RESUMEN
Growth and extensive urbanisation of the human population has been accompanied by increased manufacture and use of chemical compounds. To classify the fate and behaviour of these compounds in the environment, a series of international standardised biodegradation screening tests (BSTs) were developed over 30â¯years ago. In recent years, regulatory emphasis (e.g. REACH) has shifted from measuring biodegradation towards prioritisations based on chemical persistence. In their current guise, BSTs are ineffective as screens for persistence. The marine BST OECD 306 in particular is prone to high levels of variation and produces a large number of fails, many of which can be considered false negatives. An ECETOC funded two-day workshop of academia, industry and regulatory bodies was held in 2015 to discuss improvements to the marine BSTs based on previous research findings from the Cefic LRI ECO11 project and other foregoing studies. During this workshop, methodological improvements to the OECD 306 test were discussed, in addition to clarifying guidance on testing and interpretation of results obtained from marine BSTs (such as pass criteria, lag phases, freshwater read across and complex substances). Methodologically: (i) increasing bacterial cell concentrations to better represent the bacterial diversity inherent in the sampled environments; and (ii) increasing test durations to investigate extended lag phases observed in marine assessments, were recommended to be validated in a multi-institutional ring test.
Asunto(s)
Biodegradación Ambiental , Guías como Asunto , Agua de Mar/análisis , Contaminantes Químicos del Agua/metabolismo , Congresos como Asunto , Organización para la Cooperación y el Desarrollo EconómicoRESUMEN
Urban regions of the world are expanding rapidly, placing additional stress on water resources. Urban water bodies serve many purposes, from washing and sources of drinking water to transport and conduits for storm drainage and effluent discharge. These water bodies receive chemical emissions arising from either single or multiple point sources, diffuse sources which can be continuous, intermittent, or seasonal. Thus, aquatic organisms in these water bodies are exposed to temporally and compositionally variable mixtures. We have delineated source-specific signatures of these mixtures for diffuse urban runoff and urban point source exposure scenarios to support risk assessment and management of these mixtures. The first step in a tiered approach to assessing chemical exposure has been developed based on the event mean concentration concept, with chemical concentrations in runoff defined by volumes of water leaving each surface and the chemical exposure mixture profiles for different urban scenarios. Although generalizations can be made about the chemical composition of urban sources and event mean exposure predictions for initial prioritization, such modeling needs to be complemented with biological monitoring data. It is highly unlikely that the current paradigm of routine regulatory chemical monitoring alone will provide a realistic appraisal of urban aquatic chemical mixture exposures. Future consideration is also needed of the role of nonchemical stressors in such highly modified urban water bodies. Environ Toxicol Chem 2018;37:703-714. © 2017 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals, Inc. on behalf of SETAC.
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Organismos Acuáticos/efectos de los fármacos , Exposición a Riesgos Ambientales/análisis , Medición de Riesgo/métodos , Contaminantes Químicos del Agua/toxicidad , Ciudades , Ecotoxicología , Monitoreo del Ambiente , Residuos Industriales/análisis , Reproducibilidad de los ResultadosRESUMEN
Ecological risk assessments and risk management decisions are only as sound as the underlying information and processes to integrate them. It is important to develop transparent and reproducible procedures a priori to integrate often-heterogeneous evidence. Current weight-of-evidence (WoE) approaches for effects or hazard assessment tend to conflate aspects of the assessment of the quality of the data with the strength of the body of evidence as a whole. We take forward recent developments in the critical appraisal of the reliability and relevance of individual ecotoxicological studies as part of the effect or hazard assessment of prospective risk assessments and propose a streamlined WoE approach. The aim is to avoid overlap and double accounting of criteria used in reliability and relevance with that used in current WoE methods. The protection goals, problem formulation, and evaluation process need to be clarified at the outset. The data are first integrated according to lines of evidence (LoEs), typically mechanistic insights (e.g., cellular, subcellular, genomic), in vivo experiments, and higher-tiered field or observational studies. Data are then plotted on the basis of both relevance and reliability scores or categories. This graphical approach provides a means to visually assess and communicate the credibility (reliability and relevance of available individual studies), quantity, diversity, and consistency of the evidence. In addition, the external coherence of the body of evidence needs to be considered. The final step in the process is to derive an expression of the confidence in the conclusions of integrating the information considering these 5 aspects in the context of remaining uncertainties. We suggest that this streamlined approach to WoE for the effects or hazard characterization should facilitate reproducible and transparent assessments of data across different regulatory requirements. Integr Environ Assess Manag 2017;13:573-579. © 2017 The Authors. Integrated Environmental Assessment and Management published by Wiley Periodicals, Inc. on behalf of Society of Environmental Toxicology & Chemistry (SETAC).
