A review of toxicity and mechanisms of individual and mixtures of heavy metals in the environment

The rational for the study was to review the literature on the toxicity and corresponding mechanisms associated with lead (Pb), mercury (Hg), cadmium (Cd), and arsenic (As), individually and as mixtures, in the environment. Heavy metals are ubiquitous and generally persist in the environment, enabling them to biomagnify in the food chain. Living systems most often interact with a cocktail of heavy metals in the environment. Heavy metal exposure to biological systems may lead to oxidation stress which may induce DNA damage, protein modification, lipid peroxidation, and others. In this review, the major mechanism associated with toxicities of individual metals was the generation of reactive oxygen species (ROS). Additionally, toxicities were expressed through depletion of glutathione and bonding to sulfhydryl groups of proteins. Interestingly, a metal like Pb becomes toxic to organisms through the depletion of antioxidants while Cd indirectly generates ROS by its ability to replace iron and copper. ROS generated through exposure to arsenic were associated with many modes of action, and heavy metal mixtures were found to have varied effects on organisms. Many models based on concentration addition (CA) and independent action (IA) have been introduced to help predict toxicities and mechanisms associated with metal mixtures. An integrated model which combines CA and IA was further proposed for evaluating toxicities of non-interactive mixtures. In cases where there are molecular interactions, the toxicogenomic approach was used to predict toxicities. The high-throughput toxicogenomics combines studies in genetics, genome-scale expression, cell and tissue expression, metabolite profiling, and bioinformatics.     

Ecotoxicological testing with new kinetic Photorhabdus luminescens growth and luminescence inhibition assays in microtitration scale

Miniaturized luminescence and growth inhibition assays in microtitration plates with the terrestric enthomopathogenic nematode symbiont Photorhabdus luminescens (DSMZ 3368T) are presented and compared with standardized tests with Vibrio fischeri (DSMZ 7151/NRRLB-11177) and Pseudomonas putida (DSMZ 50026). Toxicological parameters (EC and G values) of selected reference toxicants (e.g. heavy metals and environmental samples) were obtained at different temperatures and without sodium chloride supplementation. Kinetic data recordings were compared with results of a cuvette test protocol using integral and endpoint calculation. Growth inhibition experiments with reference samples reveal similar or higher sensitivities as for the Vibrio or Pseudomonas spp. The luminescence inhibition assay shows reduced sensitivities to most of the reference samples compared with the V. fischeri standard assay. But G values obtained with other standardized aquatic assay systems with daphnids, algae and growth inhibition assays with V. fischeri and Ps. putida correspond more closely to data observed with Ph. luminescens. The test procedures are easily to perform and to evaluate and seem to be reliable alternatives to the established protocols at low osmolarities. The sample specifity of the toxic responses of the marine and the terrestric strain recommends to employ both assays to determine the toxic potential of environmental samples. This will support to reduce false positive results in future investigations.     

Effects of heavy metals on the nitrogen metabolism of the aquatic moss Fontinalis antipyretica L. ex Hedw. A 15N tracer study

The assessment of pollution in aquatic systems necessitates an accurate indication of toxicity of heavy metals for organisms and ecosystems. We used the stable nitrogen isotope 15N to estimate the influence of the heavy metals Cd, Pb and Zn on the synthesis of nitrogen-containing fractions in the aquatic moss Fontinalis antipyretica. This method permits conclusions concerning inhibitory effects of these heavy metals on the assimilation of nitrogen and the biosynthesis of amino acids and proteins. The moss was exposed to metal concentrations of 25-500 microM over a period of 5-10 days. 15N abundance of exposed plants was compared with that of control plants. Similar to a loss of vitality determined using a fluorometric assay, a decrease of the 15N abundance in the N fractions of Fontinalis antipyretica was measured in dependence on the metal concentration. Nevertheless, the individual inhibition by the distinct metals was different, so that the following order of toxicity was derived: Cd > Pb > Zn.     

