全身麻醉下小鼠幸福度的系统评价
Summary
我们制定了一个协议, 以评估小鼠的福祉在程序中使用全麻。分析了一系列的行为参数, 说明了幸福水平和糖皮质激素代谢产物。该议定书可作为一般援助, 以科学的、以动物为中心的方式估计严重程度。
Abstract
根据罗素和科里开发的3R 原理 (替换、还原、细化), 科学研究应尽可能使用动物实验的替代品。如果不能替代动物实验, 那么所使用的实验动物总数应该是获取有价值数据所需的最低数量。此外, 应采用适当的细化措施, 以尽量减少伴随实验程序的痛苦、痛苦和痛苦。用于分类疼痛、痛苦和痛苦程度的类别是不恢复的、轻度的、中等的或严重的 (欧盟指令 2010/63)。要确定哪些类别适用于个别案例, 使用科学的声音工具是至关重要的。
这里提出的幸福评估协议是为使用全麻的程序设计的。该协议的重点是家庭笼活动, 鼠标鬼脸规模, 和奢侈行为, 如穴居和巢建筑行为作为福祉的指标。它也使用自由探索范式的特质焦虑相关行为。粪便皮质酮代谢物作为急性应激指标的测量在24小时后麻醉期间。
该议定书提供了有关全麻后小鼠的福祉的科学可靠的信息。由于其简单性, 该协议可以很容易地被修改和集成在一个计划的研究。虽然它没有提供一个规模, 根据欧盟指令2010/63 分类的苦恼, 它可以帮助研究人员估计一个程序的严重性程度, 使用科学的声音数据。它提供了一种以科学、动物为中心的方式来改进对幸福的评估的方法。
Introduction
欧盟指令 2010/631规定, 每当需要动物试验时, 由罗素和2开发的3R 原理 (替换、还原、细化) 将被应用.欧盟指令的最终目标是逐步淘汰所有动物试验, 但该指令承认, 目前还需要一些动物实验来进行研究, 以保护人类和动物的健康。因此, 如果动物实验不能被任何替代方法所取代, 只有最小数量的实验动物被用来获得可靠的结果。此外, 使用适当的细化措施, 应尽量减少伴随实验程序的疼痛、痛苦和苦恼的数量。欧盟指令2010/63 规定, 过程的严重性必须前瞻性归类为非恢复、温和、适度或严重的1。由于严重性分类是根据个案来决定的, 因此有科学的声音工具来估计给定过程的严重性是很重要的。
根据莫顿和格里菲斯的建议, 评分表3是检测任何偏离正常状态的重要工具, 包括对福祉4的负面影响。评分表用于回顾性地确定由实验引起的疼痛、痛苦和苦恼, 并着重于个体动物 (例如、体重、皮毛、步态) 的物理状态的可见变化。虽然, 欧盟指令2010/63 的附件八提供了每个严重性类别的例子, 但研究人员仍然缺乏利用科学基础的数据来估计给定程序严重性程度的工具。
缺乏显示消极福祉的指标并不是确定动物状况的唯一方法;指向正福利的指示器的存在也很重要5、6、7、8。例如, 动物只在满足所有基本需求时, 才会表现出诸如挖洞和筑巢等奢侈行为。如果减少了福利, 奢侈品行为首先会下降5,7。用于评估福祉的议定书应包括指示动物生理、生理/生物化学和心理状态的指标, 以便以详细和全面的方式对其福祉进行评估9。
在改进的范围内, 制定了一项议定书, 以满足这些要求, 并评估一项涉及全麻对小鼠福祉的程序的影响10。同时, 目标是尽量减少任何额外的压力, 使协议易于集成到给定的实验中。该协议考虑穴居行为, 家庭笼行为, 如活动, 食物摄取, 和巢, 和特质焦虑相关的行为。此外, 还包括小鼠的鬼脸量表和粪便中皮质酮代谢物的无创性分析。该议定书的目的是促进以科学和动物为中心的方式评估福祉, 并提供支持严重程度分类的福祉信息。除了评分表之外, 它还可以为过程的严重性分类提供有用的信息。由于该协议易于执行且不需要广泛的设备, 因此可以将其集成到一个正在进行的实验中, 而不影响研究结果。应该指出的是, 动物研究: 报告的活体实验 (到达) 指南11将在所有涉及动物实验的研究中观察到, 目的是改进设计、分析和报告。
Protocol
Representative Results
Discussion
该议定书最初是为了评估接受单一麻醉或重复异氟醚麻醉的 C57BL/6JRj 小鼠的福祉而制定的。研究结果证实, 对奢侈品行为的测试以及其他措施 (例如自由探索范式、mg、穴居食物摄取) 都是评估幸福感的敏感方法。反复异氟醚麻醉对特质焦虑相关行为、特征和穴居行为产生短期影响。此外, 反复异氟醚麻醉似乎影响食物摄取量10。
自由探索范式表明, 减少…
Disclosures
The authors have nothing to disclose.
Acknowledgements
多亏了 Rassam 的样本收集, 伊迪丝 Klobetz–对 FCM 的分析, PD 博士. 兽医。habil。萝丝维塔梅尔协助统计分析, 并 Wiebke Gentner 校对手稿。这项研究是柏林-勃兰登堡研究平台 BB3R (www.bb3r.de) 的一部分, 由德国联邦教育部和研究部资助 (赠款号: 031A262A) (www.bmbf.de/en/index.html)。
Materials
| Isofluran | CP-Pharma Handelsgesellschaft mbH | 1214 | |
| InfraMot – Sensore Units | TSE Systems | 302015-SENS | |
| InfraMot – Control Units | TSE Systems | 302015-C/16 | |
| InfraMot – Software | TSE Systems | 302015-S | |
| Nestlet N | Ancare – Plexx | NES3600 | |
| Camera EOS 350D | Canon |
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