Toxicological response of a bioluminescent microbial assay to Zn, Pb and Cu in an artificial soil solution: relationship with total metal concentrations and free ion activities.

The relationship between toxicological response and both total concentrations and free ion activities of Pb, Cu and Zn in an artificial soil solution has been investigated using lux-marked Escherichia coli HB101 (pUCD607) as a bioassay. SO4(2-) (as K2SO4) was added as an inorganic complexing agent up to 0.01 M representing the range of ionic strengths found in soil solutions and giving a wide range of free metal ion activities. EC50 values expressed in terms of concentration, varied significantly with K2SO4 molarity for all metals. However, when EC50 values were expressed in terms of free ion activity they were not significantly different for Pb and Zn, supporting the free ion activity model. Conversely, EC50 values expressed as free Cu activity did vary significantly with K2SO4 molarity, possibly due to a greater degree of adsorption of Cu onto inactive sites on the cell surfaces than for Zn and Pb. Linear regression analysis of bioluminescence on free ion activity revealed significant correlations for each metal above the toxicity threshold. In conclusion, lux-marked E. coli is suitable for investigating the toxicity of metal ions and complexes in non saline systems although cell surface adsorption effects could be important for some metals, e.g. Cu.     

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.     

Metal availability and soil toxicity after repeated croppings of Thlaspi caerulescens in metal contaminated soils

Metal phytoextraction with hyperaccumulating plants could be a useful method to decontaminate soils, but it is not fully validated yet. In order to quantify the efficiency of Cd and Zn extraction from a calcareous soil with and without Fe amendment and an acidic soil, we performed a pot experiment with three successive croppings of Thlaspi caerulescens followed by 3 months without plant and 7 weeks with lettuce. We used a combined approach to assess total extraction efficiency (2 M HNO3-extractable metals), changes in metal bio/availability (0.1 M NaNO3-extractable metals and lettuce uptake) and toxicity (lettuce biomass and the BIOMET biosensor). The soil solution was monitored over the whole experiment. In the calcareous soil large Cu concentrations were probably responsible for chlorosis symptoms observed on T. caerulescens. When this soil was treated with Fe, the amount of extracted metal by T. caerulescens increased and metal availability and soil toxicity decreased when compared to the untreated soil. In the acidic soil, T. caerulescens was most efficient: Cd and Zn concentrations in plants were in the range of hyperaccumulation and HNO3-extractable Cd and Zn, metal bio/availability, soil toxicity, and Cd and Zn concentrations in the soil solution decreased significantly. However, a reduced Cd concentration measured in the third T. caerulescens cropping indicated a decrease in metal availability below a critical threshold, whereas the increase of dissolved Cd and Zn concentrations after the third cropping may be the early sign of soil re-equilibration. This indicates that phytoextraction efficiency must be assessed by different approaches in order not to overlook any potential hazard and that an efficient phytoextraction scheme will have to take into account the different dynamics of the soil-plant system.     

Influence of glyphosate and its formulation (Roundup) on the toxicity and bioavailability of metals to Ceriodaphnia dubia

This study examined the toxicological interaction between glyphosate (or its formulation, Roundup) and several heavy metals to a freshwater cladoceran, Ceriodaphnia dubia. We demonstrated that all binary combinations of Roundup and metals (Cd, Cu, Cr, Ni, Pb, Se and Zn) exhibited "less than additive" mixture toxicity, with 48-h LC50 toxic unit > 1. Addition of glyphosate alone could significantly reduce the acute toxicity of Ag, Cd, Cr, Cu, Ni, Pb and Zn (but not Hg and Se). The ratio between glyphosate and metal ions was important in determining the mitigation of metal toxicity by glyphosate. A bioaccumulation study showed that in the presence of glyphosate the uptake of some metals (e.g. Ag) was halted but that of others (e.g. Hg) was increased significantly. Therefore, our study strongly suggests that glyphosate and its commercial formulations can control the toxicity as well as the bioavailability of heavy metals in aquatic ecosystems where both groups of chemicals can co-occur.     

