Risk Assessment Of Mixtures Research Articles

Ecological risk assessment of mixtures of radiological and chemical stressors: Methodology to implement an msPAF approach

A main challenge in ecological risk assessment is to account for the impact of multiple stressors. Nuclear facilities can release both radiological and chemical stressors in the environment. This study is the first to apply species sensitivity distribution (SSD) combined with mixture models (concentration addition (CA) and independent action (IA)) to derive an integrated proxy of the ecological impact of combined radiological and chemical stressors: msPAF (multisubstance potentially affected fraction of species). The approach was tested on the routine liquid effluents from nuclear power plants that contain both radioactive and stable chemicals. The SSD of ionising radiation was significantly flatter than the SSD of 8 stable chemicals (namely Cr, Cu, Ni, Pb, Zn, B, chlorides and sulphates). This difference in shape had strong implications for the selection of the appropriate mixture model: contrarily to the general expectations the IA model gave more conservative (higher msPAF) results than the CA model. The msPAF approach was further used to rank the relative potential impact of radiological versus chemical stressors.

Single and mixture toxicity of abamectin and difenoconazole to adult zebrafish (Danio rerio)

Concerns have been raised in recent years on the potential risks related with pesticide mixtures that are likely to be present in agricultural edge-of-field waterbodies. Despite the high use of pesticides in tropical countries like Brazil, studies evaluating pesticide mixtures are especially scarce in the tropics. The insecticide abamectin and the fungicide difenoconazole are the main pesticides intensively used in Brazilian strawberry crop and are hence likely to occur simultaneously. The aim of the present study was therefore to evaluate the toxicity of abamectin, difenoconazole and their mixture to the tropical fish Danio rerio. Laboratory toxicity tests with the individual pesticides indicated 48 h-LC50 values of 59 μg L−1 for abamectin and 1.4 mg L−1 for difenoconazole. Mixtures of the two pesticides revealed a synergistic deviation of the independent action model. Implications of study findings for the aquatic risk assessment of pesticide mixtures, especially in tropical countries and indications for future research are discussed.

Zinc-arsenic interactions in soil: Solubility, toxicity and uptake

Arsenic (As) and zinc (Zn) are common co-contaminants in mining impacted soils. Their interaction on solubility and toxicity when present concurrently is not well understood in natural systems. The aim of this study was to observe their interaction in solubility (soil-solution), bioaccumulation (shoot uptake) and toxicity to cucumber (Cucumis sativa L) conducting 4 weeks pot study in 5 different soils spiked with As (0, 2, 4, 8 to 1024 mg kg−1) individually and with Zn at two phytotoxic doses. The As pore-water concentration was significantly reduced (df = 289, Adjusted R2 = 0.84, p < 0.01) in the presence of Zn in the whole dataset, whereas Zn and Zn2+ activity in pore-water was reduced significantly only in the two alkaline soils. This outcome may be due to adsorption/surface precipitation or tertiary bridging complexation. No homogenous precipitation of zinc arsenate could be established using electron microscopy, XRD or even equilibrium calculations. For bioaccumulation phase, no significant effect of Zn on As uptake was observed except acidic MG soil whereas, Zn uptake was significantly reduced (p < 0.05) by As in whole dataset. However, an additive response was observed mostly except acidic MG soil. The synergistic response (more than additive) was predominant in this soil for a wide range of inhibition concentration (0–80%) at both Zn EC10 and EC50 levels. Since additive response is mostly considered in risk assessment for mixtures, precautions should be implemented for assessment of toxicity for As-Zn mixture in acidic soil due to their synergistic response in some soils.

A Human Mixture Risk Assessment for Neurodevelopmental Toxicity Associated with Polybrominated Diphenyl Ethers Used as Flame Retardants.

