Single Metal Exposure Research Articles

Cadmium and lead mixtures are less toxic to the Chinese medicinal plant Ligusticum chuanxiong Hort. Than either metal alone.

Agricultural production of Ligusticum chuanxiong Hort. is often affected by heavy metal pollution in soil, especially mixtures of cadmium (Cd) and lead (Pb). We assessed metal-induced phytotoxicity in L. chuanxiong by exposing the plants to soil treated with Cd, Pb, or Cd/Pb mixtures. A combined Cd/Pb treatment alleviated the inhibition in plant growth, photosynthesis, and secondary metabolite generation seen in single-metal exposures in three of the four combinations. Most combined Cd/Pb treatments resulted in preferential uptake of magnesium, copper, and nitrogen in underground plant parts and accumulation of phosphorus and calcium in aboveground plant parts, thereby leading to improvements in photosynthetic potential. Compared with single-metal exposures, combined Cd/Pb treatment significantly decreased the contents of Cd by 16.67%-40.12% and Pb by 10.68%-21.70% in the plant, respectively. At the subcellular level, the Pb presence increased the Cd percentage associated with cell wall from 64.79% to 67.93% in rhizomes and from 32.76% to 45.32% in leaves, while Cd reduced Pb contents by 9.36%-46.39% in the subcellular fractions. A combined Cd/Pb treatment decreased the contents of water- and ethanol-extractable metal forms and increased the contents of acetic acid- and hydrochloric acid-extractable forms. The lower toxic effects of the Cd/Pb mixture in L. chuanxiong were associated with photosynthetic potential, subcellular distribution, the chemical forms of Cd and Pb, and synthesis of secondary metabolites. These findings are useful for plant production strategies in soils contaminated by heavy metals.

Influence of Relative Humidity on Germination and Metal Accumulation in Vigna radiata Exposed to Metal-based Nanoparticles

The effect of single (0.1, 1, and 10 mg L−1) and binary mixtures (0.1 + 0.1, 1 + 1, and 10 + 10 mg L−1) of metal-based nanoparticles (CdO and CuO) on the germination of Vigna radiata was studied under two humidity ranges (70% and 80%). Filter paper-based tests were conducted. The surface-sterilized seeds were exposed to CdO and CuO under controlled environmental conditions (70% and 80% humidity at 35 °C). Germination rates were scored after 24 h and 48 h. The accumulation of metals was tested in seedlings after 48 h using inductively coupled plasma mass spectrometry. Compared with 70% humidity, the germination rate was higher under 80% humidity in all tested conditions. The germination rate of the CdO + CuO treatment was less than that of the single metal exposure under both humidities (70% and 80%) at 48 h. By two-way analysis of variance (ANOVA), we found that germination was greatly influenced by humidity. The accumulation of metal was higher in the CuO test than in the CdO test. Metal accumulation was concentration and humidity dependent, except for Cd accumulation in the CdO + CuO treatment. Here we show that the germination of seeds depends on the humidity and concentration of metal oxide nanoparticles. Understanding these strategies in seeds might help to avoid environmental and chemical stress and improve crop yield.

Single and Combined Effects of Cadmium and Aroclor 1254 on Oxidative Stress in Gills of Mytilus coruscus

Both polychlorinated biphenyls (PCBs) and cadmium (Cd) can be frequently found in marine ecosystems and have detrimental effects on marine organisms, especially on filter-feeding marine mussels. Although biological responses to single metal or PCB exposure in mussels have been well-studied, information about oxidative stress is still limited, especially in different tissues in mussels. Considering the variety of contaminants existing in the actual marine environment, the exposures of the marine mussel Mytilus coruscus to Cd2+ alone (0.194, 0.388, and 0.775 mg/L) and Aroclor 1254 alone (0.005, 0.010, and 0.050 mg/L) and the co-exposures of the marine mussel Mytilus coruscus to Cd2+ (0.194 and 0.388 mg/L) and Aroclor 1254 (0.005 and 0.010 mg/L) were tested in an 8-day exposure experiment followed by a 7-day acclimation experiment. The alterations in superoxide dismutase, catalase, glutathione peroxidase, and malondialdehyde levels in the gills of the mussels were assessed. The effects of the depressed antioxidant were induced by the exposures of Cd2+ and Aroclor 1254 and their co-exposures. All exposures resulted in an initial increase and then a reduction in antioxidant enzyme activities. The range and rate of the antioxidant enzyme activities were positively correlated with stress duration and the concentration of the stress material. The effect of combined stress was stronger than that of each individual stressor. The valuable information for future investigations of stress response mechanisms, especially in relation to tissue functions in marine organisms, has been provided by the results and experimental model. The study of combined pollution effects has more scientific significance for marine pollution monitoring.

