Internet Update

September 2004

Nine Steps for Writing Effective Animal Use Protocols

Wright State's Laboratory Animal Care and Use Committee (LACUC) is charged with ensuring the proper and humane care and use of laboratory animals at the university. For all research activities involving the use of vertebrate laboratory animals, a principal investigator (PI) must prepare an animal use protocol for review and approval by the LACUC. The following suggestions for writing effective animal protocols were developed by Nicholas Reo, Ph.D. (LACUC chair); Robert Grubbs, Ph.D. (recent LACUC member); and Harold Stills, D.V.M. (WSU veterinarian).

1. Address alternatives to using animals and justify why alternatives cannot be used.

Using an alternative to animals is always preferable if it is available. The PI must make an honest effort to locate and evaluate any possible alternatives to procedures causing pain or distress, or to the use of animals in procedures where pain or distress is produced.

A literature search is required and it is to your benefit to consult with the WSU Reference Librarian. This can save time and effort for the PI, and make it easier on the LACUC reviewer. Remember, a WSU Reference Librarian serves on the LACUC.

The literature search should be current (date of search is required). OLAW policy mandates the search for these reasons: to consider alternative methods to the use of animals (i.e., computer simulations, mathematical models, etc.); to consider alternative procedures which are less painful to the animals; and to ensure that the work is not duplicative.

These factors should be considered when designing the protocol. Thus, conducting the literature search after the protocol is written defeats the purpose for this requirement!

2. Justify the number of animals to be used in your study.

This is the most common area where confusion and difficulties arise in the review of protocols. The number of animals requested must "add-up", and it is the PI's responsibility to clearly justify the sample size. A brief consultation with the WSU Statistical Consulting Center prior to submission of your protocol is highly recommended. The LACUC includes a statistical consultant.

The best way to clearly identify the number of animals required is to construct a table (see example). The table should identify the various experimental groups and other parameters that define the number of animals needed for each of these groups (i.e., number of time-point measurements, number of doses of test compounds or drugs, number of animals per group, etc.).

Justification may be based on references to similar studies in the literature, but statistical justification is preferred.

3. Specify name, volume, concentration, vehicle, route and frequency of each substance to be administered.

The PI must detail what and how much is administered. Detail the concentration of each dose, the number of doses given, and the time interval between doses.

4. Organize your protocol using tables.

This is especially useful for calculating the number of animals needed as mentioned above. You know what you are planning to do; using tables makes it easier to communicate your plan to reviewers.

5. Specify the criteria for removing animals from your study.

This is another important area where protocols commonly fail.
A 5-minute telephone consultation with the WSU veterinarian during protocol development can save much time and effort for the PI and the LACUC reviewer.

Principle VI of the "U.S. Government Principles for the Utilization and Care of Vertebrate Animals Used in Testing, Research, and Training" states: "Animals that would otherwise suffer severe or chronic pain or distress that cannot be relieved should be painlessly killed at the end of the procedure or, if appropriate, during the procedure."

Consultation with the veterinarian during protocol development is a requirement if pain and distress will be produced . It is the PI's responsibility to anticipate potential problems and define the point at which the resulting pain and distress to the animal is excessive. The LACUC closely reviews these choices.

Animals removed for this reason must be euthanized and not 'saved' for a later experiment after the animal recovers.

If you use body weight as a criterion, be sure to identify a reference point. For example, if loss of 20% body weight is a criterion for removal of an animal from an experiment, identify the reference weight and how frequently weight will be measured.

6. Provide a detailed explanation of pre- and post-operative care.

Details are essential, especially if surgery is involved. Do not assume that if a procedure is in the literature, you can do it. Again, a consultation with the WSU veterinarian is advised during protocol development.

Include details about use of analgesics (what is used, dose, frequency, etc.) to alleviate pain during the recovery period. "Analgesics as needed" is seldom an acceptable explanation to the LACUC unless you clearly state the criteria and justify the addition of a study variable (i.e., an analgesic administered to some animals but not others).

If post-surgical pain is expected and analgesics are not used, then the PI must provide scientific justification for these procedures. If multiple survival surgeries will be performed, be sure to identify the length of time between these procedures.

7. Align your animal protocol and grant proposal.

Agreement between these two documents will be checked during LACUC inspections of records. This activity occurs every 5 months.

"Just In Time" is now the PHS timeframe for requiring institutional review and approval by appropriate oversight committees. You no longer need to submit approval of the animal use protocol within 60 days of grant submission. However, confirmation of LACUC approval must be sent to PHS prior to release of funding. If your grant application receives a priority score in the fundable or near-fundable range, submit your animal use protocol to the LACUC at that time. Waiting until the confirmation of funding might result in funding being withheld until LACUC approval is received if the protocol requires extensive revisions and resubmissions to the committee.

The LACUC MUST check the protocol against the grant for which you say it applies. Discrepancies are not acceptable and you will be forced to notify your grants officer of discrepancies noted after approval of funding. If funding is awarded and the discrepancies are significant and serious, both the PI and the institution "may be subject to criminal, civil, or administrative penalties."

8. Euthanasia.

Use the preferred methods of the American Veterinary Medicine Association (AVMA) Panel on Euthanasia. WSU's Laboratory Animal Resources (LAR) can provide this information. If you need to avoid the use of anesthetics prior to euthanasia, then these procedures must be scientifically justified. Be sure to consider the objectives of your protocol as stated in section A when providing your justification. Many times the LACUC finds that the justification by itself is fine, but there doesn't seem to be any connection to the stated objectives.

9. Remember who could read your protocol.

Anyone can obtain a copy of your animal use protocol through Ohio's open records law. Inflammatory language or language that is easily misunderstood by the public should be avoided. For example, avoid using phrases like "animals are disposed of" and instead use terms such as 'euthanized' or 'sacrificed'.

Use lay language when describing your objectives and significance of the work. A 'copy and paste' of the introduction/significance for your NIH grant proposal is not a good idea. Grant proposals are typically not described in lay terms. For the protocol's lay sections, use high school-level language.

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