Environmental hazard screening of a metal-polluted site using pressurized liquid extraction and two in vitro bioassays

Rapid screening methods can improve the cost effectiveness, throughput, and quality of risk assessments of contaminated sites. In the present case study, the objective was to evaluate a combination of pressurized liquid extraction and 2 in vitro bioassays for the hazard assessment of surface soil sampled from 46 points across a pyrotechnical industrial site. Pressurized liquid extraction was used to rapidly produce soil-water extracts compatible with 2 high-capacity bioassays. Hazard assessment using combined toxicological and chemical screening revealed zones with relatively high potential risks of metal pollution. Multivariate data analysis provided indications that significant inhibition in the bioassays was correlated with levels of metals in the extracts, suggesting an elevated toxic potential from certain metals. Low pH and high concentrations of dissolved organic carbon were associated with increased cytotoxicity of extracts, indicating that these factors influence metal bioavailability. The cytotoxicity observed was more strongly correlated to metal concentrations in the extracts than in the soil, suggesting that measurements of total metal concentrations in soils do not provide good indications of the soil's potential toxicity.     

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

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

Multi-generation toxicity of zinc, cadmium, copper and lead to the potworm Enchytraeus albidus

In standard chronic terrestrial toxicity tests with invertebrates, adult organisms are exposed to the contaminants and the number of offspring is quantified. These procedures do not allow the assessment of possible effects on all life stages of the organism, which may lead to an underestimation of the toxicity of the test substance. To evaluate the importance of this issue, the potworm Enchytraeus albidus was exposed to zinc, cadmium, copper and lead for two subsequent generations. Juvenile production was assessed for both generations. Considering the variability of metal toxicity data reported in the literature, it is concluded that the two generation assay did not markedly increase the sensitivity of the standard E. albidus test for the tested metals. Therefore, toxicity data obtained with the proposed test guideline with E. albidus are protective for all life stages.     

A comparison of five rapid direct toxicity assessment methods to determine toxicity of pollutants to activated sludge

Five rapid direct toxicity assessment methods were used in three European partner countries to determine the toxicity of single toxicants, mixed toxicants and real industrial wastes. The final aim was to protect microbial degradation of organic wastes in biological treatment processes and hence enhance the quality of treated effluents to be discharged to the environment. Nitrification inhibition, Respirometry, Adenosine triphosphate luminescence and Enzyme inhibition were tested utilising activated sludge as the testing matrix. The Vibrio fischeri toxicity test was used as a surrogate to compare the various microbial bioassays. The IC50 (toxicant concentration eliciting a 50% inhibitory effect) was determined for a number of pollutants including single toxicants Cd, Cr, Cu, Zn, 3,5-dichlorophenol, toluene and linear alkylbenzenesulphonate (LAS); a standard mixture of metals and LAS; a standard mixture of organics and LAS, and 16 industrial effluents. The V. fischeri bioassay was also chosen in order to assess quality control of toxicant preparation during testing in the different laboratories of the partner countries. Comparisons of sensitivity, cost of implementation, cost per test, relevance, and ease of use were made. The most sensitive bioassays were V. fischeri and Nitrification inhibition, however, this depended in the main on the pollutant and mixtures tested. It is recommended that during assessment of wastewater toxicity a suite of tests be used rather than reliance on one particular test.     

Bioassays with Vibrio fischeri for the assessment of delayed toxicity

The standardized bioluminescence assay with Vibrio fischeri underestimates the aquatic toxicity of chemicals which interfere with metabolic pathways supporting long term processes like growth and reproduction due to its short incubation time (30 min). Therefore this short term assay was compared with two alternative bioassays with prolonged incubation times using the same test organism: the growth inhibition assay (7 h) and the long term bioluminescence assay (24 h). Two sets of compounds were selected to reflect acute and delayed toxicity. The first group comprised pentachlorophenol, dodecylpyridiniumbromide and 3,4-dichloroaniline and the second nalidixic acid, chloramphenicol and streptomycinsulfate. The effects of compounds with acute toxicity are determined with similar sensitivity in all bioassays. Substances with delayed toxicity show only minor or no toxicities in the standardized short term bioassay but severe effects in both long term bioassays independent of the parameter used. It is concluded that the standardized short term bioluminescence assay exhibits serious limitations for the assessment of aquatic toxicity. The long term bioassays, however, may help to overcome these limitations.     

