Final Report Summary - EUROECOTOX (European network for alternative testing strategies in ecotoxicology)
Executive summary:
EUROECOTOX is a two-year coordination action funded by the European Commission (EC)'s Seventh Framework Programme (FP7) that has had as main objective supporting and accelerating the development and use of alternative methods in ecotoxicology and strategies in Europe in order to replace, reduce or refine (3Rs) the use of animals, particularly those used for regulatory testing.
To accomplish its objective, the coordinating action has created the EUROECOTOX network (http://www.euroecotox.eu) serving as an umbrella to join researchers, regulators, test validators, end users and other European stakeholders with interests in advancing the 3Rs in the field of environmental risk assessment.
During the two years of the coordination action (2010-2012), EUROECOTOX has aimed to work towards:
- determining the state of the art of alternative and integrating testing approaches in Europe;
- promoting the integration, networking of efforts and the exchange of knowledge, data and expertise between all the members of the network;
- identifying and promoting new alternative ecotoxicity methods actually under research;
- promoting dialogue with validating agencies and regulators;
- dissemination of best practices for the overall process of developing regulatory valid alternative ecotoxicity tests and methods.
Major results of EUROECOTOX that will have a positive impact on the acceleration of new testing methods and thus, contribute to a future reduction in the use of animals, are outlined below:
- The creation of the European network for alternative testing strategies in ecotoxicology (EUROECOTOX).
- The EUROECOTOX online database, with up to information and data about tests methods and researchers working on the development and validation of these test methods. (http://projects.cba.muni.cz/euroecotox/).
- Publication of the EUROECOTOXREPORT as a reference book on ?Alternative approaches for ecotoxicological safety testing in Europe.
The EUROECOTOX network will continue its work further on the FP7 funding, with the support from the European Centre for Ecotoxicology and Toxicology of Chemicals (ECETOC), member of the consortium, who will take over the administration and maintenance of the EUROECOTOX network and provide support for the organisation of future activities by the network members.
Project context and objectives:
EUROECOTOX is a coordination action, funded by the FP7-ENV-2010 programme, which aims at supporting and accelerating the development and use of alternative methods in ecotoxicology and strategies in Europe in order to replace, reduce or refine (3Rs) the use of animals, particularly those used for regulatory testing. To accomplish these aims the EUROECOTOX NETWORK has been launched (http://www.euroecotox.eu). The network provides researchers, regulators, test validators, end users and other European stakeholders with interests in advancing the 3Rs in ecotoxicology and allows to promote the validation and regulatory acceptance of alternative methods in environmental risk assessment. The network is also open to non-European members, as it is important to globally address the validation and regulatory acceptance of alternative tests methods in ecotoxicology.
Current legislation in European and other industrial countries requires submission of appropriate data for risk assessment for the registration of new chemicals, pesticides, biocides and pharmaceuticals. These data include information on the toxicity at various trophic levels and require testing with vertebrates, in particular with fish and birds. As a consequence of the new programme of the European chemical policy on registration, evaluation and authorisation of chemicals (REACH) which came into force on June 2007, the number of animal experiments (mainly with fish) for environmental risk assessment is expected to increase remarkably within the next years.
Together with the need to increase the environmental safety of chemicals, animal welfare is the subject of significant societal ethical concerns and there is a strong demand to refine, reduce and replace animals by using alternative test methods. Thus, the use of animals such as fish, for environmental hazard and risk assessment testing, presents a dilemma about balancing the need to ensure these assessments are scientifically valid against concerns about animal testing.
EUROECOTOX major goal has been to contribute to accelerate the validation and regulatory acceptance of alternative test methods for ecotoxicity testing and environmental risk assessment and thus helping reduce the number of animals used for the safety assessment of chemicals. As a coordination action, EUROECOTOX has not carried out research but has had the goal of promoting and developing activities that support the coordination and networking among all stakeholders active in the field of alternative test methods in ecotoxicology.
Main objectives of the coordination action have been:
- determining the State of the Art of alternative and integrating testing approaches in Europe;
- promoting the integration, networking of efforts and the exchange of knowledge, data and expertise between all the members of the network;
- identifying and promoting new alternative ecotoxicity methods actually under research;
- networking with ongoing FP projects working in the same area;
- promoting dialogue with validating agencies and regulators;
- dissemination of best practices for the overall process of developing regulatory valid alternative ecotoxicity tests and methods.
The EUROECOTOX Network has been promoted and coordinated by a European consortium of eight organisations with an important scientific background in environmental toxicology and alternative testing, originating from academia, research centres and industry. Consortium members organisations are the SME ZF Biolabs SL (Spain) coordinating the project, the Helmholtz Centre for Environmental research UFZ (Germany), The Swiss Federal Institute of Aquatic Science and Technology -Eawag (Switzerland), the Flemish Institute for Technological Research & Centre for Advanced Research and Development of Alternative Methods VITO/CARDAM (Belgium), the University of Heidelberg (Germany), NOTOX B.V. (The Netherlands), European Centre for Ecotoxicology and Toxicology of Chemicals ? ECETOC (Belgium) and Masaryk University, Research Centre for Toxic Compounds in the Environment -RECETOX (Czech Republic).
The consortium is supported by an Advisory Board formed by representatives of relevant European and international institutions committed to the promotion, use, validation and regulatory acceptance of alternative tests and testing strategies: the joint research centre's Institute for health and consumer protection (IHCP / ECVAM), the European Chemical Industry Council (CEFIC), the Organisation for Economic Cooperation and Development (OECD), ILSI Health and Environmental Sciences Institute (HESI), the European Consensus-Platform for Alternatives (ECOPA), and the National Centre for the replacement, refinement and reduction of animals in research (NC3Rs).
