Willie J.G.M. Peijnenburg

Monitoring approaches to assess bioaccessibility and bioavailability of metals: Matrix issues

Bioavailability and bioaccessibility are complex issues that determine whether or not adverse effects are to be expected when organisms or plants are exposed to contaminants. Clearly, the determinants of bioavailability and bioaccessibility must be understood if one is to monitor or, ultimately, predict the effects of metals. On the basis of a dynamic conceptual model, this article offers an analysis of the physicochemical and biological determinants underlying bioavailability and bioaccessibility. This analysis is used as the basis for a general monitoring strategy for assessing potentially and actually available and accessible metal fractions in the environmental matrices of water, soil, and sediment. We conclude that, lack of a universal expression of bioavailable and bioaccessible metal fractions precludes the presentation of a detailed monitoring strategy that is broadly applicable. Instead, we recommend that a critical assessment of the endpoints of determination become the basis for a need-specific monitoring strategy.

Bioaccumulation of heavy metals in terrestrial invertebrates

In this literature study, accumulation data of metals in terrestrial invertebrates were collected and compared (Arthropoda and Lumbricidae). Based on total soil concentrations and body concentrations, regression equations were calculated for each metal (Cd, Cu, Pb and Zn) and each taxonomic group. We also tried to find out whether or not accumulation levels of metals in Lumbricidae are representative for all of the studied terrestrial invertebrates. Taxonomic groups could be ordered according to the extent of metal accumulation. Significant differences in accumulation levels of a factor 2-12 were found between taxonomic groups. Overall, metal concentrations were high in Isopoda and low in Coleoptera. The concentrations in Lumbricidae were in between. It should be kept in mind that the data for Lumbricidae were mainly derived from laboratory experiments, while the data for other groups were derived from field studies. The internal Pb, Cd and Cu concentration increased with the soil concentration for most taxonomic groups in the order Pb > Cd > Cu. Body concentrations of Zn were quite constant over a range of soil concentrations. The differences in accumulation level between taxonomic groups show the relevance of including detailed information on feeding behaviour in risk assessment for invertebrate-eating animals.

Correlation of the partitioning of dissolved organic matter fractions with the desorption of Cd, Cu, Ni, Pb and Zn from 18 Dutch soils

Eighteen Dutch soils were extracted in aqueous solutions at varying pH. Extracts were analyzed for Cd, Cu, Ni, Pb and Zn by ICP-AES. Extract dissolved organic carbon (DOC) was also concentrated onto a macroreticular resin and fractionation into three operationally defined fractions: hydrophilic acids (Hyd), humic acids (HA) and fulvic acids (FA). In this manner, change in absolute solution concentration and relative percentage for each fraction could be calculated as a function of extraction equilibrium pH. The soils were also analyzed for solid phase total organic carbon and total recoverable metals (EPA Method 3051). Partitioning coefficients were calculated for the metals and organic carbon (OC) based on solid phase concentrations (less the metal or OC removed by the extraction) divided by solution concentrations. Cu and Pb concentrations in solution as a function of extract equilibrium pH are greatest at low and high pH resulting in parabolic desorption/dissolution curves. While processes such as proton competition and proton promoted dissolution can account for high solution metal concentrations at low pH, these processes cannot account for higher Cu and Pb concentrations at high pH. DOC increases with increasing pH, concurrently with the increase in Cu and Pb solution concentrations. While the absolute concentrations of FA and HA generally increase with increasing pH, the relative proportional increase is greatest for HA . Variation in HA concentrations spans three orders of magnitude while FA concentrations vary an order of magnitude over the pH range examined. Correlation analysis strongly suggests that HA plays a major role in increasing the concentration of solution Cu and Pb with increasing pH in the 18 soils studied. The percentage of the OC that was due to FA was nearly constant over a wide pH range although the FA concentration increased with increasing pH and its concentration was greater than that of the HA fraction at lower pH values (pH = 3-5). Thus, in more acidic environments, FA may play a larger role than HA in governing organo-metallic interactions. For Cd, Ni, and Zn, the desorption/dissolution pattern shows high metal solution concentrations at low pH with slight increases in solution concentrations at extremely high pH values (pH>10). The results presented here suggest that the effects of dissolved organic carbon on the mobilization of Cd, Ni, and Zn may only occur in systems governed by very high pH. At high pH, it is difficult to distinguish in this study whether the slightly increased solution-phase concentrations of these cations is due to DOC or hydrolysis reactions. These high pH environments would rarely occur in natural settings.

