MS-222和丁香油对黄条麻醉效果研究*

王维鑫 崔爱君 徐永江 李海宁 柳学周王 滨 姜 燕 刘新富

王维鑫1,2崔爱君2徐永江2①李海宁3柳学周2王 滨2姜 燕2刘新富2

(1. 上海海洋大学水产与生命学院 上海 201306；2. 中国水产科学研究院黄海水产研究所青岛海洋科学与技术试点国家实验室深蓝渔业工程联合实验室 山东 青岛 266071；3. 东北林业大学生命科学学院 黑龙江 哈尔滨 150040)

1 材料与方法

1.1 实验材料

实验所需MS-222标准品(纯度98%)和丁香油均购于国药集团化学试剂有限公司。为提高丁香油的溶解率，使用时按照丁香油∶95%酒精=1∶10的比例溶解于酒精，再溶于水。

1.2 麻醉和复苏过程分期及有效质量浓度确定

根据Marking等(1985)的研究结果，本研究中麻醉剂有效质量浓度标准定义为实验鱼在3 min内进入麻醉状态(达到A6期)、5 min内完全复苏(恢复至B6期)。实验鱼进入A7期麻醉状态后再持续3 min，然后转入清洁海水复苏，成活率为100%。

1.3 不同水温下MS-222和丁香油对黄条的麻醉实验

在实验开始前，实验鱼饥饿24 h。实验容器为容积300 L的白色圆形塑料桶。设置温度为20℃和24℃，使用加热器控制水温(精确度为±1℃)，MS-222浓度梯度为40、60、80、100和120 mg/L，丁香油浓度梯度为20、40、60、80、100和120 mg/L。麻醉实验开始前，先将暂养的鱼放到预先控制好温度(20℃和24℃)的水桶中适应2 h。每组随机选取5尾鱼，用于麻醉实验，观察每尾鱼的麻醉情况，分别记录每尾鱼达到不同麻醉状态(A1～A7期)所需要的时间。麻醉时间从实验鱼放入不同麻醉剂实验组(A1期)开始计时，至A6期结束，精确到秒。麻醉后的实验鱼，转移至清洁海水中复苏，分别记录每尾鱼达到不同复苏状态(B1～B6期)所需要的时间，复苏时间从实验鱼放入清洁海水中开始计时，至实验鱼行为达B6期状态时结束，精确到秒。

1.4 血液样品采集与血清激素指标测定

表1 黄条麻醉和复苏阶段分期

表2 不同丁香油浓度和水温对黄条麻醉时间和复苏时间的影响

表3 不同MS-222浓度和水温对黄条麻醉时间和复苏时间的影响

1.5 数据处理

实验数据均使用Excel 2016和SPSS 24.0软件进行处理。利用单因素方差分析法(one-way ANOVA)和Duncan多重分析法对麻醉时间、复苏时间和血清激素和葡萄糖水平进行显著性分析，同一时间点2种不同温度(20℃和24℃)条件下血清激素和葡萄糖水平采用成对样本检验，所有结果以平均值±标准差(Mean±SD)表示，显著性水平设定为0.05，当<0.05时认为差异显著。

2 结果

2.1 不同水温下麻醉剂有效浓度的确定

2.2 不同水温下麻醉剂的效果

在同一麻醉浓度条件下，水温的升高使实验鱼进入A6期麻醉状态的时间缩短，而对复苏时间没有明显影响(表2和表3)。在2种不同水温下，丁香油浓度为20 mg/L时，实验鱼进入A6期的时间与其他各浓度均存在显著性差异(<0.05)，其他各浓度组之间无显著性差异(>0.05)，而MS-222浓度为100 mg/L和120 mg/L时，实验鱼进入A6期的时间无显著性差异(>0.05)，而与其他各浓度组之间均有显著性差异(<0.05)。MS-222浓度为40 mg/L时，实验鱼在2种水温下都不能进入A6期麻醉状态；MS-222浓度为60 mg/L时，20℃水温下实验鱼仅能达到A5期状态，而24℃水温条件下实验鱼可达A6期麻醉状态。

不同的水温下，麻醉时间与复苏时间之比随丁香油浓度的升高整体呈下降趋势，至120 mg/L降到最低。水温为20℃时，丁香油浓度为20 mg/L和40 mg/L时与其他各浓度组均有显著性差异；24℃时，仅有20 mg/L与其他各浓度组存在显著性差异(<0.05)，其他浓度组之间无显著性差异(>0.05)(图1)。不同的水温下，随着MS-222浓度的增加，麻醉时间与复苏时间之比显著降低(<0.05)，MS-222浓度为100 mg/L和120 mg/L时，二者之间无显著性差异(>0.05)，而与其他各浓度组均有显著性差异(<0.05)(图2)。

图1 不同水温与丁香油浓度下黄条的麻醉时间/复苏时间比

图2 不同水温与MS-222浓度下黄条的麻醉时间/复苏时间比

2.3 不同水温下血清激素水平对麻醉剂的响应

实验开始前，20℃下对照组实验鱼血清肾上腺素水平为(59.17±0.5006) pg/mL，24℃下对照组血清肾上腺素水平为(60.36±0.2212) pg/mL。在2种温度条件下，2种麻醉剂处理的实验鱼复苏1 h后，血清肾上腺素水平均显著升高(<0.05)(图3A、B；图4A、B)。在2种温度条件下，丁香油麻醉处理的实验鱼复苏后，实验鱼血清肾上腺素水平均在24 h达峰值，与其他各组均差异显著(<0.05)(图3A、B)。MS-222麻醉处理的实验鱼复苏后，20℃下血清肾上腺素水平在24 h达峰值，与其他各组差异显著(<0.05)(图4A)，24℃下血清肾上腺素水平在12 h达峰值，且与其他各组差异显著(<0.05)(图4B)。在2种温度条件下，丁香油和MS-222麻醉和复苏处理的实验鱼，血清肾上腺素水平呈先上升后下降的趋势，且高温度组比低温度组的肾上腺素水平上升快且恢复也快，复苏72 h后均恢复到对照组水平以下(图3A、B；图4A、B)。

