Name: using deuterium oxide as a non-invasive, non-lethal tool for assessing body composition and water consumption in mammals
Uploaded: 2020-02-20T12:00:00
Description: Watch this Scientific Journal Video about Using Deuterium Oxide as a Non-Invasive, Non-Lethal Tool for Assessing Body Composition and Water Consumption in Mammals at JoVE.com

这项研究得到了MDC合作协议（#416）、美国森林服务合作协议（16-JV-1124231-118）、美国兽医营养学院和沃尔瑟姆/皇家坎宁大学、美国赠款（赠款号：00049049）、NIH培训补助金（赠款号：T32OS011126）和密苏里大学兽医研究学者计划的支持。作者感谢香农·埃勒斯对这份手稿进行了预先审查。我们感谢罗伯特·沃库斯博士提供D2O标准并允许使用他的实验室。

这里描述的所有实验都得到密苏里大学动物护理和使用委员会的批准，并在密苏里保护部（MDC）野生动物科学采集许可证（许可证#16409和#17649）下进行。
1. 制备无菌、等位、盐渍D2O库存溶液
<ol><li>制作 9.0 g/L 盐渍 D2O 的 50 mL 库存溶液。<ol>
<li>重量为 450 mg 的制药级 NaCl，并将所有 NaCl 转移到 100 mL 的消毒烧杯中。在实验室笔记本中记录 NaCl 的确切数量到小数?...

<ol>
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自20世纪40年代以来，利用氧化氮测定TBW，并用于人类和各种家养和野生动物物种4、6、7。其他非破坏性技术已经开发，包括生物电阻抗分析 （BIA）、DXA 和定量磁共振 （QMR）。每种方法都有优点和缺点，在选择评估身体成分的特定方法之前应考虑。该协议选择使用DXA作为氧化氮的比较方法，以评估身体成分?...

身体状况评分系统和身体状况指数是评估1、2类健康状况或健康状况的常见技术。许多家养动物和动物物种都有独特的身体状况评分（BCS）系统，用于评估动物的肌肉和表面脂肪组织3。但是，BCS 评估依赖于评估者，这意味着当经过培训的评估者评估 BCS 是一个目标或半定量的测量时。在野生动物物种中，身体状况指数是常用的，而不是BCS，并且基于身体质量与体型或身体质量与前臂2的比率。身体状况的病症往往被觅食的影响混淆，可以混淆的体型，以及统计和推断问题4。
身体状况评分系统和身体状况指数的替代是使用稳定的同位素来确定身体成分。一种常用的稳定同位素是氧化铀（D2O），这是一种非放射性水，其中氢原子是铀同位素。本研究中描述的氧化氧化稀释方法可以是一种非主观、定量和可重复的技术，用于估计人类5和多种物种4、6、7的人体组成。这种技术对研究野生动物的身体组成是有利的。例如，它可用于评估身体成分的纵向变化，例如管理行动之前和之后。然而，在一些野生动物物种中，氧化氮可以高估实际含水量8。因此，在适应某一物种的技术时，通过比较氧化氧化法和非濒危物种的骨层分析来验证该方法非常重要。对于受威胁和濒危物种，应将双X射线吸收测量（DXA）等非破坏性方法视为与完全尸体分析的黄金标准破坏性方法的替代比较方法。
除了身体成分，D2O稀释技术还可用于确定单个动物的耗水量9。D2 O的这种独特应用不仅可用于回答研究问题，还可用于评估大型社会环境中个体动物的耗水量。
在这里，我们描述了D2O稀释技术的适应，用于评估昆虫、大棕色蝙蝠（Eptesus fuscus）中的身体成分，以及评估食肉动物、猫（费利斯猫）的耗水量。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>0.2 micron non-pyrogenic disk filter</td>
			<td>Argos Technologies</td>
			<td>FN32S</td>
			<td>nylon, 30mm diameter, 0.22um, sterile</td>
		</tr>
		<tr>
			<td>1.5 mL conical microcentrifuge tubes</td>
			<td>USA Scientific</td>
			<td>1415-9701</td>
			<td>1.5 ml self-standing microcentrifuge tube, natural with blue cap</td>
		</tr>
		<tr>
			<td>10 mL sterile glass vial for injection</td>
			<td>Mountainside Medical Equipment</td>
			<td>MS-SEV10</td>
			<td>clear, sterile glass injection unit</td>
		</tr>
		<tr>
			<td>10 mL syringe</td>
			<td>Becton Dickinson</td>
			<td>305219</td>
			<td>sterile 10 mL syringe individually wrapped</td>
		</tr>
		<tr>
			<td>100 mL sterile glass vial for injection</td>
			<td>Mountainside Medical Equipment</td>
			<td>AL-SV10020</td>
			<td>clear, sterile glass injection unit</td>
		</tr>
		<tr>
			<td>20 gauge needle</td>
			<td>Exel</td>
			<td>26417</td>
			<td>needles hypodermic 20g x 1&#34; plastic hub (yellow) / regular bevel</td>
		</tr>
		<tr>
			<td>22 gauge needle</td>
			<td>Exel</td>
			<td>26411</td>
			<td>needles hypodermic 22g x 1&#34; plastic hub (black) / regular bevel</td>
		</tr>
		<tr>
			<td>deuterium oxide</td>
			<td>Sigma-Aldrich</td>
			<td>151882-25G</td>
			<td>99.9 atom % D</td>
		</tr>
		<tr>
			<td>isofluorane</td>
			<td>Vetone</td>
			<td>3060</td>
			<td>fluriso isoflurane, USP</td>
		</tr>
		<tr>
			<td>OMNIC Spectra Software</td>
			<td>ThermoFisher Scientific</td>
			<td>833-036200</td>
			<td>FT-IR standard software</td>
		</tr>
		<tr>
			<td>petroleum jelly</td>
			<td>Vaseline</td>
			<td>305212311006</td>
			<td>Vaseline, 100% pure petroleum jelly, original, skin protectant</td>
		</tr>
		<tr>
			<td>plastic capillary tubes</td>
			<td>Innovative Med Tech</td>
			<td>100050</td>
			<td>sodium heparin anticoagulant, 50 &#956;L capacity, 30 mm length</td>
		</tr>
		<tr>
			<td>Sealed liquid spectrophotometer SL-3 FTIR CAF2 Cell</td>
			<td>International Crystal Laboratory</td>
			<td>0005D-875</td>
			<td>0.05 mm Pathlength</td>
		</tr>
		<tr>
			<td>sodium chloride</td>
			<td>EMD Millipore</td>
			<td>1.37017</td>
			<td>suitable for biopharmaceutical production</td>
		</tr>
		<tr>
			<td>Thermo Electron Nicolet 380 FT-IR Spectrometer</td>
			<td>ThermoFisher Scientific</td>
			<td>269-169400</td>
			<td>discontinued model, newer models available</td>
		</tr>
	</tbody>
</table>

