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Article History

Received: 22 December 2021

Accepted: 7 March 2022

First Online: 2 April 2022

Declarations

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: The guidelines established by the Administration of Affairs Concerning Animal Experimentation state that approval from the Science and Technology Bureau of China and the Department of Wildlife Administration is not necessary when the fish in question are neither rare nor near extinction (first- or second-class state protection level). Therefore, approval was not required for the experiments described in this manuscript .In this study, all experiments were approved by the Animal Care Committee of Hunan Normal University and followed the stated guidelines of the Administration of Affairs Concerning Animal Experimentation of China. Fish collect and crossing were approved by the Animal Care Committee and Protection Station of Polyploidy Fish of Hunan Normal University. All samples were raised in natural ponds, all dissections were performed with 50 mg/L MS-222 (Sigma-Aldrich, St Louis, MO, USA), and all efforts were made to minimize suffering. Specific operation methods were as follows: first, add 4 L of water to a 10-L glass container, then weigh 200 mg of MS-222 solid powder and dissolve in water. After the powder was dissolved, the experiment fish was put into the water. When the body of the experiment fish gradually lost its balance or sank to the bottom of the tank, the frequency of gill motion decreased, and the breathing was weak or stopped. At this time, the experiment fish was under deep anesthesia, so subsequent experiments could be carried out.

: Not applicable.

: The authors declare no competing interests.