and larval life forms depending on species and gestational age (Artwohl et al. 2006).

The selection of specific agents and methods for euthanasia will depend on the species involved, the animal’s age, and the objectives of the protocol. Generally, chemical agents (e.g., barbiturates, nonexplosive inhalant anesthetics) are preferable to physical methods (e.g., cervical dislocation, decapitation, use of a penetrating captive bolt); however, scientific considerations may preclude the use of chemical agents for some protocols.

Although carbon dioxide (CO2) is a commonly used method for rodent euthanasia, there is ongoing controversy about its aversive characteristics as an inhalant euthanasia agent. This is an area of active research (Conlee et al. 2005; Danneman et al. 1997; Hackbarth et al. 2000; Kirkden et al. 2008; Leach et al. 2002; Niel et al. 2008) and further study is needed to optimize the methods for CO2 euthanasia in rodents (Hawkins et al. 2006). The acceptability of CO2 as a euthanasia agent for small rodents should be evaluated as new data become available. Furthermore, because neonatal rodents are resistant to the hypoxia-inducing effects of CO2 and require longer exposure times to the agent (Artwohl et al. 2006), alternative methods should be considered (e.g., injection with chemical agents, cervical dislocation, or decapitation; Klaunberg et al. 2004; Pritchett-Corning 2009).

It is essential that euthanasia be performed by personnel skilled in methods for the species in question and in a professional and compassionate manner. Special attention is required to ensure proficiency when a physical method of euthanasia is used. Death must be confirmed by personnel trained to recognize cessation of vital signs in the species being euthanized. A secondary method of euthanasia (e.g., thoracotomy or exsanguination) can be also used to ensure death. All methods of euthanasia should be reviewed and approved by the veterinarian and IACUC.

Euthanizing animals is psychologically difficult for some animal care, veterinary, and research personnel, particularly if they perform euthanasia repetitively or are emotionally attached to the animals being euthanized (Arluke 1990; NRC 2008; Rollin 1986; Wolfle 1985). When delegating euthanasia responsibilities, supervisors should be sensitive to this issue.


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ACLAM [American College of Laboratory Animal Medicine]. 2001. Position Statement on Rodent Surgery. Available at; accessed January 7, 2010.

AORN [Association of Operating Room Nurses]. 2006. Recommended practices for traffic patterns in the perioperative practice setting. AORN J 83:681-686.

Arluke A. 1990. Uneasiness among laboratory technicians. Lab Anim 19:20-39.

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