Potentially toxic metals in ombrotrophic peat along a 400 km English-Scottish transect

Four samples of ombrotrophic peat were collected from each of 10 upland locations in a transect from the southern Pennines to the Highland Boundary Fault, a total distance of ca. 400 km. Bulk compositions and other properties were determined. Total contents of Al and heavy metals (Ni, Cu, Zn, Cd, Pb) were determined following digestion with hydrofluoric acid, and concentrations of metals extractable with dilute nitric acid were also measured. Supernatants obtained from aqueous extractions of the peat samples were analysed for pH, major cations and anions, dissolved organic carbon and dissolved metals, and concentrations of free metal ions (Al(3+), Ni(2+), etc.) were estimated by applying a chemical speciation model. Both total and HNO(3)-extractable metal concentrations varied along the transect, the highest values being found at locations close to industrial and former mining areas. The HNO(3)-extractable soil metal contents of Ni, Cu and Cd were appreciably lower than lowest-observed-effect-concentrations (LOEC) for toxicity towards microorganisms in acid, organic rich soils. However, the contents of Zn at two locations, and of Pb at five locations exceeded LOECs, suggesting that they may be exerting toxic effects in the peats. Soil solution concentrations of free heavy metal ions (Cu(2+), Zn(2+), Cd(2+), Pb(2+)) were substantially lower than LOECs for toxicity towards vascular plants, whereas concentrations of Al(3+) were near to toxic levels at two locations.     

Characterization of heavy metal concentrations in the sediments of three freshwater rivers in Huludao City, Northeast China

Wuli River, Cishan River, and Lianshan River are three freshwater rivers flowing through Huludao City, in a region of northeast China strongly affected by industrialization. Contamination assessment has never been conducted in a comprehensive way. For the first time, the contamination of three rivers impacted by different sources in the same city was compared. This work investigated the distribution and sources of Hg, Pb, Cd, Zn and Cu in the surface sediments of Wuli River, Cishan River, and Lianshan River, and assessed heavy metal toxicity risk with the application of two different sets of Sediment Quality Guideline (SQG) indices (effect range low/effect range median values, ERL/ERM; and threshold effect level/probable effect level, TEL/PEL). Furthermore, this study used a toxic unit approach to compare and gauge the individual and combined metal contamination for Hg, Pb, Cd, Zn and Cu. Results showed that Hg contamination in the sediments of Wuli River originated from previous sediment contamination of the chlor-alkali producing industry, and Pb, Cd, Zn and Cu contamination was mainly derived from atmospheric deposition and unknown small pollution sources. Heavy metal contamination to Cishan River sediments was mainly derived from Huludao Zinc Plant, while atmospheric deposition, sewage wastewater and unknown small pollution were the primary sources for Lianshan River. The potential acute toxicity in sediment of Wuli River may be primarily due to Hg contamination. Hg is the major toxicity contributor, accounting for 53.3-93.2%, 7.9-54.9% to total toxicity in Wuli River and Lianshan River, respectively, followed by Cd. In Cishan River, Cd is the major sediment toxicity contributor, however, accounting for 63.2-66.9% of total toxicity.     

Accumulation and histopathological damage in the clam Ruditapes philippinarum and the crab Carcinus maenas to assess sediment toxicity in Spanish ports