using deuterium oxide as a non-invasive, non-lethal tool for assessing body composition and water consumption in mammals

身体状况评分系统和身体状况指数是评估一个物种的健康状况或健康状况的常见技术。身体状况评分系统是评估者依赖的，具有高度主观性的潜力。身体状况指数可以通过觅食、体重的影响以及统计和推断问题来混淆。身体状况评分系统和身体状况指数的替代是使用稳定的同位素，如氧化铀来确定身体成分。氧化氧化剂稀释法是一种可重复的定量技术，用于估计人类、野生动物和国内物种的身体组成。此外，氧化铀稀释技术可用于确定单个动物的耗水量。在这里，我们描述了氧化氧化稀释技术对评估大棕色蝙蝠（Eptesus fuscus）的身体成分以及评估猫（Felis catis）的耗水量的适应。

氧化氧化稀释技术可用于评估各种物种的身体组成。为了证明适应性，我们报道了在北美昆虫蝙蝠物种Eptesicus fuscus（大棕色蝙蝠）中首次使用氧化氮稀释技术，以取得代表性的结果。通过采集D2 O注射前和后注血样本，完成定时高原，如对任何不为平衡期未知的物种进行。经测定，在非蝙蝠注射后两小时，足以进行平衡。随着平衡时间的已知，确定了13只野?...

Watch this Scientific Journal Video about Using Deuterium Oxide as a Non-Invasive, Non-Lethal Tool for Assessing Body Composition and Water Consumption in Mammals at JoVE.com

Using Deuterium Oxide as a Non-Invasive, Non-Lethal Tool for Assessing Body Composition and Water Consumption in Mammals

本文介绍了两种哺乳动物（昆虫和食肉）的氧化稀释技术，以确定全身水分、瘦体质量、身体脂肪量和耗水量。

使用氧化二氮作为评估哺乳动物体内成分和耗水量的非侵入性、非致命性工具

Body condition scoring systems and body condition indices are common techniques used for assessing the health status or fitness of a species. Body ...

