实验动物麻醉指南和药物选择

 

  • 小鼠和大鼠(以及鱼类、两栖动物、爬行动物和鸟类)
    • 使用吸入/吸收麻醉剂的简要程序
      • 无需记录
      • 简短的程序是那些只引起暂时性疼痛的程序(例如采血、尾静脉注射、用于基因分型或鼻内接种的尾巴/鳍剪断)
    • 对于上述未描述的所有其他程序
      • 需要记录
      • 如果存活,则应监测动物直至完全康复;如果不存活,则应进行安乐死
      • 记录以下内容:
        • 日期
        • 首席研究员
        • 动物协议编号
        • 动物编号
        • 物种
        • 重量
        • 程序
        • 使用的药剂、剂量、给药途径
        • 麻醉诱导时间
        • 麻醉苏醒时间或安乐死时间
  • USDA Covered Species (所有哺乳动物,  Mus属小鼠 和 Rattus属大鼠除外)
    • 任何麻醉事件
      • 需要为每只动物单独记录
      • 记录以下内容:
      • 日期
      • 首席研究员
      • 动物协议编号
      • 动物编号
      • 物种
      • 重量
      • 程序
      • 使用的药剂、剂量、给药途径
      • 麻醉诱导时间
      • 麻醉苏醒时间或安乐死时间
      • 监测(每 15 分钟一次,直到恢复或安乐死)
  • 有关手术记录信息,请参阅手术指南
  • 单击此处获取  模板麻醉记录

麻醉期间动物的支持性护理

  • 将眼药膏涂抹在双眼上以防止任何超过 5 分钟的麻醉干燥
  • 使用温暖的循环水毯、热垫和/或加热的静脉输液保持正常体温
    • 不要使用“非处方”电加热垫,因为它们容易过热
  • 在长时间麻醉期间为动物提供液体(例如,IV、IP、SQ)以保持足够的水合作用,如批准的动物协议中所述

麻醉监测和评估

  • USDA Covered Species (所有哺乳动物,  Mus属小鼠 和 Rattus属大鼠除外)
    • 监测心率、呼吸频率和体温
      • 在麻醉期间至少每 15 分钟记录一次这些参数
      • 对于啮齿动物物种,对正常心血管和呼吸功能的定性监测可能就足够了,因为很难定量评估这些参数。
      • 对于所有其他物种,预计将进行定量监测(数值心率和呼吸频率)。
      • 如果使用呼吸机,请在记录中记录通气率和潮气量
    • 监测血流动力学参数以确保充分的气体交换
      • 黏膜应该是粉红色和湿润的
      • 毛细管再填充时间应小于 2 秒
    • 应定期监测至少一项深度疼痛识别指标(踏板反射、耳廓反射等),以确保充分麻醉
      • 根据监测参数的变化调整麻醉深度,以确保持续的手术麻醉平面
  • 小鼠和大鼠
    • 定期(不超过 15 分钟的间隔)监测呼吸频率和努力、粘膜颜色和反射的眼睛颜色(白化动物)
    • 通过踏板反射(牢固的脚趾捏)评估麻醉水平,并酌情调整麻醉剂输送以保持手术平面

麻醉恢复

  • 大型动物
    • 必须从恢复笼中取出食物和水碗
    • 监测每只动物直到完全恢复(以下正常值):
      • 心率和呼吸频率
      • 保持胸骨斜卧(胸部直立)
      • 心率和呼吸频率
      • 体温
        • 建议使用循环温水加热垫
      • 水合作用通过皮肤膨胀或粘膜“粘性”来评估
    • 一旦动物可以吞咽,放气并取出插管
      • 插管到位时不要让动物无人看管
    • 偶尔重新定位卧式动物以促进更快的恢复
      • 动物不应保持同一侧朝下超过四 (4) 小时
    • 如果发生呕吐,将动物的头部略低于胸部水平以防止误吸
  • 老鼠_
    • 将啮齿动物置于温暖、清洁、干燥、安静的环境中,远离其他动物
    • 用毛巾材料覆盖或更换床上用品
      • 当动物从麻醉中恢复时,床上用品可能会粘在眼睛上或被吸入
    • 恢复期间提供温暖:
      • 市售的手术加热垫(不是“非处方药”人体加热垫)
      • 白炽灯(50-75 瓦)距离啮齿动物 12-14 英寸
        • 定位灯,以便啮齿动物可以根据需要逃离光源
        • 必须进行仔细监测以防止啮齿动物过热
      • 使用温控笼子/培养箱
    • 如果在批准的动物协议中列出,可以使用温热的无菌盐水来补充手术过程中丢失的体液
    • 必须持续监测啮齿动物,直到保持直立姿势并在笼子周围正常行走,然后再返回动物饲养室
  • 必须持续监测所有动物,直到保持直立姿势并在笼子周围正常行走,然后才能完成监测并返回动物饲养室。

