Information Resources for Institutional Animal Care and Use Committees 1985-1999 *************************

Alternatives and Database Searching

The Search for Alternatives

Tim Allen and D'Anna Jensen
U.S. Department of Agriculture, Animal Welfare Information Center
Beltsville, MD USA

Editor's note: This article originally appeared in the CAAT Newsletter, Winter 1996: Vol. 13, No. 2.

[Mention of commercial enterprises or brand names does not constitute endorsement or imply preference by the U.S. Department of Agriculture. The views expressed by the authors do not necessarily represent positions or policies of the U.S. Department o f Agriculture or any agency thereof and should not be interpreted as such.]

"Do we see some veterinarians still pursuing the methods of the ancients and perpetrating pain on a helpless subject... The result has been, and the feeling still exists, largely among the laity, that we are a hard-hearted profession..."

A modern day diatribe by animal activists. Not really. Those words were written by Dr. J.P. Turner in the Proceedings of the American Veterinary Medical Association in 1899. He was lamenting the fact that many of his colleagues were resisting the use of anesthetics to restrain animals during surgery in favor of hobbles and rope tie-downs, "the accustomed way , or the methods we were taught." While things have certainly changed, it is still not uncommon to hear "that's the way we've always done things." However, with the passage of the Improved Standards for Laboratory Animals Act in 1985, Congress let it be known that it is concerned about the use of animals in painful procedures. Under this law, scientists performing painful experiments on animals must document if there are alternative methods to the painful procedure and report this information to the Institutional Animal Care and Use Committee (IACUC) when they submit their animal use protocol form for approval. It is then the responsibility of the IACUC to determine if the alternative methods should be used. To assist IACUCs and investigators in complying with this portion of the law, Congress established the Animal Welfare Information Center (AWIC) at the National Agricultural Library. In the nex t few pages, we will look at the critical role that IACUC's play in the animal use approval process, especially the problems associated with documenting whether or not alternatives exist, and how AWIC can assist members of an IACUC and/or scientists.

The U.S. Department of Agriculture views alternatives with an eye to the 3R's concept so eloquently described by W.M.S. Russell and R.L. Burch in The Principles of Humane Experimental Technique--reduction of animal numbers, refined procedures to minimize or avoid pain, and replacement of animals with non-animal models. According to the Animal Welfare Act (AWA) regulations (9 CFR ' 2.31(d)), "the IACUC shall determine that the principle investigator has considered alternatives to procedures that may cause more than slight pain or distress to the animals, and has provided a written narrative of the methods and sources used to determine that alternatives were not available." The IACUC is also responsible for ensuring that the proposed research does not unnecessarily duplicate other research. Along with several other items, AWIC considers these the information requirements of the act (the other sections being scientific justification for withholdin g anesthetics or analgesics, or using animals in more than one major operative procedure from which s/he is allowed to recover).

While the regulations seem fairly straight forward, it has been our observation that many people are unsure exactly what an alternative is and are confused as to what information is required to show compliance. There are many opportunities to incorporate alternatives into an experimental procedure; however, many IACUCs and scientists mistakenly assume that only non-animal methods satisfy the definition of an alternative. Although outside the scope of this paper, some alternatives might include pair-ho using of rodents to alleviate the distress of isolation, proper use of analgesics in a post-procedural period, or reducing the volume of a receptor binding assay thereby reducing the amount of animal tissue needed to quantify the reaction and ultimately reducing the number of animals. The point is that IACUC's and investigators need to fully understand that identifying viable alternatives requires more than looking for non-animal models.

Animal welfare regulations require, as a minimum, that an investigator perform a search of the literature in an attempt to identify alternatives to painful procedures. Cynthia Smith, an AWIC staff member, wrote a method paper on searching for alternatives that is an excellent overview of this type of searching. But what is important to realize is that a multidatabase approach is necessary, as an alternative procedure or method may come from outside the specific discipline being studied. For example, if you concentrate on mammalian models for studying Parkinson's disease or diabetes, emerging fish models may be overlooked.

It is also important to conduct the literature search on a case by case basis. AWIC staff often are asked by an IACUC to perform a literature search on a painful procedure outside of the context of an experiment. It is impossible to look for alternatives to something as general as thoracotomies in dogs. Some of the questions that need to be addressed are why is the procedure being performed? What is the expected outcome? Is the procedure terminal? Only with complete information can a search be performed, and the IACUC properly evaluate the literature search.

Some IACUC's require attaching a literature search to the protocol with a list of the databases and strategy used to show that a good-faith effort was made to find alternatives. Many others require only that a box be checked indicating that alternatives are not available or may simply ask for a few key words and the database searched. Still others list AWIC as a source of information on the protocol form leading to many requests for information. Regardless of the system used all are fraught with problems. When an investigator contacts AWIC for help in completing an alternatives search, we commonly ask them to fax a copy of the protocol to us so that we will have all pertinent experimental information at hand. It is not uncommon to find the statement "AWIC was consulted and no alternatives were found" typed onto the protocol sheet that we are seeing for the first time. Oftentimes it is plain to see that the alternatives search is clearly an afterthought, being performed simply to comply with the law. The most common refrain is, "I'm turning in my protocol tomorrow, and I see that I have to have a literature search, can you fax that to me?" In our roles as members of Federal IACUC's, we routinely see protocol forms filled out stating that a literature search was performed, but, when we ask the investigator to provide us with a copy of the search, it usually has not been conducted. In other cases, the entire concept of alternatives is simply ignored by both the IACUC and the investigator. Are these examples the norm? Maybe not, but they occur often enough that there clearly is a problem with IACUC oversight of this particular part of the regulations. Comments made to us at meetings or workshops reveal that many scientists and IACUC members view the alternatives search as unnecessary government intrusion into the research process, and not as a resource that might enhance or improve their research. Not surprisingly, a Department of Agriculture report on enforcement of the AWA by the Animal and Plant Health Inspection Service, Regulatory Enforcement and Animal Care (ed. note: this unit is now called Animal Care), found that IACUC's do not always meet the standards of the act and that this is attributable to the fact that committee members are not always aware of the act's requirements. Two of the major deficiencies noted are failure to properly address the use of alternatives and failure to provide written assurance that activities are not unnecessarily duplicative.