The EUROECOTOX network will continue to exist administered by the EUROECOTOX partner ECETOC, who will take over the administration and maintenance of the EUROECOTOX network and provide support for the organisation of future activities by the network members.
The project started on the first of December of 2010 and has run up to the 30 November of 2012. During this two years of the project, the following activities have been carried out successfully:
- Launching of the project and of the EUROECOTOX network. As part of the dissemination work, we contacted research centres, university, industry, governmental environmental agencies, regulators, companies providing toxicology research services, animal welfare organisations, active in the field of alternative test methods and ecotoxicology to invite them to participate in EUROECOTOX. As for November 2012, the network has incorporated a total of 191 members.
- Set up of the EUROECOTOX website (http://www.euroecotox.eu). The EUROECOTOX site seeks to be the main gathering point for all interested parties in the field of alternatives to animal tests in ecotoxicological risk assessment. It is also the main dissemination tool of the project.
- Mapping of European research capacities on alternative ecotoxicological test methods. In order to obtain an actual view on European research capacities related to the development, validation and use of alternative ecotoxicological methods, a review of research groups, alternative toxicity methods under development and close to validation, major stakeholders, funding agencies, platforms and networks working in this area was carried out with the aim to provide a full state of the art analysis of the actual situation in Europe. The mapping has identified so far a total of 106 leading experts who, based mainly on research profile and scientific publications, have been identified as contributing significantly to the development of alternative assays. As for alternative test methods (in different stages of development), the mapping has gathered information on non-testing approaches, experimental alternative assays and advanced sublethal approaches addressing MOA (Mechanisms of Action).
- Design and development of a database reflecting research groups and test methods identified during the mapping exercise. Data on research groups and test methods identified during the mapping work have been organised in an online database accessible through the EUROECOXOTOX website. The EUROECOTOX database of bioassays (direct link: http://projects.cba.muni.cz/euroecotox/) has been set up with the aim to assist the identification of alternative methods to support the development, testing, application, and validation of alternatives to the use of animals in ecotoxicology testing. The database will regularly be updated when new researchers and methods are identified. In the present time (as of 29 November 2011) the database contains 85 alternative assay records, 22 standard tests (OECD, ISO and others) and 118 expert contacts. The database has been prepared in a flexible format, which allows easy modifications according to potential new requests.
- Organisation of the EUROECOTOX expert meeting in Leipzig (Germany). A two day expert meeting was organised during the 17-18 October in Leipzig (Germany) hosted by the Helmholtz Centre for Environmental Research UFZ. The meeting, attended by the members of the consortium, the advisory board and 28 invited experts, was structured in platform presentations and two blocks of break-out sessions: The first aimed to discuss alternatives approaches to main tests actually performed for environmental risk assessment and the second sessions focused on reviewing and discussing different alternative approaches. A final discussion summarised the results and discussed measures and future activities.
- Organisation of the first European Conference on the replacement, reduction and refinement of animal experiments in ecotoxicology held at eawag, Dübendorf, Switzerland, from 28-29 June 2012. The conference aimed to provide a platform for young scientists and experts from academia, industry and regulation in the field of replacement, reduction and refinement of animal tests used in environmental risk assessment. It was focused on the current state and future directions of the development, implementation and application of the 3Rs, from bench to acceptance. The programme, based on 8 invited key note speakers and presentations from submitted abstracts, consisted of four main blocks: (i) Experimental approaches, (ii) towards integration and implementation, (iii) perspectives and initiatives towards the 3Rs, (iv) Computational approaches and (v) a final plenary discussion about the future of alternative methods in ecotoxicology. Based on submitted abstracts, 26 participants have been invited to give a plenary presentation. During the open registration phase, additional 35 persons registered. Together with the consortium members, a total of 69 participants were registered. About 62 % participants were from academia, 22 % from industry and 16 % from regulators and stakeholders.
- Preparation of the EUROECOTOX report which is the main output of the EUROECOTOX project. This report, based on the work done by the consortium during the first year (the mapping work, the expert meeting and the analysis of main gaps and bottlenecks), seeks to be the most up to date reference work on alternative approaches to replace or reduce the use of animals in ecotoxicological safety testing in Europe.
Project results:
Funded under EC's FP7, the EUROECOTOX network has worked towards identifying key areas for the development of alternatives to vertebrate testing for environmental risk assessment in the European Union, and where appropriate with a global perspective. Being a coordination action, EUROECOTOX was not planned to carry out Research and development (R&D) activities but to coordinate existing research and promote discussion and exchange of knowledge between stakeholders.
Major results of EUROECOTOX that are expected to have a positive impact on the acceleration of new testing methods and thus, contribute to a future reduction in the use of animals, are described below.
1. Creation of the EUROECOTOX network
The creation of the European Network for alternative testing strategies in ecotoxicology (EUROECOTOX), open to all stakeholders (researchers, industry, regulators, test validating authorities, administration) active in alternative testing methods has been one of the objectives of the action and at the same time on of the important results. It is the tool that supports coordination and discussion and facilitates dissemination of results. The network has its own website (http://www.euroecotox.eu) and it will continue to exist supported by ECETOC, the European Centre for Ecotoxicology and Toxicology of Chemicals, partner of the consortium.
The number of network members as for the December 14th 2012 is 194 persons from 35 different countries.
The geographical distribution of members of the EUROECOTOX Network ins mainly European, with 79 % of their members coming from European countries, 13 % from America, 7 % from Asia and 1 % from Africa.
Although EUROECOTOX has been planned as a European network for alternative testing strategies in ecotoxicology, we have considered opening participation to interested non-European organisations, as it is important to globally address the development and validation of new test methods for ecotoxicity testing.
The distribution of memberships as for the type of organisations to which members of the network belong is as follows: Universities (42 %), research centre / association (29 %), Service providing companies (11 %), Governmental bodies (8 %) and industry (10 %).
