Toxicity Of Metal Mixtures Research Articles

Modeling acute toxicity of metal mixtures to wheat (Triticum aestivum L.) using the biotic ligand model-based toxic units method

The combined toxic effects of copper (Cu) and cobalt (Co) were predicted using the biotic ligand model (BLM) for different concentrations of magnesium (Mg2+) and pH levels, with parameters derived from Cu-only and Co-only toxicity data. The BLM-based toxic unit (TU) approach was used for prediction. Higher activities of Mg2+ linearly increased the EC50 of Cu and Co, supporting the concept of competitive binding of Mg2+ and metal ions in toxic action. The effects of pH on Cu and Co toxicity were related not only to free Cu2+ and Co2+ activity, respectively, but also to inorganic metal complexes. Stability constants for the binding of Cu2+, CuHCO3+, CuCO3(aq), CuOH+, Mg2+, Co2+, CoHCO3+ and Mg2+ with biotic ligands were logKCuBL 5.87, mathrm{log},{K}_{{{rm{CuHCO}}}_{3}{rm{BL}}} 5.67, mathrm{log},{K}_{{{rm{CuCO}}}_{3}{rm{BL}}} 5.44, logKCuOHBL 5.07, logKMgBL 2.93, logKCoBL 4.72, mathrm{log},{K}_{{{rm{CoHCO}}}_{3}{rm{BL}}} 5.81 and logKMgBL 3.84, respectively. The combinations of Cu and Co showed additive effects under different conditions. When compared with the FIAM-based TU model (root mean square error [RMSE = 16.31, R2 = 0.84]), the BLM-based TU model fitted the observed effects better (RMSE = 6.70, R2 = 0.97). The present study supports the BLM principles, which indicate that metal speciation and major cations competition need to be accounted for when predicting toxicity of both single metals and mixtures of metals.

Effects of exposure to multiple heavy metals on biochemical and histopathological alterations in common carp, Cyprinus carpio L.

Heavy metals are frequently encountered as mixtures of essential and non-essential elements. Therefore, evaluation of their toxic effects individually does not offer a realistic estimate of their impact on biological processes. We studied effects of exposure to mixtures of essential and toxic metals (Cr, Cd and Pb) on biochemical, immunotoxicity level and morphological characteristics of the various tissues of a biomarker freshwater fish common carp using environmentally relevant concentrations. Fish were exposed to metal mixture through tank water for 7, 15 and 30 days, under controlled laboratory conditions. Tissue accumulation of the metals was measured using Atomic Absorption Spectrophotometric techniques. Chromium, cadmium and lead accumulation in muscle, gills, liver, kidney and intestine, tissue of common carp exposed to mixture metals for 30 days increased significant compared with control group (p < 0.001). However, the activity of antioxidant enzymes such as catalase (CAT) and superoxide dismutase (SOD) levels was significant altered in various tissues of exposed fish. Besides, the lipid peroxidation (LPO) was significant (p < 0.001) increased. Moreover, the tumor necrosis factor - α (TNF-α), interleukin (IL-6), and interferon-γ (IFN-γ) contents in tissues of muscle, gills, liver, kidney and intestine were increased significant compared with control fish (p < 0.001). In addition, microscopic examination of the main alterations in general morphology of fish gills included spiking and fusion of secondary lamellae, formation of club-shaped filaments epithelium in the interlamellar regions and hepatocytes showed damage of central vein and rupture of irregular hepatic plate with more number of vacuoles in the fish exposed to metal mixture for a longer duration (30 days). These results of this study clearly demonstrate that concentration individual and mixtures of metals in aquatic systems will greatly influence the cytokine alterations may result in an immune suppression or excessive activation in the treated common carp as well as may cause immune dysfunction or reduced immunity. In conclusion, toxicity of multiple metal mixtures of Cr, Cd and Pb has antioxidant and immunotoxic effects on C. carpio.

