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1 Supporting Online Material for Harmonization of Animal Care and Use Guidance Gilles Demers,* Gilly Griffin, Guy De Vroey, Joseph R. Haywood, Joanne Zurlo, Marie Bédard *To whom correspondence should be addressed. This PDF file includes Published 5 May 2006, Science 312, 700 (2006). DOI: /science ICLAS Working Group on Harmonization of Guidelines, list of participants. "International harmonization of guidelines on humane endpoints" and "International harmonization of guidelines on euthanasia."
2 List of Participants in the International Council for Laboratory Animal Science (ICLAS) Working Group on Harmonization of Guidelines, created at the First International Meeting for the Harmonization of Guidelines on the Use of Animals in Science in Nantes, France, on 13 and 14 June 2004 Gilles Demers: International Council for Laboratory Animal Science (ICLAS) Joanne Zurlo and Harry Rozmiarek: Institute for Laboratory Animal Research (ILAR) Clément Gauthier and Gilly Griffin: Canadian Council on Animal Care (CCAC) Willem de Leeuw: Council of Europe (CoE) Harry Bloom: Federation of European Laboratory Animal Science Associations (FELASA) John Miller and Kathryn Bayne: Association for Assessment and Accreditation of Laboratory Animal Care (AAALAC) International Kathleen Conlee: Humane Society of the United States (HSUS) Tim Morris: Laboratory Animals Joseph R. Haywood: International Union of Pharmacology (IUPHAR) Patri Vergara: Sociedad española para las ciencias del animal de laboratorio [(SECAL) Spanish association for laboratory animal science] Guy Dubreuil: Association française des sciences et techniques de l animal de laboratoire [(AFSTAL), French association for laboratory animal science] Norikazu Tamaoki and Naoko Kagiyama, Japan Cecilia Carbone, Argentina Gemma Perretta, Italy Vera Baumans, The Netherlands Ronald Charbonneau: Centre hospitalier de l Université Laval (CHUL) Jim Gourdon: Cornell University Marilyn Brown: Charles River Foundation (USA) Tom Golding: GlaxoSmith Kline (UK) Guy De Vroey: Johnson & Johnson (Belgium) Other organizations, including the Organisation for Economic Co-operation and Development (OECD), the Universities Federation for Animal Welfare (UFAW), the European Centre for the Validation of Alternative Methods (ECVAM), and the Home Office (UK) signified their support of this initiative, even if they were unable to send a representative to the Nantes meeting.
3 May 9, 2005 International Harmonization of Guidelines on Humane Endpoints Because of differing legal systems and cultural backgrounds there are varying approaches to the use of animals for research, testing or training in different countries. Nonetheless, their use should be always in accord with humane practices. The varying approaches in different countries to the use of animals for biomedical purposes, and the lack of relevant legislation or of formal self regulatory mechanisms in some, point to the need for international guidelines principles elaborated as a result of international and interdisciplinary consultations. 1 On June 12-13, 2004, an International Council for Laboratory Animal Science (ICLAS) Working Group on Harmonization met in Nantes, France, to examine national/regional guidelines on humane endpoints and to make recommendations for adoption of guidelines on humane endpoints for international recognition. The Working Group agreed on general principles for the establishment of humane endpoints and recommends two documents as suitable for use as international references, namely the OECD (2000) Guidance Document on the Recognition, Assessment, and Use of Clinical Signs as Humane Endpoints for Experimental Animals Used in Safety Evaluation and the CCAC (1998) guidelines on: choosing an appropriate endpoint in experiments using animals for research, teaching and testing. Both of these documents provide general guidance on establishment of endpoints that minimize pain and distress, given the scientific goal of the study. The purpose of the OECD document is to apply the principles of the Three Rs 2, in particular refinement, to the use of animals in regulatory toxicity tests, whereas the purpose of the CCAC document is to present guidelines, also within the Three Rs framework, for selecting an endpoint that reduces animal pain and/or distress, while still satisfying the experimental design requirements for objective evaluation when animals 1 International Guiding Principles for Biomedical Research Involving Animals (CIOMS, Geneva, 1986). 2 The principles of the Three Rs Replacement, Reduction and Refinement were first enunciated by the UFAW research scholars W. M. S. Russell and R. L. Burch in 1959 and form the basis for most national systems of oversight of animal use in science. p.1/8
4 are used in biomedical research, teaching and testing. The Working Group recognizes that there is now an extensive literature available that provides further information on appropriate endpoints for particular types of studies. The Working Group also recognizes that additional research should be undertaken to further the application of performancebased standards to humane experimental endpoints so that professional judgment can be implemented in accordance with the current base of scientific knowledge. The resulting overarching principles are intended to guide investigators, ethics committees 3 and veterinarians in determining appropriate and humane conclusions to studies based on an understanding of physiological, toxicological and behavioral processes. 3 Ethics committees are referred to by a variety of titles, depending on the particularities of national jurisdictions; for example, institutional animal care and use committees, or animal care committees. p.2/8