The degree of contamination and toxicity in sediment from four Spanish ports (Cádiz, Huelva, Pasajes and Bilbao) was assessed in the present study. Two marine invertebrate species, the shore crab Carcinus maenas and the clam Ruditapes philippinarum, were exposed to the different sediments under laboratory conditions for 28 days. Relationships were developed among metal contamination (As, Cd, Cr, Cu, Fe, Hg, Mn, Ni, Pb and Zn) in sediments, metal accumulation in gill tissues and histopathological lesions in different biological tissues to assess sediment toxicity. A multivariate analysis approach was used to calculate sediment quality guidelines (SQGs) by linking metal concentration in sediments with histopathological lesions measured in the exposed organisms. The results showed significant contaminant accumulation and histopathological lesions due to As, Pb and Zn in sediments at the port of Huelva; As, Cr, Cu, Hg and Ni at the ports of Pasajes and Cádiz; and Cr and Hg at the port of Bilbao. The link between chemical concentration in sediments and histopathological lesions allowed the determination of SQGs for the chemicals As, Cd, Cu, Cr, Hg, Ni and Zn.     

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.     

Heavy metal tolerance of rice plants (Oryza sativa L.) to some metal oxides at the critical levels

The toxicity of metal oxides (CdO, ZnO, PbO) were compared with each other and the critical levels of metal uptake by rice plants were determined. The order of metal toxicity to rice plants is CdO greater than ZnO greater than PbO. The highest concentration obtained for unpolished rice was 4.97 micrograms/g at the Cd 10,000 ppm in soil. This values is higher than every other we have seen in the reports for treatment with CdO. We are able to find out that the concentration of 10,000 ppm Cd in the form of CdO in the critical one towards rice plant. By contrast, such damage was not observed at even higher levels of ZnO and PbO were considered to have low toxicity toward rice plant. Also, a negative correlation between Zn or Cu accumulation in rice plants and Cd concentration in soil was found.     

Heavy metal levels and solid phase speciation in street dusts of Delhi,India

Street dust samples were collected from three different localities (industrial, heavy traffic and rural) situated in the greater Delhi area of India. The samples analyzed for Cd, Zn, Pb, Ni, Cu, and Cr indicated remarkably high levels of Cr, Ni, and Cu in the industrial area, whilst Pb and Cd did not show any discernible variations between the three localities. A multivariate statistical approach (Principal Component Analysis) was used to define the possible origin of metals in dusts. The street dusts were sequentially extracted so that the solid pools of Cd, Zn, Pb, Ni, Cu, Cr could be partitioned into five operationally defined fractions viz. exchangeable, bound to carbonates, bound to Fe-Mn oxides, bound to organic matter and residual. Metal recoveries in sequential extractions were +/- 10% of the independently measured total metal concentrations. Cd was the only metal present appreciably (27.16%) in the exchangeable fraction and Cu was the only metal predominantly associated (44.26%) with organic fraction. Zn (45.64%) and Pb (28.26%) were present mainly in the Fe-Mn oxide fraction and the residual fraction was the most dominant solid phase pool of Cr (88.12%) and Ni (70.94%). Assuming that the mobility and bioavailability are related to the solubility of geochemical forms of the metals and decrease in order of extraction, the apparent mobility and potential metal bioavailability for these highly contaminated street dust samples is: Cd>Zn approximately equal Pb>Ni>Cu>Cr.     

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.     

Role of microbial inoculation and chitosan in phytoextraction of Cu, Zn, Pb and Cd by Elsholtzia splendens--a field case

A field experiment was carried out to study the effect of microbial inoculation on heavy metal phytoextraction by Elsholtzia splendens and whether chitosan could have a synergistic effect with the microbial inocula. The microbial inocula consisted of a consortium of arbuscular mycorrhizal fungi and two Penicillium fungi. Three treatments were included: the control, inoculation with microbial inocula, and the inoculation combined with chitosan. Microbial inoculation increased plant biomass especially shoot dry weight, enhanced shoot Cu, Zn and Pb concentrations but did not affect Cd, leading to higher shoot Cu, Zn, Pb and Cd uptake. Compared with microbial inoculation alone, chitosan application did not affect plant growth but increased shoot Zn, Pb and Cd concentrations except Cu, which led to higher phytoextraction efficiencies and partitioning to shoots of Zn, Pb and Cd. These results indicated synergistic effects between microbial inocula and chitosan on Zn, Pb and Cd phytoextraction.     

