Name: rabies necropsy techniques in large and small animals
Uploaded: 2019-07-30T13:00:00
Description: Watch this Scientific Journal Video about Rabies Necropsy Techniques in Large and Small Animals at JoVE.com

我们感谢纽约州卫生局沃兹沃斯中心对该项目的支持。我们还要感谢卫生部沃兹沃斯中心以及纽约州阿尔塔蒙特的LL牧场的艾米·威尔西和弗兰克·布莱斯德尔的支持。

所有描述的方法都通过了沃兹沃斯中心机构动物护理和使用委员会(IACUC)的批准。
1. 准备
<ol><li>唐PPE,至少护眼(眼镜或面罩),外科或N-95口罩,和非乳胶手套。</li>
	<li>准备工作区,理想情况下是生物安全柜 (BSC),带有一次性工作表面覆盖物(例如,牛皮纸或吸水垫)和清洁的尸检仪器(图 1)。</li>
	<li>将标本放在工作面上,并使用仪器操作它来评估样品的状况,包括?...

<ol>
	<li>Centers for Disease Control and Prevention (CDC). <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=West+Nile+virus+activity+-+United+States,+2009.">West Nile virus activity - United States, 2009.</a> MMWR Morbidity and Mortality Weekly Report. 59 (25), 769-772 (2010).</li><li>McDaniel, C. J., Cardwell, D. M., Moeller, R. B., Gray, G. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Humans+and+cattle:+A+review+of+bovine+zoonoses.">Humans and cattle: A review of bovine zoonoses.</a> Vector Borne and Zoonotic Diseases. 14 (1), 1-19 (2014).</li><li>Zoonotic diseases. Merck Veterinary Manual Available from: <a target="_blank" href="https://www.merckvetmanual.com/public-health/zoonoses/zoonotic-diseases">https://www.merckvetmanual.com/public-health/zoonoses/zoonotic-diseases</a> (2019)</li><li>Wenner, L., Pauli, U., Summermatter, K., Gantenbein, H., Vidondo, B., Posthaus, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Aerosol+generation+during+bone-sawing+procedures+in+veterinary+autopsies.">Aerosol generation during bone-sawing procedures in veterinary autopsies.</a> Veterinary Pathology. 54 (3), 425-436 (2017).</li><li>Green, F. H. Y., Yoshida, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characteristics+of+aerosols+generated+during+autopsy+procedures+and+their+potential+role+as+carriers+of+infectious+agents.">Characteristics of aerosols generated during autopsy procedures and their potential role as carriers of infectious agents.</a> Applied Occupational and Environmental Hygiene. 5 (12), 853-858 (1990).</li><li>Barrat, J., Meslin, F. X., Kaplan, M. M., Koprowski, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Simple+technique+for+the+collection+and+shipment+of+brain+specimens+for+rabies+diagnosis.">Simple technique for the collection and shipment of brain specimens for rabies diagnosis.</a> Laboratory techniques in Rabies 4th Edition. , 425-427 (1996).</li><li>Ness, S. L., Bain, F. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=How+to+perform+an+equine+field+necropsy.">How to perform an equine field necropsy.</a> American Association of Equine Practitioners. 55, 313-316 (2009).</li><li>. Sample requirements for TSE testing and confirmation &#8211; EURL guidance Available from: <a target="_blank" href="https://science.vla.gov.uk/tse-lab-net/documents/tse-oie-rl-samp.pdf">https://science.vla.gov.uk/tse-lab-net/documents/tse-oie-rl-samp.pdf</a> (2019)</li><li>. Rabies reports Available from: <a target="_blank" href="https://www.wadsworth.org/programs/id/rabies/reports">https://www.wadsworth.org/programs/id/rabies/reports</a> (2019)</li><li>. Protocol for postmortem diagnosis of rabies in animals by direct fluorescent antibody testing: A minimum standard for rabies diagnosis in the United States Available from: <a target="_blank" href="https://www.cdc.gov/rabies/pdf/rabiesdfaspv2.pdf">https://www.cdc.gov/rabies/pdf/rabiesdfaspv2.pdf</a> (2019)</li><li>Miller, L. D., Davis, A. J., Jenny, A. L., Fekadu, M., Whitfield, S. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Surveillance+for+lesions+of+bovine+spongiform+encephalopathy+in+U.S.+cattle.">Surveillance for lesions of bovine spongiform encephalopathy in U.S. cattle.</a> Developments in Biological Standardizations. 80, 119-121 (1993).</li><li>Andrews, C., Gerdin, J., Patterson, J., Buckles, E. L., Fitzgerald, S. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Eastern+equine+encephalitis+in+puppies+in+Michigan+and+New+York+states.">Eastern equine encephalitis in puppies in Michigan and New York states.</a> Journal of Veterinary Diagnostic Investigation. 30 (4), 633-636 (2018).</li><li>Appler, K., Brunt, S., Jarvis, J. A., Davis, A. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clarifying+indeterminate+results+on+the+rabies+direct+fluorescent+antibody+test+using+real-time+reverse+transcriptase+polymerase+chain+reaction.">Clarifying indeterminate results on the rabies direct fluorescent antibody test using real-time reverse transcriptase polymerase chain reaction.</a> Public Health Reports. 134 (1), 57-62 (2019).</li><li>Rupprecht, C. E., Fooks, A. R., Abela-Ridder, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chapter+7.+Brain+removal.">Chapter 7. Brain removal.</a> Laboratory techniques in Rabies 5th Edition. , 67-72 (2018).</li></ol>

