Key Opportunities to Replace, Reduce, and Refine Regulatory Fish Acute Toxicity Tests.

Fish acute toxicity tests are conducted as part of regulatory hazard identification and risk-assessment packages for industrial chemicals and plant protection products. The aim of these tests is to determine the concentration which would be lethal to 50% of the animals treated. These tests are therefore associated with suffering in the test animals, and Organisation for Economic Co-operation and Development test guideline 203 (fish, acute toxicity) studies are the most widely conducted regulatory vertebrate ecotoxicology tests for prospective chemical safety assessment. There is great scope to apply the 3Rs principles-the reduction, refinement, and replacement of animals-in this area of testing. An expert ecotoxicology working group, led by the UK National Centre for the Replacement, Refinement and Reduction of Animals in Research, including members from government, academia, and industry, reviewed global fish acute test data requirements for the major chemical sectors. The present study highlights ongoing initiatives and provides an overview of the key challenges and opportunities associated with replacing, reducing, and/or refining fish acute toxicity studies-without compromising environmental protection. Environ Toxicol Chem 2020;39:2076-2089. © 2020 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Parameters for assessing the aquatic environmental impact of cosmetic products.

The cosmetic industry's growing concern about the impact of its supply chain on the environment, sustainability of raw materials, and biodiversity increases the need to ensure that the final product has a lower environmental impact. The objective of this review is to summarize and compare the information available from international organizations and legislation regarding the main criteria used to assess raw materials for aquatic toxicity, as well as the most suitable alternative methods for obtaining assessment parameters. Using the literature available in databases, a review of the scientific literature and international legislation, this work discusses and compares the parameters established by international organizations such as the Environmental Protection Agency (EPA) and Cradle to Cradle (C2C), as well as European legislation, namely, European Regulation 1272/2008, for assessing environmental impact. Defining the ecotoxicity parameters of the main classes of raw materials in rinse-off cosmetic products can enable the development of products that are more environmentally sustainable, prioritizing substances with less environmental impact.

Environmental risk assessment of pesticides: state of the art and prospective improvement from science.

Pesticide risk assessment in the European regulatory framework is mandatory performed for active substances (pesticides) and the plant protection products they are constituents of. The aim is to guarantee that safe use can be achieved for the intended use of the product. This paper provides a feedback on the regulatory environmental risk assessment performed for pesticide registration at the EU and member state levels. The different steps of pesticide registration are addressed considering both exposure and hazard. In this paper, we focus on the environmental fate and behaviour in surface water together with the aquatic ecotoxicity of the substances to illustrate pesticide regulatory risk assessment performed for aquatic organisms. Current methodologies are presented along with highlights on potential improvements. For instance, as regards exposure aspects, moving from field based to landscape risk assessments is promising. Regarding ecotoxicology, ecological models may be valuable tools when applied to chemical risk assessment. In addition, interest and further developments to better take into account mitigation measures in risk assessment and management are also presented.

Regulatory ecotoxicity testing of nanomaterials - proposed modifications of OECD test guidelines based on laboratory experience with silver and titanium dioxide nanoparticles.

Regulatory ecotoxicity testing of chemicals is of societal importance and a large effort is undertaken at the OECD to ensure that OECD test guidelines (TGs) for nanomaterials (NMs) are available. Significant progress to support the adaptation of selected TGs to NMs was achieved in the context of the project MARINA ( http://www.marina-fp7.eu/ ) funded within the 7th European Framework Program. Eight OECD TGs were adapted based on the testing of at least one ion-releasing NM (Ag) and two inert NMs (TiO2). With the materials applied, two main variants of NMs (ion releasing vs. inert NMs) were addressed. As the modifications of the test guidelines refer to general test topics (e.g. test duration or measuring principle), we assume that the described approaches and modifications will be suitable for the testing of further NMs with other chemical compositions. Firm proposals for modification of protocols with scientific justification(s) are presented for the following tests: growth inhibition using the green algae Raphidocelis subcapitata (formerly: Pseudokirchneriella subcapitata; TG 201), acute toxicity with the crustacean Daphnia magna (TG 202), development toxicity with the fish Danio rerio (TG 210), reproduction of the sediment-living worm Lumbriculus variegatus (TG 225), activity of soil microflora (TGs 216, 217), and reproduction of the invertebrates (Enchytraeus crypticus, Eisenia fetida, TGs 220, 222). Additionally, test descriptions for two further test systems (root elongation of plants in hydroponic culture; test on fish cells) are presented. Ecotoxicological data obtained with the modified test guidelines for TiO2 NMs and Ag NM and detailed method descriptions are available.

