Best Practices & Definitions
Common Committee Concerns
Considering Alternatives to Animal Use
It is not enough to simply state that you need live animals for your research. In your application, state what alternatives to the use of live vertebrate animals and/or animal tissues were considered. The committee needs documentation detailing how each alternative was considered and rejected for scientific reasons. For example, describe if there are problem results from specific alternative research methods (computer models, tissue culture, clinical studies).
Detailing the Number of Animals Requested for a Breeding Protocol
Make the details regarding animal numbers easy for the committee to assess:
- Explain justifications for group size (published results, statistical analysis based on previous established difference in measurements, etc.). Detail which statistical tests are being applied (e.g., power analysis).
- Clearly define the number of animals (e.g. 5 animals/group x 3 treatments x 4 time points x 2 different sexes = 120 animals). Define the groups, including controls, treatments, time points, etc.
- When you have a complex protocol, in which the same animals or subsets of animals are used for multiple procedures, tables, flow charts, or named groups can be used to clarify the numbers.
- If you have a breeding protocol, in which animals will not be used after genotyping, then please make sure the number of animals produced and the number needed for experiments is defined.
- When describing the number of offspring used, separate your numbers into those that will be used exclusively for breeding and those that will be used in experiments.
- Make sure your numbers add up and that they are consistent with what you enter under "Animal Source" in the next section.
- When you use euthanized animals for organ culture, provide rough estimate for the number of animals (e.g. we will get 3x107 cells per liver and need 6x107 cells per experiment, for 15 experiments that means 30 animals).
Scruitiny of Committee Despite Frequent Submissions
Remember that the protocol you submit is a stand-alone document and you cannot do procedures in other protocols or publications, or assume that they are so generally used that everyone will know what you will do. The committee members do not have access to previous protocols, and references cannot take the place of procedure descriptions. Use the "IACUC Guidelines" section of the ORRP website for descriptions of commonly used procedures.
The committee is not interested in lengthy descriptions of in vitro experiments and will not read through grant applications for study procedures. You should briefly describe your aims and correlate it with your description and the justification for number of animals requested. It helps if you use common numbering systems in both sections.
The committee needs to have information on all details relating to injections of drugs, virus vectors, etc., including routes of injection, volumes, site of blood draw, number of bacteria/viruses, etc.
The endpoints for all experiments need to be clearly defined. For chronic experiments, include how long will animals survive following a procedure, as well as their final disposition (e.g., euthanasia). Justify if death is an endpoint.
Answering Pain and Distress Questions
All sections do not have to filled in if it does not apply to your specific study. For example, if it is an acute study where the animal will not recover from anesthesia, post-surgical recognition of pain does not have to be considered. If there is no surgical procedure, you do not have to fill in the "During Surgery" box. Studies involving tumor development should consider during and after surgery (implantation) signs of pain/distress.
The question is not what pain or distress will usually occur during the procedures you wish to perform, although this is clearly important, but whether you are prepared if things do not go as expected. If during your procedure pain or distress is experienced by animals, what will you do? For example: Incomplete anesthesia is recognized by xxxxx signs; it is corrected (alleviated) by increasing the anesthetic.
It is good practice to add a sentence that indicates if post-surgical pain is recognized you will consult with an attending veterinarian to discuss disposition of the animal. In some cases euthanasia may be appropriate, in others additional analgesics/antibiotics may allow you to keep the animal in the study.
Definitions & Classification
Pain or Distress
Potential Pain/Distress: Procedures are classified according to the level of potential pain or distress that the animal may experience. If more than slight or momentary pain and distress could be caused by the procedure, then relief must be provided (pain class B). If relief cannot be provided (pain class C), there must be scientific justification for withholding of relief, the justification must be included in the animal use protocol, and must be approved by the IACUC. Additional information can be found in USDA Animal Care Resource Guide, Policy #11, Painful Procedures.
USDA Pain Codes
C - No Pain or Distress
D - Pain or Distress with Relief
E - Pain or Distress Without Relief
Major Survival Surgery & Multiple Survival Surgery
Multiple Major Survival Surgery: A major survival surgery penetrates and exposes the body cavity or produces substantial impairment of physical or physiologic functions (such as laparotomy, thoracotomy, craniotomy, joint replacement, and limb amputation). Scientific justification is required in the animal use protocol if more than one major survival surgery is to be performed on an animal during the course of the protocol experiment.
Food & Fluid Restriction
Certain experimental paradigms require the use of food or water restriction in order to accomplish studies such as operant conditioning work. The IACUC is required to approve these restrictions to ensure they are scientifically justified, minimize the level of restriction, and have criteria in place to monitor the health of animals on these studies.
Animal use protocols employing food or water restriction must provide the duration of restriction, level of restriction, and justification for the restriction as part of the protocol description. Furthermore, the early removal criteria must provide methods to assess the animal's health while on restriction. Typically this will require a frequent monitoring method such as daily weighing of the animal. Note that pre-surgical food restriction is detailed in the surgery procedure as part of the animal use protocol, and does not need specific justification.