Appropriate Species And Numbers
As all scientists who use animals in their research and teaching are well aware, any and all use of animals must first be reviewed and approved by the institutional Animal Care and Use Committee (IACUC). This review is mandated by federal law, and UC Santa Cruz is legally bound to provide annual assurance to the NIH that any and all animal research, no matter what the source of support, will be reviewed by the Chancellor's Animal Research Committee (CARC). As a part of the review process, the committee needs to ascertain that the PI has provided adequate responses covering the so-called "informational requirements" under the USDA Animal Welfare Act. Much of this special information relates to the consideration of "the three Rs":
CARC's "Application for Use of Live Vertebrate Animal Subjects Involving Contact" ("Full Use" form) includes these special informational requirements under the following question numbers:
Item 10.a.
PIs must represent to the IACUC that the proposed research or
teaching activity will benefit human or animal health, the advancement
of knowledge, or the good of society. This section should be written
with sensitivity toward readers who are not scientists. Technical
text from your grant is not an acceptable substitute for a summary
in nonscientific terms. Technical terms must be made understandable
to an average member of the public with a high school education.
The issue of "relevance" presupposes that the PI has
fully considered alternatives, i.e., non-animal methods, and if not feasible, "the
three
Rs"
mentioned above. Therefore, this section
should
explain why the research design requires any animals at all, and
affirm that alternative approaches have been considered. If your
response is too vague, you will be asked to develop it further.
Substantiating references are welcome and may be requested by
the committee.
Item 10.b.
PIs must determine that the activities which they propose do not
unnecessarily duplicate previous research activities. A written
narrative must be provided showing one's familiarity with the
literature and the sources searched: i.e., databases listed
under Search Strategies, searches performed for the investigator
by others, and consultation with the Animal Welfare Information
Center (AWIC) or with specialists in the field. (See Search
Strategies
to learn about the acquisition and documentation of information.)
Item 11. PIs must establish the scientific basis for the choice
to use animals, and for the particular species and number of animals
they propose to use. (See Appropriate
Species and Numbers.)
Item 21. PIs must categorize animal use procedures as Class 'C,' 'D,' or
'E',
based upon the level of pain or distress likely to be induced in the animals.
These categories of animal
use have been established for reporting purposes under the Animal
Welfare Act and the PHS Guide, which states:
Furthermore, item IV of the "US Government Principles for the Utilization and Care of Vertebrate Animals Used in Testing, Research, and Training" states:
Animals must be claimed under the highest class involved at any point prior to euthanasia or release, except that euthanasia is not claimed as a Class D procedure. If euthanasia is the only procedure to be performed on live animals (such as in antibody production), no class is claimed under item 21.
Item 22 requires a literature-search narrative for Class D and E procedures that describes the methods and sources used to determine that there are no adequate alternatives to procedures that may cause "more than momentary or slight pain or stress to animals." (See Search Strategies to learn about the acquisition and documentation of information on alternatives.)
Item 23 will usually involve consultation with the attending veterinarian to select the most appropriate anesthetic or analgesic agents to best meet both scientific and humane requirements.
Item 24 requires an additional literature-search narrative to justify that animal pain or distress is necessary to the experiment, because analgesic or anesthetic relief would invalidate the results. (See Search Strategies to learn about the acquisition and documentation of information on alternatives.)
APPROPRIATE SPECIES AND NUMBERS
Item III of the "US Government Principles for the Utilization and Care of Vertebrate Animals Used in Testing, Research, and Training" states:
Investigators must explain their strategies in sufficient detail for CARC members to certify in good conscience that it is necessary to use animals to obtain the required data, that the appropriate species has been selected and that the investigator is using the minimum number of animals necessary. Species justification should explain both (1) why the chosen species is optimal for the project, and (2) why animals of a lower phylogenetic class are not acceptable for the project. Lower acquisition or maintenance costs should not be used as a justification. Examples of acceptable justifications might be that all of the preliminary data has been collected in this species, the technologies have developed only in this species (e.g., transgenics), the only available reagents use this species, the species has properties that model the human in certain ways, or the species is the minimum size possible on which the indicated manipulations could be performed.
To evaluate the appropriateness of animal numbers, the committee must understand their intended use. If statistical justification is necessary (consult the CARC Office if unsure), a listing of the different experimental groups and the number of animals per group may be required, and flow diagrams may be helpful. In a statistical justification, the investigator should describe the endpoint to be measured, the variance (either anticipated or based on previous experiments), the difference between experimental and control subjects that would be regarded as relevant, and the statistical tests that will be used to evaluate the data. With these numbers in hand, the investigator should ideally calculate the necessary sample size. It would be helpful to have a biostatistician aid in reviewing this portion. It is recognized that not every animal enrolled in a project provides usable data, and a modest surplus of animals may be requested to cover unforeseen problems.
Some proposals cover large projects spanning up to the maximum of three years that an application may be in effect before federal regulations require a new application, and even beyond. The committee recognizes that it may be difficult to predict with any accuracy experiments that will take place years into the future. The investigator may detail a prototype experiment with the different experimental groups, controls, group sizes, and endpoints. She/he can then indicate that particular parameters (e.g., times, doses, drug types) will be varied for an estimated "X" number of experiments. Some proposals involve novel experimental manipulations in which it is uncertain whether usable data will be obtained. The investigator may wish to break the application into phases beginning with a pilot study, to be followed by further experiments if the pilot proves successful, at which time approval can be extended to the remainder of the three years of the effective application. The number of animals requested for a particular study can be increased later by downloading the 'CARC Amendment Form' available online with the other CARC forms. Justification for additional numbers must be provided. Larger increases will require more detailed justification.
In the case of transgenics, the investigator should explain how many animals will be used to develop the transgenic, how many will be used to maintain the breeding colony, and how many will be used in experimentation. Many investigators elect to divide this into two protocols: one for development of the transgenic and one to be submitted after the transgenic is prepared detailing the experiments to be performed and the animal numbers needed.
USDA regulations [9 CFR section 2.31(d) ii-iii] under the Animal Welfare Act state:
Part (iii) is satisfied by describing a literature search under item 9b in the Full Use Form. Part (ii) requires a literature-search narrative under item 23 and/or 25 demonstrating that alternative approaches are not appropriate or available. "Alternatives" are procedures that follow "the three Rs" (See Alternative Methodologies for a general introduction, or any of the 'Three Rs' listed below):
Replacement
of vertebrate animals as model or test systems,
Reduction of animal numbers, and
Refinement of existing methodologies.
Searching for information alternatives can be challenging, but federal animal welfare inspectors regularly review CARC records and will cite UCSC for noncompliace if these requirements are not satisfactorily addressed. An adequate search often requires use of multiple sources, such as databases, scientific journals, and discussions with colleagues. Narrative descriptions must provide details on methods used and sources consulted. As a minimum, database(s) used to search the literature for alternatives and key words and title words used to perform the search must be listed and results noted. Additional pages may be attached if necessary for completeness. The committee may request to see the search record.
