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Authors
Shiferaw, Kefale Ambachew https://orcid.org/0009-0009-7877-3808
Kinde, Mebrie Zemene https://orcid.org/0000-0002-0018-778X
Mengistu, Bemrew Admassu https://orcid.org/0009-0003-9409-2902
Gessese, Abebe Tesfaye https://orcid.org/0000-0001-5674-5480
Demessie, Yitayew https://orcid.org/0000-0002-8782-2570
Wassie, Zerihun Getie https://orcid.org/0009-0005-0431-145X
Berie, Genetu Kassahun https://orcid.org/0009-0007-6686-4134
Birhanie, Merawi Kindu https://orcid.org/0009-0005-4550-0759
Shiferaw, Dejen Takele https://orcid.org/0009-0008-2539-3265
Aragaw, Zenebe Jemere https://orcid.org/0009-0002-6886-0659
Mersha, Amsalu Chanie https://orcid.org/0009-0001-6289-0358
Miliket, Addisu Workie https://orcid.org/0009-0002-6694-0014
Yesuf, Mohammed https://orcid.org/0000-0003-0708-027X
Tafere, Firdyawukal Abuhay https://orcid.org/0000-0002-7532-3969
Berihun, Asnakew Mulaw https://orcid.org/0000-0001-5804-125X
Birhan, Mastewal https://orcid.org/0000-0002-0984-5582
Tsehay, Elias Melkamu https://orcid.org/0009-0002-2098-2032
Temesgen, Abraham Belete https://orcid.org/0009-0009-5926-5568
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Article History
Received: 21 September 2025
Accepted: 20 November 2025
First Online: 24 November 2025
Declarations
:
: The authors declare no competing interests.
: All experimental procedures were conducted in accordance with international ethical standards and the ARRIVE (Animal Research: Reporting of In Vivo Experiments) guidelines to ensure animal welfare and scientific integrity. The study was approved by the Institutional Animal Care and Use Committee (IACUC) of the University of Gondar (Approval No.: CVMASc/UoG/ERRC/01/05/2025). Humane euthanasia was performed using 100% carbon dioxide (CO₂) inhalation. Each chicken was placed individually in a transparent airtight chamber (50 L capacity), and CO₂ was introduced at a controlled flow rate until loss of consciousness. The flow was then increased and maintained for at least 2 min after cessation of breathing and cardiac activity to ensure death. Chickens were continuously monitored for respiratory rate, righting reflex, and corneal reflex throughout the procedure to ensure humane and effective euthanasia.
: All authors have thoroughly examined and critically evaluated all content of this manuscript, have provided substantial revisions where necessary, and have given their explicit consent for its publication in its final form.