Uptake and distribution of Zn, Cu, Cd, and Pb in an aquatic plant Potamogeton natans

A better understanding of metal uptake and translocation by aquatic plants can be used to enhance the performance of constructed wetland systems for stormwater treatment. Specifically, this study examines whether the uptake of Zn, Cu, Cd, and Pb by Potamogeton natans is via the leaves, stems, or roots, and whether there is translocation from organs of uptake to other plant parts. Competition between the metals at uptake and at the level of the cell wall-bound part of the metals accumulated in stem and leaf tissue was also examined. The results show that Zn, Cu, Cd, and Pb were taken up by the leaves, stems, and roots, with the highest accumulation found in the roots. At the elevated metal concentrations common in stormwater the uptake of Cu, but not of Zn, Cd, or Pb, by the roots was somewhat limited at uptake due to competition with other metals. Between 24% and 59% of the metal content was bound to the cell walls of the plant. Except in the case of Pb, the cell wall-bound fraction was generally smaller in stems than in leaves. No translocation of the metals to other parts of the plant was found, except for Cd which was translocated from leaf to stem and vice versa. Dispersion of metals from sediment to water through P. natans is therefore unlikely.     

Accumulation of Al, Mn, Fe, Cu, Zn, Cd and Pb by the bryophyte Scapania undulata in three upland waters of different pH.

Measurements were made of the contents of Al, Mn, Fe, Cu, Zn, Cd and Pb in Scapania undulata in three streams (D2, D5, D11) in the English Lake District. The stream waters had average pH values of 5.35 (D2), 5.81 (D5) and 7.26 (D11), the main differences in other major chemical components being in Mg, Al, Ca and alkalinity. There was generally more metal accumulation in the older parts of the plants, but this was not significant in all cases. Extents of accumulation varied with stream pH and dissolved metal concentration. For Al, accumulation was greatest in streams D2 and D5. Mn accumulated most in D5 and Fe was without preference. Cu, Zn and Cd accumulated mostly in the plants in stream D11 and Pb accumulated more in D5 and D11. In terms of enrichment factors (amount of metal in the plants divided by stream water concentration) the sequence was Zn < Cd < Cu < Mn < Pb < Al < Fe. Laboratory experiments supported the findings of the field data, providing evidence that uptake increases with pH at constant total metal concentration. The results are interpreted qualitatively in terms of the chemical speciation of the metals in the stream water and competition between metal ions and protons at the plant-water interface. It is suggested that Al, Cu, Zn, Cd and Pb behave according to chemical complexation, whereas redox processes and/or colloidal interactions may be significant for Mn and Fe.     

Evaluating three trace metal contaminated sites: a field and laboratory investigation

Selecting guidelines to evaluate elevated metals in urban brownfields is hindered by the lack of information for these sites on ecosystem structure and function. A study was performed to compare three trace metal-contaminated sites in the metropolitan Montreal area. The goal was to obtain an idea of the organisms that may be present on urban brownfields and to measure if elevated metals alter the presence and activity of the indigenous biota. Field and laboratory studies were conducted using simple methodologies to determine the extent to which microbial activity affected by trace metal content, to assess diversity of plant and soil invertebrate communities and to measure phytoaccumulation of trace metals. It was found that microbial activity, as measured by substrate-induced respiration (SIR) and nitrification, was not affected by the levels of soil Cd, Cu, Ni, Pb and Zn recorded on the sites. Seven of the 12 invertebrate groups collected were sampled on soils with similar Cd, Cu, Ni, Pb and Zn concentrations. Diversity of plant species increased as a function of the length of time the sites had been inactive. Levels of metals in plant tissue were influenced by soil characteristics and not by total soil Cd, Cu, Ni, Pb and Zn.     