5 A. General Principles 1. There is strong scientific evidence that animals experience pain and distress in comparable situations to those which cause pain and distress for humans. 2. Death or severe pain and distress should be avoided as endpoints. 3. The earliest possible endpoint should be used, consistent with the scientific objectives of the study. 4. Studies should be designed to minimize any pain or distress likely to be experienced by the animals, while meeting the scientific objectives of the study. 5. The duration of studies involving pain and distress should be kept to a minimum. 6. Pilot studies should be encouraged as a means of determining morbidity, time course of effects, and frequency of observations required to set an earlier endpoint. 7. Before commencing the experiment, agreement should be reached on (i) appropriate endpoints for the study, and (ii) the person or persons to be responsible for making the judgment that the endpoint has been reached. 8. A team approach should be used, employing the professional judgment of the scientist, veterinarian, animal care staff, and ethics committee to agree on the appropriate endpoint for the study. 9. Research and animal care staff must be adequately trained and competent in the recognition of species-specific behavior and in particular species-specific signs of pain, distress and moribundity. 10. Animals should be monitored using behavioral, physiological and/or clinical signs at an appropriate frequency to permit timely termination of the experiment once the endpoint has been reached. p.3/8
6 B. International Reference Documents The ICLAS working group on harmonization fully supports the use of the following documents as international references: Guidelines on: choosing an appropriate endpoint in experiments using animals for research, teaching and testing (Canadian Council on Animal Care, Ottawa, ON, 1998). Guidance Document on the Recognition, Assessment, and Use of Clinical Signs as Humane Endpoints for Experimental Animals Used in Safety Evaluation, (OECD, Paris, 2000). C. Additional References These references are provided to offer guidance in implementation of the general principles on establishment of humane endpoints (see above), for specific areas of research and testing. Hendriksen, C. F. M. & Morton D. B., Eds., Humane Endpoints in Animal Experiments for Biomedical Research. Proceedings of the International Conference, November 22-25, 1998, Zeist, The Netherlands. 150pp. (Laboratory Animals Ltd., by Royal Society of Medicine Press, London, UK, 1998). The ICLAS/CCAC International Symposium on Regulatory Testing and Animal Welfare. ILAR Journal 43 (Suppl.), 136 pp. (2002). Humane Endpoints for Animals Used in Biomedical Research and Testing. ILAR Journal 41(2), 132 pp. (2000). Workman, P., Twentyman, P., Balkwill, F. et al UK Coordinating Committee on Cancer Research (UKCCCR) Guidelines for the welfare of animals in experimental neoplasia (ed. 2, July 1997). British Journal of Cancer 77, 1-10 (1998). p.4/8
7 November 8, 2004 International Harmonization of Guidelines on Euthanasia Because of differing legal systems and cultural backgrounds there are varying approaches to the use of animals for research, testing, or training in different countries. Nonetheless, their use should be always in accord with humane practices. The varying approaches in different countries to the use of animals for biomedical purposes, and the lack of relevant legislation or of formal self-regulatory mechanisms in some, point to the need for international guiding principles elaborated as a result of international and interdisciplinary consultations. 4 On June 12-13, 2004, an International Council for Laboratory Animal Science (ICLAS) Working Group on Harmonization met in Nantes, France to examine and make recommendations for adoption of guidelines on euthanasia for international recognition. The Working Group agreed on general principles for euthanasia and recommends two documents as suitable for use as international references, namely the 2000 Report of the AVMA Panel on Euthanasia and the 1996/1997 EC Recommendations for Euthanasia of Experimental Animals Parts 1&2. Although there are some inconsistencies between these two documents, Ethics Committees 5 can use the general principles to evaluate whether the given conditions of the situation meet the goal of assuring humane death. The Working Group also identified areas of inconsistency between the documents indicating where there is insufficient knowledge. The research and animal welfare community should actively promote applied research towards performance-based standards concerning laboratory animal euthanasia methods identified under Section C. These efforts could produce much needed refinements and could contribute scientific evidence that would in turn support common principles to be incorporated in future versions of 4 International Guiding Principles for Biomedical Research Involving Animals (CIOMS, Geneva, 1986). 5 Ethics committees are referred to by a variety of titles, depending on the particularities of national jurisdictions; for example, institutional animal care and use committees, or animal care committees. p.5/8
8 guidelines on euthanasia. In order to provide clarity, the Working Group recommends that any new or revised guidelines on euthanasia should report only the acceptable humane euthanasia methods and the conditions in which they are considered acceptable, as well as the life-stage of the concerned species. p.6/8