提交狂犬病尸检的标本通常具有与神经系统疾病相容的临床体征史。临床疾病的存在可能与各种疾病有关,包括人畜共患疾病,增加实验室工作人员获得感染的风险。为了减少这些风险,已经实施了减少标本处理和操作的技术。
所展示的方法代表一个尸检事件,仅从单个动物中去除所需的组织。更常见的是,多个样品在一个班次中处理,需要小心,以确保样品之间没有交叉污染。清洁?...

在狂犬病实验室的尸检地板上工作本身是危险的。有时,标本到达与嵌入豪猪毛刺,异物,包括箭头/子弹/子弹或暴露的骨头碎片,可能会穿透保护性运输包装。包装不当可能导致泄漏,危及拆包样品的个人。除了身体伤害外,尸检技术人员还冒着接触中枢神经系统中未知的人畜共体感染剂和标本体液的风险。此外,标本携带的寄生虫可能传播其他人畜共患疾病,因为跳蚤和虱子通常出现在提交的动物身上。根据地理位置和涉及的物种,所感染的疾病各不相同。如东方马脑炎病毒(EEEV)或西尼罗河病毒(WNV),由虱子传播的疾病,包括莱姆病或黄原虫病,引起Q热或结核病的细菌,以及传染性病毒,列举少数可能的危险1,2,3.
这些方法的目的是使用仪器,尽量减少气溶胶的可能性,不像电动工具或锯4,5,证明安全和有效的尸检技术。通常,狂犬病实验室中小动物的尸检需要切除颅骨肌肉,并用锤子和凿子打开6号甲鱼的胆背部分。去除这个区域的脑后暴露,包括整个小脑和颅脑干。可对颅骨的腹腔部分进行改性尸检技术,避免颅骨大颅肌肉和较厚的区域。然而,这些经过修饰的尸检技术只有在标本没有颈椎时才可能。
同样,大型动物的脑组织可以通过分离颅骨肌肉和打开头骨的颅骨背部分来去除。由于大型动物的头骨通常较厚,因此需要付出相当大的努力来暴露小脑和脑干。为了避免穿透头骨,大型动物的头部被定位,因此头骨的通风口-双孔部分面向技师。使用改良的仪器,小脑和脑干通过前兆根去除。这与TSE欧盟转运海绵状脑病(TSE)调查参考实验室推荐的样品采集方法类似。颅椎应该事先切除,以便进入前胸。
这些技术的应用有利于在狂犬病实验室中经过适当培训的技术人员。由于狂犬病实验室接收了从幼蝙蝠到成年马9的各种样本,技术人员根据个人情况有多种选择方法。为大型动物演示的方法也适合在野外进行尸检的兽医,因为运送整个大型动物头进行狂犬病检测既繁琐又昂贵。实施这些技术将降低气溶胶生产的潜力,减少试样处理并节省加工时间,从而提高安全性。然而,由于该实验室与为狂犬病检测设立的实验室没有相同的优势,因此对这些程序所做的任何修改都必须注重安全,特别是个人防护设备的使用。

