一个简单的方法来操作的动物行为观察溶解氧
Summary
本文介绍了一个简单的和可重复的协议来操作溶氧条件在实验室环境对动物行为的研究。该协议可能在教学和研究实验室的设置可用于评估在溶解氧浓度的变化无脊椎动物，鱼类，两栖类或生物体的反应。
Abstract
操纵溶解氧（DO）在实验室环境的能力有显著应用调查若干生态和生物体的行为的问题。这里所描述的协议提供了一种简单的，可重复，并且控制方法来操纵DO来研究从低氧和缺氧条件下得到的水生生物的行为反应。在执行的水脱气用氮气在实验室设置常用生态（水生）应用没有明确的方法存在于文献中，并且该协议是第一个描述的协议以degasify水来观察生物体反应。这种技术和协议是为水生无脊椎动物直接应用开发;然而，小鱼类，两栖类，和其它水生脊椎动物可以很容易被取代。它允许从2毫克/升，以11毫克/升与稳定性DO水平的容易操纵长达5分钟的动物观察期。超过5分钟的观察期水温开始上升，并在10分钟DO水平变得太不稳定维护。该协议可扩展到研究有机体，重现性好，可靠，允许快速实施到入门教学实验室和高层次的研究应用。这种技术的预期的结果应与溶解氧改变生物体的行为反应。
Introduction
溶解氧（DO）是在调解一些水生生态系统中的生物和生态过程的重要关键生理参数。暴露于急性和慢性亚致死缺氧减少某些水生昆虫的生长率，降低暴露1昆虫的生存。该协议的开发提供一个可控的方法来操纵溪水DO水平，观察动物行为的影响。由于所有的有氧水生生物的生存，为了生存和繁殖依赖于氧浓度，溶解氧浓度的变化往往反映在生物行为变化。更多的移动水生无脊椎动物和鱼类已观察到寻求语言环境具有较高的DO 2,3低氧浓度（缺氧）响应。对于流动性较差的水生生物，行为适应增加DO的摄入量可能是唯一可行的选择。 PLEC的水生底栖动物秩序的OPTera（StoneFly的）已被指出以执行“上推”运动，以增加水中的氧的流量，和吸收，在其外部鳃4 – 6。这些适应性行为已在自然的环境，并在实验室实验中观察到。
在水中溶解氧的实验室操作开辟了动物行为研究显著的机会，但在方法上部署显著存在差距。例如，一项研究使用的大型水族馆评估以下用氮气脱气大嘴低音的生理响应时间（ 加州鲈 ）缺氧的环境中，但很少详细给出的方法7。使用氮气和多孔石将气体输送到水，减少水分8 DO描述上斑马鱼（ 斑马鱼 ）进行的另一项研究。对于基于化学的应用程序，对于溶剂的脱气方法使用专门的设备9 – 11从溶剂除去氧气，但不会是适合动物行为的研究。虽然这些研究采用的方法来从水中除去氧气，没有描述的方法可以查明将允许对动物行为评价响应于DO变化。
下面描述这种方法是试图通过使用氮气充分描述了的水的DO操作的协议。此外，这种方法对观察StoneFly的行为（如俯卧撑）之间的关系发展，去做一些在大一级别的生物实验室使用。其中一个这种方法的主要好处是，它可以很容易地与常用玻璃器皿和大多数中等和高等教育机构可获得的材料在实验室内进行。该协议也很容易适应，允许个人扩展，以满足提出的研究或教学应用目标的过程。 </p>
Protocol
Representative Results
Discussion
关键步骤
这个程序提供了一个简单而有效的方式来操作DO在实验室环境对水生生物进行行为研究。我们发现有以几个关键步骤/项意识到执行该实验是直接相关的结果时的感觉。在一个试验中，它是保持室压力，以避免在水上方的气体的分压的变化的关键，和随后的DO波动。按照协议的“试实验装置的稳定”小节中所列的步骤是关键的。检查与烧瓶塞子的密封，保证真空管的完全浸没到水的1升?…
Divulgations
The authors have nothing to disclose.
Acknowledgements
The Authors would first like to acknowledge all students from the freshman Biology 121- Ecology Module lab at Juniata College for their help in generating data used in this study. We would also like to thank Dr. Randy Bennett, Chris Walls, Sherry Isenberg, and Taylor Cox for their assistance in acquiring materials necessary to develop this methodology. Additionally, we would like to thank Dr. Norris Muth and Dr. John Unger for their advice on methodological development and Dr. Jill Keeney and the Biology department for their support of this endeavor. We would also like to thank the anonymous reviewers that have helped to shape and focus this manuscript. Last but not least, I'd like to thank Hudson Grant for his help with the initial stonefly collection for use in development of this technique
Materials
| Filter flask 2 L | Pyrex | 5340 | |
| Rubber Stopper size 6 | Sigma-Aldrich | Z164534 | |
| Nalgene 180 Clear Plastic Tubing | Thermo Scienfitic | 8001-1216 | |
| Whisper 60 air pump | Tetra | N/A | |
| Standard flexible Air line tubing | Penn Plax | ST25 | |
| 0.25 inch Copper tubing | Lowes Home Improvement | 23050 | |
| Male hose barb | Grainger | 5LWH1 | |
| Female Connector | Grainger | 20YZ22 | |
| Heavy Duty Dissolved Oxygen Meter | Extech | 407510 | |
| Nitrogen gas | Matheson TRIGAS | N/A | |
| Radnor AF150-580 Regulator | Airgas | RAD64003036 |
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