With these problems fresh in mind, what can or does AWIC do to help animal care committees comply with the law? AWIC was established to provide information pertinent to employee training , to prevent unintended duplication of animal experimentation, to reduce or replace animals used in painful experimentation, or on refined methods to minimize pain to animals when no other model can be found. To help IACUC's, investigators, and animal research support people understand the alternatives section of the regulations, AWIC staff developed a two-day workshop called "Meeting the Information Requirements of the Animal Welfare Act." The workshop provides an overview of the Animal Welfare Act looking specifically at the information requirements, Federally mandated IACUC functions, criteria for granting IACUC approval for animal research, and the required contents for an institutional training program. A representative from the U.S. Department of Agriculture's REAC staff is also available for a question and answer period. The workshop also provides an overview of the "alternatives concept," multiple database resources, concepts involved in developing search strategies (but no magic formulas), and, finally, the opportunity to gain hands-on searching experience using the DIALOG database system.

The success of the workshop is measured not only by the fact that every class held at the National Agricultural Library is booked months in advance but also by the number of requests we receive to bring the workshop to offsite facilities. Even more importantly, however, are the comments received from people who have taken the training class. The most common sentiment is that the class should be required for all members of IACUC's as it addresses many of the problems common to successful IACUC functioning.

If it is true as Ben Franklin said that an investment in knowledge pays the best interest, then perhaps AWIC's greatest utility to the scientific community is the capability of providing comprehensive literature searches or other information on alternatives, animal husbandry, animal models, philosophical issues, and many other topics related to animal research. When AWIC is requested by an IACUC or an investigator to perform a literature search, the package of information they receive includes the search strategy, the databases searched, and the literature information that documents whether alternatives are available and if the research is duplicative. We may also include a copy o f one or two pertinent articles. Many IACUC's, working through institutional libraries, also maintain collections of bibliographies produced by AWIC on topics from anesthesia and analgesia to zoonoses. The AWIC staff also produces a newsletter that covers topics such as environmental enrichment, IACUC communications, alternatives, etc.

How is AWIC able to provide such a breadth of information to such a diverse audience? We owe this ability to a much underutilized resource, the National Agricultural Library (NAL), as well as new technology such as the World Wide Web, and the numerous databases available through services such as DIALOG. The NAL houses one of the largest collections of veterinary literature in the world, and is developing one of the most comprehensive collections of laboratory animal literature. These materials include NAL's AGRICOLA database, more than 400 videos and slide programs that can be used in institutional training programs, most relevant journals, codes of practice, newsletters, texts, and other published materials such as conference proceedings and abstracts relating to laboratory animals and farm animals used in biomedical research. Because of international exchange agreements, AWIC and NAL also work closely with other agencies providing information or regulatory oversight to animal care committees throughout the world. In this way, we are able to bring a broader perspective to many issues.

In 1996, AWIC is celebrating its 10th anniversary. In its brief existence, AWIC and NAL have worked hard to develop a comprehensive resource to assist IACUC's in carrying out their enormous responsibilities. Animal care committees face many problems in assuring that their institutions are complying with the Animal Welfare Act and the Animal Welfare Information Center is available to help them.

For additional information contact the staff at: Animal Welfare Information Center, U.S. Department of Agriculture, Agricultural Research Service, National Agricultural Library, 10301 Baltimore Avenue, Beltsville, MD USA 20705-2351, Tel: (301) 504-6212, Fax: (301) 504-7125, Contact us: ,


Allen, T. (1994). Meeting the information requirements of the Animal Welfare Act. A workshop. Animal Welfare Information Center Newsletter 5(3): 6.

Byrnes, C. (1995). RRReaching the A-Zone. Science and Animal Care 6(4): 1-2,4.

Code of Federal Regulations (1995), Title 9, Part 1, Subchapter A, Animal Welfare.

Russell, W.M.S. and R.L. Burch (1959). The Principles of Humane Experimental Technique. Universities Federation for Animal Welfare: Potters Bar, Hertfordshire, England, 238 pp.

Smith, C.P. (1994). AWIC tips for searching for alternatives to animal research and testing. Lab Animal March 1994: 46-48.

Stokes, W.S. and D.J.B. Jensen (1995). Guidelines for institutional animal care and use committees: consideration of alternatives. Contemporary Topics 34(3): 51-60.

Turner, J.P. (1899). A plea for the more general use of anaesthesia in veterinary surgery. Proceedings of the American Veterinary Medical Association Session of 1899, p.103-111.