2. Mapping of European research capacities on alternative ecotoxicological test methods
In order to obtain a broad view on European research capacities related to the development, validation and use of alternative ecotoxicological methods, a review of research groups, alternative toxicity methods under development and close to validation, major stakeholders, funding agencies, platforms and networks working in this area was carried out with the aim to provide a full state of the art analysis of the actual situation in Europe. The mapping has identified a total of 106 leading experts (mostly coming from universities and research centres) who, based mainly on research profile and scientific publications, have been identified as contributing significantly to the development of alternative assays. So far, 75 additional experts involved in ecotoxicological research have also been identified and included in the mapping. They may not necessarily be leaders with respect to heading established groups and / or developing novel assays. However, they may actively contribute by the use, application and / or validation of the alternative approaches. As for alternative test methods (in different stages of development), the mapping has gathered information on (i) non-testing approaches, (ii) experimental alternative assays and (iii) advanced sublethal approaches addressing mechanisms of action.
The mapping has also gathered information on main European stakeholders and relevant international R&D and institutional activities in alternative ecotoxicological test methods and strategies. A review on current European legislation requiring ecotoxicity testing with vertebrate animals was as well part of the mapping work.
A written report with the results of the mapping has been prepared and is available for downloading from the EUROCOTOX website. 3. the EUROECOTOX database of bioassays
The EUROECOTOX database has been set up with the aim to assist the identification of alternative methods to support the development, testing, application, and validation of alternatives to the use of animals in ecotoxicology testing. The database will regularly be updated when new researchers and methods are identified. In the present time (as of November 29, 2012) the database contains 85 alternative assay records, 22 standard tests (OECD, ISO and others) and 118 expert contacts. The database has been prepared in a flexible format, which allows easy modifications according to potential new requests. The database will be regularly updated.
4. The EUROECOTOX expert meeting
The EUROECOTOX meeting took place in Leipzig (Germany) during the 17-18 October 2011, hosted by the Helmholtz Centre for Environmental Research (P2). Together with the 8 members of the consortium and 6 members of the EUROECOTOX Advisory board, 29 other leading experts participated in the plenary meetings and the breakout sessions, representing universities, research institutes, industry and governmental regulatory organisations such as the OECD and the European Chemical Agency (ECHA).
Scope of the meeting
Animal experiments play an integral role in current environmental risk assessment for the registration of chemicals, pesticides, biocides, feed additives, pharmaceuticals and testing of whole effluents. As with risk assessment in human toxicology, there is a strong societal demand to replace, reduce or refine the animal experiments performed in order to protect the environment. For this purpose, a two-day expert meeting was held to provide a forum for discussion of actual regulatory requirements for ecotoxicity testing, of novel strategies and approaches to reduce animal testing, and of identification of bottlenecks for validating new methods and measures to accelerate development and validation of alternatives in environmental risk assessment. Another specific objective of the meeting was the preparation of a publicly available report and suggestions for future research needs in this area.
This two-day expert meeting served as a forum for discussion of actual regulatory requirements for ecotoxicity testing, of novel strategies and approaches to reduce animal testing, and of identification of bottlenecks for validating new methods and measures to accelerate development and validation of alternatives in environmental risk assessment.
Organisation
The meeting was organised by the Helmholtz Centre for Environmental Research, with Dr. Stefan Scholz, Department of Bioanalytical Ecotoxicology as the responsible organiser. The preparation of the meeting was supported by members of the consortium, who actively participated with suggestions for invited experts, platform lectures and breakout session topics.
The meeting was held in the KUBUS building, a modern facility at the UFZ characterised by a flexible architecture. Financially support was provided by the European Union via funds to the EUROECOTOX consortium. In addition, the Helmholtz Centre for Environmental Research had sponsored an evening reception for the participants in a scenic local restaurant.
Structure
The expert meeting consisted of platform presentations and two blocks of parallel breakout sessions. The platform presentation were dedicated to give the participants a general overview on specific approaches and problems associated with alternatives to animal experiments in environmental risk assessment such as non-testing approaches, industry perspectives, toxicogenomics and validation (see details below). The breakout sessions were intended for discussions of specific topics in detail in order to elucidate the state of the art, limitations of alternative approaches and research needs, and to provide policy/research recommendations. Each breakout session was prepared and led by one of the invited experts with particular experience in the respective field and was supported by a rapporteur from the EUROECOTOX consortium.
Breakout sessions
Four Breakout session on Tests performed for environmental risk assessment were organised:
- Session I: acute toxicity: fish and bird.
- Session II: Chronic toxicity fish and bird;
-Session III: endocrine disruption fish and amphibian.
- Session IV Bioaccumulation fish
Discussion points for each of this this breakout session:
- state of the art and current use of animal tests;
- available alternative assays/approaches, requirements for alternatives;
- provide and discuss examples (including own results);
- limitations of alternatives and priorities for validation;
- future research needs;
- regulatory acceptance;
- recommendations.
A second round of four breakout session on ?Alternative approaches were organised: Session V: QSAR.
Session VI: Non-testing approaches (e.g. refinement, extrapolation, data analysis, integrated testing, exposure waiving); Session VII ALTERNATIVE TEST SYSTEMS and Session VIII: SUBLETHAL/ ALTERNATIVE ENDPOINTS. Main discussion points for breakout sessions:
- state of the art of alternative approaches;
- provide and discuss examples (including own results);
- limitations and advantages of each approach;
- future research needs/priorities;
- regulatory acceptance;
- recommendations.
Results
Based on the expert meeting a conceptual paper titled 'A European perspective of alternatives to animal testing for environmental hazard identification and risk assessment' has been prepared with up to 30 contributing authors. The paper is currently under internal revision considering the comments of participating experts. It is planned to submit the paper the Journal of Regulatory Toxicology and Pharmacology within spring 2013.