Soil ecotoxicology in Brazil is taking its course.

Soil ecotoxicology has been motivated by the increasing global awareness on environmental issues. Northern Hemisphere has been the main driver of this science branch; however, the number and quality of contributions from the Southern Hemisphere are increasing quickly. In this case study, Brazil is taken as an example of how soil ecotoxicology has developed over the last 30 years. It starts with a brief historical overview depicting the main events on soil ecotoxicology in the country. Following, an overview on the Brazilian legislation related to soil ecotoxicology is given, covering regulations with prospective focus, mainly on the registration of pesticides. Regulations with retrospective focus in contaminated areas are also given. Then, an outline of the actors in soil ecotoxicology and examples of prospective ecotoxicological studies performed with soil organisms and plants are given by stressor groups: pesticides, pharmaceuticals, metals, and residues. Experiences from retrospective studies, mainly looking at the assessment of industrial sites, are also covered. Emphasis is given on methodological aspects, pointing to needed actions, mainly regarding the different biotic and abiotic conditions of a tropical country. Finally, the last session discusses how soil ecotoxicology could be improved in methodological adaptations as well as legal requirements.

Adverse Outcome Pathways for Regulatory Applications: Examination of Four Case Studies With Different Degrees of Completeness and Scientific Confidence.

Adverse outcome pathways (AOPs) offer a pathway-based toxicological framework to support hazard assessment and regulatory decision-making. However, little has been discussed about the scientific confidence needed, or how complete a pathway should be, before use in a specific regulatory application. Here we review four case studies to explore the degree of scientific confidence and extent of completeness (in terms of causal events) that is required for an AOP to be useful for a specific purpose in a regulatory application: (i) Membrane disruption (Narcosis) leading to respiratory failure (low confidence), (ii) Hepatocellular proliferation leading to cancer (partial pathway, moderate confidence), (iii) Covalent binding to proteins leading to skin sensitization (high confidence), and (iv) Aromatase inhibition leading to reproductive dysfunction in fish (high confidence). Partially complete AOPs with unknown molecular initiating events, such as 'Hepatocellular proliferation leading to cancer', were found to be valuable. We demonstrate that scientific confidence in these pathways can be increased though the use of unconventional information (eg, computational identification of potential initiators). AOPs at all levels of confidence can contribute to specific uses. A significant statistical or quantitative relationship between events and/or the adverse outcome relationships is a common characteristic of AOPs, both incomplete and complete, that have specific regulatory uses. For AOPs to be useful in a regulatory context they must be at least as useful as the tools that regulators currently possess, or the techniques currently employed by regulators.

Risk assessment of 'endocrine substances': guidance on identifying endocrine disruptors.

The European regulation on plant protection products (1107/2009) and other related legislation only support the marketing and use of chemical products on the basis that they do not induce endocrine disruption in humans or wildlife species. This legislation would appear to make the assumption that endocrine active chemicals should be managed differently from other chemicals presumably due to an assumed lack of a threshold for adverse effects. In the absence of agreed scientific criteria and guidance on how to identify and evaluate endocrine activity and disruption within these pieces of legislation, a European Centre for Ecotoxicology and Toxicology of Chemicals (ECETOC) task force was formed to provide scientific criteria that may be used within the context of these three legislative documents. The first ECETOC technical report and associated workshop, held in 2009, presented a science-based concept on how to identify endocrine activity and disrupting properties of chemicals for both human health and the environment. Specific scientific criteria for the determination of endocrine activity and disrupting properties that integrate information from both regulatory toxicity studies and mechanistic/screening studies were proposed. These criteria combined the nature of the adverse effects detected in studies which give concern for endocrine toxicity with an understanding of the mode of action of toxicity so that adverse effects can be explained scientifically. A key element in the data evaluation is the consideration of all available information in a weight-of-evidence approach. Both sets of data (evidence of the adverse effect in apical studies and conclusive mode of action knowledge) are essential in order to correctly identify endocrine disruption according to accepted definitions. As the legislation seeks to regulate chemicals on a mode of action rather than the more traditional approach of adverse endpoints, then conclusive evidence of the mode of action of concern should be presented. From a human safety perspective and in the absence of any compelling data that endocrine active chemicals exert their adverse effects through anything other than a threshold mechanism there is no scientific justification for not using a margin of exposure approach to risk assessment in order to best protect human health.