Antioxidant enzymes as biomarkers of Cu and Pb exposure in the ground spiders Lycosa terrestris and Pardosa birmanica.

Heavy metal exposure induces oxidative stress in terrestrial organisms, which they counteract via activation of antioxidant biomarkers. The present study investigated the effects of copper (Cu) and lead (Pb) on the total antioxidant capacity (TAC) and antioxidant enzymes such as Catalase (CAT), Glutathione reductase (GR), Superoxide dismutase (SOD) and Glutathione peroxidase (GPX) in two spider species, namely Lycosa terrestris and Pardosa birmanica. The spiders were exposed to Cu and Pb separately (10ppm) or in combination (10ppm each) via two different exposure routes (i.e. food and soil) for 10, 20 and 40 days. The results showed that metal accumulation and antioxidant biomarker responses in spiders were metal- and species-dependent. Also, the levels of all antioxidant biomarkers increased significantly with increasing exposure time and metal load in the bodies of spiders via both exposure routes. The significant inhibition of TAC and antioxidant enzyme activities was only observed in single Pb treatment through soil exposure. In L. terrestris, the activities of detoxification enzymes and TAC were significantly enhanced on single Cu exposure than Pb via both exposure routes. However, in P. birmanica consistent variation among antioxidant parameters were observed depending on the metal load and exposure routes. The combined metal exposure caused more pronounced increase in the level of antioxidants compared to single metal exposure in both species, mainly via food exposure. These results suggest that the antioxidant enzymes and TAC are sensitive to single and combined metal exposure via both uptake routes. These data show that antioxidant parameters can be used potential biomarkers of oxidative stress associated with metal exposure and for monitoring environmental health using spiders as bioindicators.

Assessment of bioaccumulation of cu and Pb in experimentally exposed spiders, Lycosa terrestris and Pardosa birmanica, using different exposure routes.

Major concerns exist regarding the environmental and human health risks caused by exposure to heavy metals. Spiders are often used as a model in ecotoxicological studies to assess soil pollution. Here, we measured the bioaccumulation of copper (Cu) and lead (Pb) in spiders, Lycosa terrestris and Pardosa birmanica, by inductively coupled plasma mass spectrometry (ICP-MS). We investigated whether Cu and Pb accumulation differs according to different spider species, single versus combined metal exposure, and routes of exposure. Spiders were exposed to 10mM CuSO4 and 10mM PbCl2 solutions separately or in combination (10mM + 10mM) through different exposure routes (spiked soil and food) for 6weeks. The effect of metals on the survival and body mass of exposed and unexposed (control) spiders was determined. We found that in both spider species, accumulation of metals increased with exposure time. In single metal exposure, Cu accumulation from food was higher than soil exposure in both spider species, whereas the opposite was observed for Pb. The simultaneous uptake of Cu and Pb significantly decreased from food and soil, respectively. Soil exposure caused more accumulation of metals in L. terrestris than P. birmanica. Metal exposure via contaminated food caused higher mortality compared to soil exposure. Body mass of both spider species was significantly decreased and negatively correlated with metal's concentration. Overall, our results show that bioaccumulation efficiency of Cu and Pb differs significantly in spiders exposed to metal's mixture compared to single metal exposure and is dependent on the exposure route, the type of metal, and spider species. More understanding of the effects of exposure to metal mixture and exposure routes is essential for designing and supporting risk assessment and ecological monitoring programs.