麻醉剂

以下是常用麻醉剂的列表。此列表不包括在内;其他麻醉剂可能会在动物方案中列出和使用。

其他可接受的适当镇痛药资源包括以下处方集:

  • Plumb 的兽药手册(Plumb)
  • 实验动物的麻醉和镇痛(ACLAM)
  • 实验动物处方集(鹰)
  • 外来动物处方集(Carpenter & Marion)
  • 兽医麻醉手册 (Muir & Hubbell)
  • 实验室里的猪(诈骗)

应与 OAR 或 IACUC 兽医协商确定合适的麻醉药物和剂量。

吸入麻醉剂

异氟醚/七氟醚 – 蒸发器

  • 当用作手术麻醉剂时,异氟醚/七氟醚必须与经过适当校准的蒸发器一起使用
  • 必须正确清除麻醉气体
    • 直接排气
    • 活性炭罐(例如 F/Air 罐)
      • 每次使用前都应称重,达到最大重量时必须丢弃
      • 必须保存重量记录
  • 蒸发器必须至少每年校准一次
    • 必须维护每个蒸发器的校准记录

异氟醚 – 滴罐法

  • 不用于七氟醚;七氟醚的浓度不能用滴罐法精确控制
  • 异氟醚可以在麻醉剂滴​​罐中给药,进行一次简短的手术
    • 不能用于任何外科手术(包括非生存和生存手术)
  • 必须正确清除麻醉气体
    • 通风柜
    • 硬管道生物安全柜
  • 动物不得直接接触液态异氟醚
  • 必须在动物之间清洁滴罐(即清除尿液/粪便)
  • 不建议将此方法与 50 mL 锥形管一起使用

异氟醚滴罐剂量:腔室的内部体积 (L) 和所需的异氟醚液体 (mL)

异氟醚浓度达到  1L 2L 3L 4L 5L
1% 0.05mL 0.10 毫升 0.15 毫升 0.20 毫升 0.26 毫升
2% 0.10 毫升 0.20 毫升 0.31 毫升 0.41 毫升 0.51 毫升
3%  0.15 毫升 0.31 毫升 0.46 毫升 0.61 毫升 0.77 毫升
4% 0.20 毫升 0.41 毫升 0.61 毫升 0.82 毫升 1.02 毫升
5% 0.26 毫升 0.51 毫升 0.77 毫升 1.02 毫升 1.28 毫升

75% 二氧化碳 / 25% 氧气

  • 75% CO2 / 25% O2 可用于单一、简短的程序
    • 不能用于任何外科手术(包括非生存和生存手术)。
  • 可从供应商处预混
  • 使用预防措施;动物很容易过量服用
  • 不适用于安乐死
  • 如果您不熟悉 CO 2 /O 2的正确使用方法,请联系动物资源办公室兽医工作人员

吸收的麻醉剂

  • 三卡因甲磺酸盐 (MS-222)
    • 青蛙:
      • 0.05-0.2%(500-2000mg/L)溶液
      • 溶液必须用碳酸氢钠缓冲至 pH 7.0-7.5
      • 将青蛙浸入溶液中 10-20 分钟
      • 麻醉水平由翻正反射的丧失、吞咽反射的丧失和对脚趾捏的退缩反应的丧失来判断
    • 鱼:
      •  0.0025-0.01%(25​​-100mg/L)溶液
      • 溶液必须用碳酸氢钠缓冲至 pH 7.0-7.5
      • 将鱼浸入溶液中,直到观察到适当的麻醉深度
      • 麻醉程度通过失去平衡、对有害刺激失去反应(捏尾巴)、鳃盖运动速度和鳃颜色来判断
    • 贮存
      • 三卡因(液体溶液)可以在室温下储存 3-5 天,如果避免暴露在光线下。
      • 如果避光保存,三卡因(液体溶液)可在 4°C(即冰箱)保存 1 个月。
      • 如果避光,三卡因(液体溶液)可在 -20°C(即冰箱)保存 1 年。
      • 如果将三卡因(粉末)储存在黑暗的容器中,则可在室温下储存长达 5 年。

