Adam C. Ryan

Influence of pH, hardness, dissolved organic carbon concentration, and dissolved organic matter source on the acute toxicity of copper to Daphnia magna in soft waters: implications for the biotic ligand model.

The influence of pH, dissolved organic carbon (DOC) concentration, water hardness, and dissolved organic matter (DOM) source on the acute toxicity of copper were investigated with standardized 48-h Daphnia magna toxicity tests. Toxicity tests were conducted according to a four-factor complete factorial design. Nominal factor levels were as follows: pH 6 and 8; DOC, 2.5 and 10 mg/L; hardness, 10, 20, and 40 mg/L as CaCO3; and two DOM sources (collected from the Black River and Edisto River, SC, USA). The experimental design resulted in 24 different factor level combinations. Results indicated that all factors had significant effects on copper toxicity. Furthermore, a strong interactive effect of DOC concentration and pH was detected. Because the biotic ligand model (BLM) has become a widely used tool for predicting toxicity and interpreting toxicity test results, its performance with these data was evaluated. Seventy percent of BLM predictions were within twofold of the observed median lethal concentrations. However, BLM parameters could be adjusted to improve model performance with this data set. This analysis suggested that in soft waters, the CuOH+ complex binds more strongly with the biotic ligand and that the competitive effect of hardness cations should be increased. The results of the present study may have implications for application of the BLM to some types of surface waters. Furthermore, a comprehensive analysis of BLM performance with all available data should be performed, and necessary updates to model parameters should be made to produce the most robust and widely applicable model.

Metal Mixture Modeling Evaluation project: 2. Comparison of four modeling approaches

As part of the Metal Mixture Modeling Evaluation (MMME) project, models were developed by the National Institute of Advanced Industrial Science and Technology (Japan), the US Geological Survey (USA), HDR|HydroQual (USA), and the Centre for Ecology and Hydrology (United Kingdom) to address the effects of metal mixtures on biological responses of aquatic organisms. A comparison of the 4 models, as they were presented at the MMME workshop in Brussels, Belgium (May 2012), is provided in the present study. Overall, the models were found to be similar in structure (free ion activities computed by the Windermere humic aqueous model [WHAM]; specific or nonspecific binding of metals/cations in or on the organism; specification of metal potency factors or toxicity response functions to relate metal accumulation to biological response). Major differences in modeling approaches are attributed to various modeling assumptions (e.g., single vs multiple types of binding sites on the organism) and specific calibration strategies that affected the selection of model parameters. The models provided a reasonable description of additive (or nearly additive) toxicity for a number of individual toxicity test results. Less-than-additive toxicity was more difficult to describe with the available models. Because of limitations in the available datasets and the strong interrelationships among the model parameters (binding constants, potency factors, toxicity response parameters), further evaluation of specific model assumptions and calibration strategies is needed.

Critical tissue copper residues for marine bivalve (Mytilus galloprovincialis) and echinoderm (Strongylocentrotus purpuratus) embryonic development: Conceptual, regulatory and environmental implications

Critical tissue copper (Cu) residues associated with adverse effects on embryo-larval development were determined for the Mediterranean mussel (Mytilus galloprovincialis) and purple sea urchin (Strongylocentrotus purpuratus) following laboratory exposure to Cu-spiked seawater collected from San Diego Bay, California, USA. Whole body no-observed-effect-residues (NOER) were similar, with means of 21 and 23 microg g(-1) dw, for M. galloprovincialis and S. purpuratus, respectively. Mean whole body median effect residues (ER50) were 49 and 142 microg g(-1) dw for M. galloprovincialis and S. purpuratus, respectively. The difference in ER50s between species was reduced to a factor of <2 when expressed as soft tissue residues. Coefficients of variation among whole body-ER50s were 3-fold lower than median waterborne effect concentrations (EC50) for both species exposed to samples varying in water quality characteristics. This suggests that tissue concentrations were a better predictor of toxicity than water concentrations. The CBRs described herein do not differentiate between the internal Cu concentrations that are metabolically available and those that are accumulated and then detoxified. They do appear, however, to be well enough related to the level of accumulation at the site of action of toxicity that they serve as useful surrogates for the copper concentration that affects embryonic development of the species tested. Results presented have potentially important implications for a variety of monitoring and assessment strategies. These include regulatory approaches for deriving saltwater ambient water quality criteria for Cu, contributions towards the development of a saltwater biotic ligand model, the conceptual approach of using CBRs, and ecological risk assessment.

