Establishing Causality between Pollution and Effects at Different Levels of Biological Organization: The VALIMAR Project

The study summarizes the objectives of the VALIMAR project and gives selected examples of biomarker responses that allow causal relationships to be established between exposure and biological effects at different levels of biological organization. In this project, active and passive biomonitoring experiments with brown trout (Salmo trutta f. fario) and stone loach (Barbatula barbatula) were performed in two small streams in southern Germany between 1995 and 1999 in parallel with investigations on contaminant mixtures in the laboratory in order to evaluate the suitability of biomarkers representing different levels of biological organization for the assessment of pollution in small streams. In addition to biomarker studies, the morphology of the test streams was characterized and limnological and chemical parameters were monitored. Early life stage tests and ecological studies of brown trout and stone loach population demography, of the fish assemblages, and the macro- and meiozoobenthos communities in the two test streams were included in the project. Several causality criteria were addressed by means of combined (1) laboratory and field studies, (2) chemical, biological, and statistical investigations, and (3) in vivo and in vitro studies that allowed establishment of cause-effect relationships at different biological levels. The comparison of results obtained at these levels allowed identification of mechanisms responsible for the respective effects (coherence of association, biological plausibility). Finally, individual responses (biomarkers, bioindicators) could be extrapolated to higher biological levels (population, community) thus addressing the criteria of ‘time order’ and ‘coherence of association’.     

Establishing Causality between Environmental Stressors and Effects on Aquatic Ecosystems

Establishing causal relationships between environmental stressors and observed effects in natural systems is difficult due to the many intrinsic environmental factors that can hinder this process and because there are no widely accepted and proven approaches for determining such relationships. Several types of approaches or combinations of approaches, each with their own sets of advantages and limitations, have been applied in a variety of ecological systems to investigate possible causal relationships between stressors and effects. These include controlled laboratory studies (including acute and chronic bioassays), experimental field manipulations, field studies based on synoptic field surveys, mathematical simulation modeling, statistical associations, various combinations of laboratory, experimental, and field studies, and the ecoepidemiological (weight or evidence) approach. The use of ecoepidemiological (“forensic toxicology”) principles is becoming increasingly attractive as a method to help establish causality because it does not involve the same limitations of other approaches and it can also be used to integrate disparate information within a logical framework so that scientifically and defensible regulatory decisions can be made. The objective of this Commentary series of papers on the issue on causality is to demonstrate the application of the ecoepidemiology approach, using a variety of case history studies, for establishing causal relationships between specific stressors and biological effects. For each case history provided in the following series of papers, the authors describe their study situation, summarize the results supporting a causal relationship, and then compare their study results against seven standard causal criteria.     

Evidence for a Causal Link between Exposure to an Insecticide Formulation and Declines in Catch of Atlantic Salmon

We report a case study showing relationships between historical insecticide applications (1973 to 1990) of an aminocarb formulation (Matacil® 1.8D ) containing 4-nonylphenol (4-NP) and catch data for Atlantic salmon (Salmo salar) populations. To address the hypothesis that Matacil® 1.8D caused reductions in Atlantic salmon catch, seven epidemiological criteria are used to evaluate causality between the stressor and effect. These criteria include: strength of association; consistency of association; specificity of association; time order; biological gradient; experimental support; and biological plausibility. We conclude that the general weight-of-evidence supporting the seven epidemiological criteria is consistent with a causal relationship between declines in salmon catch and Matacil® 1.8D exposure during parr-smolt transformation (PST). If the effects exerted by 4-NP are due to its activity as a weak estrogen, then hormonal activity stemming from other sources (e.g., domestic sewage, agricultural, industrial) might influence present day salmon populations.     

Establishing the Causal Relationship between Polycyclic Aromatic Hydrocarbon (PAH) Exposure and Hepatic Neoplasms and Neoplasia-Related Liver Lesions in English Sole (Pleuronectes vetulus)