U. S. Department of Agriculture, Office of Inspector General, Audit Report No. 33600-1-Ch, January 1995, 60 pp.


Allen, T. (1997). Online databases and the World Wide Web. What is available? What is missing? In Animal alternatives, welfare, and ethics, Proceedings of the 2nd World Congress on Alternatives and Animal Use in the Life Sciences, held in Utrecht, the Netherlands, 20-24 October 1996, L.F.M. Zutphen and M. Balls (eds.), Amsterdam, New York: Elsevier, pp. 483-490.
NAL call number: QL1.D48 v.27
Descriptors: databases, animal testing alternatives, searching for alternatives, information retrieval, terminology, scientific writing.

Allen, T. and D. Jensen (1996). IACUCs and AWIC: the search for alternatives. The Johns Hopkins Center for Alternatives to Animal Testing Newsletter 13(2): 1,3-6.
NAL call number: HV4701.J6
Descriptors: animal testing alternatives, information retrieval, information centers, committees.

Balls, M., W. de Klerck, F. Baker, M. van Beek, C. Bouillon, L. Bruner, J. Carstensen, M. Chamberlain, M. Cottin, R. Curren, and J. Dupuis (1995). Development and validation of non-animal tests and testing strategies: the identification of a coordinated response to the challenge and the opportunity presented by the Sixth Amendment to the Cosmetics Directive (76/768/EEC). Alternatives to Laboratory Animals: ATLA 23(3): 398-409.
NAL call number: Z7994.L3A5
Descriptors: animal testing alternatives, cosmetics, European Union regulations, product development, animal welfare, eyes, skin, laboratory animals, mutagenicity, injection, European Union.

Bowd, A.D. (1994). The role of animal care committees in fostering use of alternatives: A case study. In Vitro Toxicology 7(2): 169.
Descriptors: committees, alternatives to animal testing.

Dawson, M. (1996). Guidelines for the choice of an alternative to proposed animal experiments. Developments in Biological Standardization 86: 329.
Descriptors: animal testing alternatives, animal welfare, Great Britain, licensing.

Gettings, S.D., D.M. Bagley, M. Chudkowski, J.L. Demetrullas, L.C. Dipasquale, C. L. Galli, R. Gay, K.L. Hintze, J. Janus, and K.D. Marenus (1992). Development of potential alternatives to the Draize eye test: the CTFA evaluation of alternatives program. Phase II. Review of materials and methods. Alternatives to Laboratory Animals: ATLA 20 (1): 164-171.
NAL call number: Z7994.L3A5
Abstract: The CTFA Evaluation of Alternatives Program is a multi-year effort, organised by the CTFA Animal Welfare Task Force, designed to evaluate the performance of currently promising in vitro (alternative) methods to the Draize eye irritancy test. The sole criterion for inclusion of a particular test is that it shows some initial promise as an alternative to the Draize eye test, and that it is under evaluation or development by a participating CTFA member company. Tests are evaluated for their ability to rank and discriminate the ocular irritation potential of prototype cosmetic and personal care formulations compared to the Draize eye test. Test materials and in vitro methods currently under evaluation in Phase Il of the CTFA Program are described. Additional tests may be included in subsequent phases of the Program, should it be determined that they show particular promise as replacements for specific types of formulation. Conversely (at the discretion of sponsors), tests may be removed from the Program should initial promise be unfulfilled.
Descriptors: animal testing alternatives, evaluation, organizations.

Hart, L.A. (1995). The animal subjects protocol process: Applying the 3Rs. Lab Animal 24(5): 40-43.
NAL call number: QL55 A1L33
Descriptors: protocol preparation, protocol review, investigator's responsibilities, IACUC responsibilities, importance of animal wellbeing, alternatives, reducing sources of discomfort, approaches for the investigator, review of the literature, literature searching.

Hem, A., A.J. Smith, and P. Solberg (1998). Saphenous vein puncture for blood sampling of the mouse, rat, hamster, gerbil, guineapig, ferret and mink. Laboratory Animals 32(4): 364-368. This technique may be viewed at (PDF 847KB)
NAL call number: QL55 A1L3
Descriptors: blood collection, techniques, animal welfare, alternatives to retroorbital bleeding.

Hendriksen, C.F. (1996). A short history of the use of animals in vaccine development and quality control. Developments in Biological Standardization 86: 3-10.
Abstract: Man has been using animals since early times to gain an insight into health, illness and death. The oldest known medical standard work, the Corpus Hippocraticum (circa 350 BC), contains descriptions of experiments on pigs. Although the first attempt at immunoprophylaxis dates as far back as the 6th century (variolation was practised in China to protect people against smallpox), it was not until the middle of the 19th century that animal experimentation acquired full scientific status in the development and quality control of immunobiological products. It was Louis Pasteur and Robert Koch who, through studies on animals, succeeded in underpinning the causal relationship between infectious diseases and micro-organisms, thus opening the way to the discovery of effective therapeutic and prophylactic agents for a number of these diseases. In several respects, the experimental animal work carried out in the last decade of the 19th century to find an effective and reliable way of treating and preventing diphtheria determined the use of animals. Many common routine animal tests in the quality control of immunobiologicals arose from diphtheria research. Conversely, diphtheria was one of the first diseases where experimental animal research laid the foundation for effectively reducing child mortality. This had a very profound impact on the attitude of society towards animal experiments in those days and almost completely eliminated the growing influence of the antivivisection movement. The interest in the possibilities of replacement, reduction and refinement (the Three-Rs concept) of the use of laboratory animals is increasing for several reasons, including concern about animal welfare. The root of animal welfare can be traced back to the 18th century with the formulation of utilitarian ethics. One characteristic feature of these ethics was that the interests of any creature which is submitted to any procedure should be taken into consideration. This presentation sets out some major historical contributions of animal experiments to the development and quality control of immunobiologicals. Attention is also paid to the changing attitude of society towards animal experiments and its impact on the development of alternative methods. It is concluded that, although animal experiments have played an important part, a new area is now beginning in which increasing emphasis will be placed on in vitro methods.
Descriptors: animal welfare, diphtheria, poliovirus vaccine, quality control, animal testing alternatives.