注射麻醉剂

常用的注射麻醉剂

老鼠

试剂

剂量

麻醉时间

氯胺酮/甲苯噻嗪*

氯胺酮 80-100 mg/kg IP

甲苯噻嗪 10-12.5 mg/kg IP

20-30分钟

氯胺酮/甲苯噻嗪鸡尾酒*

KX 小鼠鸡尾酒 0.1mL/20g mouse wt. 知识产权

包含:

87.5 毫克/公斤氯胺酮

12.5 毫克/千克甲苯噻嗪

20-30分钟

氯胺酮/甲苯噻嗪/乙酰丙嗪

氯胺酮 60-100 mg/kg IP

甲苯噻嗪 10-15 mg/kg IP

乙酰丙嗪 2-5 mg/kg IP

60-90 分钟

戊巴比妥

50 毫克/公斤 IP

20-40 分钟

Avertin ǂ 请参阅下面的警告

240 毫克/公斤 IP

30分钟

*氯胺酮/甲苯噻嗪不与镇痛剂(阿片类药物或非甾体抗炎药)联合使用可能不足以产生手术平面麻醉。手术前给予适当的镇痛剂和/或添加乙酰丙嗪将增强氯胺酮/甲苯噻嗪的麻醉效果。

** 氯胺酮/甲苯噻嗪混合物的制备说明见下文。                 

ǂ警告:NIH 和欧洲指南不鼓励使用 Avertin。Avertin 的制备和储存要求如下所示。

指南 – 为小鼠制备氯胺酮/甲苯噻嗪鸡尾酒

  • 需要使用无菌注射小瓶(例如 redtop 采血管;商业注射小瓶)
  • 混合说明:
    • 在混合之前验证您的药物浓度
    • 对于使用 100 mg/mL 氯胺酮和 100 mg/mL 甲苯噻嗪的 10mL 小瓶,添加:
      • 1.75 毫升氯胺酮 (100 毫克/毫升)
      • 0.25 毫升甲苯噻嗪 (100 毫克/毫升)
      • 8 mL 生理盐水或无菌注射用水
  • 建议使用以下模板作为标签:
    • 小鼠麻醉剂混合物:氯胺酮/甲苯噻嗪
    • 剂量:0.1 毫升/ 20 克 IP  
    • 递送:87.5 毫克/公斤氯胺酮/12.5 毫克/公斤甲苯噻嗪
    • 浓度:17.5 mg/mL 氯胺酮/2.5 mg/mL 甲苯噻嗪

过期:____________

  • 鸡尾酒的有效期由混合日期起六个月或任何一个成分先到期(如果少于 6 个月)确定
    • 例如:稀释于 2011 年 8 月 13 日,氯胺酮于 2012 年 12 月 10 日到期,甲苯噻嗪 10/10/11 到期,无菌注射用水于 2013 年 1 月 12 日到期;鸡尾酒的有效期为 2011 年 10 月 10 日

试剂

剂量

麻醉时间

氯胺酮/甲苯噻嗪

氯胺酮 40-100 mg/kg IP

甲苯噻嗪 5-13 mg/kg IP

60-80 分钟

氯胺酮/甲苯噻嗪鸡尾酒*

KX 大鼠鸡尾酒 0.1 mL/100g 大鼠重量。知识产权

包含:

91 毫克/公斤氯胺酮

9.1 毫克/千克甲苯噻嗪

60-80 分钟

氯胺酮/甲苯噻嗪/乙酰丙嗪

氯胺酮 20-50 mg/kg IP

甲苯噻嗪 2-10 mg/kg IP

乙酰丙嗪 0.5-1.5 mg/kg IP

60-120 分钟

戊巴比妥

30-50 毫克/公斤 IP

90-120 分钟

*氯胺酮/甲苯噻嗪不与镇痛剂(阿片类药物或非甾体抗炎药)联合使用可能不足以产生手术平面麻醉。手术前给予适当的镇痛剂和/或添加乙酰丙嗪将增强氯胺酮/甲苯噻嗪的麻醉效果。