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Ecotoxicología , Política Ambiental , Contaminantes Ambientales/toxicidad , Contaminación Ambiental/legislación & jurisprudencia , Toma de Decisiones , Exposición a Riesgos Ambientales , Contaminantes Ambientales/normas , Medición de Riesgo , Gestión de RiesgosRESUMEN
A species sensitivity distribution (SSD) is a probability model of the variation of species sensitivities to a stressor, in particular chemical exposure. The SSD approach has been used as a decision support tool in environmental protection and management since the 1980s, and the ecotoxicological, statistical, and regulatory basis and applications continue to evolve. This article summarizes the findings of a 2014 workshop held by the European Centre for Toxicology and Ecotoxicology of Chemicals and the UK Environment Agency in Amsterdam, The Netherlands, on the ecological relevance, statistical basis, and regulatory applications of SSDs. An array of research recommendations categorized under the topical areas of use of SSDs, ecological considerations, guideline considerations, method development and validation, toxicity data, mechanistic understanding, and uncertainty were identified and prioritized. A rationale for the most critical research needs identified in the workshop is provided. The workshop reviewed the technical basis and historical development and application of SSDs, described approaches to estimating generic and scenario-specific SSD-based thresholds, evaluated utility and application of SSDs as diagnostic tools, and presented new statistical approaches to formulate SSDs. Collectively, these address many of the research needs to expand and improve their application. The highest priority work, from a pragmatic regulatory point of view, is to develop a guidance of best practices that could act as a basis for global harmonization and discussions regarding the SSD methodology and tools. Integr Environ Assess Manag 2017;13:664-674. © 2016 SETAC.
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Organismos Acuáticos , Ecotoxicología/métodos , Monitoreo del Ambiente/métodos , Pruebas de Toxicidad/métodos , Biota , Conservación de los Recursos Naturales , Ecología , Monitoreo del Ambiente/normas , Monitoreo del Ambiente/estadística & datos numéricos , Países Bajos , Medición de Riesgo/métodos , Sensibilidad y Especificidad , Pruebas de Toxicidad/normas , Incertidumbre , Contaminantes Químicos del AguaRESUMEN
This critical review examines the definition and implementation of environmental protection goals for chemicals in current European Union (EU) legislation, guidelines, and international agreements to which EU countries are party. The European chemical industry is highly regulated, and prospective environmental risk assessments (ERAs) are tailored for different classes of chemical, according to their specific hazards, uses, and environmental exposure profiles. However, environmental protection goals are often highly generic, requiring the prevention of "unacceptable" or "adverse" impacts on "biodiversity" and "ecosystems" or the "environment as a whole." This review aims to highlight working examples, challenges, solutions, and best practices for defining specific protection goals (SPGs), which are seen to be essential for refining and improving ERA. Specific protection goals hinge on discerning acceptable versus unacceptable adverse effects on the key attributes of relevant, sensitive ecological entities (ranging from organisms to ecosystems). Some isolated examples of SPGs for terrestrial and aquatic biota can be found in prospective ERA guidance for plant protection products (PPPs). However, SPGs are generally limited to environmental or nature legislation that requires environmental monitoring and retrospective ERA. This limitation is due mainly to the availability of baselines, which define acceptable versus unacceptable environmental effects on the key attributes of sentinel species, populations and/or communities, such as reproductive status, abundance, or diversity. Nevertheless, very few regulatory case examples exist in which SPGs incorporate effect magnitude, spatial extent, and temporal duration. We conclude that more holistic approaches are needed for defining SPGs, particularly with respect to protecting population sustainability, ecosystem function, and integrity, which are implicit in generic protection goals and explicit in the International Programme for Chemical Safety (IPCS) definition of "adverse effect." A possible solution, which the chemical industry is currently assessing, is wider application of the ecosystem services approach proposed by the European Food Safety Authority (EFSA) for the risk assessment of PPPs. Integr Environ Assess Manag 2017;13:17-37. © 2016 SETAC.