Holden, F. (1997). Alternatives committee established at Indiana. The Johns Hopkins Center for Alternatives to Animal Testing Newsletter 14(3): 6-7.
NAL call number: HV4701 J6
Descriptors: subcommittee to IACUC, communications between researchers and campus animal protectionists, monthly round table, institutional support at highest levels, membership includes--information specialists, public relations/education representative, departmental representatives, IACUC liaison, animal protectionist, veterinarian, research assistant.

Huggins, J. (1998). Co-occurring words: finding information about alternatives to animal testing. ATLA-Alternatives to Laboratory Animals 26(6): 849-856.
NAL call number: Z7994 L3A5
Descriptors: databases, indexing, skin, literature searching, utilization of terms which co-occur can enhance information retrieval, toxicology, scientific methods, endpoints measured, informatics research.

Huggins, J. (1997). Communication by keywords: Sharing information about alternatives to animal testing. Animal Welfare Information Center Newsletter 8(2): 9-11.
NAL call number: aHV4701
Descriptors: stresses need for defined vocabulary for alternatives to animal testing, need for scientists to incorporate keywords about the methods used during experiments, toxicology, endpoints, keywords for alternatives to skin irritation.

Irwin, M.H., R.J. Moffatt, and C.A. Pinkert (1996). Identification of transgenic mice by PCR analysis of saliva. Nature Biotechnology 14(September):1146-1148.
NAL call number: QH442 B5
Descriptors: alternative to surgically obtaining samples, nested primers, gene integration, DNA purification.

Jackson, L.R., L.J. Trudel, and N.S. Lipman (1999). Small-scale monoclonal antibody production in vitro: methods and resources. Lab Animal 28(3):20-30.
NAL call number: QL55 A1L33
Abstract: Monoclonal antibodies (MAbs) are valuable research tools; however, MAb production via the mouse ascites method has come under recent scrutiny, due to the pain and distress it may cause the animal. The authors present a review of in vitro production of MAbs, as well as critical considerations in selecting the appropriate technique.

Jackson M.R. (1998). Priorities in the development of alternative methodologies in the pharmaceutical industry. Archives of Toxicology 20(suppl): 61-70.
NAL call number: RA1190 A7
Abstract: Promotion of animal welfare is an underlying and laudable goal for toxicologists and there is good reason to adopt practical, focused, investigative approaches towards this aim. Pharmaceutical regulatory toxicology can be subdivided into the areas of systemic (target organ), reproductive, genetic and topical toxicology, as well as immunotoxicology and oncology. These areas can be assessed for prioritisation as to where adoption of measures to promote any or all of the 3 Rs (reduction, replacement, refinement) would lead to the most tangible benefit for animals. These measures can range, for example, from replacement of animal experimentation with alternative in vitro techniques, to adoption of regulatory protocols that reduce the number of animals required. This paper is confined to consideration of in vitro technology in terms of reducing/replacing laboratory animal use, and a suggested list of criteria for prioritisation is potential for: - Regulatory acceptability, Reducing development cost, Reducing animal numbers, Promoting welfare aspects, Elucidating toxic mechanisms, Usefulness in compound selection, Advancing the science of toxicology. Clear messages emerge from such an analysis which could influence prioritisation of the application of in vitro toxicology from the standpoints of animal welfare, feasibility and resources.
Descriptors: alternatives, animal welfare, drug industry, toxicology methods, drug approval, drug screening, Great Britain.

Jennings, M., G.R. Batchelor, P.F. Brain, A. Dick, H. Elliot, R.J. Francis, R.C. Hubrecht, J.L. Hurst, D.B. Morton, A.G. Peters, R. Raymond, G.D. Sales, C.M. Sherwin, and C. West (1998). Refining rodent husbandry: the mouse. Report of the rodent refinement working party. Laboratory Animals 32(3): 233-259.
NAL call number: QL55 A1L3
Descriptors: animal husbandry, laboratory animals, animal welfare, animal housing, hygiene, mice.

Jennings, V.M. (1995). Review of selected adjuvants used in antibody production. ILAR Journal 37 (3): 119-124.
NAL call number: QL55.A1I43.
Descriptors: adjuvants, immunostimulants, antibodies, biological production, toxicity, adverse effects, pain, granuloma, arthritis, animal welfare, laboratory animals.

Johns Hopkins University Center for Alternatives to Animal Testing (CAAT) (1997). Alternatives in monoclonal antibody production Technical Report No. 8. Baltimore: CAAT, 41 p.
Available from
Descriptors: overview, hollow fiber reactors, comparison of in vitro and in vivo techniques, cost comparisons, quality comparisons, quantity comparisons, core laboratories, regulatory issues, European perspective, IACUC responsibilities, recommendations.