Extended biotic ligand model for predicting combined Cu–Zn toxicity to wheat (Triticum aestivum L.): Incorporating the effects of concentration ratio, major cations and pH

Current risk assessment models for metals such as the biotic ligand model (BLM) are usually applied to individual metals, yet toxic metals are rarely found singly in the environment. In the present research, the toxicity of Cu and Zn alone and together were studied in wheat (Triticum aestivum L.) using different Ca2+ and Mg2+ concentrations, pH levels and Zn:Cu concentration ratios. The aim of the study was to better understand the toxicity effects of these two metals using BLMs and toxic units (TUs) from single and combined metal toxicity data. The results of single-metal toxicity tests showed that toxicity of Cu and Zn tended to decrease with increasing Ca2+ or Mg2+ concentrations, and that the effects of pH on Cu and Zn toxicity were related not only to free Cu2+ and Zn2+ activity, respectively, but also to other inorganic metal complex species. For the metal mixture, Cu–Zn interactions based on free ion activities were primarily additive for the different Ca2+ and Mg2+ concentrations and levels of pH. The toxicity data of individual metals derived by the BLM, which incorporated Ca2+ and Mg2+ competition and toxicity of inorganic metal complexes in a single-metal toxicity assessment, could predict the combined toxicity as a function of TU. There was good performance between the predicted and observed effects (root mean square error [RMSE] = 7.15, R2 = 0.97) compared to that using a TU method with a model based on free ion activity (RMSE = 14.29, R2 = 0.86). The overall findings indicated that bioavailability models that include those biochemistry processes may accurately predict the toxicity of metal mixtures.

Combined toxicity of cadmium and lead on early life stages of the Pacific oyster, Crassostrea gigas

Trace metals cause toxic effect on the early development period of marine animals, however only few studies address the toxic interactions of trace metals on bivalves. In the present study, the individual and combined toxicities of dissolved cadmium (Cd) and lead (Pb) on early life stages of the Pacific oyster Crassostrea gigas have been investigated. Embryotoxicity, larvae mortality and genotoxicity were measured under single and combined exposure of the two tested metals. For embryotoxicity, the median effective concentration (EC50) values for individual Cd, Pb and their mixture were of 272.2 μg/L, 660.3 μg/L and 373.1 μg/L, respectively. The median lethal concentrations (LC50) for 96 h larval mortality were determined to be 353.3 μg/L, 699.5 μg/L and 205.5 μg/L for individual Cd, Pb and their mixture, respectively. Moreover, the Marking-Dawson additive toxicity indices were 0.10 for embryogenesis and 1.40 for larval mortality indicating, respectively, an additive effect and a trend to synergism for the Cd and Pb combination. Furthermore, DNA strand breaks were observed in oyster embryos after individual Cd, Pb and their mixture exposure, and a significant positive correlation was demonstrated between embryotoxicity and genotoxicty. The current study suggests the toxicity of Cd is higher than that of Pb, and the Cd-Pb mixture is slightly more toxic than individual Cd or Pb to the Pacific oyster. These data will be helpful to predict the toxicity of metal mixtures, and provide biological criteria for the implementation of marine water quality standards to protect these marine organisms.

Prediction of toxicity of zinc and nickel mixtures to Artemia sp. at various salinities: From additivity to antagonism

Few studies have examined the toxicity of metal mixtures to marine organisms exposed to different salinities. The aim of the present study was to investigate the acute toxicity of zinc and nickel exposures singly and in combination to Artemia sp. under salinities of 10, 17, and 35 psu. The mixture concentrations were determined according to individual toxic units (TUs) to follow a fixed ratio design. Zinc was more toxic than nickel, and both their individual toxicities were higher at lower salinities. These changes in toxicity can be attributed to the Biotic Ligand Model (BLM) rather than to metal speciation. To analyze the mixture effect, the observed data were compared with the expected mixture effects predicted by the concentration addition (CA) model and by deviations for synergistic/antagonistic interactions and dose-level and dose-ratio dependencies. For a salinity of 35 psu, the mixture had no deviations; therefore, the effects were additive. After decreasing the salinity to 17 psu, the toxicity pattern changed to antagonism at low concentrations and synergism at higher equivalent LC50 levels. For the lowest salinity tested (10 psu), antagonism was observed. The speciations of both metals were similar when in a mixture and when isolated, and changes in toxicity patterns are more related to the organism's physiology than metal speciation. Therefore, besides considering chemical interactions in real-world scenarios, where several chemicals can be present, the influence of abiotic factors, such as salinity, should also be considered.