Asunto(s)
Monitoreo del Ambiente/normas , Biodiversidad , Conservación de los Recursos Naturales , Ecosistema , Ecotoxicología , Exposición a Riesgos Ambientales/estadística & datos numéricos , Monitoreo del Ambiente/métodos , Contaminantes Ambientales , Unión Europea , Inocuidad de los Alimentos , Medición de Riesgo/normasRESUMEN
Systematic review (SR) is a rigorous, protocol-driven approach designed to minimise error and bias when summarising the body of research evidence relevant to a specific scientific question. Taking as a comparator the use of SR in synthesising research in healthcare, we argue that SR methods could also pave the way for a "step change" in the transparency, objectivity and communication of chemical risk assessments (CRA) in Europe and elsewhere. We suggest that current controversies around the safety of certain chemicals are partly due to limitations in current CRA procedures which have contributed to ambiguity about the health risks posed by these substances. We present an overview of how SR methods can be applied to the assessment of risks from chemicals, and indicate how challenges in adapting SR methods from healthcare research to the CRA context might be overcome. Regarding the latter, we report the outcomes from a workshop exploring how to increase uptake of SR methods, attended by experts representing a wide range of fields related to chemical toxicology, risk analysis and SR. Priorities which were identified include: the conduct of CRA-focused prototype SRs; the development of a recognised standard of reporting and conduct for SRs in toxicology and CRA; and establishing a network to facilitate research, communication and training in SR methods. We see this paper as a milestone in the creation of a research climate that fosters communication between experts in CRA and SR and facilitates wider uptake of SR methods into CRA.
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Medición de Riesgo , Revisiones Sistemáticas como Asunto , Animales , Humanos , Europa (Continente) , Sustancias Peligrosas/toxicidad , Medición de Riesgo/métodosRESUMEN
Toxicological research in the 1930s gave the first indications of the link between narcotic toxicity and the chemical activity of organic chemicals. More recently, chemical activity has been proposed as a novel exposure parameter that describes the fraction of saturation and that quantifies the potential for partitioning and diffusive uptake. In the present study, more than 2000 acute and chronic algal, aquatic invertebrates and fish toxicity data, as well as water solubility and melting point values, were collected from a series of sources. The data were critically reviewed and grouped by mode of action (MoA). We considered 660 toxicity data to be of acceptable quality. The 328 data which applied to the 72 substances identified as MoA 1 were then evaluated within the activity-toxicity framework: EC50 and LC50 values for all three taxa correlated generally well with (subcooled) liquid solubilities. Acute toxicity was typically exerted within the chemical activity range of 0.01-0.1, whereas chronic toxicity was exerted in the range of 0.001-0.01. These results confirm that chemical activity has the potential to contribute to the determination, interpretation and prediction of toxicity to aquatic organisms. It also has the potential to enhance regulation of organic chemicals by linking results from laboratory tests, monitoring and modeling programs. The framework can provide an additional line of evidence for assessing aquatic toxicity, for improving the design of toxicity tests, reducing animal usage and addressing chemical mixtures.
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Organismos Acuáticos/efectos de los fármacos , Ecotoxicología , Compuestos Orgánicos/toxicidad , Animales , Exactitud de los Datos , Peces , Invertebrados/efectos de los fármacos , Modelos Teóricos , Análisis de Regresión , Estadística como Asunto , Pruebas de Toxicidad Aguda , Pruebas de Toxicidad CrónicaRESUMEN
The bioavailability of organic chemicals in soil and sediment is an important area of scientific investigation for environmental scientists, although this area of study remains only partially recognized by regulators and industries working in the environmental sector. Regulators have recently started to consider bioavailability within retrospective risk assessment frameworks for organic chemicals; by doing so, realistic decision-making with regard to polluted environments can be achieved, rather than relying on the traditional approach of using total-extractable concentrations. However, implementation remains difficult because scientific developments on bioavailability are not always translated into ready-to-use approaches for regulators. Similarly, bioavailability remains largely unexplored within prospective regulatory frameworks that address the approval and regulation of organic chemicals. This article discusses bioavailability concepts and methods, as well as possible pathways for the implementation of bioavailability into risk assessment and regulation; in addition, this article offers a simple, pragmatic and justifiable approach for use within retrospective and prospective risk assessment.