Toxicity of metals and organic chemicals evaluated with bioluminescence assays

The development of a bioluminescent sensor organism (Shk1) that was created for assessing wastewater toxicity was reported several years ago. In order to establish a test battery to better characterize wastewater toxicity, additional luminescent sensor organisms were later created. The present study focused on one promising candidate (PM6), a Pseudomonas spp. strain, because of its high level of luminescence compared to that of other newly created organisms. Using a batch toxicity testing protocol, the toxicity of 7 metals and 25 organic compounds was evaluated with the PM6 and Shk1 assays. Results indicated that the toxicity data of the PM6 and the Shk1 assays were correlated, and no assay appeared to be particularly more sensitive to a group of toxicants than the other assay. The results of the PM6 and Shk1 assays were further evaluated by comparing with the results of the Vibrio fischeri luminescence inhibition assay and activated sludge inhibition assays. Data suggested that PM6 and Shk1 more closely represented activated sludge organisms than V. fischeri. The suitability of using PM6 and Shk1 for assessing wastewater toxicity on activated sludge, both individually and in a test battery, was discussed.     

Flow cytometry assessment of cytotoxicity and reactive oxygen species generation by single and binary mixtures of cadmium, zinc and copper on populations of the ciliated protozoan Tetrahymena thermophila

In the present study, we have assessed by flow cytometry, the cytotoxicity of the heavy metals Cd, Zn and Cu on populations of the ciliated protozoan Tetrahymena thermophila. The obtained LC(50) for these metals was estimated as 0.195mgCdl(-1), 3.58mgZnl(-1) and 0.47mgCul(-1), respectively. As a result, the toxicity rank for this eukaryotic microorganism is Cd>Cu>Zn. Using the same methodology and the Concentration Addition approach, the toxicity of binary mixtures of these metals was evaluated in order to detect the type of interaction between these metals. Results indicated that antagonism is the predominant interaction but it can change to additivity or even to synergism at high metal concentrations. Besides, the concentration ratio between metals plays also a crucial role in determining the type of metallic interaction, at least in Tetrahymena. Cytotoxicity data from single and bimetallic mixtures have been compared with those from selected microalgae, other species of ciliates, fish and mammalian cell lines and metazoan. By other hand, we have detected the mitochondrial generation of peroxides induced by both single and binary treatments with Cd, Zn and Cu on populations of Tetrahymena, using the specific fluorophore dyhidrorhodamine. The nature and concentrations of metal as well as the metallic ratio were important factors in reactive oxygen species production. All results found in T. thermophila are compared with previous reports in other organisms and, some explanations and hypothesis to support results are given, including the involvement of metallothioneins as antioxidants and their role in the binding of metal cations.     

Ecotoxicological and chemical evaluation of phenolic compounds in industrial effluents

The aim of this paper was to evaluate the ecotoxicological response of industrial effluents containing phenolic compounds. All complex effluents collected from a chemical plant and then after both a chemical–physical and biological treatment were characterised with chemical analysis, biodegradability tests and four ecotoxicological tests (Daphnia magna, Artemia salina, Brachionus plicatilis and Vibrio fisheri with Microtox^®). The evaluation of the chemical and ecotoxicological data was useful for predicting the effect of the raw effluent on the treatment plant and the impact of the final treated effluent on the receiving water. Besides the toxicity of the effluent from the chemical plants, the acute toxicity of its main components was also determined. The results of the tests and toxicity data from literature were transformed in Toxic Units (TUs). Effluent toxicity was under- or over-estimated by calculating the sum of the TUs of the individual components, depending on which toxicity data and test organisms were used.     

Effects of heavy metals on sea urchin embryo development. 1. Tracing the cause by the effects

The toxicity of the polluted waters originating from a disused lead mine was evaluated using both sea urchin bioassays and heavy metal analysis. Samples from three polluted waters (a seawater and two freshwaters) were collected from the mine area and one seawater sample was taken from a non-contaminated reference site. The test waters contained higher concentrations of heavy metals such as manganese, lead, cadmium, zinc, chromium, nickel, iron, and copper than did ambient seawater. The three test waters had inhibitory effects, in a dose-dependent manner, on the first cleavage of sea urchin embryos and on pluteus formation during the development. Some malformations, such as a radialized pluteus, exo-gastrula, and spaceship Apollo-like embryos were induced by the test waters without dilution. Zinc alone also induced the same anomaly. Zinc in the test seawater was ascertained as one of the metals that caused the anomalies, but not all of the toxicity was caused by zinc. It was speculated that interactive effects, involving zinc and possibly manganese and nickel, were occurring.     