For almost 25 years our laboratory has studied the impact of PAHs and related industrial contaminants on benthic fish, following an interdisciplinary approach involving chemical exposure assessment linked to synoptic detection of various effects at several levels of biological organization. These data demonstrate a cause-and-effect relationship between neoplastic and neoplasia-related liver lesions in English sole, and exposure to PAHs, and to a lesser degree, chlorinated hydrocarbons such as PCBs. In statistical analyses of data from multiple field studies conducted since 1978, exposure to PAHs measured in various compartments has consistently been identified as a highly significant, major risk factor for neoplasms and related lesions in this species, with PCB exposure shown to be a significant, but less consistent and less strong risk factor for these lesions. A cause-and-effect relationship between PAHs and toxicopathic liver lesions in this species is further supported by the experimental induction of toxicopathic lesions identical to those observed in field-collected fish, in sole exposed in the laboratory to model carcinogenic PAHs such as BaP or to PAH-rich extracts of sediments from Eagle Harbor, a severely PAH-contaminated site in Puget Sound. More recent field studies have identified significant associations between hepatic cytochrome P4501A (CYP1A) induction and xenobiotic-DNA adduct formation, and hepatic lesion prevalences in wild subadult English sole. Field studies in Eagle Harbor subsequent to capping of the most PAH-contaminated region of this harbor with clean dredge spoils have shown a decline in exposure to PAHs as assessed by biliary fluorescent aromatic compounds (FACs) and hepatic xenobiotic-DNA adducts. This decline in PAH exposure has been accompanied by a dramatic decline in risk of occurrence of toxicopathic hepatic lesions in English sole from Eagle Harbor. Further, laboratory studies have induced lesions in English sole by injections of extracts from PAHcontaminated sediments. Overall, these findings relating to exposure to PAHs and chlorinated hydrocarbons and the occurrence of hepatic neoplasms and neoplasiarelated lesions in English sole fulfill the classic criteria for causality in epizootiological or ecological risk assessment studies, including: (1) strength of association, (2) consistency of association, (3) specificity of association, (4) toxicological and biological plausibility, (5) temporal sequence/timing (i.e., exposure precedes disease, effect decreases when the cause is decreased or removed), (6) dose-response or biological gradient, and (7) supportive experimental evidence.     

Evaluating Effects of Contaminants on Fish Health at Multiple Levels of Biological Organization: Extrapolating from Lower to Higher Levels

Effects of environmental stressors such as contaminants on the health of aquatic ecosystems usually involve a series of biological responses ranging from the biomolecular/biochemical to the population and community levels. To establish relationships and to determine the feasibility of extrapolating between higher and lower levels of biological organization, spatial patterns in fish responses to contaminant loading were investigated in a stream receiving point-source discharges of various contaminants near its headwaters. Relationships among fish responses at four major levels of biological organization (biochemical/physiological, individual, population, and community levels) were evaluated relative to patterns in contaminant loading along the spatial gradient of the stream. Both individual and integrated response analysis demonstrated that bioindicators at several levels of biological organization displayed similar downstream patterns in their response to contaminant loading within the stream. Some of the bioindicator responses at lower levels of organization appear to be useful for the ecological risk assessment process because of their sensitivity and apparent relationships to higher levels. By identifying and establishing relationships between levels of biological organization we should be better able to understand the mechanisms of stress responses in ecological systems that could ultimately result in improved predictive capability of ecological risk assessment and also allow for more informed decisions regarding remedial actions.     

Use of Biochemical Endpoints to Determine Relationships between Contaminants and Impaired Fish Health in a Freshwater Stream

Bayou Bartholomew is a low gradient stream predominantly influenced by agricultural inputs and stormwater discharge from the urban areas of Pine Bluff, Arkansas. Preliminary studies indicated induction of hepatic cytochrome P450 1A protein (CYP1A) and CYP1A-catalyzed enzyme activity (ethoxyresorufin O-deethylase-EROD) in several fish species collected in the waterway, which meanders through residential areas of Pine Bluff, and forested regions in the outskirts of the city. Largemouth bass (Micropterus salmoides) showed CYP1A repression and a lack of correlation between CYP1A and EROD activity. In addition, hepatic heme oxygenase activity was elevated in several species and demonstrated an inverse relationship with hepatic CYP1A. Hepatic metallothionein was unchanged in any species collected from these locations. Four years after this evaluation, sediments were collected and analyzed for agents that could be mechanistically consistent with the biochemical responses observed. Utilizing seven criteria for causality, these data indicated that PCBs appear to be at least one group of agents responsible for the biological effects observed in fish species from the upper Bayou Bartholomew. While this supports causality under the criteria, further studies in this waterway would strengthen the relationship between this class of compound and the effects observed in resident fish species.     