When searching databases, it is important to go beyond a simple search for "animal welfare" or "animal testing alternatives." Other key words that might be included in the search strategy are "nonanimal," "alternative," "reduction," "replacement," and "refinement." Depending on the topic and which of "the three Rs" is being addressed, PIs should consider subjects such as cell culture, toxicity, in vitro techniques, computer simulations, or statistical models. However, simply searching with a key word or title word such as "alternatives" might miss key articles, because the authors do not consider the particular term relevant to what they are doing.
Even if there is a reasonable and allowable justification for using the "more painful" procedure, you must still alleviate the pain with anesthetic, analgesic, or tranquilizing drugs, or provide an additional literature search narrative justifying why such alleviation would invalidate procedures, results, or interpretation. Note that the Animal Welfare Act defines a procedure as "painful" even if the pain is alleviated.
Melvyl's comprehensive biomedical databases, MedLine and Biosis, are generally the first place to look for animal testing alternatives. MedLine is the database most frequently cited by researchers in their search for alternatives. There are many other databases available. You may find that some of the others are more appropriate for your needs. USDA recommends that PIs include literature resources outside their discipline area. For example, AGRICOLA is often used for animal welfare issues and is currently available on the Science Library home page at http://bob.ucsc.edu/library/science. For additional help, you should consult with a librarian, particularly if your database search has not been satisfactory. For a fee, library staff will search any commercial database. The charge varies with the complexity of the search, the number of references found, and the cost of the database searched. You may request reference assistance or an on-line search by calling The Science Library at (408) 459-2886. For additional assistance, refer to AWIC Search Tips, or to AWIC Contacts for direct help in conducting searches.
Before proceeding to databases, you may wish to consult other Internet resources. There is an increasing amount of animal-related and veterinary information available on the Internet at no charge. An excellent starting point to access these resources is the server maintained by Ken Boschert, D.V.M., Washington University, http://netvet.wustl.edu. Materials from AWIC are included on this server. In addition, The Johns Hopkins Center For Alternatives To Animal Testing maintains a website describing its history, mission, and programs. Its newsletter and links to other Internet resources are also available.
The following is a partial listing of databases that you may find useful:
AGRICOLA Provider: National Agricultural Library (NAL), United States Department of Agriculture (USDA), Beltsville, MD. Description: Provides comprehensive coverage of worldwide literature of agricultural science and related topics, including animal and plant production, veterinary science, food and nutrition, rural issues and more recently, welfare aspects of animals used in exhibition, education and research. AGRICOLA contains about 3 million records dating back to 1970. The subject headings "animal testing alternatives" and "animal welfare" alone retrieve over 3000 citations. It is possible to search by AGRICOLA subject heading and title word and to combine searches using the "and" function. Online: DIALOG*: File 10 (1979-present), File 110 (1970-1978)BRS**: CAIN (1980-present), CAIB (1970-1979) Hardcopy: Bibliography of Agriculture http://bob.ucsc.edu/library/science
AGRIS Provider: National Agricultural Library (NAL), United States Department of Agriculture (USDA), Beltsville, MD Description: Provides a comprehensive inventory of world-wide agricultural literature which reflects search results, food production, and rural development. Online: DIALOG: File 203 (1975-present) Hardcopy: Agrindex, published monthly by the Food and Agriculture Organization (FAO) of the United Nations. CD-ROM: available
ASFA (Aquatic Sciences and Fisheries Abstracts) Provider: Cambridge Scientific Abstracts, Bethesda, MD, and the Food and Agriculture Organization, UN, Rome, Italy. Description: Provides comprehensive coverage of the science, technology and management of marine and freshwater environments and resources. Online: DIALOG: File 44 (1978-present) Hardcopy: Aquatic Sciences and Fisheries; Part 1: Biological Sciences and Living Resources and Part 2: Ocean Technology, Policy, and Non-Living Resources. http://bob.ucsc.edu/library/science
AVLINE Provider: National Library of Medicine (NLM), Bethesda, MD Description: An index to NLM's holdings of biomedical audio-visuals, including motion pictures, videocassettes and slides. Online: NLM***: AVLINE Hardcopy: NLM Audio-visual catalog; NLM Current Catalog Proofsheets, Audio-Visuals Section; Health Sciences Audio-visuals (on microfilm); NLM Audio-visuals Center (NMAC) Catalog for the Health Sciences (lists 16-mm films available for short term loan).
A-V ONLINE Provider: Access Innovations, Inc., Albuquerque, NM Description: Provides comprehensive coverage of non-print educational materials from pre-school to professional and graduate school. Includes all media types. Scope includes health and safety education, science and medical, and more. Online: DIALOG: File 46 (1964-present, selected earlier coverage) Hardcopy: NICEM (National Information Center for Educational Media) Film and Video Finder, NICEM Audio Cassette Finder, NICEM Film Strip and Slide Set Finder, custom catalogs. CD-ROM: available
BIOETHICSLINE Provider: National Library of Medicine (NLM), Bethesda, MD Description: Covers the literature dealing with bioethics. In August 1985, began covering the ethical issues of using animals in biomedical research. Includes selected materials in this field published prior to 1985. Online: NLM: BIOETHICSLINE Hardcopy: Bibliography of Bioethics
BIOSIS PREVIEWS Provider: BioSciences Information Service (BIOSIS), Philadelphia, PA Description: Provides comprehensive coverage of worldwide primary journals and monographs, as well as symposia, reviews, preliminary reports, and other secondary sources, on all aspects of the biosciences and medical research. Online: DIALOG: File 55 (1981-present), File 5 (1969-present) BRS: BIOL (1978-present), BIOB (1970-1977), BIOZ (1970-present) Hardcopy: Biological Abstracts, Biological Abstracts/Reports, Reviews, and Meetings Available free of charge on Melvyl
CABI Center for Agriculture and Bio-sciences International, from a British intergovernmental organization providing world-wide services to agriculture, forestry, nutrition, animal science, and the conservation of natural resources. Covers Index Veterinarius and Veterinary Bulletin. 1984-date.