Effect of Zinc on the Oxidative Stress Biomarkers in the Brain of Nickel-Treated Mice.

The overexposure to nickel due to the extensive use of it in modern technology remains a major public health concern. The mechanisms of pathological effects of this metal remain elusive. The present study was devoted to evaluate the effect of nickel on the oxidative state of the brain cells of mice and to assess whether zinc as redox state modulator could efficiently protect cells against nickel's neurotoxicity. As oxidative stress biomarkers in the present study, we have measured the concentrations of reduced glutathione, metallothioneins, and malondialdehyde and the activity of the enzyme δ-aminolevulinate dehydratase. For the single metal exposure, mice were i.p. injected once with solutions of NiCl2 and/or ZnSO4; repeated exposure was performed i.p. injecting metal salt solutions for 14 days (once a day). The control mice received i.p. injections of saline. Results of our study demonstrate that single and 14 days of Ni2+ exposure decreased reduced glutathione and increased malondialdehyde contents in the brain of mice. Repeated Ni2+ administration significantly inhibited δ-aminolevulinate dehydratase while increasing brain metallothionein concentration at both exposure periods. Zinc exhibited a protective effect against nickel-induced glutathione and lipid peroxidation in brain cells of mice at both intervals of time, while repeated exposure to this metal significantly raised the brain metallothionein content. Repeated Zn2+ pretreatment protected δ-aminolevulinate dehydratase from Ni2+-induced inhibition and significantly increased metallothionein concentration at both investigated time intervals.

Modeling of the bioaccumulative efficiency of Pistia stratiotes exposed to Pb, Cd, and Pb + Cd mixtures in nutrient-poor media

The bioaccumulation capacity of water lettuce (Pistia stratiotes) including heavy metals has been well documented. However, its bioaccumulative efficiency for Pb, Cd, and Pb + Cd mixtures in nutrient-poor media is neglected. Therefore, the present study was aimed to investigate the bioaccumulative efficiency of the water lettuce by evaluating the bioconcentration factors (BCFs) and goodness-of-fit to regression models. Plants were exposed to various concentrations of Pb, Cd, and Pb + Cd mixtures in tap water for 1, 4, and 7 days. Heavy metal accumulation in whole plant tissue was measured with ICP-MS. The highest bioaccumulation rate was recorded at 1 day exposure tests for both metals as 14.9 mg/kg for Pb and 2.21 mg/kg for Cd. The BCFs for Pb and Cd ranged between 0.13 and 2.39 and 0.08 to 1.24, respectively. The modeling of the BCFs and exposure duration indicate that all of the models developed were significant. The R2 values were higher for single metal exposure tests. The model fitted curves demonstrate the decrease in BCFs with increasing exposure duration in all groups. Our results indicate that water lettuce can be used to treat nutrient-poor media polluted with low levels of Pb, Cd, and Pb + Cd mixtures for short incubation periods.

BaP-metals co-exposure induced tissue-specific antioxidant defense in marine mussels Mytilus coruscus

Both benzo(α)pyrene (BaP) and metals are frequently found in marine ecosystem and can cause detrimental effects in marine organism, especially the filter feeder-marine mussels. Although the biological responses in mussels have been well-studied upon the single metal or BaP exposure, the information about antioxidant defense, especially in different tissues of mussels, are still limited. Considering the variety of contaminants existing in the actual marine environment, single BaP (56 μg/L) and the co-exposure with Cu, Cd and Pb (50 μg/L, 50 μg/L and 3 mg/L respectively) were applied in a 6 days exposure followed by 6 days depuration experiment. The alterations of superoxide dismutase (SOD), catalase (CAT) activities and total antioxidant capacity (TAC) level were assessed in haemolymph, gills and digestive glands of marine mussels, Mytilus coruscus. An unparalleled change in antioxidant biomarkers was observed in all cells/tissues, with the SOD activity showing higher sensitivity to exposure. A tissue-specific response showing unique alteration in gill was investigated, indicating the different function of tissues during stress responses. Depressed antioxidant effects were induced by BaP-metals co-exposure, indicating the interaction may alter the intact properties of BaP. To our knowledge, this is the first research to explore the antioxidant defense induced by combined exposure of BaP-metals regarding to tissue-specific responses in marine mussels. The results and experimental model will provide valuable information and can be utilized in the investigation of stress response mechanisms, especially in relation to tissue functions in marine organism in the future.