Johns Hopkins University, School of Hygiene and Public Health (1992). Animal Care and Use Committees and Alternatives, a Symposium Sponsored by the Johns Hopkins School of Hygiene and Public Health, Office for Research Subjects, June 18, 1992. Baltimore, Maryland: Johns Hopkins University, 110 p.
NAL call number: HV4704.A53 1992.
Descriptors: animal welfare, alternatives to animal testing, laboratory animals.

Kirchain, S. and R.P. Marini (1998). A tissue harvesting program as a method for implementing the 3Rs of biomedical research. Lab Animal 27(8): 37-39.
NAL call number: QL55 A1L33
Descriptors: reducing animal use, alternatives to animal testing, centralized database, information includes protocols, species, tissue donations and requests, sample forms, administration, IACUC approval, adoption program.

Koeter, H.B.W.M. (1993). The science and the art of regulatory toxicology: how to deal with alternative tests. In Current Trends: in vitro skin toxicology and eye irritancy testing: proceedings of the symposium, April 21-23, 1993, Radisson Hotel, Ottawa, Ontario, Canada, Ottawa : Joseph F. Morgan Research Foundation, p. 15-22.
NAL call number: RA1199.4.I5C87 1993.
Descriptors: animal testing alternatives, toxicology, regulations, guidelines, animal welfare.

Kreger, M.D. (1999). The literature search for alternatives. In The Care and Feeding of an IACUC: The Organization and Management of an Institutional Animal Care and Use Committee, M.L. Podolsky and V.S. Lukas, eds., Boca Raton, FL: CRC Press:, pp. 139-152.
Abstract: Describes the process of developing and executing a multidatabase literature search for alternatives.
Descriptors: alternatives, databases, literature search, evaluation, IACUC protocol.

Kreger, M.D. (1997). Why conduct literature searches for alternatives? ASLAP Newsletter (American Society of Laboratory Animal Practitioners) 30(3):19-23.
NAL call number: QL55 S97
Abstract: Describes the legal, ethical, and practical rational for running a multidatabase search for alternatives. Explains the usefulness of the search and gives tips to reduce online fees.
Descriptors: literature search, Policy 12, Animal Welfare Act.

Langley, G., C. Broadhead, K. Bottrill, R. Combes, R. Ewbank, P. Hawkins, R. Hubrecht, M. Jennings, C. Newman, S. Rowe, J, Southtree, M. Todd, and L. Ward (1999). Accessing information on the reduction, refinement, and replacement of animal experiments. ATLA-Alternatives to Laboratory Animals 27(2): 239-245.
NAL call number: Z7994 L3A5
Descriptors: European regulations, information resources, problems with current resources, databases, websites, proposed solutions, recommendations.

Leenaars, P.P.A.M., M.A. Koedam, P.W. Wester, V. Baumans, E. Claassen, and C.F.M. Hendriksen (1998). Assessment of side effects induced by injection of different adjuvant/antigen combinations in rabbits and mice. Laboratory Animals 32(4): 387-406.
NAL call number: QL55 A1L3
Abstract: We evaluated the side effects induced by injection of Freund's adjuvant (FA) and alternative adjuvants combined with different antigens. Rabbits and mice were injected subcutaneously, intramuscularly (rabbits) and intraperitoneally (mice) with different adjuvants (FA, Specol, RIBI, TiterMax, Montanide ISA50) in combination with several types of antigens (synthetic peptides, autoantigen, glycolipid, protein, mycoplasma or viruses). The effects of treatment on the animals' well-being were assessed by clinical and behavioural changes (POT and LABORAS assays) and gross and histopathological changes. In rabbits, treatment did not appear to induce acute or prolonged pain and distress. Mice showed behavioural changes immediately after (predominantly secondary) immunization. Injection of several adjuvant/antigen mixtures resulted in severe pathological changes, depending on adjuvant, type of antigen, animal species used and route of injection. Both rabbits and mice showed pathological changes ranging from marked to severe after injection of FA, and ranging from minimal to marked after Specol and Montanide injections. Pathological changes after RIBI injections were severe in rabbits, though slight in mice. After TiterMax injections, pathological changes were moderate in rabbits, though severe in mice. In conclusion, injection of FA according to present guidelines resulted mostly in severe pathological changes, whereas only very few clinical and behavioural signs indicated prolonged severe pain. Our findings indicate that Montanide ISA50 and Specol induce acceptable antibody titres, and cause fewer pathological changes than FA. Thus they are effective alternatives to FA.

Leeuw, W.A. de, P. de Greeve, H. Schoffl, H. Spielmann, and H.A. Tritthart (1997). Experience with the Dutch Code of Practice for the immunization of laboratory animals. Ersatz- und Erganzungsmethoden zu Tierversuchen: Forschung ohne Tierversu che 1996. Fourth Austrian International Congress on Replacement and Alternative Methods for Laboratory Animals in Biomedical Science, 24-26 September 1995, University of Linz. Wien, Austria: Springer-Verlag Wien, pp.210-217.
NAL call number: HV4913 F672 1997
Descriptors: laboratory animals, immunization, ethics, animal welfare, monoclonal antibodies, alternatives to animal testing, Netherlands.