Future activities resulting from the expert meeting
The expert meeting and its discussions were highly appreciated by the participants. The overall identification of limitations and future research need were supported by most of the participants and a high consensus among basic scientists and representatives of industry, stakeholders and regulatory bodies was recognised. Therefore, participants have agreed to jointly prepare a conceptual paper on the European perspective of alternatives to animal experiments for environmental risk assessment. A first draft of this multiple-co-author paper is already available. The manuscript is foreseen to be submitted soon to the Journal of Regulatory Pharmacology and Toxicology and would be published as open access, available from the EUROECOTOX web page or the journal's online repository, respectively.
5. First European conference on the replacement, reduction and refinement of animal experiments in ecotoxicology
EUROECOTOX organised the first. European Conference on the replacement, reduction and refinement of animal experiments in ecotoxicology. I was held at Eawag, Dübendorf, Switzerland, from 28-29 June 2012. The conference aimed to provide a platform for young scientists and experts from academia, industry and regulation in the field of replacement, reduction and refinement of animal tests used in environmental risk assessment. It was focused on the current state and future directions of the development, implementation and application of the 3Rs, from bench to acceptance.
The programme, based on invited key note speakers and presentations from submitted abstracts, consisted of four main blocks: (i) experimental approaches, (ii) towards integration and implementation, (iii) perspectives and initiatives towards the 3Rs, (iv) computational approaches and (v) a final plenary discussion about the future of alternative methods in ecotoxicology. The sections (i), (ii) and (iv) were stimulated by a key note presentation of an invited expert. A special key note lecture was given by Susanne Louhimies about the establishment of the 3Rs in European legislation on the first day in the evening. All presentations are available from the EUROCOTOX website.
Sponsored by ESTIV, the European society of toxicology in vitro, the conference was able to provide two students an award of 200 each to the best oral presentation and poster of a student (Master, Bachelor or PhD student), making thus attractive the participation of students in the conference. The jury consisted of four independent conference participants: Michelle Embry, Erika Sela, Marc Leonard and Nic Bury. The winner of the Best Young Scientist award for an oral presentation was given to Julita Stadnicka (Eawag) for her presentation Predicting chemical concentrations in a fish cell line based on measurements and models?. The award for the Young Scientist Best poster presentation was given to Melanie Knöbel (Eawag) for her poster ?Correlation of zebrafish embryo to fathead minnow acute toxicity: the relevance of physico?chemical properties, mode of toxic action, test duration and exposure concentration analysis.
6. First EUROECOTOX report: 'Alternative approaches to replace or reduce the use of animals in ecotoxicology'.
The report presents a comprehensive overview on the different aspects to be considered for developing alternatives to testing of animals in environmental risk assessment - ranging from an overview of regulations, different alternative approaches, future research needs and policy recommendations. It complements the already developed database of researchers and institutions active in the field which is available online.
It is the intention of the consortium to publish the report as book titled 'Alternatives to testing of animals in environmental risk assessment'. For this, the consortium has already submitted a book editing proposal to SETAC, (Society of environmental toxicology and chemistry), who will be willing in principle to edit and publish the book. It will be published both as open access PDF (for download from the SETAC press webpage) as well as a printed edition (for libraries, for convenient reading and use as a reference book). The first printed edition will produced with a number of 100 books. Based on the number of purchases we will order a second edition that may be financed from the revenues of the first printed edition. It is expected that the book could be available by the end of 2013, as it has to go through a peer review process before publishing it.
Support for the publishing costs will be provided in part through a grant from the EU Operational Programmes in the Czech Republic (via a European Social Fund), project 'Innovation and upgrade of the environmental education at Faculty of Science of Masaryk University' (Operational Programme OPVK: Education for Competitiveness, project No. CZ.1.07/2.2.00/15.0213)
Potential impact:
Current legislation in European and other industrial countries requires submission of appropriate data for risk assessment for the registration of new chemicals, pesticides, biocides and pharmaceuticals. These data include information on the toxicity at various trophic levels and require testing with vertebrates, in particular with fish and birds. As a consequence of the new program of REACH which came into force on June 2007, the number of animal experiments (mainly with fish) for environmental risk assessment is expected to increase remarkably within the next years.
Together with the need to increase the environmental safety of chemicals, there is a strong societal demand for reducing the number of animal tests by using alternative in vitro models. Animal welfare is the subject of significant societal ethical concerns and there is a strong European support to refine, reduce and replace animal use linked to regulations. Thus, the use of animals such as fish, for environmental hazard and risk assessment testing, presents a dilemma about balancing the need to ensure these assessments are scientifically valid against concerns about animal testing.
During the last two decades, substantial efforts have been made towards the development and international acceptance of alternative methods to safety studies using laboratory animals. An alternative toxicological test method is any method that can be used to replace, or reduce the use of animals for experimental purposes or refine procedures to make such experiments less painful of stressful (ECVAM).
EUROECOTOX aims to foster the exchange of knowledge, data, ideas and available European expertise to accelerate the R&D, (pre)validation and regulatory acceptance of alternatives. In particular, the EUROECOTOX network has worked towards promoting and supporting the intensification of alternatives for ecotoxicological testing.
EUROECOTOX has contributed to the reduction in the use of animals in ecotoxicity safety testing by promoting the exchange of knowledge and data and dissemination of novel alternative tests and methods among all European stakeholders via the EUROECOTOX networking activities carried out and through the network website. Main dissemination activities carried out are briefly commented here:
- two important meetings with experts and stakeholders we organised in Leipzig (Germany) and in Dubendorf (Switzerland);
- project newsletter with latest project news and focus articles were distributed by email to all network members;
- members of the consortium have participated in various scientific meetings and workshops presenting the network and promoting discussion and dialogue;
- a report describing in detail all dissemination activities has been presented as deliverable at the end of the project.