ECETOC Florence workshop on risk assessment of endocrine substances, including the potency concept.

The European regulation on plant protection products (1107/2009) and the Biocidal Products Regulation (EC Regulation 528/2012) only support the marketing and use of chemicals if they do not cause endocrine disruption in humans or wildlife species. Also, substances with endocrine properties are subject to authorization under the European regulation on the registration, evaluation, authorization and restriction of chemicals (REACH; 1907/2006). Therefore, the regulatory consequences of identifying a substance as an endocrine disrupting chemical are severe. In contrast to that, basic scientific criteria, necessary to define endocrine disrupting properties, are not described in any of these legislative documents. Thus, the European Center for Ecotoxicology and Toxicology of Chemicals (ECETOC) established a task force to provide scientific criteria for the identification and assessment of chemicals with endocrine disrupting properties that may be used within the context of these three legislative texts (ECETOC, 2009a). In 2009, ECETOC introduced a scientific framework as a possible concept for identifying endocrine disrupting properties within a regulatory context (ECETOC, 2009b; Bars et al., 2011a,b). The proposed scientific criteria integrated, in a weight of evidence approach, information from regulatory (eco)toxicity studies and mechanistic/screening studies by combining evidence for adverse effects detected in apical whole-organism studies with an understanding of the mode of action (MoA) of endocrine toxicity. However, since not all chemicals with endocrine disrupting properties are of equal hazard, an adequate concept should also be able to differentiate between chemicals with endocrine properties of low concern from those of higher concern (for regulatory purposes). For this purpose, the task force refined this part of their concept. Following an investigation of the key factors at a second workshop of invited regulatory, academic and industry scientists, the guidance was advanced further. For human health assessments it is based on the relevance to humans of the endocrine mechanism of toxicity, the specificity of the endocrine effects with respect to other toxic effects, the potency of the chemical to induce endocrine toxicity and consideration of exposure levels.

Identification, assessment and management of "endocrine disruptors" in wildlife in the EU substance legislation--discussion paper from the German Federal Environment Agency (UBA).

A discussion paper was developed by a panel of experts of the German Federal Environment Agency (UBA) contributing to the on-going debate on the identification, assessment and management of endocrine disruptors with a view to protect wildlife according to the EU substance legislation (plant protection products, biocides, industrial chemicals). Based on a critical synthesis of the state-of-the-art regarding regulatory requirements, testing methods, assessment schemes, decision-making criteria and risk management options, we advise an appropriate and consistent implementation of this important subject into existing chemicals legislation in Europe. Our proposal for a balanced risk management of endocrine disruptors essentially advocates transparent regulatory decision making based on a scientifically robust weight of evidence approach and an adequate risk management consistent across different legislations. With respect to the latter, a more explicit consideration of the principle of proportionality of regulatory decision making and socio-economic benefits in the on-going debate is further encouraged.

Necessity and approach to integrated nanomaterial legislation and governance.

Nanotechnology is one of the most promising technologies to emerge in recent decades. Materials that are specially engineered to have at least one dimension that is no larger than 100 nm are now continuously manufactured and incorporated as critical components of different products that people use daily. While we are taking advantage of nanomaterials (NMs) and nano-products, they may pose a risk to humans and the broader environment. Some types of fibrous NMs such as carbon nanotubes and nano-fibers may present a risk similar to that of asbestos. Some carbon or metal based NMs may threaten the environment due to their bioaccumulative nature within food webs. In order to prevent future adverse effects from products or byproducts of nanotechnology, we suggest an integrated multi-faceted approach which includes an integrated regulation that is based upon life cycle assessment, empirically derived risk assessment. Advanced research that fills the knowledge gap regarding the understanding of NMs in scientific and social norms will be helpful in a full life cycle assessment of NMs. Emphasizing nanotechnology education to the public for an increased understanding and participation associated with media coverage will finally draw governments' attention with an integrated legislation to be instituted. Developing the optimal mix of these tools, including research, public education, media coverage, integrated legislation, will be significant to proactively manage the complexity of nanotechnology and prevent any undesirable effect due to the NMs exposure.

Using mode of action information to improve regulatory decision-making: an ECETOC/ILSI RF/HESI workshop overview.