CAB ABSTRACTS Provider: Commonwealth Agricultural Bureaux (CAB) International, Forham Royal, Slough, England Description: Provides comprehensive coverage of primary journals as well as books, reports, theses, conference proceedings, patents, annual reports, and guides, in the agricultural and biological sciences, including animal breeding, animal disease, dairy science, veterinary science, and more. Online: DIALOG: File 50 (1984-present), File 53 (1972-1983)BRS: CABA (1980-present), VETR (a subset of CABA devoted to veterinary science) Hardcopy: 26 abstract journals published by CAB. CD-ROM: available
CEBA (Chemical Engineering and Biotechnology Abstracts) Provider: Royal Society of Chemistry and DECHEMA, the German Society for Chemical Equipment, Chemical Technology and Biotechnology, Cambridge, United Kingdom Description: Provides coverage on all aspects of chemical processing and reaction engineering. The file includes information on production and pilot scale studies, engineering theory, laboratory experiments, computer applications, measurement and process control, physical and chemical property data, plant and personal safety, and environmental protection. References are taken from approximately 400 core journals. Online: DIALOG: File 315 (1971-present) Hardcopy: Chemical Engineering Abstracts and Biotechnology Abstracts
CRIS/USDA (Current Research Information System) Provider: Cooperative State Research Service (CSRS), USDA, Beltsville, MD Description: Covers current research projects in agriculture and related sciences sponsored or conducted by USDA research agencies, state agricultural experiment stations, state forestry schools, and other cooperating state institutions. Includes subject access, summaries and project reports. CRIS encompasses biological, physical, social and behavioral sciences related to agriculture; fish and wildlife; environmental protection, and more. Online: DIALOG: File 60 CRIS: searches provided upon request to USDA and State Extension/Experiment personnel from: CRIS/CSRS/USDA, NAL, 5th Floor, Beltsville, MD 20705, (301) 504-3846
CRISP (Computer Retrieval of Information on Scientific Projects) Provider: Division of Research Grants, National Institutes of Health (NIH), Bethesda, MD Description: Provides comprehensive coverage of research projects supported through the various grant and contract programs of the United States Public Health Service (PHS) or conducted intramurally by NIH, the National Institute of Mental Health (NIMH), and the National Institute on Alcohol and Alcoholism (NIAA). Covers current as well as past projects, going back to 1972. Includes subject access and narrative descriptions. Online: DIALOG: through File 266 (the NTIS database FEDRIP, which also includes research projects of other federal agencies.) NIH: CRISP--searches provided upon request from: Research Documentation Section, Statistics and Analysis Branch, Division of Research Grants, NIH, Bethesda, MD 20892, (301) 496-7543 Hardcopy: Research Awards Index (does not include narrative description or intramural projects.) Current contents available on Melvyl free of charge.
CURRENT CONTENTS and UNCOVER. These databases provide coverage of the recent literature across many disciplines. They may be used to locate articles that have not yet appeared in MEDLINE or other databases. There are no subject headings and searching is primarily limited to words occurring in titles and abstracts, so it is more difficult to search comprehensively.
EMBASE Provider: Excerpta Medica, Amsterdam, Netherlands Description: Provides abstracts and citations from over 4000 worldwide biomedical journals, covering the entire field of human medicine and related disciplines. Online: DIALOG: File 72 (1982-present), File 172 (1980-1981), File 173 (June 1974-Dec. 1979) BRS: EMBED (1980-present), EMBED (1974-1987), EMBOSS (1974-present) Hardcopy: Excerpta Medica (43 specialty abstract journals and 2 literary indices.)
ERIC Provider: US Department of Education, DERI, Washington, DC, and ERIC Processing and Research Facility, Bethesda, MD Description: Consists of two subfiles: 1) Resources in Education (RIE), covering documents, and 2) Current Index to Journals in Education (CIJE), covering journals and serial publications. ERIC contains records with abstracts, and covers all levels of education and related fields. Online: DIALOG: File 1 (1966-present) BRS: ERIC (1966-present) Available free of charge on Melvyl.
FEDERAL REGISTER Provider: US Government Printing Office, Washington, DC Description: Provides full text of Federal Register-a daily publication of the US government that makes available to the public regulations and legal notices issued by Federal agencies and the President. Online: DIALOG: File 669 Hardcopy: Federal Register
FISHERIES REVIEW Based on the US Fish and Wildlife Service's print publication, this NICS product contains references (some with abstracts) on the culture, propagation, genetics, and diseases of fish, as well as limnology and oceanography. 1971-date.
LEGI-SLATE Provider: Legi-slate, Inc., a subsidiary of the Washington Post, Washington, DC Description: Provides coverage in all areas relating to Federal legislation, including Congressional Record and Federal Register text, legislation, Congress members' positions on recorded votes, biographical and geographical information on members of Congress, press briefings, and more. Online: LEGI-SLATE, 111 Massachusetts Avenue, N.W., Washington, DC 20001
LIFESCIENCES COLLECTION Provider: Cambridge Scientific Abstracts, Bethesda, MD Description: Contains abstracts of information in the fields of animal behavior, biochemistry, entomology, genetics, immunology, neurosciences, toxicology, virology, and more, from worldwide journal articles, books, conference proceedings, and report literature. Online: DIALOG: File 76 (1978-present) Hardcopy: 15 abstracting journals CD-ROM: available
MAGAZINE ASAP/MAGAZINE INDEX Provider: Information Access Company, Foster City, CA Description: Magazine Index covers approximately 500 American and Canadian general interest magazines in topics including current affairs, business and economics, education, health, medicine, environmental issues, science, technology, agriculture, and more. Magazine ASAP selectively provides the complete text and indexing of articles, editorials, columns, reviews, product evaluations and recipes from approximately 100 of the magazines listed in Magazine Index. Online: DIALOG: File 47--Magazine Index (1959-1970, 1973-present), File 647--Magazine ASAP (1983-present) BRS: MAGS (Magazine Index), MSAP (Magazine ASAP) CD-ROM: Magazine Index available on CD-ROM
MEDLINE Provider: US National Library of Medicine, Bethesda, MD Description: Provides coverage of primary journals and monographs on virtually every subject in the field of biomedicine. Online: DIALOG: File 152 (1966-1974), File 153 (1975-1982), File 154 (1983-present), File 155 (1966-present) BRS: MESH (1982-present), MESZ (1966-present) Hardcopy: MEDLINE corresponds to Index Medicus, Index to Dental Literature, and International Nursing Index. CD-ROM: available free of charge on Melvyl.
