Behavioral studies are of great importance in increasing our understanding and appreciation of animals. In addition to providing knowledge about the diversity and complexity of behavior in nature, such studies also provide information crucial to improvements in the welfare of animals maintained in laboratories, zoos, and agricultural settings. The use of animals in behavioral research and teaching does, however, raise important ethical issues. While many behavioral studies are non-invasive and involve only observations of animals in their natural habitat, some research questions cannot be answered adequately without manipulation of animals. Studies of captive animals necessarily involve keeping animals in confinement, and manipulative procedures and surgery may be necessary to achieve the aims of the research. Studies of free-living animals in their natural habitats can cause disruption, particularly if feeding, capture, marking or experimental procedures are involved.
While the furthering of scientific knowledge is a proper aim and may itself advance an awareness of human responsibility towards animal life, the investigator must always weigh the potential gain in knowledge against any adverse consequences for the animals and populations under study. This is equally true for the evaluation of animal use in animal behavior teaching activities. In fact, animal behavior courses provide an excellent opportunity to introduce students to the ethical obligations a researcher accepts when animals are studied.
In order to help their members make what are sometimes difficult ethical judgments about the procedures involved in the study of animals, the Association for the Study of Animal Behaviour and the Animal Behavior Society have formed Ethical and Animal Care committees, respectively. These committees jointly produced the following guidelines for the use of all those who are engaged in behavioral research and teaching activities involving vertebrate and invertebrate animals. These guidelines are general in scope, since the diversity of species and study techniques used in behavioral research precludes the inclusion of specific details about appropriate animal care and treatment The guidelines will be used by the Editors of Animal Behaviour in assessing the acceptability of submitted manuscripts. Submitted manuscripts may be rejected by an Editor, after consultation with the Ethical or Animal Care Committee, if the content violates either the letter or the spirit of the guidelines. These guidelines supplement the legal requirements in the country and/or state or province in which the work is carried out. They should not be considered an imposition upon the scientific freedom of individual researchers, but rather as helping to provide an ethical framework which each investigator may use in making decisions related to animal welfare.
1. Legislation

Investigators are accountable for the care and well-being of animals used in their research and teaching activities, and must therefore abide by the spirit as well as the letter of relevant legislation. For those who reside in Great Britain, a summary of the laws designed to ensure the welfare of animals is given by Crofts (1980); detailed guidance on the operation of the Animals (Scientific Procedures) Act, 1986 is provided by the Home Office (199). In the U.S.A., federal, state, and local legislation and guidelines may apply. In particular, the care and use of many vertebrate laboratory animals are regulated under the Animal Welfare Act and its amendments and regulations (code of Federal Regulations, Title 9) and/or the policies of the Public Health Service (NRC 1996; PHS 1996). Guidelines for farm animals used in research and teaching may also be applicable (Guide Development Committee 1988). In Canada, guidance can be obtained from the Guide to the Care and Use of Experimental Animals (Canadian Council on Animal Care 1992).

In Britain, lists of threatened species and laws aiming to protect them can be obtained from the International Union for the Conservation of Nature, Species Conservation Monitoring Unit (219C Huntington Road, Cambridge CB3 0D1, U.K.). In the U.S.A., information pertaining to The Endangered Species Act Of 1973 may be found in the Code of Federal Regulations (Title 50, 1973). Lists of endangered species can be obtained from the Office for Endangered Species, U.S. Department of the Interior, Fish and Wildlife Service (Rm. 430, 4401 N. Fairfax Dr., Arlington, VA 22203, phone 1-800-358-2104) or from the Committee on the Status of Endangered Wildlife in Canada, Canadian Wildlife Service (Environment Canada, Ottawa, Ontario K1A 0E7).

Investigators working in other countries must familiarize themselves with legislation both on animal welfare and on threatened and endangered species and conform with the spirit and letter of the laws. When submitting manuscripts to Animal Behaviour, all authors must confirm in their cover letter that they have adhered to the legal requirements of the country in which the study was conducted.

