Euthanasia - Confirmation of Euthanasia (Policy)

IACUC Policy:

Confirmation of Euthanasia

Policy: The IACUC has provided a set of guidance documents (Policies, Guidelines, and Informational Sheets) for use when planning animal procedures at the University of Iowa. An exception to a Policy must be described and justified in the Animal Protocol and approved by the full IACUC at a convened monthly meeting.

Purpose: This policy lists the IACUC’s requirements of confirmation of euthanasia. 

The policy requires that any animal euthanized at the University of Iowa must be subject to a confirmatory method of euthanasia.

USDA regulated Species (e.g. ferrets, Guinea pigs, swine, hamsters, etc)

• A physical confirmation of euthanasia is required following a primary method of euthanasia
• Confirmatory methods include, but are not limited to:
  • Bilateral thoracotomy
  • Sternotomy
  • Vital tissue harvest (inclusive of heart, lungs, and/or brain)
  • Decapitation

Mice and Rats

• Confirmation of euthanasia is required following a primary method of euthanasia
• Confirmatory methods for animals older than 14 days of age include, but are not limited to:
  • Cervical dislocation
    • Not acceptable for rats > 200 grams of body weight 
  • Decapitation
  • Thoracotomy
  • Vital tissue harvest (inclusive of heart, lungs, and/or brain)
  • Continued exposure to CO₂ for at least 15 minutes after respiratory arrest
• Confirmatory methods for neonates 14 days of age or younger (less than 14 days of age) include, but are not limited to:
  • Decapitation
    • Preferred confirmatory method
  • Thoracotomy
  • Vital tissue harvest (inclusive of heart, lungs, and/or brain)
  • Continued exposure to CO2
• This method is discouraged as neonates are more resistant to CO₂ hypoxia
• For isoflurane euthanasia:
  • If using a vaporizer: a physical method or continued exposure to isoflurane for at least 15 minutes after respiratory arrest is acceptable
  • If using a drop jar: a physical method (cervical dislocation, decapitation, thoracotomy or vital tissue harvest) must be used for confirmation   

Xenopus

• A physical confirmation of euthanasia is required following a primary method of euthanasia
• Confirmatory methods of euthanasia following an overdose of anesthesia:
  • Decapitation  
    • must be confirmed by double pithing
  • Cervical section
    • must be confirmed by double pithing
  • Removal of heart

Zebrafish         

• Confirmation of euthanasia is required following a primary method of euthanasia
• Confirmatory methods for Zebrafish 8 days post fertilization (dpf) and older include, but are not limited to:
  • • Observation of no opercular movements for at least 10 minutes following anesthetic overdose or chilling
    • Decapitation

• Confirmatory methods for Zebrafish 4-7 dpf include, but are not limited to:
  • • Observation of no opercular movements for at least 10 minutes following anesthetic overdose or chilling
    • Decapitation
  • Confirmatory methods for Zebrafish 0-3 dpf include, but are not limited to:
    • Exposure to a dilute (1-10%) sodium hypochlorite or calcium hypochlorite solution

References

1. " AVMA Guidelines for the Euthanasia of Animals: 2020 Edition" AVMA.org., 2020.

Last Reviewed by the IACUC 1/12/2022