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Archives of Environmental Contamination and Toxicology (v.59, #3)
Dissipation and Leaching Potential of Selected Pharmaceutically Active Compounds in Soils Amended with Biosolids by Chenxi Wu; Alison L. Spongberg; Jason D. Witter; Min Fang; Kevin P. Czajkowski; April Ames (pp. 343-351).
Biosolids land application is an important pathway introducing pharmaceuticals into the environment. In this work, laboratory column and dissipation experiments were performed using soils of varying properties in order to study the fate and transport of pharmaceutical residues introduced by the land application of biosolids. For experimentation, five pharmaceutical compounds (carbamazepine, diphenhydramine, fluoxetine, diltiazem, and clindamycin) and two metabolites (carbamazepine-10,11-epoxide and norfluoxetine) commonly found in biosolids were selected. Leaching experiments indicate that the selected pharmaceuticals have low mobility in tested soils. However, small portions of the applied pharmaceuticals were recovered in the leachates, likely attributed to sorption to dissolved organic matter. Dissipation experiments show that carbamazepine, diphenhydramine, and fluoxetine were persistent in soils, whereas the dissipation of diltiazem and clindamycin was affected by redox conditions and soil properties.
Sediment Contamination of Residential Streams in the Metropolitan Kansas City Area, USA: Part I. Distribution of Polycyclic Aromatic Hydrocarbon and Pesticide-Related Compounds by J. Tao; D. Huggins; G. Welker; J. R. Dias; C. G. Ingersoll; J. B. Murowchick (pp. 352-369).
This is the first part of a study that evaluates the influence of nonpoint-source contaminants on the sediment quality of five streams within the metropolitan Kansas City area, central United States. Surficial sediment was collected in 2003 from 29 sites along five streams with watersheds that extend from the core of the metropolitan area to its development fringe. Sediment was analyzed for 16 polycyclic aromatic hydrocarbons (PAHs), 3 common polychlorinated biphenyl mixtures (Aroclors), and 25 pesticide-related compounds of eight chemical classes. Multiple PAHs were detected at more than 50% of the sites, and concentrations of total PAHs ranged from 290 to 82,150 μg/kg (dry weight). The concentration and frequency of detection of PAHs increased with increasing urbanization of the residential watersheds. Four- and five-ring PAH compounds predominated the PAH composition (73–100%), especially fluoranthene and pyrene. The PAH composition profiles along with the diagnostic isomer ratios [e.g., anthracene/(anthracene + phenanthrene), 0.16 ± 0.03; fluoranthene/(fluoranthene + pyrene), 0.55 ± 0.01)] indicate that pyrogenic sources (i.e., coal-tar-related operations or materials and traffic-related particles) may be common PAH contributors to these residential streams. Historical-use organochlorine insecticides and their degradates dominated the occurrences of pesticide-related compounds, with chlordane and dieldrin detected in over or nearly 50% of the samples. The occurrence of these historical organic compounds was associated with past urban applications, which may continue to be nonpoint sources replenishing local streams. Concentrations of low molecular weight (LMW; two or three rings) and high molecular weight (HMW; four to six rings) PAHs covaried along individual streams but showed dissimilar distribution patterns between the streams, while the historical pesticide-related compounds generally increased in concentration downstream. Correlations were noted between LMW and HMW PAHs for most of the streams and between historical-use organochlorine compounds and total organic carbon and clay content of sediments for one of the streams (Brush Creek). Stormwater runoff transport modes are proposed to describe how the two groups of contaminants migrated and distributed in the streambed.
Sediment Contamination of Residential Streams in the Metropolitan Kansas City Area, USA: Part II. Whole-Sediment Toxicity to the Amphipod Hyalella azteca by J. Tao; C. G. Ingersoll; N. E. Kemble; J. R. Dias; J. B. Murowchick; G. Welker; D. Huggins (pp. 370-381).