<table>
	<tbody>
		<tr>
			<td>Chemistry spoon</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Curved, sharp-blunt mayo scissors</td>
			<td>Sklar</td>
			<td>14-2055</td>
			<td>Sklar Operating Scissors 5-1/2 Inch Premium OR Grade Stainless Steel Finger Ring Handle Curved Sharp/Blunt</td>
		</tr>
		<tr>
			<td>Large sharp restaurant-quality carving knife</td>
			<td>Dexter</td>
			<td>P94848</td>
			<td>8&#34; Scalloped Utility Knife, white handle</td>
		</tr>
		<tr>
			<td>Locking tumor-tenacula</td>
			<td>Diamond Scientific and Surgicals</td>
			<td>N/A</td>
			<td>Czerny Tenaculum Forcep</td>
		</tr>
		<tr>
			<td>Modified stiletto knife (6.5 inch long blade carving knife ground to 0.5 inch wide)</td>
			<td>Dexter</td>
			<td>P94848</td>
			<td>Modified 8&#34; Scalloped Utility Knife, white handle</td>
		</tr>
		<tr>
			<td>Orthopedic mallet-hammer</td>
			<td>Mortech</td>
			<td>N/A</td>
			<td>Postmortem hammer with hook</td>
		</tr>
		<tr>
			<td>Sharp councilman orthopedic bone chisel</td>
			<td>Shandon</td>
			<td>60-5</td>
			<td>Councilman&#39;s Chisel Blade: 2 in x 2.25 in standard 7 in</td>
		</tr>
		<tr>
			<td>Sharpened tablespoon or other long handled spoon</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Smooth-tipped tissue dressing forceps without teeth</td>
			<td>Shandon</td>
			<td>63-03</td>
			<td>Shandon Broad Point Dressing Thumb Forceps</td>
		</tr>
		<tr>
			<td>Powder-free non-latex gloves</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Safety glasses, goggles, or faceshield</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Surgery or N-95 mask</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Kraft paper, butcher paper, absorbent pad, etc</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

rabies necropsy techniques in large and small animals

纽约州卫生局(NYSDOH)狂犬病实验室每年接收6,000至9,000份样本,并在整个州进行狂犬病检测,纽约市除外。狂犬病实验室对各种动物进行尸检,从蝙蝠到野猪,不一而分。这些标本大多数是有神经体征的动物,然而,只有不到10%的狂犬病检测呈阳性;暗示创伤、病变或其他传染性病原体是这些症状的原因。由于未诊断的传染性病原体有气溶胶化的风险,狂犬病实验室不使用电动工具或锯子。对于头骨无法用剪刀穿透的动物,将采用三种尸检技术。实验室已实施这些技术,以减少对传染性病原体的潜在接触,消除对标本的不必要操作,并缩短加工时间。与另一种技术相反的首选技术的优点取决于经过培训的个人处理试样。

2019年1月31日至2019年2月28日期间提交的所有头骨陆地样本都有关于颈部存在和收集尸检方法的信息。在此期间,170头被尸检,有18个物种。52%(89/170)被正确斩首。其余的至少有一个椎骨连接,包括三个全身标本。腹腔法使用75%(128/170)的时间,其中,颈部存在49。用颈部提交的标本将在尸体解剖期间取出,以便尽可能采用腹腔方法。提交了三只大型动物(牛、鹿、猪),并在两种情况下使?...