Natural colloids are the dominant factor in the sedimentation of nanoparticles

Estimating the environmental exposure to manufactured nanomaterials is part of risk assessment. Because nanoparticles aggregate with each other (homoaggregation) and with other particles (heteroaggregation), the main route of the removal of most nanoparticles from water is aggregation, followed by sedimentation. The authors used water samples from two rivers in Europe, the Rhine and the Meuse. To distinguish between small (mainly natural organic matter [NOM]) particles and the remainder of the natural colloids present, both filtered and unfiltered river water was used to prepare the particle suspensions. The results show that the removal of nanoparticles from natural river water follows first-order kinetics toward a residual concentration. This was measured in river water with less than 1 mg L(-1) CeO(2) nanoparticles. The authors inferred that the heteroaggregation with or deposition onto the solid fraction of natural colloids was the main mechanism causing sedimentation in relation to homoaggregation. In contrast, the NOM fraction in filtered river water stabilized the residual nanoparticles against further sedimentation for up to 12 d. In 10 mg L(-1) and 100 mg L(-1) CeO(2) nanoparticle suspensions, homoaggregation is likely the main mechanism leading to sedimentation. The proposed model could form the basis for improved exposure assessment for nanomaterials.

Response predictions for organisms water-exposed to metal mixtures: a meta-analysis.

To develop a multimetal toxicity model requires insight into the relationships between the composition of metal mixtures and their toxicological effects on organisms. As a first step in developing such a model, quantitative data from binary and higher mixture studies of Cu, Cd, and Zn were compiled and used to assess trends in toxicological effects on various organisms. The findings of this meta-analysis show relatively little occurrence of additive effects compared with antagonistic and synergistic effects. This observation held true irrespective of test species, environmental compartment, or metal concentrations in the mixture. However, the type of effect was significantly correlated with the metal combination tested and the selected toxicological endpoint. It was also found that different methods were used for assessing deviation from additivity in the various individual studies. For robust comparison, standardization on this point is required. Toxicological responses of organisms to metal mixtures were shown to be hard to predict and were often slightly less than or slightly more than additive. The interactions observed could not be explained by metal-metal interactions alone. We therefore conclude that with current scientific knowledge it is not yet possible to predict responses to metal mixtures in individual cases; at best this is possible only in terms of general patterns. Nevertheless, in the context of environmental risk policy, the assumption of additivity produces a conservative prediction of toxicity, because toxicity of a metal mixture will be either predicted correctly or overpredicted by default in approximately 75% of all cases. The use of models based on noninteraction is satisfactory from this regulatory perspective.

A Review of the Properties and Processes Determining the Fate of Engineered Nanomaterials in the Aquatic Environment

Proper understanding of the basic processes and specific properties of engineered nanomaterials (NMs) that modify the fate and effects of NMs is crucial for NM-tailored risk assessment. This in turn requires developers of NMs and for regulators to consider the most important parameters governing the properties, behavior and toxicity of NMs. As fate and effect studies are commonly performed in laboratory settings, mimicking to a varying extent realistic exposure conditions, it is important to be able to extrapolate results of fate and effect studies in synthetic media to realistic environmental conditions. This requires detailed understanding of the processes controlling the fate and behavior of NMs in terrestrial and aquatic media, as dependent on the composition of the medium. It is the aim of this contribution to provide background reading to the NM and media specific properties and processes that affect the fate and behavior of NMs in aquatic environments, focusing on the specific properties of NMs that modulate the interactions in the aquatic environment. A general introduction on the dominant fate determining processes of NMs is supplemented by case studies on specific classes of NMs: metal NMs, stable oxides, iron oxides, and carbon nanotubes. Based on the synthesis of the current knowledge base toward essential data and information needs, the review provides a description of the particle specific properties and the water characteristics that need monitoring in order to allow for future quantification and extrapolation of fate and behavior properties of NMs in freshwater compartments of varying composition.

A European perspective on alternatives to animal testing for environmental hazard identification and risk assessment

Tests with vertebrates are an integral part of environmental hazard identification and risk assessment of chemicals, plant protection products, pharmaceuticals, biocides, feed additives and effluents. These tests raise ethical and economic concerns and are considered as inappropriate for assessing all of the substances and effluents that require regulatory testing. Hence, there is a strong demand for replacement, reduction and refinement strategies and methods. However, until now alternative approaches have only rarely been used in regulatory settings. This review provides an overview on current regulations of chemicals and the requirements for animal tests in environmental hazard and risk assessment. It aims to highlight the potential areas for alternative approaches in environmental hazard identification and risk assessment. Perspectives and limitations of alternative approaches to animal tests using vertebrates in environmental toxicology, i.e. mainly fish and amphibians, are discussed. Free access to existing (proprietary) animal test data, availability of validated alternative methods and a practical implementation of conceptual approaches such as the Adverse Outcome Pathways and Integrated Testing Strategies were identified as major requirements towards the successful development and implementation of alternative approaches. Although this article focusses on European regulations, its considerations and conclusions are of global relevance.