MICROCOMPUTER SOFTWARE GUIDE Provider: R.R. Bowker Company, New York, NY Description: Contains information on virtually every microcomputer software program and hardware system available in the United States. Records are bibliographic, and include ordering information, technical specifications, subject classifications, and a brief description. Online: DIALOG: File 278 Hardcopy: The Software Encyclopedia
PNI (Pharmaceutical News Index) Provider: UMI/Data Courier, Louisville, Kentucky. Description: Contains the latest US and international information about pharmaceutical, cosmetics, medical devices, and related health industries. Online: DIALOG: File 42 (1975- present) BRS: PNII
PHILOSOPHER'S INDEX Provider: Philosophy Documentation Center, Bowling Green State University, Bowling Green, OH Description: Provides indexing and abstracts from over 270 journals of philosophy and related interdisciplinary fields including aesthetics, epistemology, ethics, logic, and metaphysics. Online: DIALOG: File 57 (1940-present) Hardcopy: The Philosopher's Index
PIC (Primate Information Center) Provider: Primate Information Center, University of Washington, Seattle, WA Description: Provides comprehensive coverage of worldwide literature on non-human primates. Includes subject access and abstracts and allows taxonomically precise retrieval. Online: In 1985 began leasing on-site use of the entire database to institutions. Also willing to negotiate lease of subsets of the database. PIC: Searches available upon request for a low fee from: Primate Information Center, University of Washington SJ-59, Seattle, WA 98195, (206) 543-4376 Hardcopy: Primate References (a bibliographic monthly journal available by subscription), topical bibliographies
PsychINFO (Psychology collection available
at McHenry Library)
Provider: American Psychological Association, Washington, DC
Description: Records, usually including
abstracts, cover worldwide literature in psychology and related
disciplines in the behavioral sciences. Includes journals, technical
reports, conference proceedings, monographs, dissertations, and
case studies. Online: DIALOG: File
11 (1967-present) BRS: PSYC (1967-present) Hardcopy:
Psychological Abstracts (does not include all the information
in the database.) Available free
of charge on Melvyl.
TOXNET Provider: National Library of Medicine, Bethesda, MD Description: System of three non-bibliographic databases which provide numerical toxicological and biochemical data. Online: NLM: HSDB (Hazardous Substance Data Bank), TDB (Toxicology Data Bank), CCRIS (Chemical Carcinogenesis Research Information System)
WILDLIFE REVIEW Based on the print "Wildlife Review, published by US Fish and Wildlife Service, it scans the world's literature on mammals, birds, reptiles, and amphibians and includes studies of individual species and types of habitats, hunting, economics, wildlife behavior, management techniques, diseases and parasites. 1971-date.
ZOOLOGICAL RECORD ONLINE Provider:
BIOSIS, Philadelphia, PA Description:
Provides worldwide coverage of zoological literature, with particular
emphasis on systematic/taxonomic information. Includes 27 sections
devoted to various animal groups, including protozoa, nematoda,
pisces, reptilia, aves, and mammalia. Includes basic bibliographic
information as well as systematic classification of up to six
levels for the biological organisms discussed. Online:
DIALOG: File 185 (1978-present) BRS: ZREC Hardcopy: Zoological
Record
Cynthia P. Smith, M.S.
Animal Welfare Information Center
USDA, National Agricultural Library
Searching for Alternatives to Animal Research And Testing
The following guidelines were developed to assist researchers, information specialists, and Institutional Animal Care and Use Committee (IACUC) members, when conducting literature searches to determine if alternatives to the use of animals exist and whether a protocol unnecessary duplicates previous research. When searching for alternatives, the staff at the Animal Welfare Information Center (AWIC) refer to the tenets of the 3 Rs introduced by W.M.S. Russell and R.L. Burch (1959) in their book The Principles of Humane Experimental Technique.1 The 3 Rs represent reduction in the number of animals used, refinement of techniques and procedures that reduce pain and distress, and replacement of animal with non-animal techniques.
The first step in conducting a search for alternatives involves communication between the investigator and the information specialist. The specialist cannot effectively search for alternatives without a basic understanding of the type of research the investigator is proposing. The most efficient means of communicating is a direct dialogue between the investigator and the information specialist. A third party should not be used to convey information.
Investigators can assist information specialists by being prepared to give precise and specific information about their research or testing procedures. The following may serve as a guideline for the type of information the investigator may be asked to provide:
1. What is your general area of study? (e.g., cardiology, neurology, toxicology)?
2. What species are you currently working with (e.g., rats, dogs, swine)?
3. Briefly describe your experimental protocol.
4. What specific systems or parts of the anatomy are involved (e.g., central nervous system, brain stem, parabrachial nucleus)?
5. Please give correct spellings of these structures and any acronyms (e.g., CNS, PBN). European spellings are important as well.
6. If you are studying the effects of a particular hormone, enzyme, or chemical agent, please give the complete spelling of the compound as well as its trade name and acronym (e.g., bovine somatotropin, BST).
7. Do you know of any prominent authors in your area of research? Have you published any previous literature that relates to your current study?
8. What makes your study unique from previous studies (e.g., testing a new technique, investigating a new compound, further understanding of a biochemical pathway)?
9. Are you aware of any possible alternatives to your research, such as experiments conducted on alternative species, cell culture, or in vitro studies?
10. Have you had any other searches conducted for you? If so, what databases were used (e.g., MEDLINE, AGRICOLA, BIOSIS)?
a. What keywords were used (e.g., kidney, parathyroid hormone)?
b. What years were searched (e.g., 1985-present)?
As with any type of searching, success in retrieving relevant citations will depend directly on the quality of the information provided.
Search Strategy:
Once the initial exchange of information has taken place, the information specialist can begin to formulate a search strategy. Search strategies for alternatives may be divided into two phases, reduction and refinement, and replacement.
Phase I (Reduction and Refinement):
Phase I consists of a generalized database search used to retrieve citations pertinent to the investigator's field of study. Citations retrieved during this phase, should provide information on current research, alert the investigator to whether or not they are performing duplicative studies, and possibly provide information to refine experimental techniques.
During Phase I, the information specialist may find it helpful to develop search strategies using databases available on Compact Disc Read Only Memory (CD-ROM). AGRICOLA, MEDLINE, TOXLINE, and LIFE SCIENCES, are examples of several useful databases available on CD-ROM. Searching on CD-ROM allows the information specialist the freedom to experiment with keywords, explore indexes and thesauruses, and read abstracts without the pressure of being charged for on-line time. If the investigator has published previous literature this is a good time to read abstracts of his or hers previous work and become familiar with terminology used to describe the study and to note what terms were used to index the abstract. Searching on CD-ROM should provide the information specialist with a general idea of how much literature exists on a specific topic. If few relevant citations are found, the information specialist may need to broaden the search strategy or use the expanded capabilities of on-line database searching to develop the search. If hundreds of citations are retrieved using only a few years worth of bibliographic data, then it is necessary to further consult with the investigator on ways to narrow the search.
Phase II (Replacement):
Upon completion of Phase I, the information specialist should have a basic understanding of the research area including: 1) the literature published in the particular field, 2) the techniques used, and 3) the commonly used species. The information specialist is now ready to search for possible replacement alternatives.
The following questions may be used to assist in the search for replacement alternatives:
When searching for alternatives, information specialists should search multiple databases. A multidisciplinary approach to searching may yield surprising results, particularly for individuals who are not accustomed to searching the literature outside their general area of study, (e.g., Medicine). AWIC provides a fact sheet entitled Databases for Biomedical, Veterinary and Animal Science Resources2 that describes a number of useful databases.