The European Centre for Ecotoxicology and Toxicology of Chemicals (ECETOC), the International Life Sciences Institute (ILSI) Research Foundation (RF), and the ILSI Health and Environmental Sciences Institute (HESI) hosted a workshop in November 2009 to review current practice in the application of mode of action (MOA) considerations in chemical risk assessment. The aim was to provide a rationale for a more general, but flexible approach and to propose steps to facilitate broader uptake and use of the MOA concept. There was consensus amongst the workshop participants that it will require substantial effort and cooperation from the multiple disciplines involved to embrace a common, consistent, and transparent approach. Setting up a repository of accepted MOAs and associated guidance concerning appropriate data to support specific MOAs for critical effects would facilitate categorization of chemicals and allow predictions of toxicity outcomes by read-across. This should in future contribute to the reduction of toxicity testing in animals. The workshop participants also acknowledged the value and importance of human data and the importance of integrating information from biological pathway analyses into current MOA/human relevance frameworks.

Science based guidance for the assessment of endocrine disrupting properties of chemicals.

The European legislation on plant protection products (Regulation (EC) No. 1107/2009) and biocides (Directive 98/8/EC), as well as the regulation concerning chemicals (Regulation (EC) No. 1907/2006 'REACH') only support the marketing and use of chemical products on the basis that they do not induce endocrine disruption in humans or non-target species. However, there is currently no agreed guidance on how to identify and evaluate endocrine activity and disruption. Consequently, an ECETOC task force was formed to provide scientific criteria that may be used within the context of these three legislative documents. Specific scientific criteria for the determination of endocrine disrupting properties that integrate information from both regulatory (eco)toxicity studies and mechanistic/screening studies are proposed. These criteria combine the nature of the adverse effects detected in studies which give concern for endocrine toxicity with an understanding of the mode of action of toxicity so that adverse effects can be explained scientifically. The criteria developed are presented in the form of flow charts for assessing relevant effects for both humans and wildlife species. In addition, since not all chemicals with endocrine disrupting properties are of equal hazard, assessment of potency is also proposed to discriminate chemicals of high concern from those of lower concern. The guidance presented in this paper includes refinements made to an initial proposal following discussion of the criteria at a workshop of invited regulatory, academic and industry scientists.

Regulatory consideration of bioavailability for metals: simplification of input parameters for the chronic copper biotic ligand model.

The chronic Cu biotic ligand model (CuBLM) provides a means by which the bioavailability of Cu can be taken into account in assessing the potential chronic risks posed by Cu at specific freshwater locations. One of the barriers to the widespread regulatory application of the CuBLM is the perceived complexity of the approach when compared to the current systems that are in place in many regulatory organizations. The CuBLM requires 10 measured input parameters, although some of these have a relatively limited influence on the predicted no-effect concentration (PNEC) for Cu. Simplification of the input requirements of the CuBLM is proposed by estimating the concentrations of the major ions Mg2+, Na+, K+, SO4(2-), Cl- , and alkalinity from Ca concentrations. A series of relationships between log10 (Ca, mg l(-1)) and log10 (major ion, mg l(-1)) was established from surface water monitoring data for Europe, and applied in the prediction of Cu PNEC values for some UK freshwater monitoring data. The use of default values for major ion concentrations was also considered, and both approaches were compared to the use of measured major ion concentrations. Both the use of fixed default major ion concentrations, and major ion concentrations estimated from Ca concentrations, provided Cu PNEC predictions which were in good agreement with the results of calculations using measured data. There is a slight loss of accuracy when using estimates of major ion concentrations compared to using measured concentration data, although to a lesser extent than when fixed default values are applied. The simplifications proposed provide a practical evidence-based methodology to facilitate the regulatory implementation of the CuBLM.

[Toxicogenomics in hazard assessment of chemicals]. / Toksykogenomika w ocenie zagrozen substancji chemicznych.

Recent changes in the European legislation of chemicals suggest an urgent need for introduction of novel, alternative methods for testing chemical substances. Such possibility is offered by toxicogenomics--a scientific discipline combining knowledge from the field of toxicology, i.e., a science investigating the properties of toxic agents and the negative effects these agents exert on health and environment, with genomics, i.e., a science investigating the structure and function of genomes. New research strategies within the field of toxicology (transcriptomics, proteomics, metabolomics) offer conditions to assess the hazards associated with the effects of chemicals with both established and suspected toxic potentials.

Potential application of ecological models in the European environmental risk assessment of chemicals. I. Review of protection goals in EU directives and regulations.