This is the second part of a study that evaluates the influence of nonpoint sources on the sediment quality of five adjacent streams within the metropolitan Kansas City area, central United States. Physical, chemical, and toxicity data (Hyalella azteca 28-day whole-sediment toxicity test) for 29 samples collected in 2003 were used for this evaluation, and the potential causes for the toxic effects were explored. The sediments exhibited a low to moderate toxicity, with five samples identified as toxic to H. azteca. Metals did not likely cause the toxicity based on low concentrations of metals in the pore water and elevated concentrations of acid volatile sulfide in the sediments. Although individual polycyclic aromatic hydrocarbons (PAHs) frequently exceeded effect-based sediment quality guidelines [probable effect concentrations (PECs)], only four of the samples had a PEC quotient (PEC-Q) for total PAHs over 1.0 and only one of these four samples was identified as toxic. For the mean PEC-Q for organochlorine compounds (chlordane, dieldrin, sum DDEs), 4 of the 12 samples with a mean PEC-Q above 1.0 were toxic and 4 of the 8 samples with a mean PEC-Q above 3.0 were toxic. Additionally, four of eight samples were toxic, with a mean PEC-Q above 1.0 based on metals, PAHs, polychlorinated biphenyls (PCBs), and organochlorine pesticides. The increase in the incidence of toxicity with the increase in the mean PEC-Q based on organochlorine pesticides or based on metals, PAHs, PCBs, and organochlorine pesticides suggests that organochlorine pesticides might have contributed to the observed toxicity and that the use of a mean PEC-Q, rather than PEC-Qs for individual compounds, might be more informative in predicting toxic effects. Our study shows that stream sediments subject to predominant nonpoint sources contamination can be toxic and that many factors, including analysis of a full suite of PAHs and pesticides of both past and present urban applications and the origins of these organic compounds, are important to identify the causes of toxicity.
Sediment Matrix Effects in Analysis of Pyrethroid Insecticides Using Gas Chromatography–Mass Spectrometry by Dongli Wang; Jing You; Michael J. Lydy (pp. 382-392).
In the present study, we developed a gas chromatography–quadrupole mass spectrometry method for analyzing nine pyrethroid insecticides in sediment after accelerated solvent extraction and solid phase extraction cleanup. The operation was optimized, and negative chemical ionization was selected to improve analytical selectivity. The sediment matrix effects on qualification were evaluated, and matrix-matched standard solutions, along with the internal standard calibration, were used to reduce the matrix-induced chromatographic response enhancement. The method detection limits were 0.68 to 1.43 μg/kg dry weight (dw), and recoveries were 70.3 to 143.3%, 61.1 to 169.7%, and 65.7 to 118.8%, with relative SDs of 4.2% to 32.1%, 4.9 to 23.6%, and 1.5 to 23.3% at the spiked levels of 1.0, 5.0 and 20 μg/kg dw, respectively. The method was also validated by measuring pyrethroids in field-contaminated sediment samples collected in central California and southern Illinois.
Role of Lactone Ring in Structural, Electronic, and Reactivity Properties of Aflatoxin B1: A Theoretical Study by Inés Nicolás-Vázquez; Abraham Méndez-Albores; Ernesto Moreno-Martínez; René Miranda; Miguel Castro (pp. 393-406).
This study involved quantum mechanical calculations to explain the chemical behavior of the lactone ring of aflatoxin B1, which is a carcinogenic hazardous compound. The aflatoxin B1 compound, produced by the fungi Aspergillum flavus, was studied with the B3LYP/6-311+G(d,p) method; its reactivity properties were accounted for by means of the calculated geometrical and electronic parameters. The results obtained indicate that the fused A, B, C, and D rings of aflatoxin adopt a continuous planar conformation. The carbon atom of the lactone group presents a highly electrophilic character, since the population analysis yields a high positive charge for this atom, whereas high negative charges were recorded for both oxygen sites of that group. Thus, in an acidic aqueous medium, the oxygen atoms could be protonated and the carbon site may suffer a nucleophilic attack by water. Accordingly, the OC−O bond length has been lengthened substantially. So it was demonstrated that the lactonic ring of aflatoxin B1 is hydrolyzed under acidic conditions by an acid-acyl bimolecular mechanisms, AAC2, suggesting the deletion of its carcinogenic properties.
Acute Toxicity of a Shoreline Cleaner, CytoSol, Mixed With Oil and Ecological Risk Assessment of its Use on the Galician Coast by Diego Rial; Ricardo Beiras; José A. Vázquez; Miguel A. Murado (pp. 407-416).
