Introduction

Like most other biomedical research that uses living animals, experiments in neuroscience and behavior may raise concerns related to the direct physical treatment of animals (for example, surgery, injury, and veterinary care) and concerns about how animals are affected by experimental and general environmental conditions (for example, with respect to distress, well-being, and environmental enrichment). Concerns in the latter category are particularly challenging because defining and assessing such concepts as distress and well-being may not be straightforward.

In an effort to provide information and guidance on the use of mammals in neuroscience and behavioral research, the Committee on Guidelines for the Use of Animals in Neuroscience and Behavioral Research was appointed under the auspices of the Institute for Laboratory Animal Research (ILAR) of The National Academies. The committee was composed of 15 members, both researchers and laboratory animal veterinarians, and were drawn from academia and industry. This committee was funded by the National Institutes of Health and was asked to address the following four items:

  1. Identify common research themes in contemporary neuroscience and behavioral research based on input from neuroscience and behavioral researchers most familiar with current standards of practice and veterinarian specialists in laboratory animal medicine.

  2. Exercise collective, professional judgment in applying current animal care and best use practices to procedures in these areas of research.

  3. Obtain information about new scientific and responsible use developments used to maintain animals during these experiments.



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Introduction Like most other biomedical research that uses living animals, experiments in neuroscience and behavior may raise concerns related to the direct physical treatment of animals (for example, surgery, injury, and veterinary care) and concerns about how animals are affected by experimental and general environmental conditions (for example, with respect to distress, well-being, and environmental enrichment). Concerns in the latter category are particularly challenging because defining and assessing such concepts as distress and well-being may not be straightforward. In an effort to provide information and guidance on the use of mammals in neuroscience and behavioral research, the Committee on Guidelines for the Use of Animals in Neuroscience and Behavioral Research was appointed under the auspices of the Institute for Laboratory Animal Research (ILAR) of The National Academies. The committee was composed of 15 members, both researchers and laboratory animal veterinarians, and were drawn from academia and industry. This committee was funded by the National Institutes of Health and was asked to address the following four items: Identify common research themes in contemporary neuroscience and behavioral research based on input from neuroscience and behavioral researchers most familiar with current standards of practice and veterinarian specialists in laboratory animal medicine. Exercise collective, professional judgment in applying current animal care and best use practices to procedures in these areas of research. Obtain information about new scientific and responsible use developments used to maintain animals during these experiments.

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Prepare a report to serve as an informational resource to assist researchers, laboratory animal medicine veterinarians, and IACUC members in the interpretation and implementation of current standards of practice and promote the training of animal care specialists in this area. The ILAR Committee on Guidelines for the Use of Animals in Neuroscience and Behavioral Research (hereafter referred to as the authoring committee) hosted a public workshop on February 27, 2002, to obtain input from leaders in the fields of neuroscience research, behavioral research, and laboratory animal medicine. Following this workshop, the committee met three times during a nine-month period to review the literature, pertinent regulatory documents, and the many references available on the care and use of laboratory animals. After deliberating on responsible use developments and best use practices, the committee drafted this report. This report provides current information on the care and use of laboratory animals in neuroscience and behavioral research and is aimed at ensuring high-quality, humane care for laboratory animals. Because neuroscience and behavioral research is so diverse, and unique and ambiguous situations continue to arise as science advances, it is impossible for this report to provide specific guidance for every potential research situation. Further, recognizing that every potential research situation cannot be anticipated, there are few regulations or guidelines governing laboratory animal care and use that do not end with the caveat “unless a deviation is justified for scientific reasons and approved by the IACUC.” To provide such flexibility in the regulations and guidelines requires the application of professional judgment when applying these regulations and guidelines to each research situation. Often the decisions that must be made are not simple, and reaching effective solutions requires the collective judgment and cooperation of the principal investigator, veterinarian, and IACUC. Therefore, this report emphasizes that developing and evaluating an animal-use protocol requires a decision-making process, as many situations do not lend themselves to simple application of regulations and guidelines to reach a yes or no decision. It is widely held that animal-welfare regulations and guidelines are inflexible and constitute a hindrance to the conduct of high-quality research. One aim of the authoring committee was to highlight the flexibility and promote the use of professional judgment, performance standards, and the decision-making process in evaluating animal protocols, rather than indicating engineering standards. It is the responsibility of institutional animal care and use committees (IACUCs), veterinarians, and researchers to apply creativity and flexibility to balance the needs of high-quality research and humane treatment of animals. In that light, as is the case with other regulatory and guidance documents, the guidelines suggested should not be viewed as laws meant to restrict biomedical research. Rather, they should be interpreted with each unique situation. The guidelines contained in this report are deliberately general. They should be interpreted as a flexible