Toxicity of zinc oxide nanoparticles in the earthworm, Eisenia fetida and subcellular fractionation of Zn

The extensive use of nanoparticles (NPs) in a variety of applications has raised great concerns about their environmental fate and biological effects. This study examined the impact of dissolved organic matter (DOM) and salts on ZnO NP dispersion/solubility and toxicity to the earthworm Eisenia fetida. To be able to better evaluate the toxicity of NPs, exposure in agar and on filter paper was proposed for enabling a comparison of the importance of different uptake routes. A dose-related increase in mortality was observed in earthworms exposed in agar with almost 100% mortality after 96 h exposure to the highest concentration (1000 mg ZnO/kg agar). Scanning electron microscopy (SEM) showed that the addition of salts enhanced the aggregation of ZnO NPs in agar and consequently affected the dissolution behavior and biological availability of the particles. On filter paper, mortality was the highest at the lowest exposure concentration (50 mg ZnO/L) and seemed to decrease with increasing exposure levels. TEM images of ZnO showed that the solubility and morphology of NPs were changed dramatically upon the addition of humic acids (HA). The subcellular distribution pattern of Zn in earthworms after 96 h exposure in agar and on filter paper showed that the Zn taken up via dietary ZnO particles (from agar) was mainly found in organelles and the cytosol while the Zn accumulated as soluble Zn from filter paper was mainly distributed in cell membranes and tissues. Antioxidant enzymatic activities (SOD, CAT, and GSH-px) were investigated in the worms surviving the toxicity tests. A slight increase of SOD activities was observed at the lowest exposure dose of ZnO (50mg/kg), followed by a decrease at 100mg/kg in the agar cubes. Activities of both CAT and GSH-Px enzymes were not significantly influenced in the worms exposed to agar, although a slight decrease at 500 and 1000 mg ZnO/kg agar was observed. A similar change trend of SOD activities was observed for the earthworms on filter paper, but a significant decrease began at a higher ZnO NP concentration of 500 mg ZnO/L. The use of soil extracts instead of deionized water (DW) to simulate a realistic exposure system significantly reduced the toxicity of the ZnO NPs on filter paper, which increases the predictive power of filter paper toxicity tests for the environmental risk assessment of NPs.

Biological significance of metals partitioned to subcellular fractions within earthworms (Aporrectodea caliginosa)

Metal ions in excess of metabolic requirements are potentially toxic and must be removed from the vicinity of important biological molecules to protect organisms from adverse effects. Correspondingly, metals are sequestrated in various forms, defining the accumulation pattern and the magnitude of steady-state levels reached. To investigate the subcellular fractions over which Ca, Mg, Fe, Cu, Zn, Cd, Pb, Ni, and As are distributed, earthworms (Aporrectodea caliginosa) collected from the field were analyzed by isolating metal-rich granules and tissue fragments from intracellular microsomal and cytosolic fractions (i.e., heat-stable proteins and heat-denatured proteins). The fractions showed metal-specific binding capacity. Cadmium was mainly retrieved from the protein fractions. Copper was equally distributed over the protein fraction and the fraction comprising tissue fragments, cell membranes, and intact cells. Zinc, Ca, Mg, and As were mainly found in this fraction as well. Lead, Fe, and Ni were mainly isolated from the granular fraction. To study accumulation kinetics in the different fractions, three experiments were conducted in which earthworms were exposed to metal-spiked soil and a soil contaminated by anthropogenic inputs and, indigenous earthworms were exposed to field soils. Although kinetics showed variation, linear uptake and steady-state types of accumulation patterns could be understood according to subcellular compartmentalization. For risk assessment purposes, subcellular distribution of metals might allow for a more precise estimate of effects than total body burden. Identification of subcellular partitioning appears useful in determining the biological significance of steady-state levels reached in animals.

Modeling lifetime and degradability of organic compounds in air, soil, and water systems (IUPAC Technical Report)

Degradability of organic compounds in air, soil, and water is the most important factor for evaluating their environment fate as well as possible adverse effects to humans and the environment. The primary degradation process in the troposphere is the reaction with the hydroxyl radical. For water and soil compartments, the primary degradation process is biodegradation. The objectives of this report are: (i) to review published models and their evaluation studies, (ii) to perform an in-house evaluation of general models for estimating tropospheric degradation and biodegradation of organic compounds, and (iii) to recommend reliable procedures for estimating degradability of organic compounds in the environment. The extensive evaluation procedure has shown that the most accurate method for estimating tropospheric degradation is Atkinsons group contribution method. Although this method has some limitations, it seems to be a method of choice. A viable alternative to Atkinsons method is a direct calculation, performed today almost routinely, of the reaction rate constants with hydroxyl radicals. Recently, a methodology based on reliable semiempirical potential energy surfaces was developed that enables the calculation of reaction rate constants within a factor of 2 of their measured values. A partial least squares (PLS) model and a set of seven biodegradation rules have been found to be the most reliable in estimating complete biodegradation of organic compounds. However, it is recommended to use all four evaluated methods to estimate biodegradation in the environment. If their results agree, such estimates are very reliable.