Recovery in Diversity of Fish and Invertebrate Communities Following Remediation of a Polluted Stream: Investigating Causal Relationships

Spatial and temporal responses of biota to anthropogenic disturbance were measured over a 15 year period in a contaminated stream undergoing remediation and recovery. Along the spatial gradient of the stream, levels of contaminants decreased downstream along with improved responses of instream biota at several levels of biological organization. Recovery of the biota in this stream over the 15 year study period is demonstrated by the temporal relationships between levels of decreasing contaminants and the concomitant responses of the periphyton, macroinvertebrate, and fish communities and changes in the various bioindicators of individual fish health. Decreases in contaminants over a temporal scale were followed closely by an improvement in physiological and organismal-level indicators, increases in the diversity of macroinvertebrate and fish communities, and rapid increases in the chlorophyll a biomass and photosynthesis rate of the periphyton community. These results emphasize that field studies designed to assess and evaluate the effectiveness of restoration activities on stream recovery should incorporate a variety of response endpoints ranging from sensitive and short-term responses to long-term but ecological relevant indicators of change. The close spatial and temporal relationships observed between changes in physicochemical factors and positive responses in various components of the stream biota over the 15-year study period suggest a strong cause and effect relationship between remediation activities and stream recovery. Understanding causal relationships and the mechanistic processes between environmental stressors, stress responses of biota, and the recovery process is important in the effective management and restoration of aquatic ecosystems. An erratum to this article is available at .     

Onchocerca volvulus and epilepsy: A comprehensive review using the Bradford Hill criteria for causation

BackgroundThe possibility that onchocerciasis may cause epilepsy has been suggested for a long time, but thus far, an etiological link has not been universally accepted. The objective of this review is to critically appraise the relationship between Onchocerca volvulus and epilepsy and subsequently apply the Bradford Hill criteria to further evaluate the likelihood of a causal association.MethodsPubMed and gray literature published until September 15, 2020, were searched and findings from original research were synthesized. Adherence to the 9 Bradford Hill criteria in the context of onchocerciasis and epilepsy was determined to assess whether the criteria are met to strengthen the evidence base for a causal link between infection with O. volvulus and epilepsy, including the nodding syndrome.ResultsOnchocerciasis as a risk factor for epilepsy meets the following Bradford Hill criteria for causality: strength of the association, consistency, temporality, and biological gradient. There is weaker evidence supporting causality based on the specificity, plausibility, coherence, and analogy criteria. There is little experimental evidence. Considering the Bradford Hill criteria, available data suggest that under certain conditions (high microfilarial load, timing of infection, and perhaps genetic predisposition), onchocerciasis is likely to cause epilepsy including nodding and Nakalanga syndromes.ConclusionApplying the Bradford Hill criteria suggests consistent epidemiological evidence that O. volvulus infection is a trigger of epilepsy. However, the pathophysiological mechanisms responsible for seizure induction still need to be elucidated.     

Determination of Field Effects of Contaminants—Significance of Pollution-Induced Community Tolerance

The concept of pollution-induced community tolerance (PICT) consists of the phenomenon that communities in an ecosystem exhibit increased tolerance as a result of exposure to contaminants. Although a range of ‘classic’ ecological principles explains the processes that increase tolerance of a community, the value of PICT for ecological risk assessment was recognized only recently (Blanck et al. 1988). The following issues are recognized: First, regarding the question on the role of suspect compounds causing ecological effects, the PICT approach covers the issue of causality better than ‘classical’ ecological community response parameters like species densities or species diversity indices. This relates to the fact that the level of PICT is assumed to be relatively constant (compared to density and diversity), whereas the suspect compound causing the observed effect can be deduced with relative clear inference from artificial exposure experiments. Second, PICT directly addresses a level of biological organization (the community), the level of concern for many ecological risk assessment methods. Other methods for risk assessment, like toxicity testing or bioassays, focus on individual or population-level effects, and need extrapolation of the results to the field. Such extrapolation step may pose problems regarding validity of the outcome of risk assessment. The occurrence of PICT is, however, not (yet) a community endpoint that is sufficiently underpinned to trigger risk mitigation activities. This paper especially focuses on the possibility to improve risk assessment approaches by incorporation of PICT assessments, especially focusing on the issue of causality and on the ecological meaning of PICT. Despite the advantages over ‘classical’ parameters, literature analysis suggests that the PICT approach may be strengthened by determining to which degree the PICT approach relates to ecological changes, like shifts in community structure, functioning, and stability. The aim of this paper is to summarize some literature, putting the emphasis on terrestrial studies, to get insights whether PICT is a sensitive and powerful tool to quantify ecological effects in field conditions, to link them to toxicant stress, and thus to determine whether PICT may be taken into consideration in risk assessment.     