In vivo bioconcentration of a metal mixture by Danio rerio eleutheroembryos

Exposure to heavy metals has represented one of the most serious health risks of environmental pollution over the last 50 years. Most of the bioconcentration studies that have been carried out to date explored only individual contaminants, unlike the real situations that occur in the environment. In this work, zebrafish eleutheroembryos were exposed to a mixture of CH3Hg(II), iAs(III), Ag(I) and Cd(II), and new BCFs were calculated and compared with those calculated from single metal exposures. In both cases, experimental conditions meet the OECD Test 305 conditions established for aquatic systems. In addition, spatial imaging obtained by laser ablation coupled to inductively plasma mass spectrometry (LA-ICP/MS), has been directly performed in these samples providing complementary information. The new BCF's have revealed some differences compared to single metal exposures when eleutheroembryos were exposed to the metal mixture, especially for iAs(III) and Cd(II). LA-ICP/MS images are in good agreement with the BFC's found, representing an interesting approach to get spatial distribution of metals that reinforces the toxicokinetic information.

Internal versus External Dose for Describing Ternary Metal Mixture (Ni, Cu, Cd) Chronic Toxicity to Lemna minor.

Simultaneous determinations of internal dose ([M]tiss) and external doses ([M]tot, {M2+} in solution) were conducted to study ternary mixture (Ni, Cu, Cd) chronic toxicity to Lemna minor in alkaline solution (pH 8.3). Also, concentration addition (CA) based on internal dose was evaluated as a tool for risk assessment of metal mixture. Multiple regression analysis of dose versus root growth inhibition, as well as saturation binding kinetics, provided insight into interactions. Multiple regressions were simpler for [M]tiss than [M]tot and {M2+}, and along with saturation kinetics to the internal biotic ligand(s) in the cytoplasm, they indicated that Ni-Cu-Cd competed for uptake into plant, but once inside, only Cu-Cd shared a binding site. Copper inorganic complexes (hydroxides, carbonates) played a role in metal bioavailability in single metal exposure but not in mixtures. Regardless of interactions, the current regulatory approach of using CA based on [M]tot can sufficiently predict mixture toxicity (∑TU close to 1), but CA based on [M]tiss was closest to unity across a range of doses. Internal dose integrates all metal-metal interactions in solution and during uptake into the organism, thereby providing a more direct metric describing toxicity.

Toxicity of platinum, palladium and rhodium to Daphnia magna in single and binary metal exposure experiments

Mainly due to automobile traffic, but also due to other sources, the platinum group elements (PGE) platinum (Pt), palladium (Pd) and rhodium (Rh) are introduced into aquatic biotopes where they accumulate in sediments of lakes and rivers. However, the toxicity of these noble metals to aquatic organisms is not well understood and especially toxicity studies under standardized condition are lacking. Thus, the toxicity of Pt, Pd and Rh to Daphnia magna was tested in single metal exposure experiments according to OECD guideline 202. Immobility and lethality was recorded after 24 h and 48 h of exposure and EC50 and LC50, respectively, were determined. As the nominal exposure concentration of Pd differed significantly from the quantified concentration, the control of the real exposure concentration by chemical analysis is mandatory, especially for Pd.The toxicity decreased in the order Pd > Pt ≫ Rh with e.g. LC50(48 h) values of 14 μg/L for Pd, 157 μg/L for Pt and 56,800 μg/L for Rh. The exposure period had a clear effect on the toxicity of Pt, Pd and Rh. For Pt and Rh the endpoint immobility was more sensitive than the endpoint lethality whereas Pd toxicity was similar for both endpoints. The Hill slopes, which are a measure for the steepness of the concentration-response curves, showed no significant discrepancies between the different metals.The binary metal exposure to Pt and Pd revealed a more-than-additive, i.e. a synergistic toxicity using the toxic unit approach. The present study is a start to understand the toxicity of interacting PGE. The modes of action behind the synergistic effect are unclear.