Background:The European Food Safety Authority recently concluded that the exposure of small children (1–3 y old) to brominated diphenyl ether (BDE)-99 may exceed acceptable levels defined in relation to neurodevelopmental toxicity in rodents. The flame retardant BDE-209 may release BDE-99 and other lower brominated BDEs through biotic and abiotic degradation, and all age groups are exposed not only to BDE-209 and -99 but also to a cocktail of BDE congeners with evidence of neurodevelopmental toxicity. The possible risks from combined exposures to these substances have not been evaluated.Objectives:We performed a congener-specific mixture risk assessment (MRA) of human exposure to combinations of BDE-209 and other BDEs based on estimated exposures via diet and dust intake and on measured levels in biologic samples.Methods:We employed the Hazard Index (HI) method by using BDE congener-specific reference doses for neurodevelopmental toxicity.Results:Our HI analysis suggests that combined exposures to polybrominated diphenyl ethers (PBDEs) may exceed acceptable levels in breastfeeding infants (0–3 mo old) and in small children (1–3 y old), even for moderate (vs. high) exposure scenarios. Our estimates also suggest that acceptable levels of combined PBDEs may be exceeded in adults whose diets are high in fish. Small children had the highest combined exposures, with some estimated body burdens that were similar to body burdens associated with developmental neurotoxicity in rodents.Conclusions:Our estimates corroborate reports from several recent epidemiological studies of associations between PBDE exposures and neurobehavioral outcomes, and they support the inclusion of BDE-209 in the persistent organic pollutant (POP) convention as well as the need for strategies to reduce exposures to PBDE mixtures, including maximum residue limits for PBDEs in food and measures for limiting the release of PBDEs from consumer waste. https://doi.org/10.1289/EHP826

Endocrine Disruption in Human Fetal Testis Explants by Individual and Combined Exposures to Selected Pharmaceuticals, Pesticides, and Environmental Pollutants.

Background:Numerous chemicals are capable of disrupting androgen production, but the possibility that they might act together to produce effects greater than those of the most effective component in the mixture has not been studied directly in human tissues. Suppression of androgen synthesis in fetal life has been associated with testis maldescent, malformations of the genitalia at birth, and poor semen quality later in life.Objectives:Our aim was to investigate whether chemicals can act together to disrupt androgen production in human fetal testis explants and to evaluate the importance of mixture effects when characterizing the hazard of individual chemicals.Methods:We used an organotypic culture system of human fetal testes explants called FEtal Gonad Assay (FEGA) with tissue obtained at 10 and 12 gestational wk (GW 10–12), to screen 27 chemicals individually for their possible anti-androgenic effect. Based on the results of the screen, we selected 11 compounds and tested them as mixtures.Results:We evaluated mixtures composed of four and eight antiandrogens that contained the pharmaceuticals ketoconazole and theophylline and several previously untested chemicals, such as the pesticides imazalil and propiconazole. Mixtures of antiandrogens can suppress testosterone synthesis in human fetal testicular explants to an extent greater than that seen with individual chemicals. This revealed itself as a shift towards lower doses in the dose–response curves of individual antiandrogens that became more pronounced as the number of components increased from four to eight.Conclusions:Our results with the FEGA provide the foundations of a predictive human mixture risk assessment approach for anti-androgenic exposures in fetal life. https://doi.org/10.1289/EHP1014

Assessment and prediction of joint algal toxicity of binary mixtures of graphene and ionic liquids

Graphene and ionic liquids (ILs) released into the environment will interact with each other. So far however, the risks associated with the concurrent exposure of biota to graphene and ILs in the environment have received little attention. The research reported here focused on observing and predicting the joint toxicity effects in the green alga Scenedesmus obliquus exposed to binary mixtures of intrinsic graphene (iG)/graphene oxide (GO) and five ILs of varying anionic and cationic types. The isolated ILs in the binary mixtures were the main contributors to toxicity. The binary GO-IL mixtures resulted in more severe joint toxicity than the binary iG-IL mixtures, irrespective of mixture ratios. The mechanism of the joint toxicity may be associated with the adsorption capability of the graphenes for the ILs, the dispersion stability of the graphenes in aquatic media, and modulation of the binary mixtures-induced oxidative stress. A toxic unit assessment showed that the graphene and IL toxicities were additive at low concentration of the mixtures but antagonistic at high concentration of the mixtures. Predictions made using the concentration addition and independent action models were close to the observed joint toxicities regardless of mixture types and mixture ratios. These findings provide new insights that are of use in the risk assessment of mixtures of engineered nanoparticles and other environmentally relevant contaminants.

Development of a procedure for determining the mixing ratios in ecotoxicological experiments and its application in binary salt mixture experiments.

