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Article History

Received: 15 February 2026

Accepted: 11 April 2026

First Online: 2 May 2026

Declarations

:

: Ethical approval was obtained from the Animal Experiment Ethics Committee of Yangzhou University (Approval No. 202203043) on March 3, 2022. All animal procedures were performed in accordance with the guidelines and regulations of the Animal Experiment Ethics Committee of Yangzhou University, under License No. SYXK (SU) 2022-0044 for mice and SYXK (SU) 2021–0027 for chickens. Mice were anaesthetized by carbon dioxide (CO₂) inhalation prior to retro-orbital blood collection. CO₂ was introduced into the chamber at a gradual displacement rate of 20%–30% of the chamber volume per minute, allowing the animals to gradually lose consciousness and ensuring that sampling was conducted in a pain-free state. Chickens were killed via CO₂ inhalation using a gradual-fill method, in which the CO₂ concentration was steadily increased to 60–70% of the chamber volume, ensuring a humane and distress-free induction of unconsciousness.

: Not Applicable.

: The authors declare no competing interests.