"Animal testing alternatives" is a phrase used to index citations regarding alternatives in the AGRICOLA, MEDLINE, TOXLINE, and CANCERLIT databases. However it is not used to index alternative studies in other databases such as EMBASE, BIOSIS PREVIEWS, LIFE SCIENCES, and CAB Abstracts. Although useful, this phrase should never be the only strategy used to retrieve information on alternatives. Depending on the study, other terms such as tissue culture, cell culture, in vitro, simulation, model, refinement, reduction, or alternative may be used. For a listing of terms that may be helpful when conducting alternative searches information specialists may refer to Animal Welfare Information Center Scope Notes3 available from AWIC at no charge.
Keep in mind that although electronic databases are powerful resource tools, most databases do not index journals before the mid-sixties and relevant information from early studies will not be retrieved. In addition, information on alternatives is available in newsletters, books, and proceedings that not all databases index.
Sample Search for Alternatives:
The following is an actual search that was requested by an IACUC member and a description of the steps that the AWIC information specialist performed. The IACUC member requested a search for alternatives to the use of zona free hamster oocytes to test human sperm penetration, motility, and viability.
After initial information was exchanged about the protocol, a list of keywords were developed. The specialist then conducted a brief initial search on MEDLINE and AGRICOLA on CD-ROM to become familiar with abstracts in which human sperm penetration, motility, and viability were tested. The specialist was aware that extensive tests have been developed to assess semen characteristics in domestic farm species and therefore contacted a farm animal reproductive physiologist for further information. The physiologist confirmed that alternative methods exist to test bovine and human sperm penetration such as a variety of cervical mucus tests. Based on this information the specialist developed the following search strategy:
Databases Selected:
Thirty five different biological and medical databases were selected and searched simultaneously including BIOSIS PREVIEWS, AGRICOLA, CAB ABSTRACTS, CRIS, PASCAL, MEDLINE, EMBASE, PHARMACEUTICAL HEALTH CARE INDUSTRY NEWS, LIFE SCIENCES, SCISEARCH and others.
Search Strategy:
SET ITEMS1 DESCRIPTION
1 214550 SPERM OR SPERMATID? OR SPERMATOZOA? OR SEMEN
2 110475 MOTILITY OR VIABILITY OR MORPHOLOGY OR MOTILE OR VIABLE
3 45369 S1 AND S2
4 42329 S3 NOT HAMSTER
5 1384 S4 AND (MUCUS OR MUCOUS)
6 1049 S5 AND HUMAN
7 364 S6 AND PY=1988:1993
8 191 RD S7 (UNIQUE ITEMS)
9 154 S8 AND (EVALUAT? OR ANALYSIS OR TEST? OR VITRO)
10 94 S9 AND PY=1990:1993
11 2 S4 AND (COMPUTER(2N)IMAGING)
1 ITEMS = number of citations retrieved that contain search terms
Sample Titles Retrieved:
Hyaluronic acid as a medium for human sperm migration tests. Keywords: cervical mucus, spermatozoa, penetration.
Human sperm-cervical mucus interaction using bovine cervical mucus and hen's egg white in the evaluation of male infertility. Keywords: penetration, sperm motility, physiological model, in-vitro test.
The use of hen's egg white as a substitute for human cervical mucus in assessing human infertility. Keywords: penetration, semen analysis, sperm capacity, sperm motility.
Comparison of measurements of human sperm motility characteristics by the automated CELLSOFT system and time exposure photomicrography. Keywords: automated analysis, sperm motility.
In this case, consultation with an expert and review of the literature supported information available on replacement alternatives. If that had not been the case the search may have focused more on refinement and/or replacement alternatives. The IACUC may have asked, what methods are being used to superovulate and flush oocytes from the hamster, how are the hamsters anesthetized during procedures, and what are the fewest number of animals that can be used?
General Comments:
Protocols should be evaluated on a case by case basis. A perfect strategy to retrieve every citation regarding reduction, refinement, and replacement does not exist. Many factors may affect the outcome of a literature search, including the area of research, species involved, procedures used, chemical(s) tested, experimental design, and whether or not articles have been indexed. Additional factors include: 1) the degree of communication between the information specialist and the investigator, 2) the knowledge and educational background of the information specialist, and 3) time and money constraints.
References:
1. Russell, W. and R. Burch (1959) The Principles of Humane Experimental Technique Methuen and Company, London.
2.* Bielenberg, K. and D. Berry. (December 1990) Databases for Biomedical, Veterinary and Animal Science Resources AWIC Fact Sheet.
3.* Swanson, J. (March 1991) "Animal
Welfare Information Center Scope Notes." AWIC Series #6.
8 p.
Additional Resources:
*Clingerman, K., C. Dowling, and J. Swanson. (June 1990) Searching AGRICOLA for Animal Welfare STS-03. June 1990. 20 p. (*Available from AWIC)
Kreger, M. and T. Allen. (October 1993) "Electronic Information for Animal Care and Use." Lab Animal 22(10):52-53.
Snow, B. (July 1990) "Online Searching for Alternatives to Animal Testing" Online p. 94-97.
Alternatives Terminology
Alternatives to the use of animals in research, testing, and education. Any method that can be considered to reduce, replace, or refine research methods utilizing animals. Also, an in depth discussion of the appropriateness of an animal model for a particular disease can be important when considering alternatives. Such methods (for more information, refer to Alternative Methodologies) and terminology may include:
animal model(s)
animal testing alternative(s)
alternative(s)
artificial
vitro (method, model, technique)
culture (cell, tissue, organ)
isolated (cell, tissue, organ)
single-celled organism(s)
model?
plastinat?
bacteria, protozoa
invertebrate(s)
fish, cephalopod(s)
simulation(s)
simulator(s)
handling, housing, facility design
caging
train?, educat?, teach?
welfare, pain?, stress?, distress?
computer simulation
computer software
computer aided instruction
digital image(s)
interactive
virtual (surgery, reality)
artificial intelligence systems
video(s) (disk, disc, display)
mannequin (manikin)
mathematical model(s)
cadaver(s)
anesthe?, anasthe?, anaesthe?
analges?, sedative?
anxiolytic
euthanasia
assay(s), technique(s)
method(s)
environmental enrichment
behavioral enrichment
The Animal Welfare Information Center (AWIC) is located on the fifth floor of the National Agricultural Library in Beltsville, Maryland. (AWIC, NAL, 5th Floor 10301 Baltimore Blvd., Beltsville, MD 20705-2351). The AWIC office is staffed Monday through Friday from 8:00 a.m. to 4:30 p.m. An answering machine will record messages if staff members are unavailable or during non-office hours including weekends. AWIC publications and other information may be accessed via the AWIC home page (http://netvet.wustl.edu/awic.htm).