Although a plethora of models exist to describe the characteristics and risk assessment of chemical mixtures in ecotoxicology, there is no specific procedure to decide on the mixing ratios (i.e. proportions of the individual chemical substances that form the mixture) at any desired level of concentration in an ecotoxicological mixture experiment. In this study, an attempt was made to develop a procedure for determining the mixing ratios in ecotoxicological experiments. In brief, from a single salt exposure test, the relative toxic fractions, which represent the toxic effect exerted by the individual salts, are determined. Thereafter, the proportions of each individual salt at any level of concentration in the mixture are estimated by multiplying the desired concentration with the relative toxic fraction of that particular salt. The procedure was applied to ecotoxicological experiments involving four binary salt mixtures (MgCl2 + MgSO4, NaCl + Na2SO4, MgCl2 + Na2SO4 and NaCl + MgSO4) and Caridina nilotica, an indigenous South African freshwater shrimp. It is hoped that the application of this developed procedure will ensure administering the correct proportions of individual chemical substances in chemical mixtures in order to obtain the desired levels of concentration in aquatic ecotoxicological mixture experiments.

Lead substances selection using GHS approach for the classification of mixtures: Case study of painting in the work environment

We developed a lead substances selection approach based on the concept of mixture classification of UN GHS for the purpose of efficient risk assessment of mixtures consisting of multiple components. Lead substances selection methods are being actively developed in Europe, but these methods are predicated on the regulations and information sources available within Europe and are therefore not readily applicable to countries outside Europe. In this study, the features of the GHS-based approach and the risk assessment results for outdoor painting work as a specific utilization example of the GHS-based approach were described. Comparison with the DPD + method and the CCA method proposed in Europe revealed that the GHS-based approach resulted in the selection of the safest lead substances. The GHS method, like the DPD + method, is a classification-based approach. We believe that a classification-based approach based on the GHS method can be an appropriate tool to efficiently implement risk assessment of mixtures for countries outside Europe. Some tools for business operators to conduct the management of chemicals using the GHS classification have been established in Japan. We plan to propose the GHS-based approach as a standardized assessment tool.

Genetic Toxicity of Complex Mixtures of Polycyclic Aromatic Hydrocarbons: Evaluating Dose-Additivity in a Transgenic Mouse Model.

This study evaluates the risk assessment approach currently employed for polycyclic aromatic hydrocarbon (PAH)-contaminated media, wherein carcinogenic hazards are evaluated using a dose-addition model that employs potency equivalency factors (PEFs) for targeted carcinogenic PAHs. Here, MutaMouse mice were subchronically exposed to PAH mixtures (p.o.), and mutagenic potency (MP) values were determined for five tissues. Predicted dose-additive mixture MPs were generated by summing the products of the concentrations and MPs of the individual targeted PAHs; values were compared to the experimental MPs of the mixtures to evaluate dose-additivity. Additionally, the PEF-determined BaP-equivalent concentrations were compared to those determined using a bioassay-derived method (BDM) (i.e., an additivity-independent approach). In bone marrow, mixture mutagenicity was less than dose-additive and the PEF-method provided higher estimates of BaP-equivalents than the BDM. Conversely, mixture mutagenicity in site-of-contact tissues (e.g., small intestine) was generally more than dose-additive and the PEF-method provided lower estimates of BaP-equivalents than the BDM. Overall, this study demonstrates that dose-additive predictions of mixture mutagenic potency based on the concentrations and potencies of a small number of targeted PAHs results in values that are surprisingly close to those determined experimentally, providing support for the dose-additive assumption employed for human health risk assessment of PAH mixtures.

Acute effects of binary mixtures of Type II pyrethroids and organophosphate insecticides on Oreochromis niloticus.

Pyrethroid and organophosphate insecticides have been used for more than 20 years worldwide to control a variety of insect pest in different settings. These pesticides have been detected in a variety of environmental samples, including surface waters and sediments and therefore there is significant concern about their potential toxic effects on non-target organisms. Mixtures of compounds from these groups of pesticides have been found to frequently show enhanced toxicity but it has been a challenge to predict whether or not enhanced toxicity will occur for a given combination of compounds. This study therefore studied the effects of binary pyrethroid-organophosphate mixtures using cypermethrin, deltamethrin and dimethoate in an acute toxicity test system with Oreochromis niloticus. The 96 h LC50s for individual insecticides were 9.13 µg/l, 9.42 µg/l and 45.52 mg/l for cypermethrin, deltamethrin and dimethoate respectively. These showed that the pyrethroid insecticides were highly toxic to Oreochromis niloticus and were far more toxic than dimethoate. All mixtures were also more toxic than single insecticides throughout the concentration-response curve with mixtures resulting in mortality at concentrations which the individual pesticides in the mixture were below their respective NOECs. In addition, observed mixture toxicities deviated from the predicted mixture effects based either on the Concentration Addition (CA) or Independent Action (IA) models independent of mixture ratio. However, the extent of observed mixture mortality deviation was dependent on the effect level. Significant deviations (MDR > 2.0) were observed at lower concentrations indicating synergistic effects at lower and possibly environmentally relevant concentrations. This is not unexpected since organophosphate insecticides are known to inhibit acetylcholinesterase as well as inactivate esterase, resulting in reduced detoxification of pyrethroid insecticides and consequently greater toxicity than would be expected. This has important implications for risk assessment of mixtures since the risk of pyrethroid-organophosphate mixtures may be underestimated if either the CA or IA model is employed.