Modelling toxicity of metal mixtures: A generalisation of new advanced methods, considering potential application to terrestrial ecosystems

ABSTRACTNo chemical properties have been found to be a reliable predictor of metal toxicity, attributed to the influence of various specific processes on metal toxicokinetics and toxicodynamics.Metal toxicity is affected by interactions between metal ions and of metals with organisms. The present review aims to provide an outlook on recent progress in estimating toxicity of metal mixtures. Three methods applied recently have the potential of accounting for the interactions between metals as well as the interactions of metals with organisms. The methods are based on the Biotic Ligand Model (BLM), electrostatic interactions, and Windermere Humic Aqueous Model (WHAM). These methods can be generalised to a common principle that affinity of metals for binding sites on sorption surfaces (i.e., biotic ligands on organisms, plasma membrane surface, and humic acid-surrogated biological surface) is a unifying factor for the biological response. The scientific basis for these approaches was reviewed in the present study, accompanied by a preliminary assessment on the toxicity of Cu2+, Ag+, Zn2+, and their binary mixtures to lettuce, Lactuca sativa, using previously published data. The difference in the predictive performance of the modelling approaches reflects the difference in the simulation of metal-ligand binding and is related to the specific exposure conditions considered.

Toxicity models of metal mixtures established on the basis of “additivity” and “interactions”

Observed effects of metal mixtures on animals and plants often differ from the estimates, which are commonly calculated by adding up the biological responses of individual metals. This difference from additivity is commonly referred to as being a consequence of specific interactions between metals. The science of how to quantify metal interactions and whether to include them in risk assessment models is in its infancy. This review summarizes the existing predictive tools for evaluating the combined toxicity of metals present in mixtures and indicates the advantages and disadvantages of each method. We intend to provide eco-toxicologists with background information on how to make good use of the tools and how to advance the methods for assessing toxicity of metal mixtures. It is concluded that statistically significant deviations from additivity are not necessarily biologically relevant. Incorporation of interactions between metals in a model does not on forehand mean that the model is more accurate than a model developed based on additivity only. It is recommended to first use a relatively simple method for effect prediction of uninvestigated metal mixtures. To improve the reliability of toxicity modeling for metal mixtures, further efforts should focus on balancing the relationship between the significance of statistics and the biological meaning, and unraveling the toxicity mechanisms of metals and their mixtures. Open image in new window

Systematic Evaluation of Chronic Metal-Mixture Toxicity to Three Species and Implications for Risk Assessment.

Metal contamination generally occurs as mixtures. However, it is yet unresolved how to address metal mixtures in risk assessment. Therefore, using consistent methodologies, we have set up experiments to identify which mixture model applies best at low-level effects, i.e., the independent action (IA) or concentration addition (CA) reference model. The toxicity of metal mixtures (Ni, Zn, Cu, Cd, and Pb) to Daphnia magna, Ceriodaphnia dubia, and Hordeum vulgare was investigated in different waters or soils, totaling 30 different experiments. Some mixtures of different metals, each individually causing <10% inhibition, yielded much larger inhibition (up to 66%) when dosed in combination. In general, IA was most accurate in predicting mixture toxicity, while CA was the most conservative. At low-effect levels important in risk assessments, CA overestimated mixture toxicity to daphnids and H. vulgare, on average, with a factor 1.4 to 3.6. Observed mixture interactions could be related to bioavailability or by competition interactions, either for binding sites of dissolved organic carbon or for biotic ligand sites. Our study suggests that the current metal-by-metal approach in risk evaluations may not be conservative enough for metal mixtures.

Interactive toxicity of Ni, Zn, Cu, and Cd on Daphnia magna at lethal and sub-lethal concentrations

The toxicity of metal mixtures is currently of particular interest among aquatic toxicologists. To provide insight into whether the interaction of multiple metals is similar at different biological levels, the survival and feeding behavior of Daphnia magna were studied following exposure to four metals (Cd, Cu, Ni, Zn) and their binary and quaternary combinations. In terms of survival, Zn-Cu and Cu-Cd mixtures produced more-than-additive mortality, while Ni-Cd mixtures resulted in less-than-additive mortality. Regarding behavior, Zn-Cu and Zn-Cd mixtures produced a more-than-additive reduction in feeding rate. Four (i.e. Zn-Cu, Cu-Cd, Ni-Cd, and Zn-Cd) out of six binary mixtures in the present study interacted differently at the survival and behavioral levels, strengthening the emphasis on carefully selecting the toxicological endpoint when addressing metal mixture toxicity. The results of the present study demonstrated that metals are toxic to feeding behavior of D. magna at much lower concentrations (i.e. 27–63 times lower) compared to survival, suggesting that applying sub-lethal endpoints are required for producing protective regulations.