Development and application of a multimetal multibiotic ligand model for assessing aquatic toxicity of metal mixtures

A multimetal, multiple binding site version of the biotic ligand model (mBLM) has been developed for predicting and explaining the bioavailability and toxicity of mixtures of metals to aquatic organisms. The mBLM was constructed by combining information from single-metal BLMs to preserve compatibility between the single-metal and multiple-metal approaches. The toxicities from individual metals were predicted by assuming additivity of the individual responses. Mixture toxicity was predicted based on both dissolved metal and mBLM-normalized bioavailable metal. Comparison of the 2 prediction methods indicates that metal mixtures frequently appear to have greater toxicity than an additive estimation of individual effects on a dissolved metal basis. However, on an mBLM-normalized basis, mixtures of metals appear to be additive or less than additive. This difference results from interactions between metals and ligands in solutions including natural organic matter, processes that are accounted for in the mBLM. As part of the mBLM approach, a technique for considering variability was developed to calculate confidence bounds (called response envelopes) around the central concentration-response relationship. Predictions using the mBLM and response envelope were compared with observed toxicity for a number of invertebrate and fish species. The results show that the mBLM is a useful tool for considering bioavailability when assessing the toxicity of metal mixtures.

Effects of water chemistry on the chronic toxicity of lead to the cladoceran, Ceriodaphnia dubia

As the first step toward parameterization of a chronic lead (Pb) biotic ligand model (BLM) for Ceriodaphnia dubia, 7-d toxicity tests were performed in waters modified to evaluate the influences of hardness, DOM (as Suwannee River NOM and Aldrich humic acid (HA)), pH (buffered with 4 mM MOPS) and alkalinity on the chronic toxicity of Pb. Calculated EC(20)s for the control base water test and each of the most extreme modified test waters were as follows in μg L(-1) Pb (95% confidence interval): base water control=45 (14-53), 5 mM CaSO(4)=22 (12-30), 32 mg L(-1) DOM=523 (388-573), 2.5 mM NaHCO(3)=73 (21-120) and pH 6.4 buffered with MOPS=3.9 μg L(-1) Pb (1-5). Results indicate that hardness does not protect against chronic toxicity of Pb to C. dubia, whereas HA does protect at the highest concentration tested (597 μM). Additionally, our findings suggest that low pH increases the chronic toxicity of Pb whereas increased alkalinity is protective. The findings reported herein support the need for a chronic Pb BLM as an alternative approach to hardness-based regulations.

Influences of water chemistry on the acute toxicity of lead to Pimephales promelas and Ceriodaphnia dubia

The acute toxicity of lead (Pb) was examined for fathead minnows (Pimephales promelas; 96-h) and daphnids (Ceriodaphnia dubia; 48-h) in waters modified for hardness (as CaSO₄), dissolved organic carbon (DOC; as Aldrich humic acid) and alkalinity (as NaHCO₃) for parameterization of an acute freshwater biotic ligand model (BLM). Additionally, acute (96-h) and chronic (30-d) bioassays were performed for P. promelas to more clearly define the influence of pH (5.5-8.3) on Pb toxicity as modified by addition of HCl or NaOH using an automated titration system. Results indicate that Ca(2+) is protective against acute Pb toxicity to P. promelas but not C. dubia. Strong protection was afforded by DOC and NaHCO(3) against acute Pb toxicity to P. promelas, whereas milder protection was observed for C. dubia with both parameters. Dissolved Pb LC50s from the P. promelas pH bioassays revealed a complex effect of pH on Pb toxicity, likely explained in part by Pb speciation and the competitive interaction of H(+) with ionic Pb(2+). Chronic pH bioassays also demonstrated that 30-d growth is not impaired in fathead minnows at relevant Pb concentrations. The findings reported herein suggest that development of separate BLMs for P. promelas and C. dubia should be considered.

Toxicity versus accumulation for barley plants exposed to copper in the presence of metal buffers: Progress towards development of a terrestrial biotic ligand model†

Development of a terrestrial biotic ligand model (TBLM) for higher plants requires a root-Cu accumulation value that corresponds to the 50% inhibitory concentration (IC50). However, it is not yet known which of the two previously reported Cu-binding ligands on the root is associated with Cu toxicity. The aim of the present study was therefore to investigate the relationship between Cu binding to each ligand group and toxicity, so that the key toxicological site could be identified. To obtain accumulation and toxicity data that were not biased by limited Cu supply to the root, 2-d-old barley seedlings were exposed for 48 h to a range of free Cu ion activities (i.e., {Cu(2+)}) in simple exposure media buffered by nitrilotriacetic acid (NTA) or ethylenediaminetetraacetic acid (EDTA). Comparison of the amount of predicted root-bound Cu (calculated with the aqueous geochemical program PHREEQC) with root elongation data showed that toxicity likely resulted from Cu binding to low-affinity ligands, as the high-affinity ligands were approximately 99% saturated when a reduction in root elongation was first observed. For plants exposed to both NTA- and the EDTA-buffered {Cu(2+)} solutions, the root-Cu accumulation value corresponding to the IC50 was approximately 80 microg/g root dry weight, which is similar to the value obtained from previous work with wheat. The linear relationship between the amount of Cu bound to the low-affinity ligands and the percent root growth inhibition suggests that this relationship will be a robust predictor of Cu toxicity when incorporated into the TBLM, and applied to varied exposure scenarios. For the simple solutions used here, the TBLM-predicted and measured IC50 values were statistically indistinguishable.