Larval aquatic insect responses to cadmium and zinc in experimental streams.

To evaluate the risks of metal mixture effects to natural stream communities under ecologically relevant conditions, the authors conducted 30-d tests with benthic macroinvertebrates exposed to cadmium (Cd) and zinc (Zn) in experimental streams. The simultaneous exposures were with Cd and Zn singly and with Cd+Zn mixtures at environmentally relevant ratios. The tests produced concentration-response patterns that for individual taxa were interpreted in the same manner as classic single-species toxicity tests and for community metrics such as taxa richness and mayfly (Ephemeroptera) abundance were interpreted in the same manner as with stream survey data. Effect concentrations from the experimental stream exposures were usually 2 to 3 orders of magnitude lower than those from classic single-species tests. Relative to a response addition model, which assumes that the joint toxicity of the mixtures can be predicted from the product of their responses to individual toxicants, the Cd+Zn mixtures generally showed slightly less than additive toxicity. The authors applied a modeling approach called Tox to explore the mixture toxicity results and to relate the experimental stream results to field data. The approach predicts the accumulation of toxicants (hydrogen, Cd, and Zn) on organisms using a 2-pKa bidentate model that defines interactions between dissolved cations and biological receptors (biotic ligands) and relates that accumulation through a logistic equation to biological response. The Tox modeling was able to predict Cd+Zn mixture responses from the single-metal exposures as well as responses from field data. The similarity of response patterns between the 30-d experimental stream tests and field data supports the environmental relevance of testing aquatic insects in experimental streams. Environ Toxicol Chem 2017;36:749-762. Published 2016 Wiley Periodicals Inc. on behalf of SETAC. This article is a US government work and, as such, is in the public domain in the United States of America.

Metal uptake of Nerium oleander from aerial and underground organs and its use as a biomonitoring tool for airborne metallic pollution in cities.

The analysis of the airborne particulate matter-PM-incorporated to plant leaves may be informative of the air pollution in the surroundings, allowing their use as biomonitoring tools. Regarding metals, their accumulation in leaves can be the result of both atmospheric incorporation of metallic PM on aboveground plant organs and root uptake of soluble metals. In this study, the use of Nerium oleander leaves as a biomonitoring tool for metallic airborne pollution has been assessed. The metal uptake in N. oleander was assessed as follows: (a) for radicular uptake by irrigation with airborne metals as Pb, Cd, Cr, Ni, As, Ce and Zn (alone and in mixture) and (b) for direct leave exposure to urban PM. Plants showed a high resistance against the toxicity of metals under both single and multiple metal exposures. Except for Zn, the low values of translocation and bioaccumulation factors confirmed the excluder behaviour of N. oleander with respect to the metals provided by the irrigation. For metal uptake from airborne pollution, young plants grown under controlled conditions were deployed during 42days in locations of the city of Zaragoza (700,000h, NE Spain), differing in their level of traffic density. Samples of PM2.5 particles and the leaves of N. oleander were simultaneously collected weekly. High correlations in Pb concentrations were found between leaves and PM2.5; in a lesser extent, correlations were also found for Fe, Zn and Ti. Scanning electron microscopy showed the capture of airborne pollution particles in the large and abundant substomatal chambers of N. oleander leaves. Altogether, results indicate that N. Oleander, as a metal resistant plant by metal exclusion, is a suitable candidate as a biomonitoring tool for airborne metal pollution in urban areas.