使用本协议，氧化铝或D2O，一种非放射性、稳定的水同位素，可用于估计人体成分和水的消耗。这种技术是非侵入性的，因此可以用来评估濒危物种以及人类和国内物种的身体组成。氧化铝稀释技术的独特应用是能够测量自由范围野生动物的用水量，这些野生动物的回收率高，或者社会饲养的动物。虽然不需要麻醉，但对于那些不熟悉进行皮下注射以在交付氧化铝之前使动物感到窒息的人来说，它可能更容易。与莎拉·胡珀和我一起演示这个程序的将是阿曼达·埃舍尔曼和阿什利·考恩，兽医专业的学生，还有我实验室的研究生艾丽西娅·罗伊斯塔彻。要使50毫升的库存溶液每升9克氧化铝，首先重450毫克的药级氯化钠，并记录精确量到小数点四位。将所有盐转移到消毒的100毫升烧杯中，并在无菌的分级油缸中测量出大于或等于99.8%的氧化铝50克。将氧化钠的确切量记录到小数点四位，然后将氧化铝转移到氯化钠的无菌烧杯中。接下来，将一根 20 量针连接到非基因无菌盘过滤器上，该过滤器装有 10 毫升注射器筒的亚微孔，并将针头插入无菌、空的 100 毫升小瓶的隔膜中。将真空管连接到 22 米针上，并将针头插入 100 毫升小瓶的隔膜中。然后将10毫升的等尘强度氯化钠装入注射器桶中，然后慢慢打开真空，直到氧化铝溶液开始慢慢过滤到小瓶中。继续将氧化铝库存溶液倒入注射器筒中，直到过滤整个 50 毫升的体积。在确认麻醉蝙蝠对脚趾捏没有反应后，使用酒精准备垫清洁脑间静脉上的尿素，让消毒的尾膜干燥。在脑间静脉上涂抹一层薄薄的石油果冻，并使用 29 量针刺穿体间静脉的后部部分。然后使用塑料肝素钠毛细管收集100微升的血液。为确保将全血与肝素钠充分混合，在血液采集后轻轻卷起每个管，然后再贴标签。在确定以克为单位的氧化铀质量后，加载装有29量针的胰岛素注射器，并含有适当体积的氧化铝。称重装有二分量的胰岛素注射器和针头，将重量记录到小数点四位，并在麻醉蝙蝠的后臀区域下皮注射整个体积的氧化氮。我们建议使用带防静电防板的精密刻度，并确保在注射前拉回柱塞，以确保负压。注射后立即称重现在空的胰岛素注射器和针头，并记录重量到小数点四位。在采集血液后30分钟内，使用造血离心机以1万次g下旋转每个毛细管，每次5分钟。使用锋利的剪刀，将整片血液和血浆之间的塑料毛细管切割，并使用200微升移液器将血浆直接排出到有标签的500微升存储管中。平衡期后，从脑间静脉收集和储存另外100微升血液，正如刚刚证明的。对于 FTIR 分光光度测量分析，将沙浴温度设置为 60 摄氏度以方便蒸馏，并将每个等离子样品和标准 50 微升添加到单独的 1.5 毫升锥形微离心帽中。将微离心管倒置，将 1.5 毫升锥形微离心管拧到盖上，将倒置管与砂浴中的沙子接触至少 12 小时。第二天早上，在每个管子上放置一个新的清洁盖，并在微离心中脉冲微离心管10秒。将液体传输单元安装到 FTIR 光谱仪中，然后用甲醇填充电池。连接注射口，慢慢用背景水填充细胞，同时小心地取出甲醇注射器，以降低气泡的风险。将管子连接到输出端口，以便进行分析后去除样品，并打开 FTIR 光谱仪软件。根据表设置参数，并使用稀释剂 0.22 微米过滤蒸馏水收集背景样本。注入 40 微升，每百万氧化铀标准为零，并记录和保存光谱作为逗号分隔值文件。在注入并保存所有标准的光谱以创建标准曲线后，将每个蒸馏样品的 40 微升注入液体传输单元，并保存光谱。在已知的平衡时间，总身体水，瘦身体质量，和身体脂肪质量野生捕获大棕色蝙蝠和俘虏大棕色蝙蝠可以确定。负体脂肪质量可以计算由于以下一种或多个原因：不接收整个剂量的氧化铝，在平衡阶段变得发皮，具有异常大的脂肪质量和最小的瘦质量，或具有低于三至五%的身体脂肪由双X射线吸收测定确定。在这里，与双X射线吸收法相比，由氧化铝稀释技术确定的身体脂肪的代表性百分比。这两种技术关联得很好，身体脂肪质量显示出身体脂肪和体重之间的强相关性，而氧化铝稀释技术不能一致超过或低估身体脂肪质量。氧化铝稀释法在猫身上已经得到验证，并进行了改造，以便测量在实验的每个饮食区块中社会养猫的日常用水量。请记住，这种技术应该只对健康的动物执行，你应该准确地称量，记录，并交付给每个动物的氧化铝的全部剂量。可以完成确定身体成分的辅助方法，如完整的尸体分析或DEXA，以验证氧化铝技术。这项技术使我们能够识别出一只社会住的猫，它消耗的异常大量的水，而没有将每只猫分开来测量它们的用水量。

