Kathryn Heivilin, CPIA, RBP
Senior IACUC Manager
UM Biosafety Officer
Last Review Date: November 4, 2020
The purpose of this Standard Operating Procedure (SOP) is to outline the procedures on euthanasia and to ensure consistency with the AVMA Guidelines for the Euthanasia of Animals: 2020 Edition. This procedure applies to all personnel involved in the euthanasia of animals involved in research projects sanctioned by The University of Montana Institutional Animal Care and Use Committee (IACUC).
It is LAR policy to meet or exceed all federal, state, and local regulations and guidelines and to comply with all institutional policies and procedures as they apply to the use of animals in research. Personnel must attend any applicable training in animal care and use, occupational health and safety, equipment operation, and Standard Operating Procedures prior to performing activities outlined in this SOP or work under the direct supervision of a trained LAR staff member.
III. Animal Identification
It is the responsibility of the person performing euthanasia to verify accurately the identity of the animal(s) to be euthanatized and to ensure that the principal investigator (PI) or designated animal user has authorized euthanasia. It is imperative to double-check, at a minimum, animal identity before performing euthanasia.
IV. Selection of Specific Agents
The selection of specific agents and methods for euthanasia will depend on the species involved and the objectives of the protocol. Generally, inhalant (non-explosive inhalant anesthetics or CO2) or injectable chemical agents (such as barbiturates) are preferable to physical methods (such as cervical dislocation, decapitation, and use of a penetrating captive bolt). Euthanasia is to be performed by personnel who are skilled in methods for the species in question and in a professional and compassionate manner. Death should be confirmed by personnel who can recognize the cessation of vital signs in the species being euthanatized.
V. Non-Explosive Inhalant, CO2
This method is used for rodents and guinea pigs 600 grams and under.
- Sign-in using clipboard next to the CO2 unit. Fill out completely including PI name and number of boxes and number of animals.
- Place animal to be euthanatized in the chamber, inside its primary enclosure, if possible.
- Fill the euthanasia chamber with CO2 by turning the tank on.
- The regulator has been preset for low-flow volume appropriate to the chamber. DO NOT readjust the brass knob.
- When no movement is detected, turn off CO2
- Remove animal and check for
cessationof vital signs
- Use a secondary mechanical means of euthanasia to ensure death (cervical dislocation, decapitation, or creation of pneumothorax).
- When the
animalis confirmed dead, place the animalin carcass bags (ziploc bag) and label appropriately (e.g., wild-type, transgenic, contaminated)
- Discard into LAR
trash cansor appropriate carcass freezer ensuring compliance with NIH disposal guidelines
- Wipe CO2 chamber with Peroxigard spray after use
VI. IP Pentobarbital Euthanasia for Rodents
This method is used for rodents and guinea pigs weighing 600 grams and under.
- Inject animal with an IP overdose of pentobarbital
- Place used needle and syringe in sharps container (do not recap needle)
- When the animal no longer shows any vital signs, use a secondary mechanical means of euthanasia to ensure death (cervical dislocation, decapitation, or creation of pneumothorax).
- When the
animalis confirmed dead, place the animal in carcass bags (ziploc bag)
- Discard into LAR
trashcans or appropriate carcass freezer ensuring compliance with NIH disposal guidelines
VII. Injectable Chemical Agents
Chemical agents such as barbiturates are used on larger species such as rabbits, sheep, and non-human primates. Each species requires different dose levels to ensure painless death. It is required that trained personnel perform these procedures, under the direct supervision of the Attending Veterinarian (AV).
- Check with AV for proper dose requirements. Depending on the research protocol, it may be possible to administer a tranquilizer subcutaneously or intramuscularly 10-15 minutes before injection of the euthanasia solution.
- Use the
properneedle and syringe size for the species and drug dose volume
- Euthanasia solution may be obtained from the AV or authorized LAR personnel. All doses must be signed out and logged in the controlled substance log book.
- Use an easily identifiable vein for injection
- Place used needle and syringe into a
sharpscontainer (do not recap needle)
- When no movement is detected, check for
cessationof breathing and heart beatusing a stethoscope
- When the
animalis confirmed dead, place the animal in a carcass bag (ziploc bag)
- Label bag with investigator’s name, species, animal ID, date of birth (DOB), strain, date euthanized, and euthanasia drug used.
- Place the bag in the designated carcass freezer.