List of websites: http://www.euroecotox.eu
EUROECOTOX is a two-year coordination action funded by the European Commission (EC)'s Seventh Framework Programme (FP7) that has had as main objective supporting and accelerating the development and use of alternative methods in ecotoxicology and strategies in Europe in order to replace, reduce or refine (3Rs) the use of animals, particularly those used for regulatory testing.
To accomplish its objective, the coordinating action has created the EUROECOTOX network (http://www.euroecotox.eu) serving as an umbrella to join researchers, regulators, test validators, end users and other European stakeholders with interests in advancing the 3Rs in the field of environmental risk assessment.
During the two years of the coordination action (2010-2012), EUROECOTOX has aimed to work towards:
- determining the state of the art of alternative and integrating testing approaches in Europe;
- promoting the integration, networking of efforts and the exchange of knowledge, data and expertise between all the members of the network;
- identifying and promoting new alternative ecotoxicity methods actually under research;
- promoting dialogue with validating agencies and regulators;
- dissemination of best practices for the overall process of developing regulatory valid alternative ecotoxicity tests and methods.
Major results of EUROECOTOX that will have a positive impact on the acceleration of new testing methods and thus, contribute to a future reduction in the use of animals, are outlined below:
- The creation of the European network for alternative testing strategies in ecotoxicology (EUROECOTOX).
- The EUROECOTOX online database, with up to information and data about tests methods and researchers working on the development and validation of these test methods. (http://projects.cba.muni.cz/euroecotox/).
- Publication of the EUROECOTOXREPORT as a reference book on ?Alternative approaches for ecotoxicological safety testing in Europe.
The EUROECOTOX network will continue its work further on the FP7 funding, with the support from the European Centre for Ecotoxicology and Toxicology of Chemicals (ECETOC), member of the consortium, who will take over the administration and maintenance of the EUROECOTOX network and provide support for the organisation of future activities by the network members.
Project context and objectives:
EUROECOTOX is a coordination action, funded by the FP7-ENV-2010 programme, which aims at supporting and accelerating the development and use of alternative methods in ecotoxicology and strategies in Europe in order to replace, reduce or refine (3Rs) the use of animals, particularly those used for regulatory testing. To accomplish these aims the EUROECOTOX NETWORK has been launched (http://www.euroecotox.eu). The network provides researchers, regulators, test validators, end users and other European stakeholders with interests in advancing the 3Rs in ecotoxicology and allows to promote the validation and regulatory acceptance of alternative methods in environmental risk assessment. The network is also open to non-European members, as it is important to globally address the validation and regulatory acceptance of alternative tests methods in ecotoxicology.
Current legislation in European and other industrial countries requires submission of appropriate data for risk assessment for the registration of new chemicals, pesticides, biocides and pharmaceuticals. These data include information on the toxicity at various trophic levels and require testing with vertebrates, in particular with fish and birds. As a consequence of the new programme of the European chemical policy on registration, evaluation and authorisation of chemicals (REACH) which came into force on June 2007, the number of animal experiments (mainly with fish) for environmental risk assessment is expected to increase remarkably within the next years.
Together with the need to increase the environmental safety of chemicals, animal welfare is the subject of significant societal ethical concerns and there is a strong demand to refine, reduce and replace animals by using alternative test methods. Thus, the use of animals such as fish, for environmental hazard and risk assessment testing, presents a dilemma about balancing the need to ensure these assessments are scientifically valid against concerns about animal testing.
EUROECOTOX major goal has been to contribute to accelerate the validation and regulatory acceptance of alternative test methods for ecotoxicity testing and environmental risk assessment and thus helping reduce the number of animals used for the safety assessment of chemicals. As a coordination action, EUROECOTOX has not carried out research but has had the goal of promoting and developing activities that support the coordination and networking among all stakeholders active in the field of alternative test methods in ecotoxicology.
Main objectives of the coordination action have been:
- determining the State of the Art of alternative and integrating testing approaches in Europe;
- promoting the integration, networking of efforts and the exchange of knowledge, data and expertise between all the members of the network;
- identifying and promoting new alternative ecotoxicity methods actually under research;
- networking with ongoing FP projects working in the same area;
- promoting dialogue with validating agencies and regulators;
- dissemination of best practices for the overall process of developing regulatory valid alternative ecotoxicity tests and methods.
The EUROECOTOX Network has been promoted and coordinated by a European consortium of eight organisations with an important scientific background in environmental toxicology and alternative testing, originating from academia, research centres and industry. Consortium members organisations are the SME ZF Biolabs SL (Spain) coordinating the project, the Helmholtz Centre for Environmental research UFZ (Germany), The Swiss Federal Institute of Aquatic Science and Technology -Eawag (Switzerland), the Flemish Institute for Technological Research & Centre for Advanced Research and Development of Alternative Methods VITO/CARDAM (Belgium), the University of Heidelberg (Germany), NOTOX B.V. (The Netherlands), European Centre for Ecotoxicology and Toxicology of Chemicals ? ECETOC (Belgium) and Masaryk University, Research Centre for Toxic Compounds in the Environment -RECETOX (Czech Republic).
The consortium is supported by an Advisory Board formed by representatives of relevant European and international institutions committed to the promotion, use, validation and regulatory acceptance of alternative tests and testing strategies: the joint research centre's Institute for health and consumer protection (IHCP / ECVAM), the European Chemical Industry Council (CEFIC), the Organisation for Economic Cooperation and Development (OECD), ILSI Health and Environmental Sciences Institute (HESI), the European Consensus-Platform for Alternatives (ECOPA), and the National Centre for the replacement, refinement and reduction of animals in research (NC3Rs).
The EUROECOTOX network will continue to exist administered by the EUROECOTOX partner ECETOC, who will take over the administration and maintenance of the EUROECOTOX network and provide support for the organisation of future activities by the network members.