Evaluating Complex Mixtures in the Zebrafish Embryo by Reconstituting Field Water Samples: A Metal Pollution Case Study.

Accurately assessing the toxicity of complex, environmentally relevant mixtures remains an important challenge in ecotoxicology. The goal was to identify biological effects after exposure to environmental water samples and to determine whether the observed effects could be explained by the waterborne metal mixture found in the samples. Zebrafish embryos were exposed to water samples of five different sites originating from two Flemish (Mol and Olen, Belgium) metal contaminated streams: “Scheppelijke Nete” (SN) and “Kneutersloop” (K), and a ditch (D), which is the contamination source of SN. Trace metal concentrations, and Na, K, Mg and Ca concentrations were measured using ICP-MS and were used to reconstitute site-specific water samples. We assessed whether the effects that were observed after exposure to environmental samples could be explained by metal mixture toxicity under standardized laboratory conditions. Exposure to “D” or “reconstituted D” water caused 100% mortality. SN and reconstituted SN water caused similar effects on hatching, swim bladder inflation, growth and swimming activity. A canonical discriminant analysis confirmed a high similarity between both exposure scenarios, indicating that the observed toxicity was indeed primarily caused by metals. The applied workflow could be a valuable approach to evaluate mixture toxicity that limits time and costs while maintaining biological relevance.

Toxicity Assessment of Binary Metal Mixtures (Copper-Zinc) to Nitrification in Soilless Culture with the Extended Biotic Ligand Model.

Metals are always found in the environment as mixtures rather than as solitary elements. Only a limited number of studies have developed appropriate models that incorporate bioavailability to estimate the toxicity of heavy-metal mixtures. In the present study, we explored the applicability of two extended biotic ligand model (BLM) approaches-BLM-f mix and BLM-toxicity unit (TU)-to predict and interpret mixture toxicity with the assumption that interactions between metal ions obey the BLM theory. Exposure assays of single and mixed metals were performed with inoculums of an ammonia-oxidizing bacterium SD5 isolated from soil. Nitrification of the cultures was the end point used to quantify the toxic response. The results indicated that the developed BLM-f mix approach could well estimate the single toxicity of Cu2+ and Zn2+ as well as their binary mixture toxicity to nitrification with >90% of toxicity variation explained. Assuming that metal ions compete with each other for binding at a single biotic ligand, the BLM-f mix approach (root-mean-square error [RMSE]=19.66, R 2=0.8879) showed better predictive power than the BLM-TU approach (RMSE=31.12, R 2=0.6892). The present study supports the use of the accumulation of metal ions at the biotic ligands as predictor of toxicity of single metals and metal mixtures.

Development and validation of a metal mixture bioavailability model (MMBM) to predict chronic toxicity of Ni-Zn-Pb mixtures to Ceriodaphnia dubia

Recently, several bioavailability-based models have been shown to predict acute metal mixture toxicity with reasonable accuracy. However, the application of such models to chronic mixture toxicity is less well established. Therefore, we developed in the present study a chronic metal mixture bioavailability model (MMBM) by combining the existing chronic daphnid bioavailability models for Ni, Zn, and Pb with the independent action (IA) model, assuming strict non-interaction between the metals for binding at the metal-specific biotic ligand sites. To evaluate the predictive capacity of the MMBM, chronic (7d) reproductive toxicity of Ni-Zn-Pb mixtures to Ceriodaphnia dubia was investigated in four different natural waters (pH range: 7–8; Ca range: 1–2 mM; Dissolved Organic Carbon range: 5–12 mg/L). In each water, mixture toxicity was investigated at equitoxic metal concentration ratios as well as at environmental (i.e. realistic) metal concentration ratios. Statistical analysis of mixture effects revealed that observed interactive effects depended on the metal concentration ratio investigated when evaluated relative to the concentration addition (CA) model, but not when evaluated relative to the IA model. This indicates that interactive effects observed in an equitoxic experimental design cannot always be simply extrapolated to environmentally realistic exposure situations. Generally, the IA model predicted Ni-Zn-Pb mixture toxicity more accurately than the CA model. Overall, the MMBM predicted Ni-Zn-Pb mixture toxicity (expressed as % reproductive inhibition relative to a control) in 85% of the treatments with less than 20% error. Moreover, the MMBM predicted chronic toxicity of the ternary Ni-Zn-Pb mixture at least equally accurately as the toxicity of the individual metal treatments (RMSEMix = 16; RMSEZn only = 18; RMSENi only = 17; RMSEPb only = 23). Based on the present study, we believe MMBMs can be a promising tool to account for the effects of water chemistry on metal mixture toxicity during chronic exposure and could be used in metal risk assessment frameworks.

