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Use & Euthanasia Procedures for Avian Embryos

A page within IACUC

PURPOSE:  To provide guidance for the euthanasia procedures for avian embryos in various stages of development as well as to ensure euthanasia procedures are in place for inadvertently hatched chicks. This policy applies to all avian embryos used in research and teaching activities; however, the specific developmental times below pertain to the use of chickens.

BACKGROUND: Avian embryos are not considered live animals by U.S. Public Health Service (PHS) policy. However, avian embryos reaching a certain stage of development may be able to feel pain. Because there is no consensus in the scientific community on the exact point for pain perception in avian embryos and that there are potential species-specific differences in pain perception, UWL has  adopted the 2020 American Veterinary Medical Association (AVMA) Guidelines, which state that, “Bird embryos that have attained > 80% incubation demonstrate EEG activity that is sustained, with increases in amplitude suggesting the potential for pain perception in conscious embryos” and that “euthanasia of embryos should be performed based on the best available data and with attention to assuring, as best as possible, that conscious suffering does not occur.” Additionally, OLAW expects Assured institutions to have policies and procedures in place that address the care or euthanasia of birds that hatch unexpectedly. Consequently, the UWL IACUC has adopted the following guidelines that apply to all avian species. Chicken embryos, which hatch in approximately 21 days, are considered the model species and are highlighted below. If another avian species is used, the guidelines should be adjusted based on relative time to hatching for that species.

POLICY:

  • The use of avian embryos that have attained > or = 80 % incubation (embryonic day 17 (E17) and older for chicken embryos) will require submission of a full IACUC Animal Use Protocol for review and approval.
  • The use of avian embryos < 80% incubation (E16 or younger for chicken embryos) does not require an IACUC Animal Use Protocol. However, the investigator is required to submit the “Notice of Use of Avian Embryos” form to the IACUC for approval. As part of this process, investigators are required to describe their methods for care and humane euthanasia of birds that hatch unexpectedly.
  • Avian embryos < 80 % incubation (E16 or younger for chicken embryos) may be euthanized by prolonged exposure (> 20 min) to CO2, cooling (< 4°C for at least 4 hours), or placing the eggs in a -20°C freezer. Death should be confirmed by a secondary method of euthanasia such as decapitation, cervical dislocation, or membrane disruption. 
  • Completion of the "Working with Poultry in Agriculture Research Settings" training course in CITI is required for researchers working with avian embryos that have attained > or = 80% incubation and/or hatched individuals. The training course is not required for researchers working with avian embryos < 80% incubation. 
  • Researchers working with avian embryos that have attained < 80% incubation and hazardous agents (chemical, radiological, or biological) must complete the applicable appendix/appendices and attach it to the Notice for Use of Avian Embryos Form: 

EUTHANASIA:

  • Avian embryos that have attained > or = 80 % incubation (E17 and older for chicken embryos) must be euthanized by similar methods used in avian neonates such as anesthetic overdose, decapitation, or prolonged (> 20 min) exposure to CO2. Death should be confirmed by a physical secondary method of euthanasia when euthanizing with anesthetic overdose or CO2.
  • Avian embryos < 80 % incubation (E16 or younger for chicken embryos) may be euthanized by prolonged exposure (> 20 min) to CO2, cooling (< 4°C for at least 4 hours), or placing the eggs in a -20°C freezer. Death should be confirmed by a secondary method of euthanasia such as decapitation, cervical dislocation, or membrane disruption.

REFERENCES:

Adoption date: 2/7/24

Amended: 4/22/24

Reference minutes: 1/12/23, 4/22/24