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framework that can be applied to diverse research situations to guide decision-making. This publication attempts to demystify the decision-making process that IACUCs, veterinarians, and researchers step through when developing, evaluating, and implementing an animal-research protocol. This publication is not meant to supersede the guidelines put forth in the 1996 ILAR document Guide for the Care and Use of Laboratory Animals (the Guide). Instead, it is an informational document that identifies common themes in neuroscience and behavioral research and describes current best practices for animal care and use. It expands on the general guidelines provided in the Guide, the Animal Welfare Act, the US Department of Agriculture Animal Welfare Regulations (AWRs) and Policies, the Public Health Service Policy on Humane Care and Use of Laboratory Animals (PHS Policy), and the US Government Principles for the Utilization and Care of Vertebrate Animals Used in Testing, Research, and Training and discusses them as they pertain to the intricacies of neuroscience and behavioral research. This publication is separated into two main sections. The first section, “General Animal Care and Use Principles and Considerations,” contains overarching principles of animal care and use as they pertain to neuroscience and behavioral research. This section includes Chapters 1–3 and includes discussions on the basics of animal husbandry, the definitions of pain and distress, and differentiating between major and minor surgery. Individuals that are well acquainted with the Animal Welfare Act, the Guide, and PHS Policy, may wish to immediately direct their attention to the second section, Chapters 4–9, “Applications to Common Research Themes in Neuroscience and Behavioral Research.” This section is based on the six common research themes that the committee identified: survival studies, prolonged nonsurvival studies, studies of neural injury and disease, perinatal studies, agents and treatments, and behavioral studies. This second section discusses current best practices and how they apply to research situations unique to neuroscience and behavioral research. Chapter 1 covers regulatory and ethical considerations. It identifies the complex and often overlapping regulatory institutions to help neuroscientists to understand the oversight to which they are subject. Chapter 2 deals with the development of animal protocols and issues central to this process, such as euthanasia, minimization of pain and distress, and humane endpoints. The committee emphasizes the important role that the researcher and the veterinarian play in the development of animal protocols and endorses a team approach to developing protocols to prevent misunderstandings. The chapter discusses the use of pilot protocols to evaluate the appropriateness of new animal protocols and underscores that the researcher, veterinarian, and IACUC should collaborate to ensure that the maximal amount of useful preliminary information is collected from these studies. In this chapter, the committee also identifies underused or undervalued methods that researchers and veterinarians can use to assess an animal’s well-being, such as monitoring its behavior as a sensitive indicator of its physiologic status. Chapter

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3 identifies and discusses the general veterinary and programmatic elements that apply to many types of neuroscience and behavioral research, such as training and supervision of animal handlers, husbandry and nursing care, surgery and procedures, restraint, and food and water regulation. It also discusses the use of genetically modified animals, which have become important neuroscience models in recent years and for which performance-based approaches to care and use are developing constantly. These issues are discussed with emphasis on their application to neuroscience and behavioral research and with emphasis on situations for which the regulations and guidelines are unclear. Throughout Chapters 2 and 3, the text highlights how professional judgment and careful interpretation of the regulations and guidelines contribute to developing performance standards to ensure animal well-being and high-quality research. Chapters 4 through 9 cover the major experimental themes in neuroscience and behavioral research: survival studies, prolonged nonsurvival studies, studies of neural injury and disease, perinatal studies, studies of agents and treatments, and behavioral studies of neural function. Each chapter highlights the common situations in neuroscience and behavioral research that can pose difficulties for researchers, veterinarians, and IACUCs. Those situations include intended and unintended pain and/or distress, multiple major survival surgeries and modified surgical settings, implantation of devices, and the stresses associated with behavioral paradigms. Recognizing that an experimental protocol can involve elements that are addressed in more than one chapter of this publication, the authoring committee has provided extensive cross-referencing to guide the reader. Although neuroscience and behavioral research includes widely varied experimental paradigms, each with its own unique animal-welfare concerns, several general animal care and use concerns must be considered in each situation, including: Careful monitoring to identify unintended adverse effects. Ensuring care for animals that, because of experimental manipulation, may be unable to care for themselves adequately. Maintaining an appropriate environment for animals. Establishing humane endpoints in advance to avoid or minimize unintended pain and/or distress. This publication also includes appendix materials that contain information on calculating sample sizes and estimates of the numbers of animals needed to develop and maintain colonies of genetically modified animals. It is difficult for researchers to estimate necessary animal numbers in some situations and the committee included this information to disseminate it to the neuroscience and behavioral research community.