The project started on the first of December of 2010 and has run up to the 30 November of 2012. During this two years of the project, the following activities have been carried out successfully:
- Launching of the project and of the EUROECOTOX network. As part of the dissemination work, we contacted research centres, university, industry, governmental environmental agencies, regulators, companies providing toxicology research services, animal welfare organisations, active in the field of alternative test methods and ecotoxicology to invite them to participate in EUROECOTOX. As for November 2012, the network has incorporated a total of 191 members.
- Set up of the EUROECOTOX website (http://www.euroecotox.eu). The EUROECOTOX site seeks to be the main gathering point for all interested parties in the field of alternatives to animal tests in ecotoxicological risk assessment. It is also the main dissemination tool of the project.
- Mapping of European research capacities on alternative ecotoxicological test methods. In order to obtain an actual view on European research capacities related to the development, validation and use of alternative ecotoxicological methods, a review of research groups, alternative toxicity methods under development and close to validation, major stakeholders, funding agencies, platforms and networks working in this area was carried out with the aim to provide a full state of the art analysis of the actual situation in Europe. The mapping has identified so far a total of 106 leading experts who, based mainly on research profile and scientific publications, have been identified as contributing significantly to the development of alternative assays. As for alternative test methods (in different stages of development), the mapping has gathered information on non-testing approaches, experimental alternative assays and advanced sublethal approaches addressing MOA (Mechanisms of Action).
- Design and development of a database reflecting research groups and test methods identified during the mapping exercise. Data on research groups and test methods identified during the mapping work have been organised in an online database accessible through the EUROECOXOTOX website. The EUROECOTOX database of bioassays (direct link: http://projects.cba.muni.cz/euroecotox/) has been set up with the aim to assist the identification of alternative methods to support the development, testing, application, and validation of alternatives to the use of animals in ecotoxicology testing. The database will regularly be updated when new researchers and methods are identified. In the present time (as of 29 November 2011) the database contains 85 alternative assay records, 22 standard tests (OECD, ISO and others) and 118 expert contacts. The database has been prepared in a flexible format, which allows easy modifications according to potential new requests.
- Organisation of the EUROECOTOX expert meeting in Leipzig (Germany). A two day expert meeting was organised during the 17-18 October in Leipzig (Germany) hosted by the Helmholtz Centre for Environmental Research UFZ. The meeting, attended by the members of the consortium, the advisory board and 28 invited experts, was structured in platform presentations and two blocks of break-out sessions: The first aimed to discuss alternatives approaches to main tests actually performed for environmental risk assessment and the second sessions focused on reviewing and discussing different alternative approaches. A final discussion summarised the results and discussed measures and future activities.
- Organisation of the first European Conference on the replacement, reduction and refinement of animal experiments in ecotoxicology held at eawag, Dübendorf, Switzerland, from 28-29 June 2012. The conference aimed to provide a platform for young scientists and experts from academia, industry and regulation in the field of replacement, reduction and refinement of animal tests used in environmental risk assessment. It was focused on the current state and future directions of the development, implementation and application of the 3Rs, from bench to acceptance. The programme, based on 8 invited key note speakers and presentations from submitted abstracts, consisted of four main blocks: (i) Experimental approaches, (ii) towards integration and implementation, (iii) perspectives and initiatives towards the 3Rs, (iv) Computational approaches and (v) a final plenary discussion about the future of alternative methods in ecotoxicology. Based on submitted abstracts, 26 participants have been invited to give a plenary presentation. During the open registration phase, additional 35 persons registered. Together with the consortium members, a total of 69 participants were registered. About 62 % participants were from academia, 22 % from industry and 16 % from regulators and stakeholders.
- Preparation of the EUROECOTOX report which is the main output of the EUROECOTOX project. This report, based on the work done by the consortium during the first year (the mapping work, the expert meeting and the analysis of main gaps and bottlenecks), seeks to be the most up to date reference work on alternative approaches to replace or reduce the use of animals in ecotoxicological safety testing in Europe.
Project results:
Funded under EC's FP7, the EUROECOTOX network has worked towards identifying key areas for the development of alternatives to vertebrate testing for environmental risk assessment in the European Union, and where appropriate with a global perspective. Being a coordination action, EUROECOTOX was not planned to carry out Research and development (R&D) activities but to coordinate existing research and promote discussion and exchange of knowledge between stakeholders.
Major results of EUROECOTOX that are expected to have a positive impact on the acceleration of new testing methods and thus, contribute to a future reduction in the use of animals, are described below.
1. Creation of the EUROECOTOX network
The creation of the European Network for alternative testing strategies in ecotoxicology (EUROECOTOX), open to all stakeholders (researchers, industry, regulators, test validating authorities, administration) active in alternative testing methods has been one of the objectives of the action and at the same time on of the important results. It is the tool that supports coordination and discussion and facilitates dissemination of results. The network has its own website (http://www.euroecotox.eu) and it will continue to exist supported by ECETOC, the European Centre for Ecotoxicology and Toxicology of Chemicals, partner of the consortium.
The number of network members as for the December 14th 2012 is 194 persons from 35 different countries.
The geographical distribution of members of the EUROECOTOX Network ins mainly European, with 79 % of their members coming from European countries, 13 % from America, 7 % from Asia and 1 % from Africa.
Although EUROECOTOX has been planned as a European network for alternative testing strategies in ecotoxicology, we have considered opening participation to interested non-European organisations, as it is important to globally address the development and validation of new test methods for ecotoxicity testing.
The distribution of memberships as for the type of organisations to which members of the network belong is as follows: Universities (42 %), research centre / association (29 %), Service providing companies (11 %), Governmental bodies (8 %) and industry (10 %).