Evaluation of FOCUS surface water pesticide concentration predictions and risk assessment of field-measured pesticide mixtures-a crop-based approach under Mediterranean conditions.

FOCUS models are used in the European regulatory risk assessment (RA) to predict individual pesticide concentrations in edge-of-field surface waters. The scenarios used in higher tier FOCUS simulations were mainly based on Central/North European, and work is needed to underpin the validity of simulated exposure profiles for Mediterranean agroecosystems. In addition, the RA of chemicals are traditionally evaluated on the basis of single substances although freshwater life is generally exposed to a multitude of pesticides. In the present study, we monitored 19 pesticides in surface waters of five locations in the Portuguese 'Lezíria do Tejo' agricultural area. FOCUS step 3 simulations were performed for the South European scenarios to estimate predicted environmental concentrations (PECs). We verified that 44% of the PECs underestimated the measured environmental concentrations (MEC) of the pesticides, showing a non-compliance with the field data. Risk was assessed by comparing the environmental quality standards (EQS) and regulatory acceptable concentrations with their respective MECs. Risk of mixtures was demonstrated in 100% of the samples with insecticides accounting for 60% of the total risk identified. The overall link between the RA and the actual situation in the field must be considerably strengthened, and field studies on pesticide exposure and effects should be carried out to assist the improvement of predictive approaches used for regulatory purposes.

Assessment of AcuteIn VitroHuman Cellular Responses to Smoke Extracts from a Reduced Toxicant Prototype Cigarette

Abstract Assessment of novel tobacco and nicotine products for their harm reduction potential compared with cigarettes is of significance for regulators and public health. The National Research Council's report, “Toxicity Testing in the 21st Century,” outlines recent advances in molecular biology and the utility of human in vitro systems in toxicity testing. Such approaches could enable efficient, ethical, and economical risk assessment of chemicals and chemical mixtures. We investigated the potential of a simple in vitro lung model to discriminate between a commercially available cigarette (CC) and a reduced toxicant prototype (RTP) cigarette, which yielded significantly reduced toxicant emissions compared with the CC. The investigation compared cytotoxicity, oxidative stress-related and proinflammatory endpoints, associated with smoking-related disease development. Furthermore, we assessed the discriminatory potential of the model upon exposure to a selection of test matrices, which captured different f...

Mixture risk assessment due to ingestion of arsenic, copper, and zinc from milkfish farmed in contaminated coastal areas.

Human health risks associated with the consumption of metal-contaminated fish over extended periods have become a concern particularly in Taiwan, where fish is consumed on a large scale. This study applied the interaction-based hazard index (HI) to assess the mixture health risks for fishers and non-fishers who consume the arsenic (As), copper (Cu), and zinc (Zn) contaminated milkfish from As-contaminated coastal areas in Taiwan, taking into account joint toxic actions and potential toxic interactions. We showed that the interactions of As-Zn and Cu-Zn were antagonistic, whereas As-Cu interaction was additive. We found that HI estimates without interactions considered were 1.3-1.6 times higher than interactive HIs. Probability distributions of HI estimates for non-fishers were less than 1, whereas all 97.5%-tile HI estimates for fishers were >1. Analytical results revealed that the level of inorganic As in milkfish was the main contributor to HIs, indicating a health risk posed to consumers of fish farmed in As-contaminated areas. However, we found that Zn supplementation could significantly decrease As-induced risk of hematological effect by activating a Zn-dependent enzyme. In order to improve the accuracy of health risk due to exposure to multiple metals, further toxicological data, regular environmental monitoring, dietary survey, and refinement approaches for interactive risk assessment are warranted.

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.