PHONE INTERNET
AWIC Main Number (301) 504-6212 AWIC@NAL.USDA.GOV
Ruth Criscio, Secretary (301) 504-6212 RCRISCIO@NAL.USDA.GOV
Jean Larson, Coordinator (301) 504-5215 JLARSON@NAL.USDA.GOV
Information Specialists:
Tim Allen (301) 504-5174 TALLEN@NAL.USDA.GOV
Mike Kreger (301) 504-5563 MKREGER@NAL.USDA.GOV
Richard Crawford, DVM (301) 504-5173 RCRAWFOR@NAL.USDA.GOV
D'Anna Jensen (301) 504-5440 DJENSEN@NAL.USDA.GOV
Cynthia Smith CSMITH@NAL.USDA.GOV
FAX Transmissions (301) 504-7125.
Faxes can be received 24 hours per day, 7 days/week.
Animal Welfare Information Center User Fee Policy:
In accordance with the National Agricultural Library's user fee policy, user (with the exception of US Department of Agriculture personnel) will be billed for information support services in excess of one hour of staff time or $25.00 in on-line computer costs. This policy allows AWIC staff to perform comprehensive on-line literature searches for those individuals and organizations regulated by the Animal Welfare Act.
ARTICLE REPRINT: "ALTERNATIVE METHODOLOGIES"
Bennet, B.T., Brown, M.J., and Schofield, J.C. Essentials for Animal Research. A Primer for Research Personnel, second edition. USDA, National Agricultural Library, Beltsville, Md., 1994
In the regulations promulgated to implement the Animal Welfare Act as amended in 1985, the research facility must provide assurances that the principal investigators considered alternatives techniques to painful procedures and provide guidance concerning research and testing methods that limit the use of animals or minimize the animals' distress. In this chapter the reader will be introduced to the classical concept of alternatives with a brief discussion of each major category including a limited number of examples. For more in-depth coverage of the subject, the reader is encouraged to obtain the latest bibliography on alternative techniques available from the AWIC (see Chapter 8).
In recent years the term alternative techniques has come into common usage in the current controversy involving the use of animals in research, teaching and testing. It is a term that has different meanings to different people and this difference largely depends on which side of the issue one is found. To many biomedical researchers, alternative techniques refer to those which can be used in addition to the more traditional animal models. These techniques can focus on specific biological functions and in many cases reduce the numbers of animals used. Therefore these methods are an adjunct to the more commonly used animal models. To the so-called abolitionist who seeks the immediate end to all animal research, teaching and testing, the term alternative refers to those techniques which can entirely replace the use of animals. The dictionary, defines alternative as: "offering or expressing a choice." The dictionary also defines technique as "a method of accomplishing a desired aim." By combining these definitions, the term alternative technique becomes "one which offers a choice in accomplishing a desired aim."
In designing an experiment which involves the use of animals to confirm or refute a theory, one should consider all the possible techniques that could be used to gather the necessary data. From this review, choose the method which offers the best chance of generating the necessary information in the most economical manner. Economy, in this context, refers to time, actual cost and the number of animals used. By considering the choices that are available for accomplishing the desired aim of the experiment and choosing the one that offers the best chance for success, one has met the requirements of this literal definition of alternative techniques.
Since a literal definition provides a rather simplistic approach to dealing with our responsibility for reducing the potential pain and suffering of animals that must be used, it is necessary to develop a working definition of the term. In Dr. Rowan's book, Of Mice, Models & Men, he defines the term alternatives to refer to those techniques or methods that "replace the use of laboratory animals altogether, reduce the numbers of animals required, or refine an existing procedure or technique so as to minimize the level of stress endured by the animal." Since stress can be difficult to describe and quantitate, for the purpose of this manual it will be replaced by the term distress. The working definition of alternative techniques thus evolves to "those techniques which replace the actual use of animals, reduce the numbers used, and/or refine the techniques to minimize the potential for the animal to experience pain or distress."
This concept of the 3 R's is not new. It first appeared in a book by Russell and Burch published in 1959 entitled The Principles of Humane Experimental Technique. In the original work, the authors defined the 3 R's as follows:
"Replacement means the substitution for conscious living higher animals of insentient material. Reduction means reduction in the numbers of animals used to obtain information of given amount and precision. Refinement means any decrease in the incidence or severity of in-humane procedures applied to those animals which still have to be used."
In this text the authors included nonrecovery techniques in anesthetized animals, as well as tissue culture, as replacement methods. Reduction included statistical techniques which were designed to reduce the actual numbers needed in the study. The use of better animals was also encouraged as a means of reducing actual numbers used. Refinement referred to techniques that reduced the potential for pain and distress. This approach still holds today. It is the principles of Replacement, Reduction and Refinement that will be covered in this chapter. To attempt to address these issues for all the uses of animals that fall under the general rubric of research, teaching and testing is far beyond the scope of this manual. Therefore the comments that follow will address only broad issues with some specific examples for the purpose of clarification.
Prior to discussing the replacement of animals with non-animal models, the word animal must be defined. On the surface this appears an easy task. Common sense would tell us that an animal is one of the two major kingdoms of living organisms. The dictionary defines an animal as "any of a kingdom of living beings typically differing from plants in capacity for spontaneous movement and rapid motor response to stimulation." In the Definition of Terms promulgated to implement the amended Animal Welfare Act an animal is defined as:
"any live or dead dog, cat, nonhuman primate, guinea pig, hamster, rabbit, or any other warm blooded animal, which is being used or is intended for use for research, testing, experimentation, or exhibition purposes or as a pet. This term excludes: Birds, rats of the genus Rattus and mice of genus Mus bred for use in research, and horses and other farm animals such as but not limited to livestock or poultry used or intended for use as food or fiber, or livestock, or poultry used or intended for use for improving animal nutrition, breeding, management, or production efficiency, or for improving the quality of food and fiber."
The PHS Policy defines an animal as "Any live, vertebrate animal used or intended for use in research, research training, experimentation, or biological testing or for related purposes." On the other hand the Guide defines an animal as "any warm blooded vertebrate animal." For the purposes of this manual, and to be consistent with most approaches to discussing alternative techniques, an animal will be any living vertebrate, with the caveat that any model system which moves down the phylogenetic scale from the generally acceptable animal model will be considered an alternative.
Alternatives which replace animal models can be classified into the following broad general categories:
Use of Living Systems
Use of Nonliving Systems
Use of Computer Simulation
Use of Living Systems
In Vitro Techniques - The most commonly recognized nonanimal living systems are those which fall into the broad category of in vitro methods such as organ, tissue and cell culture. These techniques afford the investigator the greatest control of the "test subject's" environment. Since these systems will not work when the incorrect combination of atmosphere, humidity, temperature, pH and nutrients are provided, they tend to minimize the effects that nonexperimental variables can have on the final outcome of a study. Generally, when suboptimal environments are provided for an in vitro system, the problem becomes one of loss of all experimental results and not just the production of compromised results. The most commonly used of the in vitro methods are cell culture techniques for monoclonal antibody production, virus vaccine production, vaccine potency testing, screening for the cytopathic effects of various compounds and studying the function and make up of cell membranes. The potential uses of in vitro techniques are almost limitless and will continue to expand as more is learned about the various organs and their component tissues and cells, and as the technology of maintaining in vitro environments improves.