Mixture toxicity of copper, cadmium, and zinc to barley seedlings is not explained by antioxidant and oxidative stress biomarkers.

The analysis of metal mixture toxicity to plants is complicated by mutual interactions. In the present study, mixture effects of zinc (Zn), cadmium (Cd), and copper (Cu) on barley (Hordeum vulgare L.) root elongation were analyzed using oxidative stress parameters. The hypothesis was that toxic mixture effects on plant growth are better explained by biochemical parameters than by exposure information, because the former excludes interactions among metals for root uptake. Barley seedlings were exposed for 5 d or 14 d to these metals in nutrient solutions, added in isolation and as mixtures. Root elongation in Cu+Cd mixtures was well predicted from free metal ion concentrations in solution, using concentration addition (CA) or independent action (IA) reference models. In contrast, Zn acted antagonistically when combined with Cu and/or Cd, relative to both CA and IA. This protective effect of Zn correlated with the biomarkers measured in the long-term experiment; oxidative stress (indicated by malondialdehyde level, for example) decreased after addition of Zn. In addition, it was found that some biomarkers were sensitive to both Cu and Cd dosed in isolation, but not to Cu+Cd mixtures. Overall, the exposure explained mixture effects better than most of the 16 measured biomarkers (i.e., the biochemical effects). It is concluded that these biomarkers are not robust indicators for metal mixture toxicity, potentially because different metals have different parallel modes of action on growth that are insufficiently indexed by the biomarkers. Environ Toxicol Chem 2017;36:220-230. © 2016 SETAC.

Joint toxic action of binary metal mixtures of copper, manganese and nickel to Paronychiurus kimi (Collembola)

The joint toxic effects of binary metal mixtures of copper (Cu), manganese (Mn) and nickel (Ni) on reproduction of Paronhchiurus kimi (Lee) was evaluated using a toxic unit (TU) approach by judging additivity across a range of effect levels (10–90%). For all metal mixtures, the joint toxic effects of metal mixtures on reproduction of P. kimi decreased in a TU-dependent manner. The joint toxic effects of metal mixtures also changed from less than additive to more than additive at an effect level lower than or equal to 50%, while a more than additive toxic effects were apparent at higher effect levels. These results indicate that the joint toxicity of metal mixtures is substantially different from that of individual metals based on additivity. Moreover, the close relationship of toxicity to effect level suggests that it is necessary to encompass a whole range of effect levels rather than a specific effect level when judging mixture toxicity. In conclusion, the less than additive toxicity at low effect levels suggests that the additivity assumption is sufficiently conservative to warrant predicting joint toxicity of metal mixtures, which may give an additional margin of safety when setting soil quality standards for ecological risk assessment.

Mixture toxicity and interactions of copper, nickel, cadmium, and zinc to barley at low effect levels: Something from nothing?

Metal contamination is mostly a mixture of different metals, and these multicomponent mixtures can produce significant mixture effects. The present study was set up to investigate the toxicity of multiple metal mixtures of Cu, Ni, Cd, and Zn to plants at metal doses individually causing low-level phytotoxic effects. Barley (Hordeum vulgare L.) root elongation toxicity tests were performed in resin-buffered nutrient solutions to control metal speciation. Treatments included single-metal concentrations and binary, ternary, and quaternary mixtures. Mixtures of different metals at free ion concentrations, each causing <10% inhibition of root elongation, yielded significant mixture effects, with inhibition reaching up to 50%. The independent action (IA) model predicted mixture toxicity statistically better than the concentration addition (CA) model, but some synergisms relative to the IA model were observed. These synergisms relative to IA were most pronounced in quaternary mixtures and when the dose-response curves had steep slopes. Generally, antagonistic interactions relative to the CA model were observed. Increasing solution Zn concentrations shifted metal interactions (CA based) from additive or slightly synergistic at background Zn concentrations to antagonistic at higher Zn concentrations, suggesting a protective effect of Zn. Overall, the present study shows that the CA model can be used as a conservative model to predict metal mixture toxicity to barley. Environ Toxicol Chem 2016;35:2483-2492. © 2016 SETAC.