Contaminant-Adaptation and Community Tolerance in Ecological Risk Assessment: Introduction

Contaminant tolerance, either at the level of the community or an adaptation within populations, has important implications to the risk assessment field. Such tolerance has alternatively been described as a nuisance variable, complicating the extrapolation of toxicity data to field conditions, or as a ‘good weather indicator’, suggesting environmental resilience to a contaminant. These and other issues are explored in this set of invited papers, in which experienced workers from the field of contaminant tolerance have been invited to comment on the relationship between tolerance and the analysis of environmental risk. In addition, recent decades have seen the use of tolerance as a tool for assessing contaminant stress, particularly when establishing causality between specific contaminant exposure and significant ecological impact. The paradigm suggests that an increased tolerance to a contaminant is powerful causal evidence that this contaminant has exerted significant stress. Review, commentary and original data contributions within this Debate and Commentary section explore both the complicating and advantageous aspects of tolerance in risk assessment. The papers conclude that complications associated with tolerance demand careful consideration during risk assessments, and that while population adaptation does not appear to be a promising tool, community-level resistance might be a powerful instrument in ecological risk assessment.     

Development of an expert system for the integration of biomarker responses in mussels into an animal health index

Biomarkers on sentinel organisms are utilised worldwide in biomonitoring programs. However, the lack of effective interpretational capacity has hampered their uptake for use for assessment of risk in environmental management. The aim of the present study was to develop and test an objective decision-support or expert system capable of integrating biomarker results into a five-level health-status index. The expert system is based on a set of rules derived from available data on responses to natural and contaminant-induced stress of marine mussels. Integration of parameters includes: level of biological organization; biological significance; mutual inter-relationship; and qualitative trends in a stress gradient. The system was tested on a set of biomarker data obtained from the field and subsequently validated with data from previous studies. The results demonstrate that the expert system can effectively quantify the biological effects of different levels of pollution. The system represents a simple tool for risk assessment of the harmful impact of contaminants by providing a clear indication of the degree of stress syndrome induced by pollutants in mussels.     

Development of an expert system for the integration of biomarker responses in mussels into an animal health index

Biomarkers on sentinel organisms are utilised worldwide in biomonitoring programs. However, the lack of effective interpretational capacity has hampered their uptake for use for assessment of risk in environmental management. The aim of the present study was to develop and test an objective decision-support or expert system capable of integrating biomarker results into a five-level health-status index. The expert system is based on a set of rules derived from available data on responses to natural and contaminant-induced stress of marine mussels. Integration of parameters includes: level of biological organization; biological significance; mutual inter-relationship; and qualitative trends in a stress gradient. The system was tested on a set of biomarker data obtained from the field and subsequently validated with data from previous studies. The results demonstrate that the expert system can effectively quantify the biological effects of different levels of pollution. The system represents a simple tool for risk assessment of the harmful impact of contaminants by providing a clear indication of the degree of stress syndrome induced by pollutants in mussels.     

Forensic visualization of foreign matter in human tissue by near-infrared spectral imaging: methodology and data mining strategies.

BACKGROUND: Rapidity of data acquisition, high image fidelity and large field of view are of tremendous value when looking for chemical contaminants or for the proverbial "needle in the haystack" - in this case foreign inclusions in histologic sections of biopsy or autopsy tissues. Near infrared chemical imaging is one of three chemical imaging techniques (NIR, MIR and Raman) based on vibrational spectroscopy, and provides distinct technical advantages for this application. METHODS: We have chosen to utilize and evaluate near infrared (NIR) imaging for studies of foreign materials in tissue because the experimental configuration is relatively simple, data collection is rapid, and large sample areas can be screened with high image fidelity and spatial resolution. RESULTS: We have shown that NIR imaging can readily find and identify silicone gel inclusions in biological tissue samples. Additionally, preliminary results indicate that spectral signatures in the data set are also potentially sensitive to structural changes in the surrounding tissue that may be induced by the foreign body. CONCLUSIONS: NIR chemical imaging is a powerful, non-destructive tool for localization and identifying foreign contaminants in biological tissue. Preliminary results indicate that NIR imaging is also sensitive enough to differentiate tissue types (perhaps based on collagen structural differences), and provide data on the spatial localization of these components.     