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Compuestos Orgánicos/química , Disponibilidad Biológica , Medición de Riesgo , Suelo/química , Contaminantes del Suelo/análisisRESUMEN
Tests with vertebrates are an integral part of environmental hazard identification and risk assessment of chemicals, plant protection products, pharmaceuticals, biocides, feed additives and effluents. These tests raise ethical and economic concerns and are considered as inappropriate for assessing all of the substances and effluents that require regulatory testing. Hence, there is a strong demand for replacement, reduction and refinement strategies and methods. However, until now alternative approaches have only rarely been used in regulatory settings. This review provides an overview on current regulations of chemicals and the requirements for animal tests in environmental hazard and risk assessment. It aims to highlight the potential areas for alternative approaches in environmental hazard identification and risk assessment. Perspectives and limitations of alternative approaches to animal tests using vertebrates in environmental toxicology, i.e. mainly fish and amphibians, are discussed. Free access to existing (proprietary) animal test data, availability of validated alternative methods and a practical implementation of conceptual approaches such as the Adverse Outcome Pathways and Integrated Testing Strategies were identified as major requirements towards the successful development and implementation of alternative approaches. Although this article focusses on European regulations, its considerations and conclusions are of global relevance.
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Alternativas a las Pruebas en Animales , Contaminantes Ambientales/toxicidad , Sustancias Peligrosas/toxicidad , Alternativas a las Pruebas en Animales/legislación & jurisprudencia , Alternativas a las Pruebas en Animales/métodos , Alternativas a las Pruebas en Animales/tendencias , Animales , Contaminantes Ambientales/química , Unión Europea , Regulación Gubernamental , Guías como Asunto , Sustancias Peligrosas/química , Proyectos de Investigación , Medición de RiesgoRESUMEN
To use and implement an assessment scheme for the evaluation of endocrine disrupting properties of chemicals in ecotoxicology, the types of effect need to be agreed. Effects that merit further consideration in this context should fulfil the following three criteria: caused by an endocrine mode of action, be adverse, and be relevant at the population level to reflect the protection goal of ecotoxicological assessments. Thereafter, a comparison of effect values, regardless of the causative mechanisms, should be made, firstly to determine if endocrine toxicity generates the lowest endpoint within a taxon, and secondly if it is the lowest endpoint compared to that of other taxa living in the same compartment. These comparisons inform on two levels of specificity and determine if endocrine-mediated side-effects determine the ecotoxicological profile of a chemical. Various quantitative measures for the assessment of potency are also presented, which could assist in determining how to handle substances in the risk assessment when a regulatory concern is identified. Finally, derogation criteria should be defined for compounds that were designed as endocrine disruptors for non-vertebrates and those for which there is 'negligible exposure'. This paper discusses and provides proposals on how to apply these concepts for assessment of substances.
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Ecotoxicología/métodos , Disruptores Endocrinos/toxicidad , Plaguicidas/toxicidad , Medición de Riesgo/métodos , Animales , Relación Dosis-Respuesta a Droga , Ecotoxicología/legislación & jurisprudencia , Ecotoxicología/tendencias , Disruptores Endocrinos/administración & dosificación , Exposición a Riesgos Ambientales , Unión Europea , Humanos , Agencias Internacionales , Legislación de Medicamentos , Plaguicidas/normas , Medición de Riesgo/legislación & jurisprudencia , Medición de Riesgo/tendencias , Terminología como AsuntoRESUMEN
The Sox family of transcription factors are involved in a variety of developmental processes including sex determination and gonadal differentiation. Sox19 is a particularly interesting member of this family that has been found only in fish, though mammals have a very diverged orthologue that is designated Sox15 and assigned to a different Sox family subgroup. Here we describe the cloning and characterisation of sox19 from the European sea bass (Dicentrarchus labrax), an important aquaculture species in which sex ratios skewed in favour of males are frequently encountered. The sea bass sox19 gene contains a single intron, encodes a protein of 309 amino acids, has multiple transcription start sites and may produce a truncated splice variant. Sox19 mRNA is present in many adult tissues, with the highest expression in the brain and gonads. Interestingly, the gene is strongly upregulated in the differentiation of the ovary but not the testis, suggesting a role in ovarian differentiation.