Martin, B.J., J.B.Watkins, 3rd, and J. Ramsey (1998). Venipuncture in the medical physiology laboratory. American Journal of Physiology 274(6 Pt 2): S62-7.
Abstract: Medical physiology laboratories, traditionally devoted to animal experimentation, face unprecedented difficulties linked to cost, staffing, instrumentation, and the use of animals. At the same time, laboratory experiences with living creatures play a unique role in medical education. In this article we describe the use of venipuncture and subsequent blood analysis, with medical students serving as both subjects and experimenters, in a sequence of first-year physiology laboratories. These experiments are safe, robust, inexpensive, and time efficient, and they teach the principles of cardiovascular, respiratory, renal, nutritional, and gastrointestinal physiology. In addition, they enhance medical education in several other important dimensions. First, they teach safe venous blood collection and handling, a training appropriate for students at this level. Second, by serving each week as subjects as well as experimenters, students experience aspects of both sides of the doctor-patient relationship. Third, the laboratories can be used to teach fundamentals of research design and analysis. Finally, because blood analysis is central to medicine, and because the student's own blood data are discussed, students are enthusiastic and cooperative, and the clinical relevance of the data is clear.
Descriptors: medical education, phlebotomy, physiology education, technology, medical laboratory education, glucose tolerance test, hematocrit, hemoglobins-analysis, hemostasis-physiology, kidney-physiology, metabolism-physiology, nutrition, respiration-physiology, teaching.

McArdle, J. (1998). Alternatives to ascites production of monoclonal antibodies. Animal Welfare Information Center Newsletter 8(3-4): 1-2, 15-18.
NAL call number: aHV4701 A9522
Descriptors: ascites, monoclonal antibodies, laboratory animals, animal welfare, methods, mice.

Morton, D.B. (1998). The importance of non-statistical design in refining animal experiments. ANZCCART News 11(2): 1-12(Insert).
NAL call number: SF405.5 A3
Descriptors: laboratory animals, animal experiments, animal welfare, ethics, pain, experimental design.

Orlans, F.B. (1996). The three Rs in the research and education: a long road ahead in the United States. Alternatives to Laboratory Animals: ATLA 24 (2): 151-158.
NAL call number: Z7994.L3A5.
Abstract: Attitudes toward the Three Rs concept of refinement, reduction and replacement in the United States in research and education are widely divergent. Positive responses have come from several sources, notably from four centres established to disseminate information about alternatives. Funding sources to support work in the Three Rs have proliferated. The activities of institutional oversight committees have resulted in the nationwide implementation of important refinements. In the field of education, student projects involving pain or death for sentient animals have declined, and the right of students to object to participation in animal experiments on ethical grounds has been widely established. However, there is still a long way to go. Resistance to alternatives is deep-seated within several of the scientific disciplines most closely associated with animal research. The response of the National Institutes of Health to potentially important Congressional directives on the Three Rs has been unsatisfactory. The prestigious National Association of Biology Teachers, which at first endorsed the use of alternatives in education, later rescinded this policy, because of opposition to it. An impediment to progress is the extreme polarisation of viewpoints between the biomedical community and the animal protectionists.
Descriptors: animal testing alternatives, animal experiments, education, animal welfare.

Pakes, S.P. (1990). Contributions of the laboratory animal veterinarian to refining animal experiments in toxicology. Fundamentals of Applied Toxicology 15(1): 17-24.
Descriptors: animal pain, psychology, measurement, research design, alternatives, trends, veterinarians, ACUC.

Pavletic, M.M., A. Schwartz, J. Berg, D. Knapp (1994). An assessment of the outcome of the alternative medical and surgical laboratory program at Tufts University [School of Veterinary Medicine]. Journal of the American Veterinary Medical Association 205(1): 97-100.
Descriptors: animal welfare, cadavers, dogs, euthanasia, veterinary education, alternatives.

Purchase, I.F., P.A. Botham, L.H. Bruner, O.P. Flint, J.M. Frazier, and W.S. Stokes (1998). Workshop overview: scientific and regulatory challenges for the reduction, refinement, and replacement of animals in toxicity testing. Toxicology Science 43(2): 86-101.
Abstract: Public concern for animal welfare has been expressed through legislative control of animal use for experimental purposes since the first legislation was introduced in 1876 in the United Kingdom. Legislative control of animal use has been introduced in virtually every developed country, with major initiatives in Europe (1986) and the United States (1966 and 1985). Advances in scientific thinking resulted in the development of the concept of the three Rs--refinement, reduction, and replacement--by Russell and Burch in 1959. The field has expanded substantially since, with specialist scientific journals dedicated to alternatives, World Congresses organized to discuss the scientific and philosophical issues, and European and U.S. validation organizations being launched. Current scientific attention is focused on validation of alternative methods. The underlying scientific principles of chemical toxicity are complicated and insufficiently understood for alternative methods for all toxicity endpoints of importance in protecting human health to be available. Important lessons have been learned about how to validate methods, including the need to have prediction models available before the validation is undertaken, the need to understand the variability of the animal-based data which is to be used as the validation standard, and the need to have well-managed validation programs. Future progress will depend on the development of novel methods, which can now be validated through international collaborative efforts.
Descriptors: animal testing, alternatives, regulations, legislation, education, Europe, Great Britain, United States reproducibility of results, toxicology.