Using Delaunay triangulation and Voronoi tessellation to predict the toxicities of binary mixtures containing hormetic compound

Concentration addition (CA) was proposed as a reasonable default approach for the ecological risk assessment of chemical mixtures. However, CA cannot predict the toxicity of mixture at some effect zones if not all components have definite effective concentrations at the given effect, such as some compounds induce hormesis. In this paper, we developed a new method for the toxicity prediction of various types of binary mixtures, an interpolation method based on the Delaunay triangulation (DT) and Voronoi tessellation (VT) as well as the training set of direct equipartition ray design (EquRay) mixtures, simply IDVequ. At first, the EquRay was employed to design the basic concentration compositions of five binary mixture rays. The toxic effects of single components and mixture rays at different times and various concentrations were determined by the time-dependent microplate toxicity analysis. Secondly, the concentration-toxicity data of the pure components and various mixture rays were acted as a training set. The DT triangles and VT polygons were constructed by various vertices of concentrations in the training set. The toxicities of unknown mixtures were predicted by the linear interpolation and natural neighbor interpolation of vertices. The IDVequ successfully predicted the toxicities of various types of binary mixtures.

Comparison of four methods for bioavailability‐based risk assessment of mixtures of Cu, Zn, and Ni in freshwater

Although chemical risk assessment is still mainly conducted on a substance-by-substance basis, organisms in the environment are typically exposed to mixtures of substances. Risk assessment procedures should therefore be adapted to fit these situations. Four mixture risk assessment methodologies were compared for risk estimations of mixtures of copper (Cu), zinc (Zn), and nickel (Ni). The results showed that use of the log-normal species sensitivity distribution (SSD) instead of the best-fit distribution and sampling species sensitivities independently for each metal instead of using interspecies correlations in metal sensitivity had little impact on risk estimates. Across 4 different monitoring datasets, between 0% and 52% of the target water samples were estimated to be at risk, but only between 0% and 15% of the target water samples were at risk because of the mixture of metals and not any single metal individually. When a natural baseline database was examined, it was estimated that 10% of the target water samples were at risk because of single metals or their mixtures; when the most conservative method was used (concentration addition [CA] applied directly to the SSD, i.e., CASSD ). However, the issue of metal mixture risk at geochemical baseline concentrations became relatively small (2% of target water samples) when a theoretically more correct method was used (CA applied to individual dose response curves, i.e., CADRC ). Finally, across the 4 monitoring datasets, the following order of conservatism for the 4 methods was shown (from most to least conservative, with ranges of median margin of safety [MoS] relative to CASSD ): CASSD > CADRC (MoS = 1.17-1.25) > IADRC (independent action (IA) applied to individual dose-response curves; MoS = 1.38-1.60) > IASSD (MoS = 1.48-1.72). Therefore, it is suggested that these 4 methods can be used in a general tiered scheme for the risk assessment of metal mixtures in a regulatory context. In this scheme, the CASSD method could serve as a first (conservative) tier to identify situations with likely no potential risk at all, regardless of the method used (the sum toxic unit expressed relative to the 5% hazardous concentration [SumTUHC5 ] < 1) and the IASSD method to identify situations of potential risk, also regardless of the method used (the multisubstance potentially affected fraction of species using the IASSD method [msPAFIA,SSD ] > 0.05). The CADRC and IADRC methods could be used for site-specific assessment for situations that fall in between (SumTUHC5 > 1 and msPAFIA,SSD < 0.05). Environ Toxicol Chem 2017;36:2123-2138. © 2017 SETAC.

Combined ecotoxicological risk assessment in the frame of European authorization of pesticides.

Organisms are frequently exposed to mixtures of chemical contaminants in the environment, causing a potential "cocktail effect", or combined effect. The joint action of different molecules with similar or different modes of action could result in a potentially unlimited number of additives, synergistic or antagonistic combinations. Since the large number of contaminants makes it impossible to perform ecotoxicity tests for each potential mixture, a robust approach for prospective environmental risk assessment of chemical mixtures is needed. A number of recent publications by the European Commission and the authorities in charge prove the increasing interest that is spreading in the European community towards the topic of the assessment of chemical mixtures. The current EU regulation for Plant Protection Products authorization (Reg. 1107/2009 EC) explicitly requires the evaluation of the potential combined effects of active substances. We reviewed current methods and limitations of mixture assessment of pesticides (7 fungicides and 4 herbicides) through the analysis of the approaches adopted to investigate possible risks for different non-target organisms. The Concentration Addition (CA) approach was the most used approach to predict multiple toxicity to non-target organisms. The guidance for birds and mammals first introduced standard procedures to assess the multiple toxicity based on on CA concept. The recent aquatic EFSA guidance introduced some requirements to evaluate potential mixture toxicity, while the current guidance requirements for terrestrial organisms still lack clear indications on how to conduct the assessment. Moreover, new indications come from the draft guidance for the assessment of terrestrial plants and in-soil organisms. However, the approval and implementation of these new guidelines are still at a developmental stage. Some final considerations are drawn on the future possibilities to improve risk assessment procedures so as to identify harmful effects of pesticides mixtures on non-target organisms.