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Lubina/crecimiento & desarrollo , Lubina/genética , Proteínas de Peces/genética , Regulación del Desarrollo de la Expresión Génica , Factores de Transcripción SOX/genética , Diferenciación Sexual/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Femenino , Proteínas de Peces/química , Masculino , Ratones , Datos de Secuencia Molecular , Especificidad de Órganos , Ovario/crecimiento & desarrollo , Ovario/metabolismo , Factores de Transcripción SOX/química , Caracteres Sexuales , Testículo/crecimiento & desarrollo , Testículo/metabolismoRESUMEN
The European regulation on plant protection products (1107/2009) (EC, 2009a), the revisions to the biocides Directive (COM[2009]267) (EC, 2009b), and the regulation concerning chemicals (Regulation (EC) No. 1907/2006 'REACH') (EC.2006) only support the marketing and use of chemical products on the basis that they do not induce endocrine disruption in humans or wildlife species. In the absence of agreed 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 resulting ECETOC technical report (ECETOC, 2009a) and the associated workshop (ECETOC, 2009b) presented a science-based concept on how to identify endocrine activity and disrupting properties of chemicals for both human health and the environment. The synthesis of the technical report and the workshop report was published by the ECETOC task force (Bars et al., 2011a,b). Specific scientific criteria for the determination of endocrine activity and disrupting properties that integrate information from both regulatory (eco)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. However, to be able to discriminate chemicals with endocrine properties of low concern from those of higher concern (for regulatory purposes), the task force recognised that the concept needed further refinement. Following a discussion of the key factors at a second workshop of invited regulatory, academic and industry scientists (ECETOC, 2011), the task force developed further guidance, which is presented in this paper. For human health assessments these factors include the relevance to humans of the endocrine mechanism of toxicity, the specificity of the endocrine effects with respect to other potential toxic effects, the potency of the chemical to induce endocrine toxicity and consideration of exposure levels. For ecotoxicological assessments the key considerations include specificity and potency, but also extend to the consideration of population relevance and negligible exposure. It is intended that these complement and reinforce the approach originally described and previously published in this journal (Bars et al., 2011a,b).
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Control de Medicamentos y Narcóticos , Disruptores Endocrinos/toxicidad , Pruebas de Toxicidad/normas , Toxicología/normas , Comités Consultivos , Animales , Monitoreo del Ambiente , Unión Europea , Agencias Gubernamentales , Regulación Gubernamental , Guías como Asunto , Humanos , Agencias Internacionales , Medición de Riesgo , Toxicología/legislación & jurisprudenciaRESUMEN
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.