** 氯胺酮/甲苯噻嗪混合物的制备说明见下文。

指南 – 为大鼠制备氯胺酮/甲苯噻嗪混合物

  • 需要使用无菌注射小瓶(例如 redtop 采血管;商业注射小瓶)
  • 混合说明:
    • 在混合之前验证您的药物浓度
    • 对于使用 100 mg/mL 氯胺酮和 100 mg/mL 甲苯噻嗪的 10mL 小瓶,添加:
      • 10 毫升氯胺酮 (100 毫克/毫升)
      • 1 毫升甲苯噻嗪 (100 毫克/毫升)
  • 建议使用以下模板作为标签:
    • 大鼠麻醉剂混合物: 氯胺酮/甲苯噻嗪
    • 用量:0.1 毫升/100 克 IP
    • 递送:91 mg/kg 氯胺酮,9.1 mg/kg 甲苯噻嗪
    • 浓度:91 mg/mL 氯胺酮,9.1 mg/mL 甲苯噻嗪
    • 过期:____________
    • 鸡尾酒的有效期由混合日期起六个月或任何一个成分先到期(如果少于 6 个月)确定
      • 例如:稀释于 2011 年 8 月 13 日,氯胺酮于 2012 年 10 月 12 日到期,甲苯噻嗪 11 年 10 月 10 日到期,注射用无菌水 2013 年 1 月 12 日到期,鸡尾酒的有效期为 2011 年 10 月 10 日

兔子

试剂 剂量
氯胺酮/甲苯噻嗪 氯胺酮 22-50 毫克/公斤 IM

甲苯噻嗪 2.5-10 mg/kg IM

戊巴比妥20-60 毫克/公斤静脉注射

试剂 剂量
氯胺酮/甲苯噻嗪 氯胺酮 20 毫克/千克 IM

甲苯噻嗪 2 毫克/千克 IM

特拉唑/氯胺酮替拉唑 4.4 mg/kg

氯胺酮 2.2 毫克/公斤

戊巴比妥20-40 毫克/公斤静脉注射

试剂 剂量
氯胺酮/甲苯噻嗪 5-15 mg/kg IM 氯胺酮

0.05-0.2 mg/kg IM甲苯噻嗪

硫喷妥钠10-16 毫克/公斤静脉注射

雪貂

试剂 剂量
氯胺酮/甲苯噻嗪 10-25 mg/kg IM 氯胺酮

0.25-0.5 mg/kg IM甲苯噻嗪

其他物种或麻醉剂:
请联系爱荷华大学临床 兽医(链接发送电子邮件) 艾维汀

(三溴乙醇)的制备和储存指南

  • Avertin 是一种速效、非药物级* 麻醉剂,用于小鼠的短期外科手术。
    • 注意:  根据第 8 版《实验动物护理和使用指南》,需要在《动物议定书》中描述和科学证明非药物级化学品或物质的使用。
  • 防范措施:
    • 如果您有以下情况,请勿服用 Avertin:
      • 非无菌溶液
      • 过时的解决方案
      • 更浓缩的解​​决方案
      • 高于推荐剂量
    • 由于导致胃肠道刺激,阿佛汀只能给药一次(不得重复给药)
  • 使用 Avertin 的缺点:
    • 组织刺激,特别是在高剂量、高浓度或重复剂量下
    • 在热或光存在下降解产生有毒副产物,这些副产物可能具有肾毒性和肝毒性
    • 注射后数周可引起肠梗阻
    • 对 16 天以下的小鼠或碳水化合物代谢改变的小鼠(例如,用作糖尿病或肥胖模型的小鼠品系)产生不可预测的影响
    • 当阿佛汀用作麻醉剂时,一些欧洲期刊拒绝接受研究手稿
  • 原料:
    • 2.5 克 2,2,2 三溴乙醇
    • 5 毫升 2-甲基-2-丁醇(水合戊烯,叔戊醇)
    • 200 毫升蒸馏水 – 中性 pH 值(无菌)
  • 制备(12.5 mg/ml 溶液):
    • 将 2.5 克三溴乙醇溶解在 5 毫升水合戊二烯中。  
    • 将溶解的溶液加热至 40°C,同时剧烈搅拌。
      • 不要超过 40°C。
    • 加入蒸馏水,不断搅拌,直至最终体积为 200 ml。
    • 通过 Millipore 过滤器(0.5 微米)过滤除菌
    • 贮存:
      • 将最终溶液过滤到红盖采血管或琥珀色(棕色)无菌玻璃容器中
      • 溶液容​​器必须用铝箔包裹以保护溶液避光
      • 溶液容​​器必须标明内容物和制备日期
      • 存放在冰箱或冰柜中
  • 满足以下任一条件即为过期:
    • 如果存放在冰箱中,有效期为两周
    • 如果存放在冰箱中,有效期为一年
    • 溶液结晶
    • 溶液颜色变黄