Watch this Scientific Journal Video about Rabies Necropsy Techniques in Large and Small Animals at JoVE.com

Rabies Necropsy Techniques in Large and Small Animals

该协议的目的是在小型和大型动物中展示安全尸体解剖技术,以获得令人满意的组织样本进行狂犬病检测。

大、小动物狂犬病的解剖技术

The New York State Department of Health (NYSDOH) Rabies Laboratory receives between 6,000 to 9,000 specimens annually and performs rabies testing for the ...

狂犬病检测的组织收集具有明显和不明显的危险。技师必须使用最安全的方法，并穿戴适当的个人防护设备。这些协议提供安全的尸检技术，而无需使用电动工具或锯，最大限度地减少了气溶胶的生产。大型动物协议也可以在野外执行。在野外使用大型动物样本检索方法，消除了将重达45公斤的动物头运送到实验室的挑战。大型动物方法类似于欧洲可传播海绵状脑病样本检索方案，可用于使用脑干和小脑进行的任何研究。要通过腹法取回小脑和脑干，请将标本心室侧向上放置，鼻子朝生物安全柜的背面倾斜。持正形锤在主导手和议员凿在另一只手，定位凿子在45度的角度与角指向凿子在V形开口之间的时间骨和腹骨的右侧。用锤子打凿子的顶部，直到两根骨头分开。并在基础骨骼附近做一个切口。如刚刚演示的，将标本左侧的时骨和腹骨分开。并使用凿子向下弯曲头骨的 V 区域，以暴露整个大脑中的菱形脑区域。然后，用剪刀和钳子挖出脑干和小脑，必要时从头骨上取出剩余的组织碎片。如果试样不允许腹体取回位置，则样品的背对背，鼻子朝生物安全柜的背面倾斜。并使用肿瘤十角细胞抓住左时间肌肉与牙脑的牙齿。挤压手柄将牙齿锁在组织周围，用锋利的雕刻刀将时间肌肉切到骨骼。用十纳库拉和刀子旋转标本180度，并在对面的时间肌肉上重复切口，露出头骨。将头部返回其原始方向，将凿子以 45 度角放置，角点到头骨中心处的凿子，在腹骨和腹骨的交汇处。用锤子打凿子的顶部，直到头骨上半部分的骨骼上半部分有一个水平开口。旋转标本180度，并在头骨的对面打开，正如刚刚证明的。根据需要将头部插入第一个切口的末尾，以 90 度角向水平开口插入，然后击打凿子，直到开口到达腹骨。滚动标本，以相同方式在相反的腹骨上做一个开口。头骨的开口应该像一个倒置的U.插入头骨的底部，并拉回骨头，露出脑和小脑的骨端。然后用剪刀作为勺子从腔内撬出整个小脑。并使用组织钳从前人中分出脑干。对于大型动物小脑和脑干检索，放置标本，使头骨的后部分与尸检表面接触。头骨的骨部分和前置巨无霸面对生物安全柜的前面。用钳子抓住脑干。将经过改良的高跟鞋刀插入脊髓和脊髓脑膜之间的前体，并在脊髓周围冲刷，将小脑和脑干与脊髓脑膜分离。然后，用钳子将茎从前人中拉出。当刀通过前脑大物插入时，轻轻角刀沿头骨尽可能跟随，并插入一个金属铲或刀与足够长的手柄，以达到大脑下的神经组织和脊柱脑膜之间的空间。在小脑周围探针，以确保与脊柱脑膜的连接被切断。当脑膜被完全切断了前进的铲子玫瑰花和 dorsally 铲起小脑。然后用铲子挖出小脑。2019 年 1 月 31 日至 2019 年 2 月 28 日之间提交的所有带头骨的地面样本都有关于颈部存在和采集尸检方法的信息。在此期间，170头被尸检，18个物种代表。52% 被正确斩首。其余的至少有一个椎骨附着，包括三个全身标本。75%的时间使用腹体方法。其中，颈部有49个。提交了三个大型动物，在两种情况下，使用了大型动物协议。一只松鼠被提交的一个粉碎的头骨，只是切断皮肤暴露脑组织，所以没有使用任何演示的方法。在处理多种动物时，用于制作显微镜幻灯片和或从分子方法中提取的组织使用新的手套仪器和干净的工作区避免交叉污染。执行这些协议时要小心，因为样本可能感染了狂犬病病毒或其他人畜共和疾病。请注意，样品中也可能存在尖锐物体。