A test of the additivity of acute toxicity of binary-metal mixtures of ni with Cd, Cu, and Zn to Daphnia magna, using the inflection point of the concentration-response curves.

Mixtures of metals are often present in surface waters, leading to toxicity that is difficult to predict. To provide data for development of multimetal toxicity models, Daphnia magna neonates were exposed to individual metals (Cd, Cu, Ni, Zn) and to binary combinations of those metals in standard 48-h lethality tests conducted in US Environmental Protection Agency moderately hard reconstituted water with 3 mg dissolved organic carbon (DOC)/L added as Suwannee River fulvic acid. Toxicity tests were performed with mixtures of Ni and 1) Cd, which is considerably more toxic than Ni; 2) Cu, which is less toxic than Cd but more toxic than Ni; and 3) Zn, which has a toxicity threshold similar to Ni. For each combination of metals in the binary mixtures, the concentration of 1 metal was held constant while the second metal was varied through a series that ranged from nonlethal to lethal concentrations; then the roles of the metals were reversed. Inflection points of the concentration-response curves were compared to test for additivity of toxicity. Sublethal concentrations of Ni caused less-than-additive toxicity with Cd, slightly less-than-additive toxicity with Zn, and greater-than-additive toxicity with Cu. One explanation of these results might be competition among the metals for binding to biological ligands and/or dissolved organic matter. Therefore, models might have to incorporate sometimes competing chemical interactions to accurately predict metal-mixture toxicity. Environ Toxicol Chem 2016;35:1843-1851. © 2015 SETAC.

Ecological risk assessment of boreal sediments affected by metal mining: Metal geochemistry, seasonality, and comparison of several risk assessment methods.

The mining industry is a common source of environmental metal emissions, which cause long-lasting effects in aquatic ecosystems. Metal risk assessment is challenging due to variations in metal distribution, speciation, and bioavailability. Therefore, seasonal effects must be better understood, especially in boreal regions in which seasonal changes are large. We sampled 4 Finnish lakes and sediments affected by mining for metals and geochemical characteristics in autumn and late winter, to evaluate seasonal changes in metal behavior, the importance of seasonality in risk assessment, and the sensitivity and suitability of different risk assessment methods. We compared metal concentrations in sediment, overlying water, and porewater against environmental quality guidelines (EQGs). We also evaluated the toxicity of metal mixtures using simultaneously extracted metals and an acid volatile sulfides (SEM-AVS) approach together with water quality criteria (US Environmental Protection Agency equilibrium partitioning benchmarks). Finally, site-specific risks for 3 metals (Cu, Ni, Zn) were assessed using 2 biotic ligand models (BLMs). The metal concentrations in the impacted lakes were elevated. During winter stratification, the hypolimnetic O2 saturation levels were low (<6%) and the pH was acidic (3.5-6.5); however, abundant O2 (>89%) and neutral pH (6.1-7.5) were found after the autumnal water overturn. Guidelines were the most conservative benchmark for showing an increased risk of toxicity in the all of the lakes. The situation remained stable between seasons. On the other hand, SEM-AVS, equilibrium partition sediment benchmarks (ESBs), and BLMs provided a clearer distinction between lakes and revealed a seasonal variation in risk among some of the lakes, which evidenced a higher risk during late winter. If a sediment risk assessment is based on the situation in the autumn, the overall risk may be underestimated. It is advisable to carry out sampling and risk assessment during periods in which metals are assumed to be the most environmentally harmful. Integr Environ Assess Manag 2016;12:759-771. © 2015 SETAC.

Reproductive toxicity of binary and ternary mixture combinations of nickel, zinc, and lead to Ceriodaphnia dubia is best predicted with the independent action model.