2. Choice of Species and Non-animal Alternatives

Investigators should choose a species for study that is well suited for investigation of the questions posed. Choosing an appropriate subject usually requires knowledge of a species' natural history and phylogenetic level. Knowledge of an individual animal's previous experience, such as whether or not it has spent a lifetime in captivity, is also important. When research or teaching involves procedures or housing conditions that may cause pain, discomfort or stress to the animal, and when alternative species can be used, the researcher should employ the species which, in the opinion of the researcher and other qualified colleagues, is least likely to suffer (OTA 1986). Live animal subjects are generally essential in behavioral research, but non-animal alternatives such as video records from previous work or computer simulations can sometimes be used (Smyth 1978). Material of this kind also exists for teaching purposes and can be used instead of live animals to expand the range of behavioral subjects available to students.

3. Number of Individuals

The researcher should use the smallest number of animals necessary and sufficient to accomplish the research goals, especially in studies which involve manipulations that are potentially detrimental to the animal or the population. The number of animals used in an experiment can often be dramatically reduced by pilot studies, good experimental design and the use of statistical tests that enable several factors to be examined simultaneously. Hunt (1980), Still (1982) and McConway? (1992) discuss ways of reducing the number of animals used in experiments through alternative designs. Useful reference works are Cox (1958) and Cochran & Cox (1996).

4. Procedures

Investigators are encouraged to discuss with colleagues both the scientific value of their research proposals as well as possible ethical considerations. There are several models for evaluating animal research which can be of use when making ethical decisions (Bateson 1986; Orlans 1987; Shapiro & Field 1988; Donnelly & Nolan 1990; Porter 1992). If procedures used in research or teaching involve exposure to painful, stressful or noxious stimuli, the investigator must consider whether the knowledge that may be gained is justified. Bateson (1991) discusses the assessment of pain and suffering. Additional information can be obtained from the U.S. National Academy of Sciences Publication, "Recognition and Alleviation of Pain and Distress in laboratory Animals" (NRC 1996), and from the American Veterinary Medical Association panel report on animal pain and distress (AVMA 1987). Researchers are urged to consider the use of alternative procedures before employing techniques that are likely to cause physical or psychological discomfort to the animal. Pain or suffering should be minimized both in duration and magnitude to the greatest extent possible under the requirements of the experimental design. Attention should be given to proper pre- and post-operative care in order to minimize preparatory stress and residual effects. Unless specifically contraindicated by the experimental design, procedures that are likely to cause pain or discomfort should be performed only on animals that have been adequately anaesthetized. Analgesics should be used after such procedures to minimize pain and distress whenever possible (Flecknell 1985; Benson et al. 1990).
The following more specific points may be of use:

1. Fieldwork. Investigators studying free-living animals must take precautions to minimize interference with individuals as well as the populations and ecosystems of which they are a part. Capture, marking, radio-tagging, collection of physiological data such as blood or tissue samples or field experiments may not only have immediate effects on the animal, but may also have consequences such as a reduced probability of survival and reproduction. Investigators should consider the effects of such interference and use less disruptive techniques, such as individual recognition by the use of natural features rather than marking (Scott 1978) where possible. Cuthill (1991) discusses the ethical issues associated with field experiments and recommends pilot investigations to assess potential environmental disruption and follow-up studies to detect and minimize persistent effects. Investigators should weigh the potential gain in knowledge from field studies against the adverse consequences of disruption for the animals used as subjects and also for other animals and plants in the ecosystem. When an experimental protocol requires that animals be removed from the population either temporarily or on a long-term basis, investigators should ensure that suffering or discomfort are minimized not only for the removed animals but for others dependent on them (e.g., dependent offspring). Removed individuals and their dependents must be housed and cared for appropriately. Sources of further information of field techniques are the books edited by Stonehouse (1978) and Amlaner & Macdonald (1980).
2. Aggression, Predation and Intraspecific Killing. The fact that the agent causing harm may be another non-human animal does not free the experimenter from the normal obligations to experimental animals. Huntingford (1984) and Elwood (1991) discuss the ethical issues involved and suggest ways to minimize suffering. Wherever possible, field studies of natural encounters should be used in preference to staged encounters. Where staged encounters are necessary, the use of models should be considered, the number of subjects should be kept to the minimum needed to accomplish the experimental goals, and the experiments made as short as possible. Suffering can also be reduced by continuous observation with intervention to stop aggression at predefined levels, and by providing protective barriers and escape routes for the subjects.
3. Aversive Stimulation and Deprivation as Motivational Procedures. Aversive stimulation or deprivation can cause pain or distress to animals. To minimize suffering, the investigator should ascertain that there is no alternative way of motivating the animal and that the levels of deprivation or aversive stimulation used are no greater than necessary to achieve the goals of the experiment. Alternatives to depravation include the use of highly preferred foods and other rewards which may motivate even satiated animals. Use of minimal levels requires a knowledge of the technical literature in the relevant area; quantitative studies of aversive stimulation are reviewed by Church (1971) and Rushen (1986) and the behavior of satiated animals is considered by Morgan (1974). Further comments on reducing distress due to motivational procedures are to be found in Lea (1979) and Moran (1975).
4. Social Deprivation, Isolation and Crowding. Experimental designs that require keeping animals in over-crowded conditions, or that involve social deprivation or isolation, may be extremely stressful to the animals involved. Because the degree of stress experienced by the animal can vary with species, age, sex, reproductive condition, developmental history and social status, the natural social behavior of the animals concerned and their previous social experience must be considered in order to minimize such stress.
5. Deleterious Conditions. Studies aimed at inducing deleterious conditions in animals are sometimes performed in order to gain scientific knowledge of value to human or animal problems. Such conditions include inducing diseases, increasing parasite loads, and exposing animals to pesticides or homeostatic stressors. Where feasible, studies inducing a deleterious condition in animals should address the possible treatment or alleviation of the condition induced. Animals exposed to deleterious conditions that might result in suffering or death should be monitored frequently and, whenever possible, considering the aims of the research, humanely killed as soon as they show signs of distress. If the goals of the research allow it, the investigator should also consider experimental designs in which the deleterious condition is removed (e.g., removing rather than adding parasites as the experimental treatment) or in which naturally occurring instances of deleterious conditions are observed.