英文版

Recordkeeping for Anesthesia/Sedation

  • Applies to all survival and non-survival procedures performed under anesthesia
  • Mice and Rats (as well as fish, amphibians, reptiles and birds)
    • Brief procedures using inhalant/absorbed anesthetics
      • No record required
      • Brief procedures are those that cause no more than momentary pain (e.g. blood collection, tail vein injection, tail/fin snip for genotyping or intranasal inoculations)
    • For all other procedures not described above
      • A record is required
      • Animals should be monitored until full recovery if survival or euthanasia if non-survival
      • Document the following:
        • Date
        • Principal Investigator
        • Animal Protocol number
        • Animal ID
        • Species
        • Weight
        • Procedure
        • Agent(s) used, dosage, route of administration
        • Time of induction of anesthesia
        • Time of recovery from anesthesia or time of euthanasia
  • USDA Covered Species (all mammals except for mice of the genus Mus and rats of the genus Rattus)
    • Any anesthetic event
      • An individual record for each animal is required
      • Document the following:
      • Date
      • Principal Investigator
      • Animal Protocol number
      • Animal ID
      • Species
      • Weight
      • Procedure
      • Agent(s) used, dosage, route of administration
      • Time of induction of anesthesia
      • Time of recovery from anesthesia or time of euthanasia
      • Monitoring (every 15 minutes until recovery or euthanasia)
  • See surgery guidelines for surgery record information
  • Click here for  template anesthesia records

Supportive Care of Animals During Anesthesia

  • Apply ophthalmic ointment to both eyes to prevent desiccation for any anesthesia longer than 5 minutes
  • Maintain normal body temperature using a warm circulating water blanket, thermal pads, and/or warmed IV fluids
    • DO NOT use an “over the counter” electric heating pad as these are prone to overheating
  • Provide fluids (e.g., IV, IP, SQ) to animals during prolonged anesthesia to maintain adequate hydration as described in the approved Animal Protocol

Monitoring and Assessment of Anesthesia

  • USDA Covered Species (all mammals except for mice of the genus Mus and rats of the genus Rattus)
    • Monitor heart rate, respiratory rate, and temperature
      • Document these parameters at least every 15 minutes during anesthesia
      • For rodent species, qualitative monitoring for normal cardiovascular and respiratory function may be sufficient, as it is difficult to assess these parameters quantitatively.
      • For all other species, quantitative monitoring (numerical heart and respiratory rates) is expected.
      • If using a ventilator, note ventilation rate and tidal volume in the records
    • Monitor hemodynamic parameters to assure adequate gas exchange
      • Mucous membranes should be pink and moist
      • Capillary refill time should be less than 2 seconds
    • Monitoring for at least one indicator of deep pain recognition (pedal reflex, pinna reflex, etc.) should be performed regularly to ensure adequate anesthesia
      • Adjust the depth of anesthesia as dictated by changes in the monitored parameters to ensure continued surgical plane of anesthesia
  • Mice and Rats
    • Monitor respiratory rate and effort, color of mucous membranes, and reflected eye color (in albino animals) at regular intervals (no longer than 15 minute intervals)
    • Assess level of anesthesia by pedal reflex (firm toe pinch) and adjust anesthetic delivery as appropriate to maintain surgical plane

Anesthetic Recovery

  • Large Animals
    • Food and water bowls must be removed from the recovery cage
    • Monitor each animal until fully recovered (normal values for the following):
      • Heart and respiratory rate
      • Maintaining sternal recumbency (lying upright on chest)
      • Heart and respiratory rate
      • Body temperature
        • A circulating warm water heating pad is recommended
      • Hydration is assessed by skin turgor or mucous membrane “tackiness”
    • Deflate and remove the intubation tube once animal can swallow
      • Do not leave animal unattended while intubation tube is in place
    • Occasionally reposition recumbent animals to promote a quicker recovery
      • Animals should not remain with the same side down for more than four (4) hours
    • Lower animal’s head slightly below chest level to prevent aspiration if vomiting occurs
  • Mice and Rats
    • Place rodent in warm, clean, dry, quiet environment away from other animals
    • Cover or replace bedding material with toweling material
      • Bedding can stick to eyes or be inhaled while animals are recovering from anesthesia
    • Provide warmth during recovery:
      • Commercially-available surgical heating pad (NOT an “over the counter” human heating pad)
      • Incandescent lamp (50-75 watt) 12-14 inches away from rodent
        • Position lamp so that rodent can escape the light sources if desired
        • Attentive monitoring must be performed to prevent overheating of rodent
      • Use of a temperature-controlled cage/incubator
    • If listed in approved Animal Protocol, warm sterile saline can be administered to replace body fluids lost during surgery
    • Rodents must be continuously monitored until maintaining upright posture and walking normally about the cage before return to the animal housing room
  • All animals must be continuously monitored until maintaining upright posture and walking normally about the cage before completion of monitoring and return to the animal housing room.