图3 不同温度下丁香油麻醉后黄条血清内激素含量变化

图4 不同温度下MS-222麻醉后黄条血清内激素含量变化

3 讨论

3.1 麻醉剂的有效浓度和麻醉效果

麻醉剂的主要作用是抑制大脑皮质的中枢神经系统，从而对鱼体产生一定的麻痹作用，在面对外界环境胁迫的刺激时，降低鱼体应激反应，达到生理保护作用(Devi, 2019)。因此，麻醉剂的浓度直接影响鱼类的麻醉效果，浓度过低起不到抗应激作用，浓度过高则容易导致鱼体受到生理胁迫伤害，甚至休克死亡。因此，在对鱼类进行不同的生产操作时需要明确不同的给药剂量，合适的麻醉浓度有利于生产操作，还能起到保活运输作用(Chanseau, 2002)。

3.2 水温对黄条麻醉效果的影响

3.3 MS-222和丁香油对黄条血清生理指标的影响

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Anesthetic Effect of MS-222 and Clove Oil on Yellowtail Kingfish

WANG Weixin1,2, CUI Aijun2, XU Yongjiang2①, LI Haining3, LIU Xuezhou2, WANG Bin2, JIANG Yan2, LIU Xinfu2

(1. School of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China; 2. Joint Laboratory for Deep Blue Fishery Engineering of Pilot National Laboratory for Marine Science and Technology (Qingdao), Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao, Shandong 266071, China; 3. College of Life Science, Northeast Forestry University, Harbin, Heilongjiang 150040, China)

Yellowtail kingfishis a globally distributed, circumtropical pelagic fish of importance in both commercial fisheries and aquaculture. It swims fast and has fierce behavior under artificial culture conditions. To develop a suitable manipulation technique with low stress for yellow tail kingfish, the anesthetizing effects of two anesthetics, MS-222 and clove oil, on one-year-old yellowtail kingfish under different temperature conditions (20℃and 24℃) were studied. The optimal anesthesia times, recovery times, and anesthesia dosages of the two anesthetics were determined based on the discrimination of swimming behavior and investigation of serum hormones levels, including cortisol, adrenaline, and glucose. According to the behavioral characteristic changes of the yellowtail kingfish during anesthesia and recovery, the anesthesia process was divided into seven stages, whereas the recovery process was divided into six stages. The results showed that the optimal concentrations for MS-222 for anesthetizing yellowtail kingfish were 100～120 mg/L and 100 mg/L at water temperatures of 20℃ and 24℃, respectively. For clove oil, the optimal concentration for anesthetizing yellowtail kingfish was 40 mg/L at both temperatures. All the experimental yellowtail kingfish were anaesthetized within three minutes and recovered within five minutes. By increasing the anesthetic concentration, the time for the yellowtail kingfish to enter anesthesia was reduced and the recovery time prolonged. The increase in water temperature reduced the time to enter anesthesia, but had no obvious effect on the recovery time. When anesthetized with 40 mg/L clove oil at 20℃and 24℃, serum cortisol significantly peaked at 6 h and 12 h, respectively (<0.05). At 24℃, the serum adrenaline level significantly peaked at 12 h (<0.05) in yellowtail kingfish anesthetized with 100 mg/L of MS-222. The levels of serum adrenaline, glucose, and cortisol in the other experimental groups all significantly peaked after 24 h (<0.05) in fish anesthetized using both anesthetics. Under the two temperature conditions, the two anesthetics caused the three serum hormone levels to all decrease significantly (<0.05) to low levels that were lower than the initial levels after 72 h of recovery, indicating that yellowtail kingfish could be physiologically adaptive to the optimal treatment methods of clove oil and MS-222 under different temperatures. The results from the present study provide theoretical and technical support for the development of standard experimental and farming management technology for yellowtail kingfish.

Yellowtail kingfish; MS-222; Clove oil; Anesthetic effect; Physiological response

XU Yongjiang, E-mail: xuyj@ysfri.ac.cn

S948

A

2095-9869(2021)06-0205-10

10.19663/j.issn2095-9869.20210224001

http://www.yykxjz.cn/

WANG W X, CUI A J, XU Y J, LI H N, LIU X Z, WANG B, JIANG Y, LIU X F. Anesthetic effect of MS-222 and clove oil on yellowtail kingfish. Progress in Fishery Sciences, 2021, 42(6): 205–214

徐永江，研究员，E-mail: xuyj@ysfri.ac.cn

2021-02-24,

2021-03-31

*山东省支持青岛海洋科学与技术试点国家实验室重大科技专项(2018SDKJ0303-1)、国家重点研发计划项目(2019YFD0900901; 2018YFD0901204)、中国水产科学研究院创新团队(2020TD47)和财政部和农业农村部：国家现代农业产业技术体系(CARS-47)共同资助[This work was supported by Marine S&T Fund of Shandong Province for Pilot National Laboratory for Marine Science and Technology (Qingdao) (2018SDKJ0303-1), National Key R&D Program of China (2019YFD0900901; 2018YFD0901204), Science and Technology Innovation Team of Chinese Academy of Fishery Sciences (2020TD47), and China Agriculture Research System of MOF and MARA（CARS-47）].王维鑫，E-mail: wwxshou@163.com

(编辑 冯小花)