Decoding P4-ATPase substrate interactions

Cellular membranes display a diversity of functions that are conferred by the unique composition and organization of their proteins and lipids. One important aspect of lipid organization is the asymmetric distribution of phospholipids (PLs) across the plasma membrane. The unequal distribution of key PLs between the cytofacial and exofacial leaflets of the bilayer creates physical surface tension that can be used to bend the membrane; and like Ca²⁺, a chemical gradient that can be used to transduce biochemical signals. PL flippases in the type IV P-type ATPase (P4-ATPase) family are the principle transporters used to set and repair this PL gradient and the asymmetric organization of these membranes are encoded by the substrate specificity of these enzymes. Thus, understanding the mechanisms of P4-ATPase substrate specificity will help reveal their role in membrane organization and cell biology. Further, decoding the structural determinants of substrate specificity provides investigators the opportunity to mutationally tune this specificity to explore the role of particular PL substrates in P4-ATPase cellular functions. This work reviews the role of P4-ATPases in membrane biology, presents our current understanding of P4-ATPase substrate specificity, and discusses how these fundamental aspects of P4-ATPase enzymology may be used to enhance our knowledge of cellular membrane biology.     

Establishing Causality between Exposure to Metals and Effects on Fish

This study evaluates causal relationships between chronic exposure of fish to metals and effects at different levels of biological organization based on a weight-of-evidence approach. Criteria for evaluation of causality were strength, consistency, and specificity of the association, as well as biological gradient and plausibility. Field sampling was conducted three times between 1998 and 2000, in Furnas Stream, impacted by an abandoned lead mine, and in three other locations, including two reference and one impacted sites. Levels of Pb, Zn, Cd, and Ag in sediments from the Furnas Stream exceeded background levels, and their concentrations were above sediment quality guidelines. Residual levels of metals in fish tissue were high enough to indicate reduced growth, reproduction and/or survival according to toxicological benchmarks. Lead-induced biochemical changes (ALA-D activity depletion) were observed in two species of siluriform catfish. The condition factor of a predatory catfish was reduced, and the percentage of prey generalists was higher in Furnas than at the noncontaminated sites. Reduction in fish community diversity and density was observed. Integration of data provided supporting evidence that observed effects on fish from the Furnas Stream resulted from long-term exposure to metals, however influences from other stressors cannot be ruled out.     

Biological monitoring of water quality in Australia: Workshop summary and future directions

Abstract Biological methods are widely accepted in water quality monitoring programmes worldwide; however, some concern remains over their effectiveness in predicting the effects of contaminants on aquatic ecosystems. While the so-called‘early warning’ approaches, such as bioassays and biomarkers, have been used in Australia to demonstrate mechanisms of toxic action and exposure to contaminants, as elsewhere, little attempt has been made to link observed effects at these lower levels of biological organization to real impacts on aquatic systems. The ecological consequences of exposure to contaminants is undoubtedly best studied at higher levels of biological organization (i. e. at the population or community level). However, monitoring aquatic communities is labour intensive and inadequate for the early detection of impacts. Research is needed to identify links between the bioassessment measures used, so that changes at the lowest biological level (e. g. using biomarkers and bioassays) can be translated into likely‘real’ impacts on the aquatic system, as measured at the population or community level. Monitoring the genetic structure of populations of aquatic organisms, particularly invertebrates, may provide a potential link between subtle effects observed in bioassay tests and subsequent changes in population density and/or community structure. A streamlined approach to monitoring changes at the community level needs to be developed to improve predictive ability and to make this approach more responsive to the early detection and prevention of unacceptable impacts. In addition, research on the use of ecosystem level parameters, such as production/respiration ratios or community metabolism, should be undertaken to determine their suitability for routine biomonitoring of water quality in Australian inland waters.     