5. Endangered Species

All research on endangered or locally rare species must comply with relevant legislation and be coordinated with official agencies responsible for the conservation effort for the particular species under study. Legislation and sources of help in identifying endangered species have been outlined in Section A. Members of threatened species should not be placed at risk except as part of a serious attempt at conservation. Observation alone can result in serious disturbance, including higher predation rates on nests or young, or their abandonment, and should only be undertaken after careful consideration of techniques and of alternative species. Investigators should also consider further adverse consequences of their work, such as opening up remote areas for subsequent access or teaching techniques of anesthetization and capture which might be misused (e.g., by poachers).

6. Procurement of Animals

When it is necessary to procure animals either by purchase or donation from outside sources, only reputable suppliers should be used. For workers in the U.K., advice about purchasing animals may be obtained from the Laboratory Animal Breeder's Association, Charles River (U.K.) Ltd., Manson Research Centre, Manson Road, Margate, Kent CT9 4LP. In the U.S.A., information on licensed animal dealers can be obtained from the local office of the U.S. Department of Agriculture (U.S.D.A.). Other sources of information on laboratory animal suppliers in North America are the American Association for Laboratory Animal Science (M524 Biological Sciences Building, University of Alberta, Edmonton, Alberta T6G 3E9). If animals are procured by capture in the wild, this must be done in as painless and humane a manner as possible and must comply with any relevant legislation. Individuals of endangered species or populations should not be taken from the wild unless they are part of an active conservation program. So far as is possible, the investigator should ensure that those responsible for handling purchase, donated or wild-caught animals en route to the research facilities provide adequate food, water, ventilation and space, and do not impose undue stress.

7. Housing and Animal Care

The researcher's responsibilities extend also to the conditions under which the animals are kept when not being studied. Caging conditions and husbandry practices must meet, at the very least, minimal recommended requirements of the country in which the research is carried out. Guidance can be obtained from the Universities Federation for Animal Welfare (U.F.A.W.) handbook (Poole, 1987), the National Research Council (NRC 1985) guide, the U.S.D.A. Animal Welfare Act Regulations (Code of Federal Regulations), the Guide for the Care and Use of Agricultural Animals in Agricultural Research and Teaching (Guide Development Committee 1988), and the Canadian Council on Animal Care's Guide to the Care and Use of Experimental Animals (1992).