Anesthetic Agents

The following is a list of commonly used anesthetic agents.  This list is not inclusive; other anesthetic agents may be listed and used in an Animal Protocol.

Other accepted resources for appropriate analgesics include the following formularies:

  • Plumb’s Veterinary Drug Handbook (Plumb)
  • Anesthesia and Analgesia in Laboratory Animals (ACLAM)
  • Formulary for Laboratory Animals (Hawk)
  • Exotic Animal Formulary (Carpenter & Marion)
  • Handbook of Veterinary Anesthesia (Muir & Hubbell)
  • Swine in the Laboratory (Swindle)

Appropriate anesthetic drugs and dosage(s) should be determined in consultation with an OAR or IACUC veterinarian.

Inhaled Anesthetic Agents

ISOFLURANE/SEVOFLURANE – VAPORIZER

  • Isoflurane/sevoflurane must be administered with a properly calibrated vaporizer when used as an anesthetic agent for surgery
  • Anesthetic gases must be scavenged properly
    • Direct exhaust
    • Activated charcoal canister (e.g. F/Air canister)
      • Should be weighed before each use and must be discarded when maximum weight is achieved
      • Weight records must be maintained
  • Vaporizers must be calibrated at least yearly
    • Calibration records for each vaporizer must be maintained

ISOFLURANE – DROP JAR METHOD

  • NOT for sevoflurane use; the concentration of sevoflurane cannot be accurately controlled with the drop jar method
  • Isoflurane can be administered in an anesthetic drop jar for a single, brief procedure
    • Cannot be used for any surgical procedures (includes non-survival and survival surgeries)
  • Anesthetic gases must be scavenged properly
    • Fume hood
    • Hard-ducted biosafety cabinet
  • Animals must not  come into direct contact with liquid isoflurane
  • Drop jar must be cleaned (i.e. removal of urine/feces) between animals
  • Use of this method with a 50 mL conical tube is not recommended

Drop jar dosing for Isoflurane: Internal Volume of Chamber (L) and isoflurane liquid required (mL)

Isoflurane Concentration achieved  1L 2L 3L 4L 5L
1% 0.05mL 0.10 mL 0.15 mL 0.20 mL 0.26 mL
2% 0.10 mL 0.20 mL 0.31 mL 0.41 mL 0.51 mL
3% 0.15 mL 0.31 mL 0.46 mL 0.61 mL 0.77 mL
4% 0.20 mL 0.41 mL 0.61 mL 0.82 mL 1.02 mL
5% 0.26 mL 0.51 mL 0.77 mL 1.02 mL 1.28 mL

75% CO2 / 25% O2

  • 75% CO2 / 25% O2 can be used for single, brief procedures
    • Cannot be used for any surgical procedures (includes non-survival and survival surgeries).
  • Available pre-mixed from vendors
  • Use precaution; animals can be easily overdosed
  • Not intended for euthanasia
  • Contact a member of the Office of Animal Resources veterinary staff if you are unfamiliar with the proper use of CO2/O2