9 A. General Principles 1. Whenever an animal s life is to be taken it should be treated with the highest degree of respect. 2. Euthanasia should place emphasis on making the animal s death painless and distressfree. Therefore, the method likely to cause the least pain and distress to the animals should be used whenever possible. 3. Euthanasia techniques should result in rapid loss of consciousness, followed by cardiac or respiratory arrest and ultimate loss of brain function 4. Techniques should require minimum restraint of the animal and should minimize distress and anxiety experienced by the animal, prior to loss of consciousness 5. Techniques used should be appropriate for the species, age and health of the animal 6. Death must be verified following euthanasia and prior to disposal of the animal 7. Personnel responsible for carrying out the euthanasia techniques should be trained: a. To carry out euthanasia in the most effective and humane manner b. To recognize signs of pain, fear and distress in relevant species c. To recognize and confirm death in relevant species 8. Human psychological responses to euthanasia should be taken into consideration when selecting the method of euthanasia, but should not take precedence over animal welfare considerations. 9. Ethics committees should be responsible for approval of the method of euthanasia (in line with any relevant legislation) for any study involving the use of animals. This should include euthanasia as part of the experimental protocol as well as euthanasia for animals found to be experiencing unanticipated pain and distress. 10. A veterinarian experienced with the species in question should be consulted when selecting the method of euthanasia, particularly when little species-specific euthanasia research has been done. p.7/8
10 B. International Reference Documents The ICLAS working group on harmonization supports the use of the following documents as international references: Report of the AVMA Panel on Euthanasia, Journal of the American Veterinary Medical Association 218(5), (2001). This document was prepared at the request of the American Veterinary Medical Association Council on Research, by a Panel on Euthanasia convened in 1999 to review and make necessary revisions to the fifth Panel Report, published in In the 2000 Report, the panel updated information on euthanasia of animals in research and animal care and control facilities; expanded information on ectothermic, aquatic and fur-bearing animals; added information on horses and wildlife; and deleted methods or agents considered to be unacceptable. 2. Recommendations for euthanasia of experimental animals Part 1. Laboratory Animals 30, (1996) and Recommendations for euthanasia of experimental animals Part 2. Laboratory Animals 31, 1-32 (1997). These documents were prepared for DGXI of the European Commission to be used with Directive 86/609/EEC of 24 November 1986, on the approximation of laws, regulations and administrative provisions of the Member States regarding the protection of animals used for experimental and other scientific purposes (No L 358, ISSN ). It refers especially to Article 2(1) published by the European Commission in October 1995 which defines humane method of killing as the killing of an animal with a minimum of physical and mental suffering, depending on the species. The ICLAS Working Group on Harmonization recommends that institutions carefully consider any new information on specific euthanasia techniques that may refine methods supported in the above documents, in order to improve welfare outcomes for the animals. C. Recommendations for research where scientific knowledge is lacking Issues that still need further research include mass animal euthanasia, euthanasia of fetuses and neonates, euthanasia of cold-blooded animals, proper use of CO 2 according to species, decapitation with or without prior anesthesia, cervical dislocation, use of N 2 and/or argon gas, p.8/8
11 May 9, 2005 International Harmonization of Guidelines on Humane Endpoints Because of differing legal systems and cultural backgrounds there are varying approaches to the use of animals for research, testing or training in different countries. Nonetheless, their use should be always in accord with humane practices. The varying approaches in different countries to the use of animals for biomedical purposes, and the lack of relevant legislation or of formal self regulatory mechanisms in some, point to the need for international guidelines principles elaborated as a result of international and interdisciplinary consultations. 1 On June 12-13, 2004, an International Council for Laboratory Animal Science (ICLAS) Working Group on Harmonization met in Nantes, France, to examine national/regional guidelines on humane endpoints and to make recommendations for adoption of guidelines on humane endpoints for international recognition. The Working Group agreed on general principles for the establishment of humane endpoints and recommends two documents as suitable for use as international references, namely the OECD (2000) Guidance Document on the Recognition, Assessment, and Use of Clinical Signs as Humane Endpoints for Experimental Animals Used in Safety Evaluation and the CCAC (1998) guidelines on: choosing an appropriate endpoint in experiments using animals for research, teaching and testing. Both of these documents provide general guidance on establishment of endpoints that minimize pain and distress, given the scientific goal of the study. The purpose of the OECD document is to apply the principles of the Three Rs 2, in particular refinement, to the use of animals in regulatory toxicity tests, whereas the purpose of the CCAC document is to present guidelines, also within the Three Rs framework, for selecting an endpoint that reduces animal pain and/or distress, while still satisfying the experimental design requirements for objective evaluation when animals 1 International Guiding Principles for Biomedical Research Involving Animals (CIOMS, Geneva, 1986). 2 The principles of the Three Rs Replacement, Reduction and Refinement were first enunciated by the UFAW research scholars W. M. S. Russell and R. L. Burch in 1959 and form the basis for most national systems of oversight of animal use in science. p.1/8
