Reduction, Refine= ment, Replacement and, Responsibility in Animal Research

We are all familiar with the 3 “R”’s, Reduction, Refinement, and Replacement, but there is a fourth “R̶= 1;; that of Responsibility. There= have been many papers, seminars, suggestions and opinions on how best to accompl= ish and capture the true intentions of Russell and Burch when they first develo= ped the concept of responsible use of laboratory animals with an eye toward compassion and care. It has long been known that animals that are humanely treated and well cared for give a more accurate picture of physiological data. Animals under stress th= rough overcrowding or substandard conditions often produce inaccurate data and therefore are not good research subjects. That is, of course, unless you are studying the effects of stress due to adverse environmental conditions. It = is in all of our best interests to produce that highest level of valid and statistically sound, and accurate research data we are capable of by using = our laboratory animals wisely, ethically, and humanely; never forgetting our responsibility to the scientific community, the animals, and mankind. With this in mind, I would like to submit to you the following:

Principles of Animal Research

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§ Association for the Assessment and Accreditation of Laboratory Animal Care

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§IAC= UC approved animal use protocols ALWAYS REQUIRED regardless of

l&nb= sp; the source of funding

= • = intramural or extramural

l&nb= sp; the source of the animals

= • = purchased, obtained free, or bred in-house

l&nb= sp; the number of animals used

= • = pilot studies, learning techniques

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§Literature Se= arch

§§§§§§§§§§ These should be clearly defined and strictly adhered to.

§ It is important to note that even = if the animal will be used in an acute procedure only for tissue collection, it mu= st be entered into category C. All animals used exclusively for breeding purpo= ses would be category B. As noted in the chart, there is no category A.

USDA Pain Levels:

USDA Category B

USDA Category C

USDA Category D

USDA Category E

Breeding or Holding Colony Protocols

No more than momentary or slight pain or distress and no use of pain-relievi= ng drugs, or no pain or distress. For example: euthanatized for tissues; just observed under normal conditions; positive reward projects; routine procedures; injections; and blood sampling.

Pain or distress appropriately relieved with anesthetics, analgesics and/or tranquilizer drugs or other methods for relieving pain or distress.

Pain or distress or potential pain or distress that is not relieved with anesthetics, analgesics and/or tranquilizer drugs or other methods for relieving pain or distress.

(Note: there is no USDA Pain Category A.)

Guidelines for determining USDA classification in protocols involving tissue collection before/after euthanasia and/or animal perfusion:

If an animal will be euthanatized = by an approved physical or chemical method of euthanasia solely for the collec= tion of tissues (after the animal's death), the procedure should be classified as USDA C.

If an animal will be anesthetized so that non-vital tis= sues can be collected (liver or skin biopsy), and the animal will then be allowe= d to recover, the procedure should be classified as USDA D (survival surgery).

If an animal will be anesthetized so that non-vital tis= sues can be collected (liver or skin biopsy, etc.); and the animal will then be euthanatized, the procedure should be classified as USDA D (non-survival surgery). In this scenario, it is necessary to justify why the animal could= n't be euthanatized (USDA category C) rather than anesthetized.

If an animal will be anesthetized so that vital tissues= can be collected (heart, both kidneys or lungs, whol= e liver, etc.), the animal will obviously succumb to the procedure. To determine whe= ther this will be euthanasia or non-survival surgery, we must consider the definition of euthanasia. A critical component of this definition is "rapid unconsciousness followed by loss of cardiac, respiratory and br= ain function". Based on this definition, procedures which require tissue manipulation or other prolonged techniques prior to the animals death (more than a few minutes) should be classified as non-survival surgery (USDA D). Similarly, if an animal will be anesthetized so that the tissue can be collected in the "freshest" possible state (i.e. heart) and the tissues will be rapidly excised, the procedure s= hould be classified as euthanasia (USDA C). (Note: In this scenario, it is diffic= ult to justify why the animal couldn't be euthanatized rather than anesthetized= .)

If an animal will be anesthetized so that it can be chemically perfused, the same "test of time" applies (i.e.: long, technical manipulations should be classified as USDA D; while rapid intravascular injection of the perfusate without other manipulations should= be classified as USDA C).

NOTE: Because the USDA classification system is based on the "potential for pain, distress or discomfort," the anesthetic/euthanasia drug dose becomes a critical concern. For example, if= a known "euthanasia dose" of pentobarbital will be administered, dr= ug irreversibility is assumed. Thus, once the animal is confirmed to be in an anesthetic plane (toe pinch response, etc.), tissues can be collected/ procedures can be performed without the concern about what the animal will = be perceiving. This procedure would then be classified as= USDA C. The Committee recommends using a euthanizing dose whenever possible. Oth= er methods may be appropriate with proper scientific justification.

Responsibility in Animal Research

&= nbsp; The use of animals in biomedical research may be summed up in only few words, “it is a privilege, not a right”. We, in the research community have a responsibility to the animals we work with. It is our responsibility= to treat these animals with respect and acknowledge and address the basic issu= es of humane care and thoughtful stewardship of their use. We must be ever min= dful of the rules and regulations that govern the use of animals in all types of research including, behavioral, medical and psychological studies. The introduction of genetic research demands more attention be paid to the need= s of genetically altered animals since the needs of these animals may have chang= ed and so also the care it takes to maintain them. Consequently, all of the responsib= ility and ethical decision making rests with us.= Everyone involved in any phase of animal research, from the daily ca= re of the husbandry technician, the lab personnel who process the data, the investigator who designs and submits the protocol, and the member of the Institutional Animal Care and Use Committee who approves the study has an interest in the responsible, humane, and ethical care and use of our most valuable and venerable laboratory animals. We owe them a debt for their invaluable service to the advancement of science.