Ray, S. (1998). An alternative to water deprivation techniques in animal learning studies. Animal Technology 49(3): 113-120.
NAL call number: QL55 I5
Abstract: Many laboratories use a period of water deprivation to motivate animals on a variety of water reinforced learning paradigms including aversive conditioning and maze learning tasks. Such procedures can lead to long periods without water and increase inter-animal variability in learning performance. Reported is an alternative procedure using sucrose rich drinks, or sucrose solutions, as a reward in maze and discrimination learning procedures with no prior water deprivation.
An initial experiment compares performance over trials of a water deprived group of rats learning to negotiate a Y maze, and a group of genetically matched animals running an identical maze with no water deprivation. Both groups negotiating the maze for a sucrose reward. Results show that non-deprived animals showed teaming that was equally as good as the water deprived animals. Similar results were confirmed in a Lashley jump stand discrimination task. The ability to study learning in non-deprived animals may be of great interest in studies of learned behaviour after lesion or other surgical interventions, when periods of dehydration may affect the animal's health. Further, the development of non-deprivation motivated techniques will reduce the severity of many commonly employed rodent learning paradigms.
These results may also offer a useful heuristic to explore learning paradigms without food or water deprivation schedules in other species.

Rowan, A.N. (1995). The third R: refinement. Alternatives to Laboratory Animals: ATLA 23(3): 332-346.
NAL call number: Z7994.L3A5
Abstract: This review attempts to provide an introduction to the complicated subject of refinement, the third R in the concept of alternatives. It starts with a brief discussion of what refinement means and the lack of specific attention paid to this third R. This is followed by an analysis of the conceptual underpinnings of pain, distress and suffering, and the problems of both definition and measurement which must be faced if we are to be objective and consistent in our search for refinement. The review then touches upon husbandry, care and handling issues as they affect animal discomfort and distress. Antibody production, both polyclonal and monoclonal, is discussed as an example of the refinement of research techniques Finally, a few brief comments are offered on the refinement of a variety of other experimental techniques, including those used in toxicology, cancer research and behavioural research.
Descriptors: animal experiments, pain, anxiety, stress factors, animal welfare, mice, inflammation.

Schwetz, B and D. Gaylor (1998). Alternative tests: carcinogenesis as an example. Environmental Health Perspective 106(Suppl 2): 467-71.
NAL call number: RA565 A1E54 v.106 suppl.2
Abstract: Acceptance of new tests that are alternatives to currently used toxicology tests is a topic of considerable importance in the field of toxicology. Carcinogenicity testing today normally includes 2-year studies in rats and mice of both sexes, following widely accepted procedures for husbandry; selection of dose levels; pathology and toxicity observations; and statistical interpretation of tumor data. These studies are usually preceded by tests for genetic toxicity and subchronic toxicity studies to select dose levels for the 2-year studies. Although these data are used for quantitative risk assessment, the mechanistic basis for effects is usually unknown. The series of studies is very expensive and requires 5 years or more to conduct. Alternative approaches are being developed that would provide more mechanistic information and hopefully would permit decisions to be made about carcinogenic potential without the need to conduct 2-year studies in rats and mice of both sexes. Decisions could be based on a profile of data rather than on the result of one test. Procedures for regulatory acceptance of new approaches for carcinogenicity testing are critical to future progress.
Descriptors: alternatives, carcinogenicity tests, methods, toxicity, animal welfare, decision making, government, mice, public policy, rats, research design, trends, time factors.

Selection and use of replacement methods in animal experimentation. (1998). Herts, UK: Universities Federation for Animal Welfare, 32 p.
Copies are available from: FRAME, Russell and Burch House, 96-98 North Sherwood Street, Nottingham NG1 4EE, UK, tel: +44 0115 958 4740, fax: +44 0115 950 3570, email: or UFAW, The Old School, Brewhouse Hill, Wheathampstead, Herts AL4 8AN, UK, tel: +44 0158 283 1818, fax: +44 0158 283 1414, e-mail:
Descriptors: legislative requirements (UK), introduction to the 3Rs, improved use of information, physical and chemical methods, mathematical and computer models, SAR, in vitro techniques, lower organisms, human tissues and volunteers, sources of tissues, sources of information, organizations, databases, on-line services.

Smaje, L.H., J.A. Smith, R.D. Combes, R. Ewbank,-R., J.A. Gregory,-J.A., M. Jennings, G.J. Moore, and D.B. Morton (1998). Advancing refinement of laboratory animal use. Laboratory Animals 32(2): 137-142.
NAL call number: QL55.A1L3
Abstract: Whatever view is taken of the morality of using animals in scientific research and safety testing, it can generally be agreed that so long as such use continues, every effort should be made to keep animal suffering to a minimum. This is the thinking behind the 'Three Rs' of refinement, reduction and replacement of laboratory animal use. This paper concerns refinement. We recognize that the Three Rs are taken very seriously in many countries of the world [see for example a recent editorial in the journal Science (Goldberg et al. 1996)] and, although we have written this paper from our own perspective in the UK, its principles are generally applicable.

Snow, B. (1990). On-line searching for alternatives to animal testing. Online (July): 94-97.
NAL call number: QA76.55 O6
Descriptors: developing search strategies, boolean operators, databases, category codes, terminology.