Invertebrate Animals - Invertebrates are another type of living system which can be used to replace the more commonly used laboratory animals. Over 90 percent of the animal species thus far identified are invertebrates. An invertebrate which has long been used in biomedical research is the fruit fly, Drosophila melanogaster -- a classic model for the study of genetics. This species also can be used for detecting mutagenicity, teratogenicity and reproductive toxicity. The marine invertebrates represent different species which have not been widely investigated. However in neurobiology a number of different marine species have been well characterized and used to study the physiology of the nervous system.
Micro-Organisms - The micro-organisms represent a third system which has been used to replace traditional animal models. The Ames mutagenicity/carcinogenicity test uses Salmonella typhimurium cultures to screen compounds that formerly required the use of animals. Such systems allow for an almost limitless number of compounds to be tested which can create an interesting dilemma. The more compounds that can undergo screening, the more compounds that will be potentially available to test in animals. Alternative techniques can replace the number of animals at a given step in the screening process. However, use of alternatives may increase the number of compounds that must be finally tested in intact animals.
Plants - Plants offer another alternative living system which can be used to replace animals in studies of basic molecular mechanisms. There is very little morphological and functional difference between the organelles isolated from plants and those isolated from animals. The rigid cell wall of plants, however limits their applicability for use as undisrupted cells.
Use of Nonliving Systems
Chemical Techniques - The most widely used nonliving model system involves the use of modern chemical techniques. This is particularly true of the analytical techniques which can be used to identify substances and to determine their concentration or potency. Immunochemical techniques use the binding capacity of highly specific antibodies to seek out minute quantities of antigen. A classical example of this technique can be demonstrated by the currently used techniques for identifying bacterial toxins. Toxin identification previously required the injection of as many as several hundred mice with supernatant from cultures of suspected contaminating bacteria.
These new antibody techniques save animals and speed up confirmation of a tentative diagnosis. By adding a color marker to the Enzyme Linked Immunosorbent Assay system (ELISA), the whole process becomes a commercially available test kit such as those used in home pregnancy detection. A test that previously required the use of a rabbit now can be performed using an over-the-counter test kit. There are a variety of chemical techniques that can be used to determine the presence of a particular chemical reaction or the presence of an enzyme necessary for a specific reaction. At the most basic level, the identification of a particular chemical structure in a compound can provide a great deal of insight into the potential reactivity and thus the resulting toxicity of a given substance.
Physical and/or Mechanical Systems - The use of physical and/or mechanical systems to replace living animals of even the highest order has application in teaching specific skills and/or reactions to a well defined set of predetermined circumstances. The use of computer-linked mannequins in teaching basic principles of medicine and applied techniques can be best illustrated by the mannequins used to train people in cardiopulmonary resuscitation.
Historical data can be used for analyses in a variety of databases commonly used in the field of epidemiology. However, while the body of potentially useful information that already exists in a variety of sources is immense, it may not always be in a format which permits ready accessibility for evaluation. For this reason, retrospective epidemiological studies are often the subject of fairly heated debates. Yet with the increasing access to historical data available on existing computer programs, this problem may to a large extent be overcome in the future.
Use of Computer Simulation
The standout in the alternative techniques controversy is the claim made for computer simulation as a means of virtually replacing the use of living animals. In order for a biological phenomena to be adapted to a computer model, the basic processes must be expressed in a mathematical formula. Once a formula is developed then an enormous number of variables can be introduced and swiftly processed. The key element for success is the generation of a program from the mathematical formula. The more complete the formula, the more useful the program. The problem is that many of the questions being asked of an animal model are not defined well enough to develop the necessary mathematical model. As the core knowledge of the biological processes expands so will the opportunities to use computer simulation to replace the number of live animals being used.
In discussing the ways to reduce the numbers of animals used, the definition of an animal and the principle of moving down the phylogentic scale must also be kept in mind. The four broad categories for reducing the number of animals used are:
Animal Sharing
Improved Statistical Design
Phylogenetic Reduction
Better Quality Animals
Animal Sharing
Sharing of animals can significantly reduce the number of animals used within a given institution. Between institutions, sharing is more difficult, but can be effective as demonstrated with the Primate Supply Information Clearinghouse, Regional Primate Research Center (SJ-50) University of Washington, Seattle, WA 98195. This service has reduced the total number of primates used by helping to optimize the usage of those already in facilities throughout the country.
Sharing can be as simple as allowing someone to practice a surgical approach on an animal that has been, or is to be euthanatized for other purposes, or providing organs or tissues at the time of necropsy. Sharing becomes more complicated when attempting to maximize the use of control animals, but it can significantly reduce the number used within an institution. If two studies involve the need to perform a sham operation, the administration of compounds by identical routes, the use of standard control diets or the need to condition animals to a particular environment, control animals could be shared within the institution. Animal sharing would require some form of centralized clearing process within the Institutional Animal Care Program to match the needs of the various investigators and their studies.
Improved Statistical Design
Anyone who has ever taken a course in experimental design or applied statistics has been bombarded with the importance of consulting with the statistician during the design phase of the experiment and not when the data collected needs to be analyzed. Improper design of experimental protocols and/or the failure to use appropriate statistical methods can result in the usage of an inappropriate number of experimental animals. A variety of design strategies are available which can reduce the number of animals needed in a given study. Experimental protocols which utilize serial sacrifice, group sequential testing, and crossover designs can significantly reduce the numbers of animals required.
The availability of low cost statistical packages for almost every computer on the market permits more and more investigators access to sophisticated data management and analysis. This accessibility makes possible the use of design criteria and complicated statistical analysis which heretofore have been largely confined to institutions with large statistical support units. With this ability at their finger tips, investigators should be able to maximize the analysis of the data generated from each animal used, thus reducing the total numbers of animals necessary for a particular set of data.
Phylogenetic Reduction
Projects which can be designed to use one of the myriad of invertebrate species instead of a non-human primate species represent a type of phylogenetic reduction which was discussed as a replacement technique. Such broad jumps across the phylogenetic scale are not always possible, but less dramatic shifts can significantly reduce the numbers of higher species being used in research, teaching and testing. In many instances, the theory of phylogenetic reduction has been blurred by a species's use as a companion animal with little regard for phylogenetic ranking. The animals chosen for project usage should be the least advanced from a phylogenetic standpoint that will provide the necessary data.