The application of embryo–larval bioassay with the purple sea urchin Paracentrotus lividus and the mussel Mytilus galloprovincialis at 48 hours, and with neonates of the mysid Siriella armata at 96 hours, was used to evaluate the acute toxicities of the following preparations: (1) the shoreline cleaning agent CytoSol; (2) the water-accommodated fraction of CytoSol plus a light crude oil; and (3) the runoff from a pilot-scale treatment with CytoSol of a rocky coastal substrate impregnated with residues from the Prestige oil spill (which occurred on November 19, 2002). The mussel was the most sensitive organism to CytoSol and runoff effects (EC50 = 8.0 μL/L and 64.3 mL/L, respectively), and the mysid was the least sensitive to the runoff (EC50 > 200 mL/L). The predicted no-effect environmental concentration (PNEC) was calculated from the no observed–effect concentration of the species most sensitive to the runoff. The predicted environmental concentration (PEC) was estimated from a simple and reasonable dilution model, and the PEC/PNEC ratio was calculated according to the area treated and the values of the variables considered in the model. Implications for the management of the treatment operations are discussed.
Fungal Growth Inhibition of Regenerated Cellulose Nanofibrous Membranes Containing Quillaja Saponin by Vivechana Dixit; Jagdish Tewari; S. Kay Obendorf (pp. 417-423).
Antifungal properties were introduced in nonwoven regenerated cellulose (RC) nanofibrous membrane using Quillaja saponin. To generate cellulose membranes, deacetylation of electrospun cellulose acetate (CA) nanofibrous membranes was performed using 0.05 M NaOH and ethanol for membranes both loaded and unloaded with Quillaja saponin. Chemical and physical properties of nonwoven CA and RC nanofibrous membrane were characterized using scanning electron microscopy, attenuated total reflectance–Fourier transform infrared spectroscopy, differential scanning calorimetry, and tensile properties. Our results showed that saponin doping did not affect the morphology of the resulting fibers and that the membrane structure was maintained during deacetylation. The antifungal properties of saponin-loaded fabric were determined at 0 and 24 h against two household fungi, Penicillium roguefortii and Aspergillus ochraceus, and compared with control samples. Our findings show that after 24 h the saponin-loaded fabrics had spores kill of 80.4% and 53.6% for P. roguefortii and A. ochraceus, respectively. Fabric containing Quillaja saponin has potential for household applications and could be evaluated further for medical applications.
Relative Tolerance of a Range of Australian Native Plant Species and Lettuce to Copper, Zinc, Cadmium, and Lead by Dane T. Lamb; Hui Ming; Mallavarapu Megharaj; Ravi Naidu (pp. 424-432).
The tolerance of wild flora to heavy-metal exposure has received very little research. In this study, the tolerance of four native tree species, four native grass species, and lettuce to copper (Cu), zinc (Zn), cadmium (Cd), and lead (Pb) was investigated in a root-elongation study using Petri dishes. The results of these studies show a diverse range of responses to Cu, Zn, Cd, and Pb amongst the tested plant species. Toxicity among metals decreased in the following order: Cd ~ Cu > Pb > Zn. Metal concentrations resulting in a 50% reduction in growth (EC50) varied considerably, ranging from (μM) 30 (Dichanthium sericeum) to >2000 (Acacia spp.) for Cu; from 260 (Lactuca sativa) to 2000 (Acacia spp.) for Zn; from 27 (L. sativa) to 940 (Acacia holosericea) for Cd; and from 180 (L. sativa) to >1000 (Acacia spp.) for Pb. Sensitive native plant species identified included D. sericeum, Casuarina cunninghamiana, and Austrodanthonia caespitosa. However, L. sativa (lettuce) was also among the most sensitive to all four metals. Acacia species showed a high tolerance to metal exposure, suggesting that the Acacia genus shows potential for use in contaminated-site revegetation.
Changes in Bioturbation of Iron Biogeochemistry and in Molecular Response of the Clam Ruditapes decussates upon Perkinsus olseni Infection by Márcio F. Simão; Ricardo B. Leite; Carlos Rocha; M. Leonor Cancela (pp. 433-443).
A series of artificial microcosms was used to test the effect of clam density on benthic iron biogeochemistry and, subsequently, if the response of clam Ruditapes decussatus to infection with Perkinsus olseni, a common opportunistic parasite known to be iron dependent, was correlated with the dynamics of iron sediment pore waters within the chambers. Three series of benthic microcosms were used in the experiment, comparing similar densities of clams (none, one, two, three, or four individuals/chamber) between a control set (no deliberate infection) and two parallel sets of clams that were deliberately infected with the parasite after 10 days of incubation. Fifteen chambers were used simultaneously and the experiment was conducted for 35 days. In order to avoid spurious effects of differential organic loading and clam feeding efficiency on the oxidative state of the sediment, the iron balance was tentatively shifted during incubation toward decreased dissolved iron in pore water. This was done by applying a constant flow of air to all chambers and refraining from supplying extra organic matter during the experimental run, which led to the reduction of benthic oxygen demand as the experiment progressed. Results showed that microcosms bearing both higher clam densities and lower infection levels were able to exert a quantitative influence in iron biogeochemistry through bioturbation activity. This effect was significantly depressed in chambers hosting clams with high infection levels. In addition, analysis of molecular markers responsive to iron and parasite stress revealed an upper regulation of HSP70 and ferritin in infected clams, thus suggesting a role of those molecules on both host protection and response to parasite presence by limiting iron availability. Together, these findings suggest a correlation between the expression of clam molecular iron/stress markers and iron bioavailability, which can be modified by the presence or absence of Perkinsus infection. In turn, we propose that clam lethargy in response to parasite invasion might help to combat infection by reducing iron mobilization in the surrounding sediment through a decrease in bioturbation activity, thus reducing its availability to the parasite.