Cadmium-induced olfactory dysfunction in rainbow trout: Effects of binary and quaternary metal mixtures

A functioning olfactory response is essential for fish to be able to undertake essential behaviors. The majority of work investigating the effects of metals on the olfactory response of fish has focused on single-metal exposures. In this study we exposed rainbow trout to cadmium, copper, nickel, zinc, or a mixture of these four metals at or below the current Canadian Council of Ministers of the Environment guidelines for the protection of aquatic life. Measurement of olfactory acuity using an electro-olfactogram demonstrated that cadmium causes significant impairment of the entire olfactory system, while the other three metals or the mixture of all four metals did not. Binary mixtures with cadmium and each of the other metals demonstrated that nickel and zinc, but not copper, protect against cadmium-induced olfactory dysfunction. Testing was done to determine if the protection from cadmium-induced olfactory dysfunction could be explained by binding competition between cadmium and the other metals at the cell surface, or if the protection could be explained by an up-regulation of an intracellular detoxification pathway, namely metallothionein. This study is the first to measure the effects of binary and quaternary metal mixtures on the olfactory response of fish, something that will aid in future assessments of the effects of metals on the environment.

Esterase activity (EA), total oxidant status (TOS) and total antioxidant capacity (TAC) in gills of Mytilus galloprovincialis exposed to pollutants: Analytical validation and effects evaluation by single and mixed heavy metal exposure

The aims of the present study were to optimize and validate methods for esterase activity (EA), total oxidant status (TOS) and total antioxidant capacity (TAC) determination in mussel' gills, and to establish the relationships between these biomarkers and Pb, Cd and Cu pollution, in single form and ternary mixture. Two different buffers for sample homogenization, the need of ultracentrifugation, and analytical validation were evaluated. Coefficients of variation, when buffer without additives and ultracentrifugation were used, were <15%, and recovery were 97%–109% in all cases. The EA response tends to decrease with treatments, TOS decreased significantly in Cd and ternary groups, while TAC tended to increase in treatments with Pb, Cd and ternary groups. In conclusion, the methods for EA, TOS and TAC measurements in gills of mussel were precise and accurate and could be interesting resources in biomonitoring programmes.

Perinatal and Childhood Exposure to Cadmium, Manganese, and Metal Mixtures and Effects on Cognition and Behavior: A Review of Recent Literature.

Lead (Pb) and mercury (Hg) neurotoxicity is well established. In recent years, a growing body of evidence suggests that environmental exposure to other metals including arsenic (As), cadmium (Cd), and manganese (Mn) and their mixtures also poses public health threats. In this paper, we summarize the recent literature examining the relationship of prenatal and childhood environmental metal exposures with cognitive and behavioral outcomes in children. We conducted a literature search to identify epidemiologic studies that examined the relationship of Cd, Mn, and metal mixtures with children's neurodevelopmental/cognitive and behavioral outcomes. We restricted the search to peer-reviewed studies published in English between January 2009 and March 2015. We identified a total of 31 articles of which 16, 17, and 16 studies examined the effects of Cd, Mn, or metal mixtures, respectively. Based on our review, there is suggestive evidence that prenatal/childhood Cd exposure may be associated with poorer cognition, but additional research is clearly needed. We found little evidence of behavioral effects of early life Cd exposure, and no studies found a significant relationship with attention deficit hyperactivity disorder. Studies of early life Mn exposure consistently reported negative impacts on both cognition and behavior. There is also growing evidence that co-exposure to multiple metals can result in increased neurotoxicity compared to single-metal exposure, in particular during early life. Few studies have evaluated behavioral effects related to metal co-exposure.

