Effectiveness of Rapid Cooling as a Method of Euthanasia for Young Zebrafish (Danio rerio).
Effectiveness of Rapid Cooling as a Method of Euthanasia for Young Zebrafish (Danio rerio).
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发表时间：
2018
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通讯作者：
Chelsea K Wallace;L. Bright;J. Marx;Robert P Andersen;M. Mullins;A. Carty
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作者：
Chelsea K Wallace;L. Bright;J. Marx;Robert P Andersen;M. Mullins;A. Carty
尽管在生物医学研究中越来越多地使用斑马鱼（Danio rerio），但有关适当安乐死方法的一致信息，特别是对于胚胎，很少。目前的文献表明，快速冷却是成年斑马鱼安乐死的有效方法，但对于6个月以下的斑马鱼，尚无一致的指南。进行本研究以区分快速冷却是斑马鱼安乐死的有效方法的年龄和使用该方法可靠地安乐死斑马鱼所需的暴露时间。将受精后3、4、7、14、16、19、21、28、60和90 d的斑马鱼置于冰水浴中5、10、30、45或60 min（n = 12 - 40/组）。另外，将斑马鱼置于冰水中12 h（年龄≤14 dpf）或30 s（年龄≥14 dpf）。在快速冷却后，将鱼转移到回收槽中，并记录从冰水中取出后1、4和12-24小时的活鱼数量。当从冰水中取出后的任何时间点观察到恢复证据时，将安乐死定义为失败。结果表明，年轻的鱼需要长期暴露于快速冷却有效的安乐死，与所需的暴露时间减少鱼的年龄。虽然年轻的鱼需要长时间的接触，动物成为固定后立即暴露在冷水中，和疼痛或痛苦的行为指标很少发生。我们的结论是，斑马鱼14 dpf和年轻的需要长达12小时，那些16至28 dpf的年龄需要5分钟，和那些年龄大于28 dpf需要30秒的最小暴露于快速冷却可靠的安乐死。
Despite increased use of zebrafish (Danio rerio) in biomedical research, consistent information regarding appropriate euthanasia methods, particularly for embryos, is sparse. Current literature indicates that rapid cooling is an effective method of euthanasia for adult zebrafish, yet consistent guidelines regarding zebrafish younger than 6 mo are unavailable. This study was performed to distinguish the age at which rapid cooling is an effective method of euthanasia for zebrafish and the exposure times necessary to reliably euthanize zebrafish using this method. Zebrafish at 3, 4, 7, 14, 16, 19, 21, 28, 60, and 90 d postfertilization (dpf) were placed into an ice water bath for 5, 10, 30, 45, or 60 min (n = 12 to 40 per group). In addition, zebrafish were placed in ice water for 12 h (age ≤14 dpf) or 30 s (age ≥14 dpf). After rapid cooling, fish were transferred to a recovery tank and the number of fish alive at 1, 4, and 12-24 h after removal from ice water was documented. Euthanasia was defined as a failure when evidence of recovery was observed at any point after removal from ice water. Results showed that younger fish required prolonged exposure to rapid cooling for effective euthanasia, with the required exposure time decreasing as fish age. Although younger fish required long exposure times, animals became immobilized immediately upon exposure to the cold water, and behavioral indicators of pain or distress rarely occurred. We conclude that zebrafish 14 dpf and younger require as long as 12 h, those 16 to 28 dpf of age require 5 min, and those older than 28 dpf require 30 s minimal exposure to rapid cooling for reliable euthanasia.