Organochlorine Pesticides in Consumer Fish and Mollusks of Liaoning Province, China: Distribution and Human Exposure Implications by Zheng Liu; Hongmei Zhang; Minhui Tao; Shaobin Yang; Liwei Wang; Ying Liu; Dandan Ma; Zhiming He (pp. 444-453).
Fish and mollusk samples were collected from markets located in 12 cities in Liaoning province, China, during August and September 2007, and 22 organochlorine pesticides (OCPs) were detected. DDT, HCH, endosulfan, chlordane, and HCB were the dominating OCPs, with mean concentrations and ranges of, respectively, 15.41 and 0.57 to 177.56 ng/g, 0.84 and below detection limit (BDL) to 22.99 ng/g, 1.31 and BDL to 13.1 ng/g, 1.05 and BDL to 15.68 ng/g, and 0.63 and BDL to 9.21 ng/g in all fish and mollusk samples. The concentrations of other OCPs generally were low and were detectable in a minority of samples, reflecting the low levels of these OCPs in the study region. In general, OCP concentrations were obviously higher in fish than in mollusks, and higher in freshwater fish than in marine fish, which indicated, first, that freshwater fish are more easily influenced than seawater fish and mollusks by OCP residues in agricultural areas and, second, that there are different biota accumulation factors for OCPs between fish and mollusk. To learn the consumption of fish and mollusk, 256 questionnaires were sent to families in 12 cities of Liaoning province. Using the contamination data, average estimated daily intakes of OCPs via fish and mollusk consumption were calculated, which were used for exposure assessment. The public health risks caused by exposure to OCPs in the course of fish and mollusk consumption were compared to noncancer benchmarks and cancer benchmarks.
Antioxidant Responses Versus DNA Damage and Lipid Peroxidation in Golden Grey Mullet Liver: A Field Study at Ria de Aveiro (Portugal) by M. Oliveira; I. Ahmad; V. L. Maria; M. Pacheco; M. A. Santos (pp. 454-463).
The present work aimed to investigate golden grey mullet (Liza aurata) liver protection versus damage responses at a polluted coastal lagoon, Ria de Aveiro (Portugal), as a tool to evaluate the human impacts on environmental health at five critical sites in Ria de Aveiro (Portugal) in comparison to a reference site (Torreira; TOR). Protection was evaluated by measuring non-enzymatic [total glutathione (GSHt) and non-protein thiols (NPT)] and enzymatic [catalase (CAT), glutathione peroxidase (GPx), glutathione-S-transferase (GST), and glutathione reductase (GR)] antioxidant defenses. Damage was assessed as DNA integrity loss and lipid peroxidation (LPO). No significant differences were found between sites in terms of non-enzymatic defenses (GSHt and NPT). CAT did not display significant differences among sites. However, GPx at Barra (BAR, associated with naval traffic), Gafanha (GAF, harbor and dry-dock activities area), Laranjo (LAR, metal contaminated associated with chlor–alkali plant), and Vagos (VAG, contaminated by polycyclic aromatic hydrocarbons) was significantly lower than the reference site. GST was lower at GAF, Rio Novo do Príncipe (RIO, pulp mill effluent area), LAR, and VAG, whereas GR was lower at RIO. The loss of antioxidant defenses was paralleled by higher LPO levels only at GAF and VAG. However, no DNA integrity loss was found. Results highlight the importance of the adopted multibiomarkers as applied in the liver of L. aurata in coastal water pollution monitoring. The integration of liver antioxidant defense and damage responses can improve the aquatic contamination assessment.