Absorbed Anesthetic Agents

  • Tricaine Methanesulfonate (MS-222)
    • ​Frogs:
      • 0.05-0.2% (500-2000mg/L) solution
      • Solution must be buffered with sodium bicarbonate to a pH of 7.0-7.5
      • Immerse frog in solution for 10-20 minutes
      • Level of anesthesia is judged by loss of righting reflexes, loss of gulping reflex and loss of withdrawal response to toe pinch
    • Fish:
      •  0.0025-0.01% (25-100mg/L) solution
      • Solution must be buffered with sodium bicarbonate to a pH of 7.0-7.5
      • Immerse fish in solution until appropriate anesthetic depth is observed
      • Level of anesthesia is judged by loss of equilibrium, loss of response to noxious stimuli (pinching base of tail), rate of opercular movement and gill color
    • Storage
      • Tricaine (liquid solution) can be stored at room temperature for 3-5 days if protected from exposure to light.
      • Tricaine (liquid solution) can be stored at 4°C (i.e. – the refrigerator) for 1 month if stored if protected from light.
      • Tricaine (liquid solution) can be stored at -20°C (i.e. – the freezer) for 1 year if protected from the light.
      • Tricaine (powder) can be stored at room temperature for up to 5 years if stored in a dark container.

Injectable Anesthetic Agents

COMMONLY USED INJECTABLE ANESTHETIC AGENTS

MOUSE

Agent

Dosage

Duration of anesthesia

Ketamine/xylazine*

ketamine 80-100 mg/kg IP

xylazine 10-12.5 mg/kg IP

20-30 minutes

Ketamine/xylazine cocktail*

KX mouse cocktail 0.1mL/20g mouse wt. IP

Contains:

87.5 mg/kg Ketamine

12.5 mg/kg Xylazine

20-30 minutes

Ketamine/xylazine/acepromazine

ketamine 60-100 mg/kg IP

xylazine 10-15 mg/kg IP

acepromazine 2-5 mg/kg IP

60-90 minutes

Pentobarbital

50 mg/kg IP

20-40 minutes

Avertinǂ See warning below

240 mg/kg IP

30 minutes

*Ketamine/xylazine without combination with an analgesic agent (opioid or NSAID) may be insufficient to produce a surgical plane of anesthesia. Administration of appropriate analgesic agents prior to surgery and/or addition of acepromazine will augment the anesthetic effect of ketamine/xylazine.

** Preparation instructions for the ketamine/xylazine cocktail may be found below.                 

ǂ WARNING: NIH and European guidelines discourage the use of Avertin.  Preparation and storage requirements for Avertin may be found below.

GUIDELINES – PREPARATION OF KETAMINE/XYLAZINE COCKTAIL FOR MICE

  • Use of a sterile injection vial is required (e.g. redtop blood collection tube; commercial injection vial)
  • Mixing instructions:
    • Verify the concentration of your drugs prior to mixing
    • For a 10mL vial using ketamine 100 mg/mL and xylazine 100 mg/mL add:
      • 1.75mL ketamine (100 mg/mL)
      • 0.25 mL xylazine (100 mg/mL)
      • 8 mL saline or sterile water for injection
  • Use of the following template for a label is recommended:
    • Mouse Anesthetic Mix: Ketamine/Xylazine
    • Dosage: 0.1 ml/ 20gm IP
    • Delivers: 87.5 mg/kg Ketamine/12.5 mg/kg Xylazine
    • Concentration:  17.5 mg/mL Ketamine/2.5 mg/mL Xylazine

Expires: ____________

  • The expiration date for the cocktail is determined by either six months from the mixing date, or whichever of the components expires first (if less than 6 months)
    • E.g.: Diluted on 8/13/11, ketamine expires 12/10/2012, xylazine expires 10/10/11 and sterile water for injection expires 1/12/2013; the expiration date for the cocktail is 10/10/11

RAT

Agent

Dosage

Duration of anesthesia

Ketamine/xylazine

ketamine 40-100 mg/kg IP

xylazine 5-13 mg/kg IP

60-80 minutes

Ketamine/xylazine cocktail*

KX rat cocktail 0.1 mL/100g rat wt. IP

Contains:

91 mg/kg Ketamine

9.1 mg/kg Xylazine

60-80 minutes

Ketamine/xylazine/acepromazine

ketamine 20-50 mg/kg IP

xylazine 2-10 mg/kg IP

acepromazine 0.5-1.5 mg/kg IP

60-120 minutes

Pentobarbital

30-50 mg/kg IP

90-120 minutes

*Ketamine/xylazine without combination with an analgesic agent (opioid or NSAID) may be insufficient to produce a surgical plane of anesthesia. Administration of appropriate analgesic agents prior to surgery and/or addition of acepromazine will augment the anesthetic effect of ketamine/xylazine.