Research

JoVE Journal

Neuroscience

Electrode Fabrication and Implantation in Aplysia californica for Multi-channel Neural and Muscular Recordings in Intact, Freely Behaving Animals

Electrode Fabrication and Implantation in Aplysia californica for Multi-channel Neural and Muscular Recordings in Intact, Freely Behaving Animals

电极的制作及植入海兔californica完整的，自由的行为的动物多通道的神经和肌肉的录音

Recording from key nerves and muscles of Aplysia during feeding behavior allows us to study the patterns of neural control in an intact animal.  ...

科研

神经科学

Efficient Derivation of Human Neuronal Progenitors and Neurons from Pluripotent Human Embryonic Stem Cells with Small Molecule Induction

人类神经祖细胞和神经元小分子诱导人类胚胎干细胞的多能干高效的推导

There is a large unfulfilled need for a clinically-suitable human neuronal cell source for repair or regeneration of the damaged central nervous system ...

Large-scale Recording of Neurons by Movable Silicon Probes in Behaving Rodents

移动硅探针的神经行为鼠害的大型记录

A major challenge in neuroscience is linking behavior to the collective activity of neural assemblies. Understanding of input-output relationships of ...

Design and Assembly of an Ultra-light Motorized Microdrive for Chronic Neural Recordings in Small Animals

设计和组装的超轻型电动微型硬盘（Microdrive）慢性神经录音的小动物

The ability to chronically record from populations of neurons in freely behaving animals has proven an invaluable tool for dissecting the function of ...

Assessing Spatial Learning and Memory in Small Squamate Reptiles

评估空间学习记忆小鳞类爬行动物

Clinical research has leveraged a variety of paradigms to assess cognitive decline, commonly targeting spatial learning and memory abilities. However, ...

High-Resolution 3D Imaging of Rabies Virus Infection in Solvent-Cleared Brain Tissue

溶剂清除脑组织狂犬病病毒感染的高分辨率3D成像

The visualization of infection processes in tissues and organs by immunolabeling is a key method in modern infection biology. The ability to observe and ...

Multi-Fiber Photometry to Record Neural Activity in Freely-Moving Animals

多纤维光测量记录自由移动动物的神经活动

Recording the activity of a group of neurons in a freely-moving animal is a challenging undertaking. Moreover, as the brain is dissected into smaller and ...

Generation of Oligodendrocytes and Oligodendrocyte-Conditioned Medium for Co-Culture Experiments

共培养实验的奥里戈丁细胞和奥里戈丁酸基质的生成

In the central nervous system, oligodendrocytes are well-known for their role in axon myelination, that accelerates the propagation of action potentials ...

Real-Time fMRI Brain Mapping in Animals

动物实时功能磁共振成像大脑映射

Dynamic fMRI responses vary largely according to the physiological conditions of animals either under anesthesia or in awake states. We developed a ...

Subretinal Transplantation of Human Embryonic Stem Cell-Derived Retinal Tissue in a Feline Large Animal Model

猫大型动物模型中人类胚胎干细胞来源的视网膜组织的视网膜下移植

Retinal degenerative (RD) conditions associated with photoreceptor loss such as age-related macular degeneration (AMD), retinitis pigmentosa (RP) and ...