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Disruptores Endocrinos/toxicidad , Pruebas de Toxicidad/normas , Toxicología/normas , Comités Consultivos , Animales , Ecotoxicología/legislación & jurisprudencia , Ecotoxicología/normas , Europa (Continente) , Regulación Gubernamental , Guías como Asunto , Humanos , Agencias Internacionales , Medición de Riesgo , Toxicología/legislación & jurisprudenciaRESUMEN
Fish full life cycle (FFLC) tests are increasingly required in the ecotoxicological assessment of endocrine active substances. However, FFLC tests have not been internationally standardized or validated, and it is currently unclear how such tests should best be designed to provide statistically sound and ecologically relevant results. This study describes how the technique of multi-criteria decision analysis (MCDA) was used to elicit the views of fish ecologists, aquatic ecotoxicologists and statisticians on optimal experimental designs for assessing the effects of endocrine active chemicals on fish. In MCDA qualitative criteria (that can be valued, but not quantified) and quantitative criteria can be used in a structured decision-making process. The aim of the present application of MCDA is to present a logical means of collating both data and expert opinions on the best way to focus FFLC tests on endocrine active substances. The analyses are presented to demonstrate how MCDA can be used in this context. Each of 3 workgroups focused on 1 of 3 species: fathead minnow (Pimephales promelas), Japanese medaka (Oryzias latipes), and zebrafish (Danio rerio). Test endpoints (e.g., fecundity, growth, gonadal histopathology) were scored for each species for various desirable features such as statistical power and ecological relevance, with the importance of these features determined by assigning weights to them, using a swing weighting procedure. The endpoint F1 fertilization success consistently emerged as a preferred option for all species. In addition, some endpoints scored highly in particular species, such as development of secondary sexual characteristics (fathead minnow) and sex ratio (zebrafish). Other endpoints such as hatching success ranked relatively highly and should be considered as useful endpoints to measure in tests with any of the fish species. MCDA also indicated relatively less preferred endpoints in fish life cycle tests. For example, intensive histopathology consistently ranked low, as did measurement of diagnostic biomarkers, such as vitellogenin, most likely due to the high costs of these methods or their limited ecological relevance. Life cycle tests typically do not focus on identifying toxic modes and/or mechanisms of action, but rather, single chemical concentration-response relationships for endpoints (e.g., survival, growth, reproduction) that can be translated into evaluation of risk. It is, therefore, likely to be an inefficient use of limited resources to measure these mechanism-specific endpoints in life cycle tests, unless the value of such endpoints for answering particular questions justifies their integration in specific case studies.
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Técnicas de Apoyo para la Decisión , Ecotoxicología/métodos , Disruptores Endocrinos/toxicidad , Determinación de Punto Final/métodos , Peces/crecimiento & desarrollo , Estadios del Ciclo de Vida/efectos de los fármacos , Pruebas de Toxicidad/métodos , Animales , Femenino , MasculinoRESUMEN
Animal alternatives research has historically focused on human safety assessments and has only recently been extended to environmental testing. This is particularly for those assays that involve the use of fish. A number of alternatives are being pursued by the scientific community including the fish embryo toxicity (FET) test, a proposed replacement alternative to the acute fish test. Discussion of the FET methodology and its application in environmental assessments on a global level was needed. With this emerging issue in mind, the ILSI Health and Environmental Sciences Institute (HESI) and the European Centre for Ecotoxicology and Toxicology of Chemicals (ECETOC) held an International Workshop on the Application of the Fish Embryo Test as an Animal Alternative Method in Hazard and Risk Assessment and Scientific Research in March, 2008. The workshop included approximately 40 scientists and regulators representing government, industry, academia, and non-governmental organizations from North America, Europe, and Asia. The goal was to review the state of the science regarding the investigation of fish embryonic tests, pain and distress in fish, emerging approaches utilizing fish embryos, and the use of fish embryo toxicity test data in various types of environmental assessments (e.g., hazard, risk, effluent, and classification and labeling of chemicals). Some specific key outcomes included agreement that risk assessors need fish data for decision-making, that extending the FET to include eluethereombryos was desirable, that relevant endpoints are being used, and that additional endpoints could facilitate additional uses beyond acute toxicity testing. The FET was, however, not yet considered validated sensu OECD. An important action step will be to provide guidance on how all fish tests can be used to assess chemical hazard and to harmonize the diverse terminology used in test guidelines adopted over the past decades. Use of the FET in context of effluent assessments was considered and it is not known if fish embryos are sufficiently sensitive for consideration as a surrogate to the sub-chronic 7-day larval fish growth and survival test used in the United States, for example. Addressing these needs by via workshops, research, and additional data reviews were identified for future action by scientists and regulators.