12 are used in biomedical research, teaching and testing. The Working Group recognizes that there is now an extensive literature available that provides further information on appropriate endpoints for particular types of studies. The Working Group also recognizes that additional research should be undertaken to further the application of performancebased standards to humane experimental endpoints so that professional judgment can be implemented in accordance with the current base of scientific knowledge. The resulting overarching principles are intended to guide investigators, ethics committees 3 and veterinarians in determining appropriate and humane conclusions to studies based on an understanding of physiological, toxicological and behavioral processes. 3 Ethics committees are referred to by a variety of titles, depending on the particularities of national jurisdictions; for example, institutional animal care and use committees, or animal care committees. p.2/8
13 A. General Principles 1. There is strong scientific evidence that animals experience pain and distress in comparable situations to those which cause pain and distress for humans. 2. Death or severe pain and distress should be avoided as endpoints. 3. The earliest possible endpoint should be used, consistent with the scientific objectives of the study. 4. Studies should be designed to minimize any pain or distress likely to be experienced by the animals, while meeting the scientific objectives of the study. 5. The duration of studies involving pain and distress should be kept to a minimum. 6. Pilot studies should be encouraged as a means of determining morbidity, time course of effects, and frequency of observations required to set an earlier endpoint. 7. Before commencing the experiment, agreement should be reached on (i) appropriate endpoints for the study, and (ii) the person or persons to be responsible for making the judgment that the endpoint has been reached. 8. A team approach should be used, employing the professional judgment of the scientist, veterinarian, animal care staff, and ethics committee to agree on the appropriate endpoint for the study. 9. Research and animal care staff must be adequately trained and competent in the recognition of species-specific behavior and in particular species-specific signs of pain, distress and moribundity. 10. Animals should be monitored using behavioral, physiological and/or clinical signs at an appropriate frequency to permit timely termination of the experiment once the endpoint has been reached. p.3/8
14 B. International Reference Documents The ICLAS working group on harmonization fully supports the use of the following documents as international references: Guidelines on: choosing an appropriate endpoint in experiments using animals for research, teaching and testing (Canadian Council on Animal Care, Ottawa, ON, 1998). Guidance Document on the Recognition, Assessment, and Use of Clinical Signs as Humane Endpoints for Experimental Animals Used in Safety Evaluation, (OECD, Paris, 2000). C. Additional References These references are provided to offer guidance in implementation of the general principles on establishment of humane endpoints (see above), for specific areas of research and testing. Hendriksen, C. F. M. & Morton D. B., Eds., Humane Endpoints in Animal Experiments for Biomedical Research. Proceedings of the International Conference, November 22-25, 1998, Zeist, The Netherlands (Laboratory Animals Ltd., by Royal Society of Medicine Press, London, UK, 1998), 150 pp. The ICLAS/CCAC International Symposium on Regulatory Testing and Animal Welfare. ILAR Journal 43 (Suppl.), 136 pp. (2002). Humane Endpoints for Animals Used in Biomedical Research and Testing. ILAR Journal 41(2), 132 pp. (2000). Workman, P., Twentyman, P., Balkwill, F. et al UK Coordinating Committee on Cancer Research (UKCCCR) Guidelines for the welfare of animals in experimental neoplasia (ed. 2, July 1997). British Journal of Cancer 77, 1-10 (1998). p.4/8
15 November 8, 2004 International Harmonization of Guidelines on Euthanasia Because of differing legal systems and cultural backgrounds there are varying approaches to the use of animals for research, testing, or training in different countries. Nonetheless, their use should be always in accord with humane practices. The varying approaches in different countries to the use of animals for biomedical purposes, and the lack of relevant legislation or of formal self-regulatory mechanisms in some, point to the need for international guiding principles elaborated as a result of international and interdisciplinary consultations. 