** Preparation instructions for the ketamine/xylazine cocktail may be found below.

GUIDELINES – PREPARATION OF KETAMINE/XYLAZINE COCKTAIL FOR RATS

  • Use of a sterile injection vial is required (e.g. redtop blood collection tube; commercial injection vial)
  • Mixing instructions:
    • Verify the concentration of your drugs prior to mixing
    • For a 10mL vial using ketamine 100 mg/mL and xylazine 100 mg/mL add:
      • 10 mL ketamine (100 mg/mL)
      • 1 mL xylazine (100 mg/mL)
  • Use of the following template for a label is recommended:
    • Rat Anesthetic Mix: Ketamine/Xylazine
    • Dosage: 0.1 ml/ 100gm IP
    • Delivers: 91 mg/kg Ketamine, 9.1 mg/kg Xylazine
    • Concentration:  91 mg/mL Ketamine, 9.1 mg/mL Xylazine
    • Expires: ____________
    • The expiration date for the cocktail is determined by either six months from the mixing date, or whichever of the components expires first (if less than 6 months)
      • E.g.: Diluted on 8/13/11, ketamine expires 12/10/2012, xylazine expires 10/10/11 and sterile water for interjection expires 1/12/2013, the expiration date for the cocktail is 10/10/11

RABBIT

Agent Dosage
Ketamine/xylazine ketamine 22-50 mg/kg IM

xylazine 2.5-10 mg/kg IM

Pentobarbital20-60 mg/kg IV

PIG

Agent Dosage
ketamine/xylazine ketamine 20 mg/kg IM

xylazine 2 mg/kg IM

Telazol/ketaminetelazol 4.4 mg/kg

ketamine 2.2 mg/kg

Pentobarbital20-40 mg/kg IV

SHEEP

Agent Dosage
Ketamine/xylazine 5-15 mg/kg IM ketamine

0.05-0.2 mg/kg IM xylazine

Thiopental10-16 mg/kg IV

FERRET

Agent Dosage
Ketamine/xylazine 10-25 mg/kg IM ketamine

0.25-0.5 mg/kg IM xylazine

Other species or anesthetic agents:
Please contact a University of Iowa clinical veterinarian(link sends e-mail) for consultation

GUIDELINES FOR PREPARATION AND STORAGE OF AVERTIN (TRIBROMOETHANOL)

  • Avertin is a quick-acting, non-pharmaceutical grade* anesthetic that is used for short duration surgical procedures in mice.
    • NOTE:  Per the Guide for the Care and Use of Laboratory Animals 8th edition, the use of non-pharmaceutical grade chemicals or substances needs to be described and scientifically justified in the Animal Protocol.
  • Precautions:
    • Do not administer Avertin if you have a/an:
      • Non-sterile solutions
      • Outdated solutions
      • More concentrated solutions
      • Higher dosages than recommended
    • Avertin should only be administered one time (no redosing) due to resultant gastrointestinal irritation
  • Disadvantages of the use of Avertin:
    • Tissue irritation, especially at high dosages, high concentrations or repeated doses
    • Degrades in the presence of heat or light to produce toxic byproducts which can be both nephrotoxic and hepatotoxic
    • Can cause intestinal ileus several weeks after injection
    • Unpredictable effects in mice under 16 days of age or in mice with altered carbohydrate metabolism (e.g., mouse strains used as diabetes or obesity models)
    • Some European journals are rejecting research manuscripts when Avertin  is used as an anesthetic
  • Ingredients:
    • 2.5 grams 2,2,2 Tribromoethanol
    • 5 ml 2-methy-2-butanol (amylene hydrate, tertiary amyl alcohol)
    • 200 ml distilled water – neutral pH (sterile)
  • Preparation (12.5 mg/ml solution):
    • Dissolve 2.5 grams Tribromoethanol in 5 ml amylene hydrate.
    • Heat dissolved solution to 40°C while stirring vigorously.
      • Do not exceed 40°C.
    • Add distilled water, stirring continuously, up to a final volume of 200 ml.
    • Filter sterilize through a Millipore filter (0.5 micron)
    • Storage:
      • Filter final solution into red-cap blood collection tubes or amber (brown) colored sterile glass containers
      • Solution container must be wrapped in aluminum foil to protect solution from light
      • Solution container must be labeled with contents and date of preparation
      • Store in refrigerator or freezer
  • Expiration has occurred if any one of the following conditions are met:
    • Two week expiration date if stored in refrigerator
    • One year expiration date if stored in freezer
    • Crystallization of solution
    • Solution has turned yellow in color

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