Development and application of a biotic ligand model for predicting the chronic toxicity of dissolved and precipitated aluminum to aquatic organisms

Aluminum (Al) toxicity to aquatic organisms is strongly affected by water chemistry. Toxicity-modifying factors such as pH, dissolved organic carbon (DOC), hardness, and temperature have a large impact on the bioavailability and toxicity of Al to aquatic organisms. The importance of water chemistry on the bioavailability and toxicity of Al suggests that interactions between Al and chemical constituents in exposures to aquatic organisms can affect the form and reactivity of Al, thereby altering the extent to which it interacts with biological membranes. These types of interactions have previously been observed in the toxicity data for other metals, which have been well described by the biotic ligand model (BLM) framework. In BLM applications to other metals (including cadmium, cobalt, copper, lead, nickel, silver, and zinc), these interactions have focused on dissolved metal. A review of Al toxicity data shows that concentrations of Al that cause toxicity are frequently in excess of solubility limitations. Aluminum solubility is strongly pH dependent, with a solubility minimum near pH 6 and increasing at both lower and higher pH values. For the Al BLM, the mechanistic framework has been extended to consider toxicity resulting from a combination of dissolved and precipitated Al to recognize the solubility limitation. The resulting model can effectively predict toxicity to fish, invertebrates, and algae over a wide range of conditions. Environ Toxicol Chem 2018;37:70-79.

Evaluating the effects of pH, hardness, and dissolved organic carbon on the toxicity of aluminum to freshwater aquatic organisms under circumneutral conditions

Although it is well known that increasing water hardness and dissolved organic carbon (DOC) concentrations mitigate the toxicity of aluminum (Al) to freshwater organisms in acidic water (i.e., pH < 6), these effects are less well characterized in natural waters at circumneutral pHs for which most aquatic life regulatory protection criteria apply (i.e., pH 6-8). The evaluation of Al toxicity under varying pH conditions may also be confounded by the presence of Al hydroxides and freshly precipitated Al in newly prepared test solutions. Aging and filtration of test solutions were found to greatly reduce toxicity, suggesting that toxicity from transient forms of Al could be minimized and that precipitated Al hydroxides contribute significantly to Al toxicity under circumneutral conditions, rather than dissolved or monomeric forms. Increasing pH, hardness, and DOC were found to have a protective effect against Al toxicity for fish (Pimephales promelas) and invertebrates (Ceriodaphnia dubia, Daphnia magna). For algae (Pseudokirchneriella subcapitata), the protective effects of increased hardness were only apparent at pH 6, less so at pH 7, and at pH 8, increased hardness appeared to increase the sensitivity of algae to Al. The results support the need for water quality-based aquatic life protection criteria for Al, rather than fixed value criteria, as being a more accurate predictor of Al toxicity in natural waters. Environ Toxicol Chem 2018;37:49-60.

Chronic toxicity of aluminum, at a pH of 6, to freshwater organisms: Empirical data for the development of international regulatory standards/criteria

The chemistry, bioavailability, and toxicity of aluminum (Al) in the aquatic environment are complex and affected by a wide range of water quality characteristics (including pH, hardness, and dissolved organic carbon). Data gaps in Al ecotoxicology exist for pH ranges representative of natural surface waters (pH 6-8). To address these gaps, a series of chronic toxicity tests were performed at pH 6 with 8 freshwater species, including 2 fish (Pimephales promelas and Danio rerio), an oligochaete (Aeolosoma sp.), a rotifer (Brachionus calyciflorus), a snail (Lymnaea stagnalis), an amphipod (Hyalella azteca), a midge (Chironomus riparius), and an aquatic plant (Lemna minor). The 10% effect concentrations (EC10s) ranged from 98 μg total Al/L for D. rerio to 2175 μg total Al/L for L. minor. From these data and additional published data, species-sensitivity distributions (SSDs) were developed to derive concentrations protective of 95% of tested species (i.e., 50% lower confidence limit of a 5th percentile hazard concentration [HC5-50]). A generic HC5-50 (not adjusted for bioavailability) of 74.4 μg total Al/L was estimated using the SSD. An Al-specific biotic ligand model (BLM) was used to develop SSDs normalized for bioavailability based on site-specific water quality characteristics. Normalized HC5-50s ranged from 93.7 to 534 μg total Al/L for waters representing a range of European ecoregions, whereas a chronic HC5 calculated using US Environmental Protection Agency aquatic life criteria methods (i.e., a continuous criterion concentration [CCC]) was 125 μg total Al/L when normalized to Lake Superior water in the United States. The HC5-50 and CCC values for site-specific waters other than those in the present study can be obtained using the Al BLM. Environ Toxicol Chem 2018;37:36-48.