The principle of phylogenetic reduction is generally well accepted as a way to reduce the number of animals used, but it often brings many hidden difficulties. As one descends the phylogenetic scale, the available information on the maintenance and use of these animals in a biomedical setting often becomes difficult, if not impossible, to obtain. When choosing a study model, it is critical that the principal investigator take into account the ability of the institution to provide appropriate care for the species chosen. Phylogenetic reduction is an important means of decreasing the number of animals used, but should be practiced carefully and with the full knowledge of the requirements of the species chosen.
Better Quality Animals
It is a rare study in which the initial cost of the animals to be used represents the single most expensive aspect of the study. For this reason it can often be false economy to select the source of the animal based on cost alone. When purchasing laboratory animals, it is important to keep in mind that cost and quality are usually directly correlated. By choosing the best quality animal in terms of health status, the possibility that animals will be lost or data compromised by the intrusion of a concurrent disease condition is minimized, if not eliminated. Choosing the best quality animals, in terms of genetic status, will virtually insure the consistency of animals from study to study. This requires an institutional commitment to the use of animals of defined health status and limits the investigators to the animal sources approved by the institution. Mixing of animals of different health status is a disaster waiting to happen and may negate all the benefits derived from the use of quality animals.
The role of the investigator and staff in assuring the integrity of an animal colony cannot be overemphasized. In choosing a source of animals, a veterinarian should be consulted to insure that the best animals that can be effectively maintained in the institution are purchased. Animals of different or unknown health status should never share the same environment nor common equipment in the animal facility or in the research laboratory.
Refinement refers to techniques which reduce the pain and distress to which an animal is subjected. For the purpose of this manual these techniques can be classified into the following broad categories:
Decreased Invasiveness
Improved Instrumentation
Improved Control of Pain
Improved Control of Techniques
Decreased Invasiveness
A hallmark of most of the new diagnostic and therapeutic techniques used in human medicine is the minimal degree of invasiveness that is required to successfully perform a procedure to obtain a given set of data. In many instances these techniques are applicable in the research environment and can be adopted for use in animals. A sophisticated example could be the use of Magnetic Resonance Imaging for results that formerly required euthanasia of multiple animals along a time curve to obtain assay tissue. Today one animal can provide all the information along a given curve. A less dramatic example is the vascular access device which permits repeated samples or injections in a single animal instead of using several animals. Invasiveness reduction methods are available in almost every area of biomedical research, and in project design, it is important to identify and use these methods wherever possible. Not only do they represent an alternative technique, but they generally provide much more consistent and reproducible data.
Improved Instrumentation
Monitoring Animals - In this age of microelectronics, fiber optics and laser instrumentation, the potential for refining techniques used in animal experimentation seems almost limitless. Improved instrumentation can minimize animal distress by reducing the level of restraint and/or manipulation necessary to obtain biological samples. Included in this category are the use of tethers in a variety of species to allow continuous access to the various organ systems, while permitting the animal virtually unrestricted movement within its primary enclosure. The advantages of these systems are numerous, not the least of which is minimizing a variety of nonexperimental variables associated with prolonged restraint.
Analyzing Samples - Once obtained, samples
can be analyzed in very small volumes for a multitude of parameters.
Examples of this can be found in the commercially available diagnostic
laboratory equipment which require only microliter blood samples
to perform a variety of diagnostic tests. The use of smaller sample
sizes permits the use of smaller animal species and prevents the
need to euthanatize many of these species to obtain the necessary
volume of blood. It is now possible to obtain serial blood samples
from small laboratory rodents which reduces the number of animals
necessary to obtain data over the length of the study.
Improved Control of Pain
The Animal Welfare Act requires "that the principal investigator consider alternatives to any procedure likely to produce pain or distress in an experimental animal" and in any practice which could cause pain to animals that a doctor of veterinary medicine is consulted in the planning of such procedures for the use of tranquilizers, analgesics and anesthetics. Since appropriate anesthetic and analgesic agents can minimize the potential pain and distress experienced by animals, an entire chapter of this manual is devoted to the principles of using these agents. Suffice it to say, that of all the possible ways that the 3 R's can be utilized this is an area where the laboratory animal veterinarian can often be of most help to the investigator.
Improved Control of Techniques
Proficiency in the handling and restraint of animals makes it easier to perform a variety of routine procedures with minimal or no pain or distress to the animals involved. Animals are creatures of habit and when proper handling is part of their regular routine, the degree of distress caused by the procedures is minimized. Animals can be trained or conditioned to accept a variety of procedures which if suddenly forced upon them can be distressful. Almost every animal commonly used in the laboratory responds positively to a little tender loving care. It's inexpensive, readily portable, safe even at the highest doses and spreads rapidly through the staff. To develop the proper techniques and gain confidence in their use requires training by someone with appropriate experience. This can be the veterinarian, a member of the animal care staff or a fellow investigator. Whomever it may be should be sought out before a new species or technique is incorporated into the study. This will reduce the potential distress of all animals involved in the study up to and including the principal investigator.
In this chapter, the use of alternative techniques has been defined in terms of the present regulatory requirements and the principles of Replacement, Reduction and Refinement were introduced. In summary, the reader should consider a fourth R--Responsibility. The use of animals in teaching and research brings with it a responsibility to minimize animal pain and distress. The adoption of the 3 Rs as part of the process of planning and conducting projects using laboratory animals will go a long way toward implementing Responsibility--the fourth R.
Animal Welfare Act (Title 7 U.S.C. 2131-2156) as amended by PL 99-198, December 23, 1985.
Guide for the Care and Use of Laboratory Animals, NIH Publication No. 86-23.
Models for Biomedical Research: A New Perspective, l985. National Academy Press, Wash., DC; l985.
Navian, J.B. Animal Models in Dental Research. The University of Alabama Press.
Paton, William. Man & Mouse Animals in Medical Research. Oxford University Press, New York, 1984.
Public Health Service Policy on Humane Care and Use of Laboratory Animals. Revised September 1986.
Public Law 99-198. Code of Federal Regulations, Title 9, Subchapter A, Animal Welfare, 1989.
Rowan, A.N. Of Mice, Models, & Men: A Critical Evaluation of Animal Research. State Univ. of N., 1984.
Russel, W.M.S. and Burch, R.L. The Principles of Humane Experimental Technique, Methuen & Co, Ltd., London, 1959.
U. S. Congress, Office of Technology Assessment. Alternatives to Animal Use in Research, Testing, and Education. (OTA-BA-273, Feb. 1986)
Webster's Ninth New Collegiate Dictionary, Merriam-Webster, Inc., Springfield, MA; 1986.
Wessler, S. 1976. Animal Models of Thrombosis and Hemorrhagic Diseases, NIH Publication No. 76-982.