A framework for the use of single-chemical transcriptomics data in predicting the hazards associated with complex mixtures of polycyclic aromatic hydrocarbons

The assumption of additivity applied in the risk assessment of environmental mixtures containing carcinogenic polycyclic aromatic hydrocarbons (PAHs) was investigated using transcriptomics. MutaTMMouse were gavaged for 28 days with three doses of eight individual PAHs, two defined mixtures of PAHs, or coal tar, an environmentally ubiquitous complex mixture of PAHs. Microarrays were used to identify differentially expressed genes (DEGs) in lung tissue collected 3 days post-exposure. Cancer-related pathways perturbed by the individual or mixtures of PAHs were identified, and dose–response modeling of the DEGs was conducted to calculate gene/pathway benchmark doses (BMDs). Individual PAH-induced pathway perturbations (the median gene expression changes for all genes in a pathway relative to controls) and pathway BMDs were applied to models of additivity [i.e., concentration addition (CA), generalized concentration addition (GCA), and independent action (IA)] to generate predicted pathway-specific dose–response curves for each PAH mixture. The predicted and observed pathway dose–response curves were compared to assess the sensitivity of different additivity models. Transcriptomics-based additivity calculation showed that IA accurately predicted the pathway perturbations induced by all mixtures of PAHs. CA did not support the additivity assumption for the defined mixtures; however, GCA improved the CA predictions. Moreover, pathway BMDs derived for coal tar were comparable to BMDs derived from previously published coal tar-induced mouse lung tumor incidence data. These results suggest that in the absence of tumor incidence data, individual chemical-induced transcriptomics changes associated with cancer can be used to investigate the assumption of additivity and to predict the carcinogenic potential of a mixture.

Toward a conceptual approach for assessing risks from chemical mixtures and other stressors to coastal ecosystem services.

Growth of human populations and increased human activity, particularly in coastal areas, increase pressure on coastal ecosystems and the ecosystem services (ES) they provide. As a means toward being able to assess the impact of multiple stressors on ES, in the present study we propose an 8-step conceptual approach for assessing effects of chemical mixtures and other stressors on ES in coastal areas: step A, identify the relevant problems and policy aims; step B, identify temporal and spatial boundaries; step C, identify relevant ES; step D, identify relevant stressors (e.g., chemicals); step E, translate impacts into ES units; step F, assess cumulative risk in ES units; step G, rank stressors based on their contribution to adverse effects on ES; and step H, implement regulation and management as appropriate and necessary. Two illustrative case studies (Swedish coastal waters and a coastal lagoon in Costa Rica) are provided; one focuses on chemicals that affect human food supply and the other addresses pesticide runoff and trade-offs among ES. The 2 cases are used to highlight challenges of such risk assessments, including use of standardized versus ES-relevant test species, data completeness, and trade-offs among ES. Lessons learned from the 2 case studies are discussed in relation to environmental risk assessment and management of chemical mixtures. Integr Environ Assess Manag 2017;13:376-386. © 2016 SETAC.

Regulatory assessment of chemical mixtures: Requirements, current approaches and future perspectives

This paper reviews regulatory requirements and recent case studies to illustrate how the risk assessment (RA) of chemical mixtures is conducted, considering both the effects on human health and on the environment. A broad range of chemicals, regulations and RA methodologies are covered, in order to identify mixtures of concern, gaps in the regulatory framework, data needs, and further work to be carried out. Also the current and potential future use of novel tools (Adverse Outcome Pathways, in silico tools, toxicokinetic modelling, etc.) in the RA of combined effects were reviewed.The assumptions made in the RA, predictive model specifications and the choice of toxic reference values can greatly influence the assessment outcome, and should therefore be specifically justified. Novel tools could support mixture RA mainly by providing a better understanding of the underlying mechanisms of combined effects. Nevertheless, their use is currently limited because of a lack of guidance, data, and expertise. More guidance is needed to facilitate their application. As far as the authors are aware, no prospective RA concerning chemicals related to various regulatory sectors has been performed to date, even though numerous chemicals are registered under several regulatory frameworks.