Although these publications provide general guidance, the housing and care regimes established for the commonly used laboratory animals are not necessarily suitable for wild animals or for individuals of wild species born in captivity. Special attention may be required to enhance the comfort and safety of these animals. Normal maintenance of captive animals should incorporate, as much as possible, aspects of the natural living conditions deemed important to welfare and survival. Consideration should be given to providing features such as natural materials, refuges, perches, and dust and water baths. Companions should be provided for social animals where possible, providing that this does not lead to suffering or injury. Frequency of cage cleaning should represent a compromise between the level of cleanliness necessary to prevent diseases and the amount of stress imposed by frequent handling and exposure to unfamiliar surroundings, odors, and bedding. Researchers in the United States should also ensure that the requirements outlined under the 1985 Amendment to the Animal Welfare Act to provide exercise for laboratory-housed dogs and to ensure the psychological well-being of captive non-human primates are met.

The nature of human-animal interactions during routine care and experimentation should be considered by investigators. Depending upon species, rearing history and the nature of the interaction, animals may perceive humans as conspecifics, predators or symbionts (Estep & Hetts 1992). Special training of animal care personnel can help in implementing procedures that foster habituation of animals to caretakers and researchers and minimize stress. Stress can also be reduced by training animals to cooperate with handlers and experimenters during routine husbandry and experimental procedures (Biological Council 1992).

8. Final Disposition of Animals

When research projects or teaching exercises using captive animals are completed, it may sometimes be appropriate to distribute animals to colleagues for further study or breeding, if permitted by local legislation. However, if animals are distributed care must be taken to ensure that the same animals are not used repeatedly in stressful or painful experiments and that they continue to receive a high standard of care. Animals should never be subjected to more than one major surgery unless it is an unavoidable element of a single experiment. Except as prohibited by national, federal, state, provincial, or local laws, researchers may release field-trapped animals if this is practical and feasible, especially if it is critical to conservation efforts. However, the researcher should assess whether releases into the wild might be injurious or detrimental both to the released animal and to existing populations in the area. Animals should be released at the same site where they were trapped (unless conservation efforts dictate otherwise), and only when their ability to survive in nature has not been impaired and when they do not constitute a health or ecological hazard to existing populations. If animals must be killed subsequent to a study this must be done as humanely and painlessly as possible; death of the animals should be confirmed before their bodies are discarded. A veterinarian should be consulted for advice on methods of euthanasia that are appropriate for the particular species being used. Additional information on euthanasia methods can be found in the report of the AVMA panel on euthanasia (AVMA 1993).

9. Obtaining Further Information

There are a number of organizations that provide publications and detailed information about the care and use of animals. These include The Canadian Council on Animal Care (1105-151 Slater Street, Ottawa, Ontario, K1P 5H3, Canada), the Scientists Center for Animal Welfare (7833 Walker Dr. , suite 340, Greenbelt, MD 20770, U.S.A., scaw@erols.com), AAALAC International -Association for the Assessment and Accreditation of Laboratory Animal Care (1-800-926-0066, http://www.aaalac.org), NetVet? (http://netvet.wustl.edu/), National Academy of Sciences (http://www.nas.edu/homepage/pubs/pubs.html), National Academy Press (http://www.nap.edu/readingroom/) and the Universities Federation for Animal Welfare (8 Hamilton Close, South Mimms, Potters Bar, Hertfordshire EN6 3QD, U.K.). The Animal Welfare Information Center at the National Library (Room 205, Beltsville, MD 20705, U.S.A.) publishes a series of bibliographies on special topics and can also provide individualized database searches for investigators on potential alternatives, including techniques for replacement with non-animal models or alternative species, methods for reducing the total number of animals necessary to address the research question, and experimental refinements which can reduce pain and stress. For those with access to it, the Internet provides a wealth of information on animal care and welfare issues: Many of these are government web pages, particularly those of NIH and USDA. Good starting places are: http://www.nih.gov/ and http://www.aphis.usda.gov/. Additional information at the APHIS site can be found at /reac/reachome.html. On the NIH site is /grants/oppr/library from which one can access the 1996 Institute of Laboratory Animal Resources (ILAR) Guide for Care and Use of Laboratory Animals (published by the National Academy Press) in its entirety, as well as get information on the IACUC Guidebook published by ARENA (Applied Research Ethics National Association). The Animal Welfare Information Center (AWIC) at the National Agricultural Library (NAL) can be accessed via the USDA home page, and they also have available a Compendium of Animal Resources (CARE) CD ROM. For more information, contact Michael Kreger at the NAL: email: mkreger@nal.usda.gov or write to AWIC, National Agricultural Library, 5th floor, 10301 Baltimore Ave., Beltsville, MD 20705.

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