Trace Elements and Carbon and Nitrogen Stable Isotopes in Organisms from a Tropical Coastal Lagoon by A. A. Pereira; B. van Hattum; J. de Boer; P. M. van Bodegom; C. E. Rezende; W. Salomons (pp. 464-477).
Trace elements (Fe, Mn, Al, Zn, Cr, Cu, Ni, Pb, Cd, Hg, and As) and stable isotope ratios (δ13C and δ15N) were analyzed in sediments, invertebrates, and fishes from a tropical coastal lagoon influenced by iron ore mining and processing activities to assess the differences in trace element accumulation patterns among species and to investigate relations with trophic levels of the organisms involved. Overall significant negative relations between trophic level (given by 15N) and trace element concentrations in gastropods and crustaceans showed differences in internal controls of trace element accumulation among the species of different trophic positions, leading to trace element dilution. Generally, no significant relation between δ15N and trace element concentrations was observed among fish species, probably due to omnivory in a number of species as well as fast growth. Trace element accumulation was observed in the fish tissues, with higher levels of most trace elements found in liver compared with muscle and gill. Levels of Fe, Mn, Al, and Hg in invertebrates, and Fe and Cu in fish livers, were comparable with levels in organisms and tissues from other contaminated areas. Trace element levels in fish muscle were below the international safety baseline standards for human consumption.
Lethal Effect and In Vivo Genotoxicity of Profenofos to Chinese Native Amphibian (Rana spinosa) Tadpoles by Xianbin Li; Shaonan Li; Shaoying Liu; Guonian Zhu (pp. 478-483).
Amphibian populations are decreasing in size due to environmental stressors in most areas of southern China. Pesticides are known to be a group of potential stressors to amphibians, especially in agricultural ecosystems. Profenofos, an organophosphate insecticide and acaricide, is widely used for controlling insect pests in China. The aim of this study is to evaluate the acute lethality and genotoxicity of profenofos to amphibian under controlled conditions. Results showed that profenofos was highly lethal to tadpoles of Rana spinosa, with 50% lethal concentration (LC50) values of 1.59, 1.14, 0.77, and 0.58 mg l−1 at 24, 48, 72, and 96 h, respectively. DNA damage of erythrocytes was observed by alkaline single-cell gel electrophoresis assay at all tested sublethal concentrations. The study also showed, by micronucleus test, that profenofos at moderate to high sublethal concentration might have genotoxicity to the tadpole after 96 h exposure. Furthermore, based on our results, it is suggested that the alkaline single-cell gel electrophoresis assay could be used as a screening tool for biomonitoring of pesticide contamination in aquatic systems or agricultural ecosystems.
Columba livia as a Sentinel Species for the Assessment of Urban Air Genotoxicity by Matteo Sicolo; Maria Tringali; Pietro Fumagalli; Angela Santagostino (pp. 484-491).
This study explored the comet assay as a possible tool to assess genotoxicity in erythrocytes of Columba livia to detect genotoxic effects induced by exposure to urban air pollution. Fieldwork was conducted between June 2004 and June 2005 in the city of Milan, Italy, by sampling pigeons in different areas almost twice a week. Six air contaminants—CO, PM10, NO2, O3 (ozone), SO2, and C6H6—plus polycyclic aromatic hydrocarbons (PAHs) in fine particles, temperature, and ultraviolet index, were considered. Genotoxicity levels, expressed as %DNA migrated, tail moment, and damage index (DI), were always higher in wild pigeons than in pigeons living indoors (controls). Animals exposed to urban air showed significant differences from season to season, and the genotoxic parameters presented the highest values in summer (45.30% ± 1.40% %DNA migrated, 12.73 ± 0.80 tail moment, and 22.30 ± 0.15 × DI × 10−1); regression analyses showed a positive relation between DI and O3 concentrations (P < 0.001). The use of the comet assay DI parameter as a rapid assessment of incipient genotoxic risk by pollution, as measured in C. livia living in urban areas, is also discussed.
Polybrominated Diphenyl Ethers (PBDEs), Polychlorinated Biphenyles (PCBs), Hydroxylated and Methoxylated-PBDEs, and Methylsulfonyl-PCBs in Bird Serum from South China by Juan Liu; Xiao-Jun Luo; Le-Huan Yu; Ming-Jing He; She-Jun Chen; Bi-Xian Mai (pp. 492-501).