Research

JoVE Journal

Environment

Laboratory-determined Phosphorus Flux from Lake Sediments as a Measure of Internal Phosphorus Loading

实验室确定的磷通量湖泊沉积物作为内部磷载入一个测量

Eutrophication is a water quality issue in lakes worldwide, and there is a critical need to identify and control nutrient sources. Internal phosphorus (P) ...

科研

环境科学

Use of Chironomidae (Diptera) Surface-Floating Pupal Exuviae as a Rapid Bioassessment Protocol for Water Bodies

Use of Chironomidae Diptera Surface-Floating Pupal Exuviae as a Rapid Bioassessment Protocol for Water Bodies

使用摇蚊科（双翅目）的表面浮蛹蜕为快速生物议定书水体

Rapid bioassessment protocols using benthic macroinvertebrate assemblages have been successfully used to assess human impacts on water quality. ...

EPA Method 1615. Measurement of Enterovirus and Norovirus Occurrence in Water by Culture and RT-qPCR. Part III. Virus Detection by RT-qPCR

EPA方法1615测量肠道病毒和诺瓦克病毒发生水由文化和RT-qPCR的了。第三部分。病毒检测用RT-qPCR的

EPA Method 1615 measures enteroviruses and noroviruses present in environmental and drinking waters. This method was developed with the goal of having a ...

Removal of Trace Elements by Cupric Oxide Nanoparticles from Uranium In Situ Recovery Bleed Water and Its Effect on Cell Viability

Removal of Trace Elements by Cupric Oxide Nanoparticles from Uranium In Situ Recovery Bleed Water and Its Effect on Cell Viability

去除微量元素的氧化铜纳米粉体制备铀<em&gt;原位</em&gt;恢复泌水及其对细胞活力的影响

In situ recovery (ISR) is the predominant method of uranium extraction in the United States. During ISR, uranium is leached from an ore body and extracted ...

Soil Lysimeter Excavation for Coupled Hydrological, Geochemical, and Microbiological Investigations

土壤蒸渗仪开挖加之水文，地球化学和微生物调查

Studying co-evolution of hydrological and biogeochemical processes in the subsurface of natural landscapes can enhance the understanding of coupled ...

Screening for Endocrine Activity in Water Using Commercially-available In Vitro Transactivation Bioassays

Screening for Endocrine Activity in Water Using Commercially-available In Vitro Transactivation Bioassays

在用水筛查内分泌活动市售的<em&gt;体外</em&gt;式激活生物测定

In vitro transactivation bioassays have shown promise as water quality monitoring tools, however their adoption and widespread application has been ...

Protocol for Assessing the Relative Effects of Environment and Genetics on Antler and Body Growth for a Long-lived Cervid

评估环境和遗传因素对鹿茸和长寿命的鹿的身体生长的相对影响的议定书 》

Cervid phenotype can be placed into one of two categories: efficiency, which promotes survival over extravagant morphometric growth, and luxury, which ...

Composition and Properties of Aquafaba: Water Recovered from Commercially Canned Chickpeas

Aquafaba 的成分和性质: 从商用罐装鹰嘴中回收的水

Chickpea and other pulses are commonly sold as canned products packed in a thick solution or a brine. This solution has recently been shown to produce ...

A Simple Planting Technique for Re-establishing Trees Where Frequent Inundation Occurs

常淹发生的重构树简易造林技术

Many of the Everglades tree islands have lost elevation over the past century and most of their trees have died such that they are now covered with ...

Optimized Procedure for Determining the Adsorption of Phosphonates onto Granular Ferric Hydroxide using a Miniaturized Phosphorus Determination Method

用小型化磷测定法测定膦酸在粒状氢氧化铁上吸附的优化方法

This paper introduces a procedure to investigate the adsorption of phosphonates onto iron-containing filter materials, particularly granular ferric ...