<Emphasis Type="Italic">Cylindrospermopsis raciborskii</Emphasis> exudate–Cu complexes: impact on copper dynamics and bioavailability in an aquatic food chain

INTRODUCTION: The increasing contamination of aquatic environments motivates studies on the interactions among natural dissolved organic matter, metals, and the biota. This investigation focused on the organic exudates of the toxic cyanobacteria Cylindrospermopsis raciborskii as a Cu carrier through a three-level aquatic trophic chain (bacteria, protozoa, and copepod). DISCUSSION: The effects of bacteria activity and growth on the metal-organic complexes were evaluated through changes in free Cu(2+) ions, total dissolved, and total particulate Cu. To be sure that the added copper would be complexed to the exudates, its complexing properties were previously determined. The cyanobacteria exudate-Cu complexes were furnished to bacteria that were further used as a food source to the protozoan Paramercium caudatum. This was then furnished as food to the copepod Mesocyclops sp. The results showed that, in general, the cyanobacterial exudates decreased Cu bioavailability and toxicity to the first trophic level (bacteria), but because the heterotrophic bacteria accumulated Cu, they were responsible for the transference for the otherwise low availability metal form. Both the bacteria and protozoan organisms accumulated Cu, but no metal accumulation was detected in the copepods.     

Two new growth inhibition tests with the filamentous algae Ceramium strictum and C. tenuicorne (Rhodophyta)

Two growth inhibition tests using the red marine macroalgae Ceramium strictum and the brackish water relative C. tenuicorne have been developed. Besides using phenol as a reference substance, the toxicity of a metal, a flame retardant and a complex effluent water were assayed. The two methods are reliable and repeatable bioassays for salinities between 4 and 30 per thousandth. The coefficients of variation (CV) for toxicity of the reference substance phenol were 15% for the Stereo Microscope Analysis test and between 24 and 51% for the Computer Image Analysis test (n=5). Ceramium spp. are common and important primary producers in temperate coastal waters and are thus relevant as test organisms. Both algae grow well in laboratorial conditions and tests can be performed all year around.     

Cadmium and copper toxicity in three marine macroalgae: evaluation of the biochemical responses and DNA damage

Marine macroalgae have evolved a different mechanism to maintain physiological concentrations of essential metal ions and non-essential metals. The objective of the present work was to evaluate the antioxidant response and DNA damage of copper and cadmium ions in three halophytes, namely, Acanthophora spicifera, Chaetomorpha antennina, and Ulva reticulata. Accumulation of copper was significantly higher (P?P?U. reticulata > A. spicifera > C. antennina. DNA damage index analysis supported that copper was significantly (P?相似文献   

Guidelines for copper in sediments with varying properties

A major weakness of sediment quality guidelines (SQGs) is their poor ability to predict how toxicity thresholds change for different sediment types. Using species sensitivity distributions (SSDs) of copper effects data, new guidelines were derived for copper in non-sulfidic marine sediments in which organic carbon (OC) and particle size strongly influence copper bioavailability. The derived SQGs varied in a predictable manner with changes in sediment particle size and organic carbon (OC), and were shown to offer a significant improvement on the existing ‘single value’ SQG. Adequate protection for all benthic organisms is expected to be achieved for a OC-normalised copper concentration of 3.5 mg Cu g⁻¹ OC in the <63 μm sediment fraction. For short-term exposures, the equivalent acute guideline is 11 mg < 63 μm Cu g⁻¹ OC. The new SQGs incorporate a high degree of conservatism owing to the use of copper-spiked sediments and laboratory-based bioassays that were expected to result in greater metal exposure of organisms to bioavailable copper than would be expected for field-contaminated sediments with similar total copper concentrations. SQGs that vary with sediment properties were prepared in an easily referenced tabular format.     

Urease inhibition: a tool for toxicity identification in sediment elutriates

A method for the detection and confirmation of heavy metal toxicants in sediment elutriates based on a urease inhibition assay, ICP-AES analysis and EDTA chelation in the frame of toxicity identification evaluation (TIE) is presented. Zinc was identified as the major toxicant in pHstat elutriates of sediments of the river Saale (Germany). Implications of natural and anthropogenic chelating agents, which are frequently present in environmental samples, on toxicity confirmation of heavy metals based on the toxic unit approach are discussed.     