2. Mapping of European research capacities on alternative ecotoxicological test methods
In order to obtain a broad view on European research capacities related to the development, validation and use of alternative ecotoxicological methods, a review of research groups, alternative toxicity methods under development and close to validation, major stakeholders, funding agencies, platforms and networks working in this area was carried out with the aim to provide a full state of the art analysis of the actual situation in Europe. The mapping has identified a total of 106 leading experts (mostly coming from universities and research centres) who, based mainly on research profile and scientific publications, have been identified as contributing significantly to the development of alternative assays. So far, 75 additional experts involved in ecotoxicological research have also been identified and included in the mapping. They may not necessarily be leaders with respect to heading established groups and / or developing novel assays. However, they may actively contribute by the use, application and / or validation of the alternative approaches. As for alternative test methods (in different stages of development), the mapping has gathered information on (i) non-testing approaches, (ii) experimental alternative assays and (iii) advanced sublethal approaches addressing mechanisms of action.
The mapping has also gathered information on main European stakeholders and relevant international R&D and institutional activities in alternative ecotoxicological test methods and strategies. A review on current European legislation requiring ecotoxicity testing with vertebrate animals was as well part of the mapping work.
A written report with the results of the mapping has been prepared and is available for downloading from the EUROCOTOX website. 3. the EUROECOTOX database of bioassays
The EUROECOTOX database has been set up with the aim to assist the identification of alternative methods to support the development, testing, application, and validation of alternatives to the use of animals in ecotoxicology testing. The database will regularly be updated when new researchers and methods are identified. In the present time (as of November 29, 2012) the database contains 85 alternative assay records, 22 standard tests (OECD, ISO and others) and 118 expert contacts. The database has been prepared in a flexible format, which allows easy modifications according to potential new requests. The database will be regularly updated.
4. The EUROECOTOX expert meeting
The EUROECOTOX meeting took place in Leipzig (Germany) during the 17-18 October 2011, hosted by the Helmholtz Centre for Environmental Research (P2). Together with the 8 members of the consortium and 6 members of the EUROECOTOX Advisory board, 29 other leading experts participated in the plenary meetings and the breakout sessions, representing universities, research institutes, industry and governmental regulatory organisations such as the OECD and the European Chemical Agency (ECHA).
Scope of the meeting
Animal experiments play an integral role in current environmental risk assessment for the registration of chemicals, pesticides, biocides, feed additives, pharmaceuticals and testing of whole effluents. As with risk assessment in human toxicology, there is a strong societal demand to replace, reduce or refine the animal experiments performed in order to protect the environment. For this purpose, a two-day expert meeting was held to provide a forum for discussion of actual regulatory requirements for ecotoxicity testing, of novel strategies and approaches to reduce animal testing, and of identification of bottlenecks for validating new methods and measures to accelerate development and validation of alternatives in environmental risk assessment. Another specific objective of the meeting was the preparation of a publicly available report and suggestions for future research needs in this area.
This two-day expert meeting served as a forum for discussion of actual regulatory requirements for ecotoxicity testing, of novel strategies and approaches to reduce animal testing, and of identification of bottlenecks for validating new methods and measures to accelerate development and validation of alternatives in environmental risk assessment.
Organisation
The meeting was organised by the Helmholtz Centre for Environmental Research, with Dr. Stefan Scholz, Department of Bioanalytical Ecotoxicology as the responsible organiser. The preparation of the meeting was supported by members of the consortium, who actively participated with suggestions for invited experts, platform lectures and breakout session topics.
The meeting was held in the KUBUS building, a modern facility at the UFZ characterised by a flexible architecture. Financially support was provided by the European Union via funds to the EUROECOTOX consortium. In addition, the Helmholtz Centre for Environmental Research had sponsored an evening reception for the participants in a scenic local restaurant.
Structure
The expert meeting consisted of platform presentations and two blocks of parallel breakout sessions. The platform presentation were dedicated to give the participants a general overview on specific approaches and problems associated with alternatives to animal experiments in environmental risk assessment such as non-testing approaches, industry perspectives, toxicogenomics and validation (see details below). The breakout sessions were intended for discussions of specific topics in detail in order to elucidate the state of the art, limitations of alternative approaches and research needs, and to provide policy/research recommendations. Each breakout session was prepared and led by one of the invited experts with particular experience in the respective field and was supported by a rapporteur from the EUROECOTOX consortium.
Breakout sessions
Four Breakout session on Tests performed for environmental risk assessment were organised:
- Session I: acute toxicity: fish and bird.
- Session II: Chronic toxicity fish and bird;
-Session III: endocrine disruption fish and amphibian.
- Session IV Bioaccumulation fish
Discussion points for each of this this breakout session:
- state of the art and current use of animal tests;
- available alternative assays/approaches, requirements for alternatives;
- provide and discuss examples (including own results);
- limitations of alternatives and priorities for validation;
- future research needs;
- regulatory acceptance;
- recommendations.
A second round of four breakout session on ?Alternative approaches were organised: Session V: QSAR.
Session VI: Non-testing approaches (e.g. refinement, extrapolation, data analysis, integrated testing, exposure waiving); Session VII ALTERNATIVE TEST SYSTEMS and Session VIII: SUBLETHAL/ ALTERNATIVE ENDPOINTS. Main discussion points for breakout sessions:
- state of the art of alternative approaches;
- provide and discuss examples (including own results);
- limitations and advantages of each approach;
- future research needs/priorities;
- regulatory acceptance;
- recommendations.
Results
Based on the expert meeting a conceptual paper titled 'A European perspective of alternatives to animal testing for environmental hazard identification and risk assessment' has been prepared with up to 30 contributing authors. The paper is currently under internal revision considering the comments of participating experts. It is planned to submit the paper the Journal of Regulatory Toxicology and Pharmacology within spring 2013.
