used in combination with appropriate analgesics and anesthetics to provide balanced anesthesia and to minimize stress associated with perioperative procedures. Neuromuscular blocking agents (e.g., pancuronium) are sometimes used to paralyze skeletal muscles during surgery in which general anesthetics have been administered (Klein 1987); because this paralysis eliminates many signs and reflexes used to assess anesthetic depth, autonomic nervous system changes (e.g., sudden changes in heart rate and blood pressure) can be indicators of pain related to an inadequate depth of anesthesia. It is imperative that any proposed use of neuromuscular blocking drugs be carefully evaluated by the veterinarian and IACUC to ensure the well-being of the animal. Acute stress is believed to be a consequence of paralysis in a conscious state and it is known that humans, if conscious, can experience distress when paralyzed with these drugs (NRC 2008; Van Sluyters and Oberdorfer 1991). If paralyzing agents are to be used, the appropriate amount of anesthetic should first be defined on the basis of results of a similar procedure using the anesthetic without a blocking agent (NRC 2003, 2008, 2009a).


Euthanasia is the act of humanely killing animals by methods that induce rapid unconsciousness and death without pain or distress. Unless a deviation is justified for scientific or medical reasons, methods should be consistent with the AVMA Guidelines on Euthanasia (AVMA 2007 or later editions). In evaluating the appropriateness of methods, some of the criteria that should be considered are ability to induce loss of consciousness and death with no or only momentary pain, distress, or anxiety; reliability; irreversibility; time required to induce unconsciousness; appropriateness for the species and age of the animal; compatibility with research objectives; and the safety of and emotional effect on personnel.

Euthanasia may be planned and necessary at the end of a protocol or as a means to relieve pain or distress that cannot be alleviated by analgesics, sedatives, or other treatments. Criteria for euthanasia include protocol-specific endpoints (such as degree of a physical or behavioral deficit or tumor size) that will enable a prompt decision by the veterinarian and the investigator to ensure that the endpoint is humane and, whenever possible, the scientific objective of the protocol is achieved (see Chapter 2).

Standardized methods of euthanasia that are predictable and controllable should be developed and approved by the AV and IACUC. Euthanasia should be carried out in a manner that avoids animal distress. Automated systems for controlled and staged delivery of inhalantsmay offer advantages for species killed frequently or in large numbers, such as rodents (McIntyre et al. 2007). Special consideration should be given to euthanasia of fetuses

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