Light at Night Co‐distributes with Incident Breast but not Lung Cancer in the Female Population of Israel

Recent studies of shift‐working women have reported that excessive exposure to light at night (LAN) may be a risk factor for breast cancer. However, no studies have yet attempted to examine the co‐distribution of LAN and breast cancer incidence on a population level with the goal to assess the coherence of these earlier findings with population trends. Coherence is one of Hill's “criteria” (actually, viewpoints) for an inference of causality. Nighttime satellite images were used to estimate LAN levels in 147 communities in Israel. Multiple regression analysis was performed to investigate the association between LAN and breast cancer incidence rates and, as a test of the specificity of our method, lung cancer incidence rates in women across localities under the prediction of a link with breast cancer but not lung cancer. After adjusting for several variables available on a population level, such as ethnic makeup, birth rate, population density, and local income level, a strong positive association between LAN intensity and breast cancer rate was revealed (p<0.05), and this association strengthened (p<0.01) when only statistically significant factors were filtered out by stepwise regression analysis. Concurrently, no association was found between LAN intensity and lung cancer rate. These results provide coherence of the previously reported case‐control and cohort studies with the co‐distribution of LAN and breast cancer on a population basis. The analysis yielded an estimated 73% higher breast cancer incidence in the highest LAN exposed communities compared to the lowest LAN exposed communities.     

Establishing Causality between Population Genetic Alterations and Environmental Contamination in Aquatic Organisms

Contaminant-induced alterations in genetic diversity or allele/genotype frequencies can occur via genetic bottlenecks, selection, or increased mutation rate, and may affect population growth, sustainability, and adaptability. Determination of causality of genetic effects requires demonstration of some or all of the following criteria: (1) Strength of association: use of multiple reference and contaminated populations, and demonstration of effects that cannot otherwise be explained by evolutionary theory; (2) Consistency of association: effects corroborated by other studies, in other species, or with multiple genetic markers; (3) Specificity of association: concordance of genetic effects with exposure/effect bioindicators, genotypedependant fitness and biomarkers, and consideration of confounding factors; (4) Temporality of effects: use of phylogenetics and analysis of genetic diversity using different methodologies to differentiate historical vs. recent events; (5) Biological gradients: sampling sites that are known to have differing levels of contamination; (6) Experimental evidence: exposure of small populations to contaminants in laboratories, mesocosms, or in situ cages, or measurement of genotype-dependant biomarkers; (7) Biological plausibility: existence of contaminants at levels great enough to affect fitness, recruitment, or mutation rates, or a demonstrated mechanism for selection. Application of these criteria to population genetic studies is illustrated by case studies involving RAPD analysis of mosquitofish populations.     

Integrating effects of contaminants across levels of biological organization: an overview

Effects of contaminants may occur at all levels oforganization, from molecular to ecosystem-levelresponses. While biochemical and physiologicalalterations in organisms may occur rapidly and areoften stressor-specific, the ecological relevance ofthese suborganismal indicators is uncertain.Alterations in populations and communities havegreater ecological relevance, but a firm mechanisticunderstanding of these responses is often lacking.Developing mechanistic linkages across levels ofbiological organization would greatly improve ourunderstanding of how organisms are affected bycontaminants in nature. The papers in this seriespresent several innovative approaches for integratingeffects of contaminants across levels of biologicalorganization. Authors were asked to describe theecological consequences of responses at lower levelsof organization (biochemical, physiological,individual) and to speculate on the underlyingmechanisms associated with population and communityalterations. The most consistent finding of the fivepapers in this series is that there is no singlespatiotemporal scale or level of biologicalorganization at which ecotoxicological investigationsshould be conducted.     

Bioavailability of Soil-Borne Chemicals: Abiotic Assessment Tools

The abiotic tools that are available, or under development, for evaluating the oral and dermal bioavailability of contaminants from soils are described in this article. These tools generally rely on one of two approaches: (1) characterizing the form of the contaminant and the chemical binding of the contaminant to the soil matrix, and (2) chemical extractions intended to evaluate the fraction of the chemical that would be liberated in biological fluids (gastrointestinal fluid or sweat). For the purpose of human health risk assessment, abiotic methods to estimate the bioavailability of inorganic contaminants in soil are considered generally to be “screening” level tools at this time. Development work for physiologically based extraction tests (PBETs) is ongoing for many inorganic contaminants, and these methods hold great promise for eventual use in making quantitative bioavailability adjustments in risk assessment. The availability of abiotic tools to evaluate the bioavailability of organic contaminants from soils lags behind that for metals, due to the difficulty in conducting in vivo bioavailability studies with organic compounds and their complex interactions with soil. However, considerable research is being conducted in this field, and new assessment tools are being validated for use in human health risk assessment.