4 On June 12-13, 2004, an International Council for Laboratory Animal Science (ICLAS) Working Group on Harmonization met in Nantes, France to examine and make recommendations for adoption of guidelines on euthanasia for international recognition. The Working Group agreed on general principles for euthanasia and recommends two documents as suitable for use as international references, namely the 2000 Report of the AVMA Panel on Euthanasia and the 1996/1997 EC Recommendations for Euthanasia of Experimental Animals Parts 1&2. Although there are some inconsistencies between these two documents, Ethics Committees 5 can use the general principles to evaluate whether the given conditions of the situation meet the goal of assuring humane death. The Working Group also identified areas of inconsistency between the documents indicating where there is insufficient knowledge. The research and animal welfare community should actively promote applied research towards performance-based standards concerning laboratory animal euthanasia methods identified under Section C. These efforts could produce much needed refinements and could contribute scientific evidence that would in turn support common principles to be incorporated in future versions of 4 International Guiding Principles for Biomedical Research Involving Animals (CIOMS, Geneva, 1986). 5 Ethics committees are referred to by a variety of titles, depending on the particularities of national jurisdictions; for example, institutional animal care and use committees, or animal care committees. p.5/8
16 guidelines on euthanasia. In order to provide clarity, the Working Group recommends that any new or revised guidelines on euthanasia should report only the acceptable humane euthanasia methods and the conditions in which they are considered acceptable, as well as the life-stage of the concerned species. p.6/8
17 A. General Principles 1. Whenever an animal s life is to be taken it should be treated with the highest degree of respect. 2. Euthanasia should place emphasis on making the animal s death painless and distressfree. Therefore, the method likely to cause the least pain and distress to the animals should be used whenever possible. 3. Euthanasia techniques should result in rapid loss of consciousness, followed by cardiac or respiratory arrest and ultimate loss of brain function 4. Techniques should require minimum restraint of the animal and should minimize distress and anxiety experienced by the animal, prior to loss of consciousness 5. Techniques used should be appropriate for the species, age and health of the animal 6. Death must be verified following euthanasia and prior to disposal of the animal 7. Personnel responsible for carrying out the euthanasia techniques should be trained: a. To carry out euthanasia in the most effective and humane manner b. To recognize signs of pain, fear and distress in relevant species c. To recognize and confirm death in relevant species 8. Human psychological responses to euthanasia should be taken into consideration when selecting the method of euthanasia, but should not take precedence over animal welfare considerations. 9. Ethics committees should be responsible for approval of the method of euthanasia (in line with any relevant legislation) for any study involving the use of animals. This should include euthanasia as part of the experimental protocol as well as euthanasia for animals found to be experiencing unanticipated pain and distress. 10. A veterinarian experienced with the species in question should be consulted when selecting the method of euthanasia, particularly when little species-specific euthanasia research has been done. p.7/8
18 B. International Reference Documents The ICLAS working group on harmonization supports the use of the following documents as international references: Report of the AVMA Panel on Euthanasia, Journal of the American Veterinary Medical Association 218(5), (2001). This document was prepared at the request of the American Veterinary Medical Association Council on Research, by a Panel on Euthanasia convened in 1999 to review and make necessary revisions to the fifth Panel Report, published in In the 2000 Report, the panel updated information on euthanasia of animals in research and animal care and control facilities; expanded information on ectothermic, aquatic and fur-bearing animals; added information on horses and wildlife; and deleted methods or agents considered to be unacceptable. 2. Recommendations for euthanasia of experimental animals Part 1. Laboratory Animals 30, (1996) and Recommendations for euthanasia of experimental animals Part 2. Laboratory Animals 31, 1-32 (1997). These documents were prepared for DGXI of the European Commission to be used with Directive 86/609/EEC of 24 November 1986, on the approximation of laws, regulations and administrative provisions of the Member States regarding the protection of animals used for experimental and other scientific purposes (No L 358, ISSN ). It refers especially to Article 2(1) published by the European Commission in October 1995 which defines humane method of killing as the killing of an animal with a minimum of physical and mental suffering, depending on the species. The ICLAS Working Group on Harmonization recommends that institutions carefully consider any new information on specific euthanasia techniques that may refine methods supported in the above documents, in order to improve welfare outcomes for the animals. C. Recommendations for research where scientific knowledge is lacking Issues that still need further research include mass animal euthanasia, euthanasia of fetuses and neonates, euthanasia of cold-blooded animals, proper use of CO 2 according to species, decapitation with or without prior anesthesia, cervical dislocation, use of N 2 and/or argon gas, p.8/8
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