Several European directives and regulations address the environmental risk assessment of chemicals. We used the protection of freshwater ecosystems against plant protection products, biocidal products, human and veterinary pharmaceuticals, and other chemicals and priority substances under the Water Framework Directive as examples to explore the potential of ecological effect models for a refined risk assessment. Our analysis of the directives, regulations, and related guidance documents lead us to distinguish the following 5 areas for the application of ecological models in chemical risk assessment: 1) Extrapolation of organism-level effects to the population level: The protection goals are formulated in general terms, e.g., avoiding "unacceptable effects" or "adverse impact" on the environment or the "viability of exposed species." In contrast, most of the standard ecotoxicological tests provide data only on organism-level endpoints and are thus not directly linked to the protection goals which focus on populations and communities. 2) Extrapolation of effects between different exposure profiles: Especially for plant protection products, exposure profiles can be very variable and impossible to cover in toxicological tests. 3) Extrapolation of recovery processes: As a consequence of the often short-term exposures to plant protection products, the risk assessment is based on the community recovery principle. On the other hand, assessments under the other directives assume a more or less constant exposure and are based on the ecosystem threshold principle. 4) Analysis and prediction of indirect effects: Because effects on 1 or a few taxa might have consequences on other taxa that are not directly affected by the chemical, such indirect effects on communities have to be considered. 5) Prediction of bioaccumulation within food chains: All directives take the possibility of bioaccumulation, and thus secondary poisoning within the food chain, into account.

An Integrated Assessment Scheme for assessing the adequacy of (eco)toxicological data under REACH.

REACH requires all available (eco)toxicological information, whether protocol studies, other experiments, or non-testing approaches such as read-across or (Q)SAR, to be collected and evaluated. However, guidance documents only limitedly address how adequacy of (eco)toxicological information can be assessed consistently and transparently. We propose an Integrated Assessment Scheme (IAS) for the evaluation of (eco)toxicological data. The IAS consists of three modules: (i) the reliability of the data; (ii) the validity of the methods the data are generated from and; (iii) the regulatory need of the data. Each module is assessed and documented using adjusted OECD principles for the validation of (Q)SARs. These adjusted principles provide a harmonised set of criteria for the evaluation of all types of (eco)toxicological data. Assessment codes, similar to Klimisch codes, are assigned to the evaluated information in each module. The coherent combination of the assessment codes of all three modules determines the overall adequacy of information for fulfilling the information requirement in REACH, and can serve as a weight in a Weight of Evidence procedure as mentioned in REACH Annex XI.

Toxicity assessment and public policies: an urgent need for research.

The new European regulation on chemicals triggers a huge number of new testing. However, more than 2 years after the enforcement of this policy, toxicity assessment and risk assessment are still using single species tests that deliver little information. As it is often the case, the link between science and policy seems to be disrupted. However, policy makers need more than ever information on the fate and effects of chemicals on living systems. Without relevant knowledge for decision making, the application of the precautionary principle is the only reasonable way to manage risks. It is necessary to develop new risk assessment strategies using the last innovations from biology: the omics tools, ecology, ecosystem modeling, chemistry, and computing. This article highlights some of the recent trends in ecotoxicology and calls for a new research strategy. This strategy implies research to be funded by its users.

Integrating omic technologies into aquatic ecological risk assessment and environmental monitoring: hurdles, achievements, and future outlook.

BACKGROUND: In this commentary we present the findings from an international consortium on fish toxicogenomics sponsored by the U.K. Natural Environment Research Council (Fish Toxicogenomics-Moving into Regulation and Monitoring, held 21-23 April 2008 at the Pacific Environmental Science Centre, Vancouver, BC, Canada). OBJECTIVES: The consortium from government agencies, academia, and industry addressed three topics: progress in ecotoxicogenomics, regulatory perspectives on roadblocks for practical implementation of toxicogenomics into risk assessment, and dealing with variability in data sets. DISCUSSION: Participants noted that examples of successful application of omic technologies have been identified, but critical studies are needed to relate molecular changes to ecological adverse outcome. Participants made recommendations for the management of technical and biological variation. They also stressed the need for enhanced interdisciplinary training and communication as well as considerable investment into the generation and curation of appropriate reference omic data. CONCLUSIONS: The participants concluded that, although there are hurdles to pass on the road to regulatory acceptance, omics technologies are already useful for elucidating modes of action of toxicants and can contribute to the risk assessment process as part of a weight-of-evidence approach.