Stokes, W.S. and D.J.B. Jensen (1995). Guidelines for institutional animal care and use committees: consideration of alternatives. Contemporary Topics in Laboratory Animal Science 34 (3):51-55, 58-60.
NAL call number: SF405.5.A23.
Descriptors: animal testing alternatives, committees, guidelines, information services, training, regulations.

Van der Kamp, M.D.O. (1994). Ways of replacing, reducing, or refining the use of animals in the quality control of veterinary vaccines.Lelystad, The Netherlands: Instituut voor Veehouderij en Diergezondheid, 107 p.
NAL call number: HV4915 K36 1994
Descriptors: overview of veterinary vaccines, quality control, regulatory climate, alternatives to animal testing, feasibility of alternatives, recommendations.

Zeller, W., H. Weber, B. Panoussis, T. Burge, and R. Bergmann (1998). Refinement of blood sampling from the sublingual vein of rats. Laboratory Animals 32(4): 369-376.
NAL call number: QL55 A1L3
Descriptors: blood sampling, stress, laboratory animals, blood specimen collection, animal welfare, pain.

Zutphen, L.F.M.van and M. Balls (eds.) (1997). Animal alternatives, welfare, and ethics, Proceedings of the 2nd World Congress on Alternatives and Animal Use in the Life Sciences, held in Utrecht, the Netherlands, 20-24 October 1996 Amsterdam, New York : Elsevier, 1260 p.
NAL call number: QL1.D48 v.27.
Descriptors: alternatives to animal testing, animal experimentation, animal welfare, laboratory animals, databases, literature searching.

Zutphen, B.F.M. van and J.B.F. van der Valk (1995). Education and training: a basis for the introduction of the three Rs alternatives into animal research. Alternatives to Laboratory Animals: ATLA 23(1): 123-127.
NAL call number: Z7994.L3A5
Abstract: Education is a highly effective way of promoting the introduction of alternatives into the everyday practice of biomedical research and testing. In some countries, specific requirement for the education of persons involved in animal experimentation have been made compulsory by law. In The Netherlands, young scientists must take a course on laboratory animal science as part of, or in addition to, their biomedical graduate programme. This course provides information on the proper design of animal exp eriments, but also covers alternatives animal welfare issues and ethical aspects of animal experimentation. The Three RB of Russell & Burch are the guiding principles of the course, during which participants are challenged to seek methods or techniques that can replace, reduce or refine the use of animals. Since 1985 more than 2500 people in The Netherlands have taken the course, and evaluations have indicated that a large majority of the participants appreciated this education as a contribution to both the quality of experiments and the welfare of the animals, and considered the course to be indispensable for those who are responsible for the design and performance of animal experiments.
Descriptors: animal testing alternatives, animal experiments, educational courses, training, laboratory animals, animal husbandry.

Useful World Wide Web Sites

A site for news, information, discussion, and resources from the field of alternatives to animal testing. This site is a collaborative effort funded by the Alternatives Research & Development Foundation, the Doerenkamp-Zbinden Foundation, the Humane Society of the United States, the Office for Protection from Research Risks at the National Institutes of Health, and the Procter & Gamble Company. It is being developed by the Center for Alternatives to Animal Testing at Johns Hopkins University, in collaboration with the Altweb Project Team (which includes AWIC), to serve academic, industrial and government scientists, educators, the media, and the general public.

Animal Welfare Information Center
Articles and other resources concerning alternatives

Association of Veterinarians for Animal Rights
This site will give you access to Alternatives in Education Database, a comprehensive listing of videos, computer simulations, and other media that can be incorporated into educational curricula from high school on through medical, veterinary, or graduate school.

Center for Alternatives to Animal Testing (CAAT)
The Johns Hopkins Center for Alternatives to Animal Testing (CAAT) is a global resource for the development of replacement, reduction and refinement alternatives for research and testing.

ECVAM : European Centre for the Validation of Alternative Methods
Validating methods and strategies to reduce or replace the use of live animals in laboratory studies.

Fund for the Replacement of Animals in Medical Education
FRAME advocates the Three Rs approach to animal experimentation through the development, validation and acceptance of replacement alternative methods.

Guide to Searching for Alternatives to the Use of Laboratory Animals
This guide assumes no previous knowledge of search techniques nor of the facilities available for obtaining information from the Internet.

ICCVAM: Interagency Coordinating Committee for the Validation of Alternative Methods
ICCVAM and its supporting center, NICEATM (the National Toxicology Program Interagency Center for the Evaluation of Alternative Toxicological Methods), coordinate the development, validation, acceptance, and harmonization of alternative toxicological test methods throughout the U.S. Federal government. Another great resource provided by the U.S. Government.

The Netherlands Centre Alternatives to Animal Use
The Netherlands Centre Alternatives to Animal (NCA) is the central point in the Netherlands for coordinating research and disseminating information on alternatives to animal experiments. One of its important tasks is to support the Alternatives to Animal Experiments Platform, in which the Dutch government, industry, and animal protection organizations collaborate.

The Norwegian Reference Centre for Laboratory Animal Science & Alternatives
Knutepunktet for forsøksdyrlære og alternativer til dyreforsøk
Links to the Norina database (Norwegian Inventory of Audiovisuals (NORINA) of alternatives and other alternativesdatabases

University of California Center for Animal Alternatives
The Center places special emphasis on disseminating information concerning models, computer programs, and other animal alternatives in education through every level of public and private education.

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Last updated June 18, 2014