Polybrominated diphenyl ethers (PBDEs), polychlorinated biphenyls (PCBs), and their derivatives, hydroxylated (OH) and methoxylated (MeO) PBDEs and methylsulfonylated (MeSO2) PCBs, were measured in sera of eight bird species collected from an e-waste recycling region in South China. Concentrations of ∑PCBs, ranging from 38 to 1700 ng/g lipid weight (lw), were one to two orders of magnitude higher than concentrations of ∑PBDEs (0.64–580 ng/g lw). The significantly positive relationship between PCB and PBDE concentrations suggested a similar pathway of exposure to these compounds. Compared with muscle in birds, serum might prefer to accumulate and/or retain less brominated/chlorinated congeners. 3-OH-BDE47 and 2′-OH-BDE68 were detected in more than 80% of the collected bird serum samples (range: not detectable (nd) to 13 and nd to 7.8 ng/g lw, respectively). The other three OH-PBDE congeners (4′-OH-BDE-17, 6-OH-BDE47, and 4′-OH-BDE-49) and two MeO-PBDE congeners (3-MeO-BDE47 and 6-MeO-BDE47) were occasionally detected in bird sera at concentrations ranging from nd to 2.5 ng/g lw. Both natural sources and metabolic transformation of PBDEs could contribute to the presence of these PBDE derivatives in the birds. The two MeSO2-PCB congeners (4-MeSO2-CB49 and 4-MeSO2-CB101) under investigation were detected at respective concentration ranges of nd to 12 and nd to 0.68 ng/g lw. 4-MeSO2-CB101 exhibited the highest concentration among the nine PCB and PBDE derivatives studied, indicating that biotransformation via the mercapturic acid pathway of PCBs might have occurred in the studied bird species.
Are Great Tits (Parus major) Inhabiting the Vicinity of a Pulp Mill Healthy? Impacts on Physiology and Breeding Performance by Ana Cláudia Norte; B. C. Sheldon; J. P. Sousa; P. C. Tavares; M. E. Pereira; A. C. Duarte; J. A. Ramos (pp. 502-512).
The evaluation of pollution effects on wildlife provides biologically meaningful information concerning environmental quality and possible impacts on populations and can be used as an early warning of environmental change. We aimed to assess the potential effects of exposure to atmospheric pollution from pulp mill emissions on the morphology, physiology, and reproductive performance of the great tit (Parus major). Great tits from a population inhabiting the vicinity of a pulp mill had significantly higher feather mercury levels and were physiologically distinct from other birds not under the direct influence of the pollution source. This distinction, due to significantly higher levels of red blood cell–glutathione peroxidase antioxidant activity, in birds from the vicinity of the pulp mill, was observed both in autumn and winter and was repeatable between years. No detectable effects were observed on breeding performance or nestling morphology and physiology. The effects of pollution in this study seem to be related to increased levels of oxidative stress.
Metallothionein Gene Expression in Liver of Rats Exposed to Cadmium and Supplemented with Zinc and Selenium by Mohamed Banni; Imed Messaoudi; Lamia Said; Jihen El Heni; Abdelhamid Kerkeni; Khaled Said (pp. 513-519).
Cadmium (Cd), one of the most widely distributed heavy metals, is highly toxic to humans and animals. It is well known that zinc (Zn) and selenium (Se) administration reduce the Cd-induced toxicity and that metallothioneins can have a protective effect to mitigate Cd toxicity in biological systems. In this study we report the expression analysis of the two metallothioneines gene classes MT-1 and MT-2 as well as the total metalloprotein content in the liver of rats exposed to Cd (200 ppm), Cd + Zn (200 ppm + 500 ppm), Cd + Se (200 ppm + 0.1 ppm) or Cd + Zn + Se (200 ppm + 500 ppm + 0.1 ppm) in their drinking water for 35 days. Metals accumulation was quantified in rat liver. Cd decreased significantly the hepatic concentrations of Se and increased those of Zn. The treatment of Cd-exposed rats with Se alone or combined with Zn reversed the Cd-induced depletion of Se concentrations in the liver. However, Zn or Zn + Se administration significantly increased the liver Cd uptake and had no effect on the Cd-induced increase in hepatic concentrations of Zn. The molecular assay showed a decreasing trend of MT-1 relative gene expression levels in animals supplemented with Zn (6.87-fold), Se (3.58-fold), and their combination (1.69-fold) when compared to Cd-treated animals (16.22-fold). Upregulation of the MT-2 expression were recorded in all conditions, although fold induction levels were less pronounced than MT-1 expressions. Our data suggest that the well-established protective effect of Zn and Se against Cd-induced toxicity passes through non-MT gene expression mechanisms, being more dependent on the oxidative stress status of the cell.