Speciation of Cd and Zn in contaminated soils assessed by DGT-DIFS, and WHAM/Model VI in relation to uptake by spinach and ryegrass

A pot experiment was carried out to investigate the impact of Cd and Zn extractability in soil and speciation in pore water of industrial contaminated soils, on metal concentration in a metal sensitive species like spinach (Spinacia oleracea) and a more metal tolerant species like Italian ryegrass (Lolium multiflorum). For chemical speciation of Cd and Zn in pore water, WHAM/Model VI version 6.0 was used. The DGT technique was used to determine the effective concentration, C(E), of Cd and Zn in soils. The free ion activity in pore water correlated well with the contents in plants, and there was a linear relationship between the C(E) values and the concentration of Cd and Zn in both spinach and ryegrass in the non-toxic range. However, the C(E) values usually overestimated the plant contents when plants, particularly the spinach plants, were subjected to toxic concentration in the pore water. Metal uptake decreased in plants affected by toxicity, whereas metal binding to the Chelex resin did not. Thus, we found no linear relationship between the C(E) and metal contents in spinach, whereas a linear relationship was found between C(E)-Zn and the Zn concentration in ryegrass (r2=0.96, p<0.001). For Cd in ryegrass this relationship was weak (r2=0.53, p=0.18). This study indicates that the transport of metals from labile metal pools to the DGT-resin is linearly related to plant uptake only when plants are growing well, and that the applicability of DGT as an indicator for plant uptake seems species dependent.     

The solid-solution partitioning of heavy metals (Cu,Zn, Cd,Pb) in upland soils of England and Wales

Ninety-eight surface soils were sampled from the uplands of England and Wales, and analysed for loss-on-ignition (LOI), and total and dissolved base cations, Al, Fe, and trace heavy metals (Cu, Zn, Cd, Pb). The samples covered wide ranges of pH (3.4-8.3) and LOI (9-98%). Soil metal contents measured by extraction with 0.43 mol l-1 HNO3 and 0.1 mol l-1 EDTA were very similar, and generally lower than values obtained by extraction with a mixture of concentrated nitric and perchloric acids. Total heavy metal concentrations in soil solution depend positively upon soil metal content and [DOC], and negatively upon pH and LOI, values of r2 ranging from 0.39 (Cu) to 0.81 (Pb). Stronger correlations (r2=0.76-0.95) were obtained by multiple regression analysis involving free metal ion (Cu2+, Zn2+, Cd2+, Pb2+) concentrations calculated with the equilibrium speciation model WHAM/Model VI. The free metal ion concentrations depend positively upon MHNO3 and negatively upon pH and LOI. The data were also analysed by using WHAM/Model VI to describe solid-solution interactions as well as solution speciation; this involved calibrating each soil sample by adjusting the content of "active" humic matter to match the observed soil pH. The calibrated model provided fair predictions of total heavy metal concentrations in soil solution, and predicted free metal ion concentrations were in reasonable agreement with the values obtained from solution-only speciation calculations.     

Earthworms as biological monitors of cadmium, copper, lead and zinc in metalliferous soils

Earthworms (Lumbricus rebellus and Dendrodrilus rubidus) were sampled from one uncontaminated and fifteen metal-contaminated sites. Significant positive correlations were found between the earthworm and 'total' (conc. nitric acid-extractable) soil Cd, Cu, Pb and Zn concentrations (data log1) transformed). The relationships were linear, and the accumulation patterns for both species were similar when a single metal was considered, even though there were species difference in mean metal concentrations. Generally, the earthworm Cd concentration exceeded that of the soil; by contrast, the worm Pb concentration was lower than the soil Pb concentration in all but one (acidic, low soil Ca) site. Our observations suggest that Cu and Zn accumulation may be physiologically regulated by both species. Total-soil Cd explained 82-86% of the variability (V2) in earthworm Cd concentration; 52-58% of worm Pb and worm Zn concentrations were explained by the total-soil concentrations of the respective metals. Total-soil Cu explained only 11-32% of the worm Cu concentration. The effect of soil pH, total Ca concentration, cation-exchange capacity (CEC) and organic carbon on metal accumulation by L. rubellus and D. rubidus was investigated by multiple regression analysis. Soil pH (coupled with CEC) and soil Ca had a major influence on Pb accumulation (V2 of worm Pb increased to 77-83%), and there was some evidence that Cd accumulation may be suppressed in extremely organic soils. The edaphic factors investigated had no effect on Cu or Zn accumulation by earthworms. In the context of biomonitoring, it is proposed that earthworms have a potential in a dual role: (1) as 'quantitative' monitors of total-soil metal concentrations (as shown for Cd); and (2) as estimators of 'ecologically significant' soil metal, integrating the effects of edaphic factors (as shown for Pb).     