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Alternativas a las Pruebas en Animales/métodos , Embrión no Mamífero/efectos de los fármacos , Monitoreo del Ambiente/métodos , Peces/embriología , Pruebas de Toxicidad/métodos , Animales , Medición de Riesgo/métodosRESUMEN
To test the hypothesis that THs play an important role in the larval to juvenile transition in the marine teleost model, sea bream (Sparus auratus), key elements of the thyroid axis were analysed during development. Specific RT-PCR and Taqman quantitative RT-PCR were established and used to measure sea bream iodothyronine deiodinases and thyroid hormone receptor (TR) genes, respectively. Expression of deiodinases genes (D1 and D2) which encode enzymes producing T3, TRs and T4 levels start to increase at 20-30 days post-hatch (dph; beginning of metamorphosis), peak at about 45 dph (climax) and decline to early larval levels after 90-100 dph (end of metamorphosis) when fish are fully formed juveniles. The profile of these different TH elements during sea bream development is strikingly similar to that observed during the TH driven metamorphosis of flatfish and suggests that THs play an analogous role in the larval to juvenile transition in this species and probably also in other pelagic teleosts. However, the effect of T3 treatment on deiodinases and TR transcript abundance in sea bream is not as clear cut as in larval flatfish and tadpoles indicating divergence in the responsiveness of TH axis elements and highlighting the need for further studies of this axis during development of fish.
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Regulación del Desarrollo de la Expresión Génica , Yoduro Peroxidasa/genética , Yoduro Peroxidasa/metabolismo , Receptores de Hormona Tiroidea/genética , Receptores de Hormona Tiroidea/metabolismo , Dorada , Animales , Regulación del Desarrollo de la Expresión Génica/efectos de los fármacos , Regulación del Desarrollo de la Expresión Génica/genética , Larva/enzimología , Larva/genética , Larva/crecimiento & desarrollo , Larva/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Dorada/genética , Dorada/crecimiento & desarrollo , Dorada/metabolismo , Tiroxina/genética , Triyodotironina/genética , Triyodotironina/farmacologíaRESUMEN
Sox genes participate in several developmental processes, including sex determination and differentiation. In this study, the genomic structure of sox17 was characterized in the sea bass (sb). Two transcripts, one producing a normal protein (sb Sox17) and another producing a truncated protein (sb t-Sox17) were detected. A third, novel transcript, originated by intron retention (sb i-sox17) was also observed. Sb sox17 was widely distributed, whereas sb i-sox17 was predominantly found in skin and brain. In gonads, sb sox17 expression first increased at 150 days of age, coinciding with the onset of sex differentiation. At 250 days and onwards, sb sox17 expression was significantly higher in females, and mRNA levels correlated with those of gonadal aromatase. Thus, this study provides the first evidence for the presence of alternative splicing by intron retention in a Sox17 gene, and for sex-related differences in expression, implicating sox17 in ovarian development and function in fish.
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Lubina/genética , Factores de Transcripción SOXF/genética , Diferenciación Sexual/genética , Secuencia de Aminoácidos , Animales , Aromatasa/genética , Aromatasa/metabolismo , Secuencia de Bases , Secuencia Conservada , Femenino , Perfilación de la Expresión Génica , Regulación de la Expresión Génica , Gónadas/enzimología , Masculino , Datos de Secuencia Molecular , Filogenia , Regiones Promotoras Genéticas/genética , ARN Mensajero/genética , ARN Mensajero/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Factores de Transcripción SOXF/metabolismo , Alineación de Secuencia , Temperatura , Vertebrados/genéticaRESUMEN
Flatfish such as the Atlantic halibut (Hippoglossus hippoglossus) undergo a dramatic metamorphosis that transforms the pelagic, symmetric larva into a benthic, cranially asymmetric juvenile. In common with amphibian metamorphosis, flatfish metamorphosis is under endocrine control with thyroid hormones being particularly important. In this report we confirm that tri-iodothyronine (T(3)) levels peak at metamorphic climax during halibut metamorphosis. Moreover, we have isolated cDNA clones of TRalpha and TRbeta genes and confirmed the presence in halibut of two TRalpha isoforms (representing the products of distinct genes) and two TRbeta isoforms (generated from a single gene by alternative splicing). Real-time PCR was used to assess expression of these genes during metamorphosis. TRbeta shows the most dramatic expression profile, with a peak occurring during metamorphic climax.