Future activities resulting from the expert meeting
The expert meeting and its discussions were highly appreciated by the participants. The overall identification of limitations and future research need were supported by most of the participants and a high consensus among basic scientists and representatives of industry, stakeholders and regulatory bodies was recognised. Therefore, participants have agreed to jointly prepare a conceptual paper on the European perspective of alternatives to animal experiments for environmental risk assessment. A first draft of this multiple-co-author paper is already available. The manuscript is foreseen to be submitted soon to the Journal of Regulatory Pharmacology and Toxicology and would be published as open access, available from the EUROECOTOX web page or the journal's online repository, respectively.
5. First European conference on the replacement, reduction and refinement of animal experiments in ecotoxicology
EUROECOTOX organised the first. European Conference on the replacement, reduction and refinement of animal experiments in ecotoxicology. I was held at Eawag, Dübendorf, Switzerland, from 28-29 June 2012. The conference aimed to provide a platform for young scientists and experts from academia, industry and regulation in the field of replacement, reduction and refinement of animal tests used in environmental risk assessment. It was focused on the current state and future directions of the development, implementation and application of the 3Rs, from bench to acceptance.
The programme, based on invited key note speakers and presentations from submitted abstracts, consisted of four main blocks: (i) experimental approaches, (ii) towards integration and implementation, (iii) perspectives and initiatives towards the 3Rs, (iv) computational approaches and (v) a final plenary discussion about the future of alternative methods in ecotoxicology. The sections (i), (ii) and (iv) were stimulated by a key note presentation of an invited expert. A special key note lecture was given by Susanne Louhimies about the establishment of the 3Rs in European legislation on the first day in the evening. All presentations are available from the EUROCOTOX website.
Sponsored by ESTIV, the European society of toxicology in vitro, the conference was able to provide two students an award of 200 each to the best oral presentation and poster of a student (Master, Bachelor or PhD student), making thus attractive the participation of students in the conference. The jury consisted of four independent conference participants: Michelle Embry, Erika Sela, Marc Leonard and Nic Bury. The winner of the Best Young Scientist award for an oral presentation was given to Julita Stadnicka (Eawag) for her presentation Predicting chemical concentrations in a fish cell line based on measurements and models?. The award for the Young Scientist Best poster presentation was given to Melanie Knöbel (Eawag) for her poster ?Correlation of zebrafish embryo to fathead minnow acute toxicity: the relevance of physico?chemical properties, mode of toxic action, test duration and exposure concentration analysis.
6. First EUROECOTOX report: 'Alternative approaches to replace or reduce the use of animals in ecotoxicology'.
The report presents a comprehensive overview on the different aspects to be considered for developing alternatives to testing of animals in environmental risk assessment - ranging from an overview of regulations, different alternative approaches, future research needs and policy recommendations. It complements the already developed database of researchers and institutions active in the field which is available online.
It is the intention of the consortium to publish the report as book titled 'Alternatives to testing of animals in environmental risk assessment'. For this, the consortium has already submitted a book editing proposal to SETAC, (Society of environmental toxicology and chemistry), who will be willing in principle to edit and publish the book. It will be published both as open access PDF (for download from the SETAC press webpage) as well as a printed edition (for libraries, for convenient reading and use as a reference book). The first printed edition will produced with a number of 100 books. Based on the number of purchases we will order a second edition that may be financed from the revenues of the first printed edition. It is expected that the book could be available by the end of 2013, as it has to go through a peer review process before publishing it.
Support for the publishing costs will be provided in part through a grant from the EU Operational Programmes in the Czech Republic (via a European Social Fund), project 'Innovation and upgrade of the environmental education at Faculty of Science of Masaryk University' (Operational Programme OPVK: Education for Competitiveness, project No. CZ.1.07/2.2.00/15.0213)
Potential impact:
Current legislation in European and other industrial countries requires submission of appropriate data for risk assessment for the registration of new chemicals, pesticides, biocides and pharmaceuticals. These data include information on the toxicity at various trophic levels and require testing with vertebrates, in particular with fish and birds. As a consequence of the new program of REACH which came into force on June 2007, the number of animal experiments (mainly with fish) for environmental risk assessment is expected to increase remarkably within the next years.
Together with the need to increase the environmental safety of chemicals, there is a strong societal demand for reducing the number of animal tests by using alternative in vitro models. Animal welfare is the subject of significant societal ethical concerns and there is a strong European support to refine, reduce and replace animal use linked to regulations. Thus, the use of animals such as fish, for environmental hazard and risk assessment testing, presents a dilemma about balancing the need to ensure these assessments are scientifically valid against concerns about animal testing.
During the last two decades, substantial efforts have been made towards the development and international acceptance of alternative methods to safety studies using laboratory animals. An alternative toxicological test method is any method that can be used to replace, or reduce the use of animals for experimental purposes or refine procedures to make such experiments less painful of stressful (ECVAM).
EUROECOTOX aims to foster the exchange of knowledge, data, ideas and available European expertise to accelerate the R&D, (pre)validation and regulatory acceptance of alternatives. In particular, the EUROECOTOX network has worked towards promoting and supporting the intensification of alternatives for ecotoxicological testing.
EUROECOTOX has contributed to the reduction in the use of animals in ecotoxicity safety testing by promoting the exchange of knowledge and data and dissemination of novel alternative tests and methods among all European stakeholders via the EUROECOTOX networking activities carried out and through the network website. Main dissemination activities carried out are briefly commented here:
- two important meetings with experts and stakeholders we organised in Leipzig (Germany) and in Dubendorf (Switzerland);
- project newsletter with latest project news and focus articles were distributed by email to all network members;
- members of the consortium have participated in various scientific meetings and workshops presenting the network and promoting discussion and dialogue;
- a report describing in detail all dissemination activities has been presented as deliverable at the end of the project.
List of websites: http://www.euroecotox.eu