Survey of heavy metal pollution and assessment of agricultural soil in Yangzhong district, Jiangsu Province, China

We investigated concentrations of Hg, Cd, Pb, Zn, Cu, As, Ni, and Cr in samples of soil, cereal, and vegetables from Yangzhong district, China. Compared to subsoils, the sampled topsoils are enriched in Hg, Cd, Cu, Pb, Zn, and As. High levels of Cd and Hg are observed in most agricultural soils. Concentrations of Cr and Ni show little spatial variation, and high Cu, Pb, and Zn contents correspond well to areas of urban development. High As contents are primarily recorded at the two ends of the sampled alluvion. The contents of Cd, Hg, and total organic carbon (TOC) increase gradually to maximum values in the upper parts of soil profiles, while Cr and Ni occur in low concentrations within sampled profiles. As, Pb, Cu, and Zn show patterns of slight enrichment within the surface layer. Compared to data obtained in 1990, Cd and Hg show increased concentrations in 2005; this is attributed to the long-term use of agrochemicals. Cr and Ni contents remained steady over this interval because they are derived from the weathering of parent material and subsequent pedogenesis. The measured As, Cu, Pb, and Zn contents show slight increases over time due to atmospheric deposition of material sourced from urban anthropogenic activity. Low concentrations of heavy metals are recorded in vegetables and cereals because the subalkaline environment of the soil limits their mobility. Although the heavy metal concentrations measured in this study do not pose a serious health risk, they do affect the quality of agricultural products.     

Glutathione levels and enzyme activity in the tissues of bank vole Clethrionomys glareolus chronically exposed to a mixture of metal contaminants

The biochemical response to chronic heavy metal exposure was studied in tissues of bank voles Clethrionomys glareolus. Animals were collected from three sites located 4, 8 and 30km from a zinc-lead smelter, the area's main source of metal contamination. Concentrations of Cd, Pb, Zn and Fe were measured in the liver, kidneys and gonads to assess the level of metal intoxication. In response to intoxication, organisms activate detoxification mechanisms which can protect animals from metals' toxicity. Glutathione plays an important role in toxic substance detoxification. Total glutathione (tGSH) and glutathione disulfide (GSSG) were measured in the tissues. Also, the activity of glutathione reductase (GR), glutathione peroxidase (GPX), and glutathione-S-transferase (GST) was measured in the studied tissues. Results indicate that levels of all studied parameters were tissue and site-dependent. Evidence indicates that the most sensitive parameter of metal toxicity for animals living in a chronically contaminated environment is the GSH/GSSG ratio. In our study, the GSH/GSSG ratio was decreased in the liver of animals with high Cd levels. However, the relationship between Pb and the GSH/GSSG ratio was positive in the gonads. Cadmium and lead negatively influenced GPX activity in the liver; this was probably connected with inhibition of the Se-dependent fraction. The relationship between iron and GR activity in the kidney was also negative, but other correlations for iron both in liver and kidney were not significant. Positive correlations between Zn levels and GST and GR activity were found in the gonads of bank voles.