, Pb, Cd) in sediments at quarry site after its closure. The decrease in precipitation rate, lowering the level of water and the dilution of industrial discharges and decrease in water pH led most probably to the desorption of metals from sediments into the water.     

Evaluation of the significance of metal-binding proteins in the gastropod Littorina littorea

Measurements of metals in Littorina littorea from clean and contaminated sites in Great Britain confirmed that body concentrations of Ag, Cd and Hg vary according to environmental contamination, while the essential elements Cu and Zn are regulated. Investigation of the cytosolic distribution of metals in L. littorea, using Sephadex G-75 gel-permeation chromatography, revealed important distinctions in the partitioning of elements. In contrast to Ag and Hg, which were mainly associated with high molecular weight ligands, Cd was bound predominantly to an intermediate molecular weight, soluble protein (CdBP-I, apparent mol. wt=20 000). However, although the main function of CdBP-I, both in clean as well as contaminated individuals, was sequestration of Cd, significant quantities of other non-essential metals (Ag, Hg) were similarly bound. Laboratory experiments confirmed the role of CdBP-I in complexing cadmium, and also revealed the induction of a second cadmium-binding protein (CdBP-II) in response to high cadmium levels. The apparent molecular weight (10 000), absorbance characteristics and high (inducible)-SH content of CdBP-II suggest similarities with metallothionein. A dose-related increase in the-SH content of very low molecular weight (>3 000) fractions was also observed in cadmium-exposed L. littorea, although no cadmium was associated with these ligands. A detoxifying role is tentatively proposed for the metal-binding proteins CdBP-I and II in L. littorea. However, some spillover of cadmium to the high molecular weight protein pool was observed in individuals exposed to cadmium in the field and laboratory. The use of L. littorea in biochemical assays of environmental contamination is discussed.     

Influence of cadmium on the storage and metabolism of HCH in rats

Male albino rats were administered Cd and HCH daily for six weeks to study the effect of Cd on storage and metabolism of HCH. The results indicated a marked inhibition of the metabolism of HCH in the group dosed with HCH + Cd. The metabolic rate of HCH in the group dosed with HCH + Cd was also significantly lower than that in the group dosed with HCH alone. The hepatic content of Cd and Zn was significantly increased whereas a marked depletion of Cu and Fe was observed in the groups exposed to Cd and HCH + Cd. A high level of free Cd and Zn or a low level of Fe and Cu in liver seemed to play an important role in the metabolic inhibition observed in this study.     

Effect of Pseudomonas fluorescens on metal phytoextraction from contaminated soil by Brassica juncea

The present study deals with metal uptake by Brassica juncea in the presence of Pseudomonas fluorescens Pf 27 for Zn, Cu and Cd removal from brass and electroplating-industry effluent-contaminated soil. Inoculation of P. fluorescens significantly (p<0.05) increased water soluble (Ws) and exchangeable (Ex) metal content in contaminated soil in laboratory conditions and also enhanced plant biomass by 99% and chlorophyll content by 91% as compared to uninoculated plants in the greenhouse. The metal uptake by B. juncea followed the pattern Zn>Cu>Cd and increased with increasing plant growth duration. P. fluorescens inoculation increased root and shoot uptake of Zn by 3.05 and 2.69, Cu by 3.19 and 2.82 and Cd by 3.11- and 2.75-fold, respectively. BCF value for each metal was>1 and increased by 44%, 42% and 38% for Zn, Cu and Cd, respectively, in inoculated conditions, whereas TF remained unaffected and followed the order Zn>Cd>Cu. P. fluorescens inoculation also enhanced Ws fraction of Zn, Cu and Cd by 99%, 77% and 90% and Ex by 107%, 70% and 93%, respectively. Results depicted that association of B. juncea with P. fluorescens could be a promising strategy for enhancing soil metal bioavailability and plant growth for successful phytoremediation of heavy metal contaminated soils.     

Comparative changes in absorption,distribution and toxicity of copper and cadmium chloride in toads during the hibernation and the role of vitamin C against their toxicity

The aim of this study was to determine the bioavailability and adverse effects of cadmium (Cd) and copper (Cu) on hibernating Egyptian toads and whether ascorbic acid (vitamin C) blocked Cd- and Cu-induced effects during hibernation. The oxidative status of liver, kidney, and intestine of Bufo regularis to Cd, Cu, and/or a combination of both metals administered orally for 2 weeks was determined. In the protection studies, vitamin C was given for 1?h prior to administration of Cu, Cd, and/or metal combination for 2 weeks. Treatment with Cu, Cd, and a combination of both metals produced a reduction in red blood count cells and hemoglobin content, while white blood count cells showed an increase in numbers during these treatments. After 2 weeks exposure, Cd and Cu increased significantly in all the tissues studied. Cu storage presented the following sequence: liver?>?intestine?>?kidney. Cd storage presented the following sequence: intestine?>?kidney?>?liver. When exposed to both metals, Cu and Cd storage presented the following sequence: liver?>?intestine?>?kidney. Histopathological examination of the liver revealed marked alterations including loss of hepatic cell architecture, and some cells exhibited distinct cytoplasmic vacuoles. The majority of blood vessels exhibited a marked dilatation and congestion with infiltration of blood cells, prominent hyperemia of hepatic veins, and significant proliferation of bile ductules. Histopathological changes in the kidney showed destruction and degeneration of both renal tubule cells and glomerular with infiltration of leukocytes and inflammatory cells. Histopathological alterations in the intestine were restricted to the innermost mucosal epithelium with marked degeneration of the villi and submucosa and an extensive fragmentation of mucosal epithelium as well as atrophy of goblet cells. The administration of vitamin C 1?h prior to administration of Cd, Cu, and metal combination did not protect against hepatic, renal, and intestinal damage. However, parental vitamin C given alone increased tissue toxicity.     

Metallothioneins and lysosomes in metal toxicity and accumulation in marine mussels: the effect of cadmium in the presence and absence of phenanthrene

It has been confirmed that metallothioneins play an important role in the accumulation of cadmium (Cd) in the digestive gland cells of mussels (Mytilus galloprovincialis Lam.). The content of Cd in the tissue of mussels exposed for 9 d to the metal (estimated dosage of 180 g Cd mussel^-1 d^-1) was 66.2 ppm. This value is about the same as the metal content found in the digestive gland of Cd-exposed mussels kept in clean water for a recovery period of 28 d. At the end of the recovery period, however, the Cd bound to thionein had increased by approximately 250%. Our data demonstrate that the stability of lysosomes, a biological parameter adopted as a cellular stress index, is extremely low in mussels exposed to Cd for 9 d, but returns to control values in the digestive gland cells of mussels allowed to recover for 28 d in uncontaminated sea water. At this point most of the Cd present in the cytosol is bound to thionein. These data demonstrate the importance of metallothionein induction in the reduction of the cytotoxic effects exerted by high levels of Cd accumulation. The results of tests designed to clarify the reasons for the long biological half-life of Cd demonstrated that, in the digestive gland of mussels, the lysosomes are not able to eliminate Cd either bound to insoluble thionein polymers or to lipid peroxidation products such as lysosomal lipofuscin, both of which are apparently involved in the elimination of copper. The absence of these two mechanisms of metal sequestration and elimination via excretion of residual bodies (tertiary lysosomes) is in agreement with the persistence of cadmium in the digestive gland of mussels. Finally, the results also demonstrate that simultaneous exposure of mussels to Cd and phenanthrene, an established lysosomal membrane destabilizer, did not significantly alter the accumulation of Cd or the kinetics of the metal in mussels.