다양한 독성법을 이용한 중금속, 나노입자 및 금속오염 토양 평가

Toxicity results of metals, nanoparticles (NPs), and soils contaminated with metals were introduced on this review. Following methods were used: seed germination, bioluminescence, enzyme activity, and mutation. In general, different sensitivities were observed, depending on types of bioassays and pollutants. Among tested seeds, sensitivities of Lactucus and Raphanus were greater than others. Of single metal exposure, effect by As(III) was greater than others, and high revertant mutation ratio (5.1) was observed at 1 mg/L arsenite, indicating high mutagenicity. No general pattern was observed on the effect of metal mixture, but synergistic effect was observed with seeds. In case of soils, no correlation was observed between total metal contents and toxicity. Toxicity of NPs was observed as follows: CuO > ZnO > NiO > TiO2, Fe2O3, Co3O4. Especially, no considerable effects were observed by TiO2, Fe2O3, and Co3O4 under tested concentration (max. 1,000 mg/L). The evaluation results of interactive toxic effects using various bioassays may comprise a useful tool for the bioassessment of various environmental pollutants.

Health Risk Assessment using in vitro digestion model in assessing bioavailability of heavy metal in rice: A preliminary study

Little is known about the bioavailability of heavy metal contamination and its health risks after rice ingestion. This study aimed to determine bioavailability of heavy metal (As, Cd, Cu, Cr, Co, Al, Fe, Zn and Pb) concentrations in cooked rice and human Health Risk Assessment (HRA). The results found Zn was the highest (4.3±0.1mg/kg), whereas As showed the lowest (0.015±0.001mg/kg) bioavailability of heavy metal concentration in 22 varieties of cooked rice. For single heavy metal exposure, no potential of non carcinogenic health risks was found, while carcinogenic health risks were found only for As. Combined heavy metal exposures found that total Hazard Quotient (HQtotal) values for adult were higher than the acceptable range (HQTotal<1), whereas total Lifetime Cancer Risk (LCRTotal) values were higher than the acceptable range (LCRTotal values >1×10−4) for both adult and children. This study is done to understand that the inclusion of bioavailability heavy metal into HRA produces a more realistic estimation of human heavy metal exposure.

Metal mixture modeling evaluation project: 3. Lessons learned and steps forward.

A comparison of 4 metal mixture toxicity models (that were based on the biotic ligand model [BLM] and the Windermere humic aqueous model using the toxicity function [WHAM-FTOX ]) was presented in a previous paper. In the present study, a streamlined version of the 4 models was developed and applied to multiple data sets and test conditions to examine key assumptions and calibration strategies that are crucial in modeling metal mixture toxicity. Results show that 1) a single binding site on or in the organism was a useful and oftentimes sufficient framework for predicting metal toxicity; 2) a linear free energy relationship (LFER) for bidentate binding of metals and cations to the biotic ligand provided a good first estimate of binding coefficients; 3) although adjustments in metal binding coefficients or adjustments in chemical potency factors can both be used in model calibration for single-metal exposures, changing metal binding coefficients or chemical potency factors had different effects on model predictions for metal mixtures; and 4) selection of a mixture toxicity model (based on concentration addition or independent action) was important in predicting metal mixture toxicity. Moving forward, efforts should focus on reducing uncertainties in model calibration, including development of better methods to characterize metal binding to toxicologically active binding sites, conducting targeted exposure studies to advance the understanding of metal mixture toxicity, and further developing LFERs and other tools to help constrain the model calibration.

Modeling and interpreting biological effects of mixtures in the environment: introduction to the metal mixture modeling evaluation project.

The fate and biological effects of chemical mixtures in the environment are receiving increased attention from the scientific and regulatory communities. Understanding the behavior and toxicity of metal mixtures poses unique challenges for incorporating metal-specific concepts and approaches, such as bioavailability and metal speciation, in multiple-metal exposures. To avoid the use of oversimplified approaches to assess the toxicity of metal mixtures, a collaborative 2-yr research project and multistakeholder group workshop were conducted to examine and evaluate available higher-tiered chemical speciation-based metal mixtures modeling approaches. The Metal Mixture Modeling Evaluation project and workshop achieved 3 important objectives related to modeling and interpretation of biological effects of metal mixtures: 1) bioavailability models calibrated for single-metal exposures can be integrated to assess mixture scenarios; 2) the available modeling approaches perform consistently well for various metal combinations, organisms, and endpoints; and 3) several technical advancements have been identified that should be incorporated into speciation models and environmental risk assessments for metals.