Heavy metal binding capacity (HMBC) of municipal solid waste landfill leachates

This research describes the use of a toxicity assay for the identification of metal toxicity, bioavailability and heavy metal binding capacity (HMBC) of municipal solid waste (MSW) landfill leachates. MetPLATE, an assay specific for heavy metal toxicity, was used to determine the HMBC of MSW leachates collected from 14 sites in Florida, with a wide range of chemical and physical characteristics. The leachates displayed a low toxicity which was attributed to the site-specific parameters, including, high concentrations of both organic and inorganic ligands. The HMBC test was undertaken to measure the effect of these site-specific parameters on metal toxicity. The potential for MSW leachate to bind and, thus, detoxify heavy metals was investigated with copper, zinc, and mercury. The HMBC values obtained ranged from 3 to 115, 5 to 93 and 4 to 101 for HMBC-Cu+2, HMBC-Zn+2, and HMBC-Hg+2, respectively. Additionally, the high strength leachates displayed the highest binding capacities, although the landfills sampled represented a wide range of characteristics. For comparison, the HMBC values reported with local lake water, Lake Alice and Lake Beverly, and a wastewater treatment plant effluent were all below 3. A partial fractionation of MSW leachate samples from sites 1, 5 and 8, was conducted to further investigate the influence of selected site-specific physico-chemical parameters on metal binding. The fractionation revealed that the HMBC of the leachate samples was heavily influenced by the concentration of solids, organics and hardness.     

Comparison of heavy metal toxicity in life stages (spermiotoxicity, egg toxicity, embryotoxicity and larval toxicity) of Hydroides elegans

A toxicity test was developed to examine the effects of heavy metal contaminants on the early life stages of the marine polychaete. We have studied the effects of metals on fertilization and early development of marine polychaete Hydroides elegans. These heavy metals have often been found in polluted ground and water near industrial discharges, and have therefore been detected from time to time in the food chain. They have been reported to alter various reproduction functions in various animals including marine populations. The toxic effect of mercury, cadmium, lead, nickel and zinc on sperm viability, fertilization, embryogenesis and larvae of H. elegans was examined. We observed that the rate of fertilization decreased when the sperm was incubated with heavy metals. Treatment of eggs with each metal did not prevent fertilization, but delayed or blocked the first mitotic divisions, and altered early embryonic development. All these effects were observed at relatively high concentrations. However, bio-accumulation in sediments and aquatic organisms have been reported. Polychaete eggs may then be in contact with very high concentrations of these heavy metals in areas where these metals are not handled or stocked properly, and then develop into abnormal embryos. In addition to bivalves and sea-urchins, polychaete embryos can provide biological criteria for seawater quality standards taking into account the sensitivity of the invertebrates and their contribution in detection of harmful chemicals with no marked effect on the species. Our results indicate that the early development of H. elegans is highly sensitive to heavy metals and this polychaete can be routinely employed as a test organism for ecotoxicity bioassays in tropical and subtropical regions.     

Interaction between components of electroplating industry wastes. Influence of the receiving water on the toxicity of the effluent

A copper-cadmium-nickel-zinc mixture was assessed in seven different river waters to study metal toxicity to the ciliate protozoan Colpidium campylum, the interactions occurring between metals, and the influence of the receiving water on toxicity. In the range of concentrations tested, which are representative of electroplating industry wastes, the main part of the toxicity can be attributed to copper and to cadmium-copper synergy. A classification of waters, based on a principal component analysis (PCA), was used to examine the main parameters of the water, which can affect the toxicity of metal mixtures. It appears that the mineralization of the water, more than the total organic carbon (TOC), is an important parameter for the expression of toxicity. A strategy for the estimation of ecotoxicological hazard assessment, based on a simplified factorial experiment is proposed. It enables one to study, in a two-step bioassay, the toxicity of an effluent, the influence of river water on its toxicity, and the effects of contact time and dilution. By applying PCA to data from very different waters, it may be possible to estimate the ecotoxicological risk associated with the discharge of an effluent, on the basis of the chemistry of the receiving water.