Metals occur as mixtures in the environment. Risk assessment procedures for metals currently lack a framework to incorporate chronic metal mixture toxicity. In the present study, the toxicity of binary and ternary mixture combinations of Ni, Zn, and Pb was investigated in 3 large-scale experiments using the standard chronic (7-d) Ceriodaphnia dubia reproductive toxicity test. These metals were selected because of anticipated differences in mode of action. The toxicity of the metals in most mixtures, expressed as either free metal ion activities or as dissolved metal concentrations, were antagonistic relative to the concentration addition model, whereas no significant (p < 0.05) interactive effects were observed relative to the independent action model. The only exception was the binary Pb-Zn mixture, for which mixture effects were noninteractive based on the dissolved concentrations, but antagonistic based on free ion activities all relative to the independent action model. Overall, the independent action model fitted the observed toxicity better than the concentration addition model, which is consistent with the different modes of action of these metals. The concentration addition model mostly overestimated toxicity. Finally, the present study warns against extrapolation of the type of interactive effects between species, even when they are closely related. Environ Toxicol Chem 2016;35:1796-1805. © 2015 SETAC.

Application of biotic ligand and toxicokinetic–toxicodynamic modeling to predict the accumulation and toxicity of metal mixtures to zebrafish larvae

Predicting the accumulation and toxicity of mixtures of metals to aquatic organisms is a key challenge in ecotoxicological studies. In this study, the accumulation and toxicity of mixed essential (Cu) and nonessential (Cd and Pb) metals in zebrafish larvae exposed to a binary mixture of these elements at environmentally relevant concentrations were predicted using a refined toxicokinetic (TK)–toxicodynamic (TD) model aided with biotic ligand model (BLM) and toxic equivalent factor (TEF) approach. Competitive inhibition and non-competitive interaction/inhibition were observed in bio-uptake. Both Pb and Cd behaved as competitive inhibitors of Cu uptake at high Cu concentrations (>0.1 μM). By contrast, Cu uptake was independent of Cd or Pb when the Cu concentrations were below 10−7 M. Furthermore, low concentrations of Cu had an adiaphorous effect on Cd or Pb uptake. Cd uptake was inhibited by Pb, and the Pb uptake rates consistently decreased in the presence of Cd. The accumulation processes of Cd–Pb, Cu–Cd, and Cu–Pb were accurately predicted by the BLM-aided TK models. The traditional TD model could successfully predict the toxicity of Cd–Pb mixtures, but not those of Cu–Cd or Cu–Pb mixtures. The revised TD model, which considered the possible different killing rates (Kk) above or below the threshold, offered better prediction for the toxicity of Cu–Cd or Cu–Pb mixtures. The overall findings may be of key significance in understanding and predicting metal uptake, accumulation, and toxicity in binary or multiple metal exposure scenarios.

Combined toxicity of heavy metal mixtures in liver cells.

With rapid industrialization, China is now facing great challenges in heavy metal contamination in the environment. Human exposure to heavy metals through air, water and food commonly involves a mixture consisting of multiple heavy metals. In this study, eight common heavy metals (Pb, Cd, Hg, Cu, Zn, Mn, Cr, Ni) that cause environmental contamination were selected to investigate the combined toxicity of different heavy metal mixtures in HL7702 cells. Toxicity (24 h LC50 ) of each individual metal on the cells ranked Hg > Cr = Cd > Cu > Zn > Ni > Mn > Pb; toxicity of the different mixtures ranked: M5 > M3PbHgCd > M5+Mn > M5+Cu > M2CdNi > M4A > M8-Mn > M8 > M5+Zn > M4B > M8-Cr > M8-Zn > M8-Cu > M8-Pb > M8-Cd > M8-Hg > M8-Ni > M3PbHgNi > M3CuZnMn. The cytotoxicity data of individual metals were successfully used to build the additive models of two- to eight-component metal mixtures. The comparison between additive model and combination model or partly additive model was useful to evaluate the combined effects in mixture. Synergistic, antagonistic or additive effects of the toxicity were observed in different mixtures. These results suggest that the combined effects should be considered in the risk assessment of heavy metal co-exposure, and more comprehensive investigations on the combined effects of different heavy metal mixtures are needed in the future. Copyright © 2016 John Wiley & Sons, Ltd.