Chapter 5 - IACUC Protocol Submission Form: Issues for Investigators


Since both the Animal Welfare Act and the PHS Policy mandate review of all research/teaching/demonstration projects involving use of vertebrate animals, in recent years it has fallen upon the IACUC to develop a mechanism that will facilitate this review, allowing us to meet regulatory requirements without imposing an undue burden on individual investigators. Publication of revised guidelines, consultation with USDA and AAALAC inspectors, as well as the Office of Laboratory Animal Welfare (OLAW), and responses to our Animal Welfare Assurance, have led us to make several revisions in our animal study protocol application (and it is likely that further revisions will follow in coming years). The most current version of the Animal Study Protocol Application can be found on the Office of Research webpage.

There are several issues related to protocol submissions that have been particularly difficult for the committee to deal with (this has been true nationally), and the pursuant revision of the submission forms has also, no doubt, caused some frustration and aggravation on the part of researchers/teachers as well. For guidance of individuals preparing protocol submission forms, the following are provided in explanation of the purpose, intent, and expectations of the IACUC in requiring such detailed information.

One general question that has been widely debated is the responsibility of the IACUC to review projects for scientific merit. A general consensus has emerged among IACUCs at universities doing animal research that discipline-based merit review (gauging the merit of the project regarding contributions to the knowledge base of the discipline) is beyond the scope and the capability of most IACUCs. However, at a more fundamental level, IACUCs can and should form basic judgments about: 1) specific scientific objectives of the project; 2) the potential value/significance of the project to human or animal health, the good of society, or the advancement of knowledge; 3) the rationale for the use of animals, including species and numbers; 4) the training and experience of investigators and research staff; and 5) appropriateness of experimental design.


Both the Animal Welfare Act Regulations and the PHS Policy require that IACUCs review animal study proposals to insure that there has been adequate consideration of alternative methods and approaches to the use of animals. Specifically, the Animal Welfare Act requires the IACUC to determine that “the principal investigator has considered alternatives to procedures that may cause more than momentary or slight pain or distress to the animals (even if that pain or distress is relieved by analgesics or anesthetics), and has provided a written narrative description of the methods and sources used to determine that alternatives were not available”. The PHS Policy requires that the institution insure adherence to the U.S. Government Principles for the Utilization and Care of Vertebrate Animals Used in Testing, Research and Training (PHS Policy on Humane Care and Use of Laboratory Animals). One of these principles is as follows: “The animals selected for a procedure should be of an appropriate species and quality and the minimum number required to obtain valid results. Methods such as mathematical models, computer simulations and in vitro biological systems should be considered.” In addition, the Guide for the Care and Use of Laboratory Animals states “Scientists have developed, and should continue to develop and use, scientifically valid, adjunctive or alternative methods to animal experimentation. The guidelines stated herein acknowledge these responsibilities.”

The term “alternative” is now generally accepted to refer to research, testing, or training methods that result in the reduction of number of animals required, that incorporate refinements of procedures which result in the lessening of pain or distress, or that provide for replacement of animals with non-whole animal systems, or the replacement of one animal species with another, particularly if the substitute species is non-mammalian or invertebrate (National Research Council, Use of Laboratory Animals in Biological, Medical and Behavioral Research, National Academy Press, Washington, D.C., 1988). This concept of defining “alternatives” in terms of reduction, replacement, and refinement of animal use is commonly referred to as “the 3 R’s”. Therefore, when the IACUC asks for a written narrative from the investigator regarding consideration of alternatives, it is appropriate to address any or all of these three approaches.

The Animal Study Protocol Application requires that the investigator provide, according to federal law, “a written narrative description of the methods and sources used to determine that alternatives were not available.” The law does not specify what methods and sources should be utilized, although the mention of the Animal Welfare Information Center implies that literature searches of relevant databases would be one way to fulfill this requirement. It is important that the submission form provide evidence that the investigator has considered whether alternatives are available, and if so, why these were not utilized. In addition, there must be an indication of the methods and sources utilized to make this determination, whether this is a bibliography, a relevant data base, the investigator’s experience in the field (appropriately documented), consultation with an expert in the field (appropriately documented), a published article, etc.

The following websites may be helpful in performing an alternatives search:

  1. The Norwegian Reference Centre for Laboratory Animal Science and Alternatives-NORINA (Norwegian Inventory of Alternatives) - This website provides information and links to many other alternative sites.
  2. Animal Welfare Information Center (AWIC) - This USDA website contains information and searchable databases on alternatives.
  3. Johns Hopkins Center for Alternatives to Animal Testing (AltWeb) - Extremely helpful site and search engine.
  4. University of California Center for Animal Alternatives - UCSD vet school site with multiple links and resources available.
  5. Fund for the Replacement of Animals in Medical Experiments (FRAME) - Good description of 3R’s and some useful links.

NOTE: If you claim to have searched a scientific literature database, you should keep a record of the key words utilized, the date of the search, and the dates of the literature searched, as well as the results of such.

1.  Replacement - Use of Non-Animal Alternatives

Categories of replacement approaches that might be envisioned for research or teaching:

  1. Information helps to prevent unnecessary duplication
  2. In vitro methods, including cell, tissue, or organ culture
  3. Mathematical models, audiovisual technologies, or computer simulations
  4. Physiochemical techniques, combined with structure-activity relationships can be used to predict biological effects of chemicals
  5. Replacement of the animal species with “lower” species especially non-mammalian or invertebrates species

2.  Reduction - Use of the Minimum Number of Animals Necessary

Researchers should strive to use the fewest number of animals that will allow them to obtain valid results. Reduction of animal numbers may be achieved in several ways including:

  1. Increase the ability to determine the effect of interest by increasing the size of the effect, decreasing extraneous variability, optimizing sensitivity of detection method changing the scale of the outcome variable (e.g., continuous vs. dichotomous)
  2. Minimizing the number of treatment groups (e.g., Do you really need six dose levels?)
  3. Wise use of controls (e.g., cross-over design vs. separate control groups)
  4. Repeated measures, replicative sample designs
  5. Statistical analysis
  6. Performing several terminal procedures per animal, tissue sharing with other investigators

3.  Refinement - Reducing Pain or Distress

Modifying procedures can facilitate reduction of pain and distress:

  1. Administration of anesthetics or analgesics where pain or distress are unavoidable
  2. Selection of less stressful endpoints (e.g., a predetermined level of body weight loss rather than death of the animal)
  3. Careful monitoring to detect pain/stress as early as possible
  4. Careful attention to housing and access to food and water in animals that are impaired (e.g., restraint or inability to ambulate)
  5. Environmental enrichment programs
  6. Humane methods of euthanasia

These issues relating to “Alternatives” are to be addressed by the investigators on the Animal Study Protocol Application. Please contact the Attending Veterinarian or the IACUC if you need some guidance in filling out these sections.

Duplication of Research

Another controversial issue in the new regulations regards the request for the investigator’s assurance regarding duplication of research. The regulations require that the IACUC determine that “The principal investigator has provided written assurance that the activities do not unnecessarily duplicate previous experiments” (Animal Welfare Act, 2.31(d) (1) (iii)).

A key word in this regulation is “unnecessarily.” The investigator should provide an explanation of how his project differs from previous work. If it is duplicative, justification of why this is necessary must be included. This information is requested on the protocol application form.