Name: safety precautions and operating procedures in an (a)bsl-4 laboratory: 3. aerobiology
Uploaded: 2016-10-03T11:00:00
Description: Watch this Scientific Journal Video about Safety Precautions and Operating Procedures in an ABSL-4 Laboratory: 3. Aerobiology at JoVE.com

The content of this publication does not necessarily reflect the views or policies of the US Department of Health and Human Services (DHHS) or of the institutions and companies affiliated with the authors. This work was funded in part through Battelle Memorial Institute's prime contract with the US National Institute of Allergy and Infectious Diseases (NIAID) under Contract No. HHSN272200700016I. J.K.B., K.J., M.R.H., D.P., L.B., and J.W. performed this work as employees of Battelle Memorial Institute. Subcontractors to Battelle Memorial Institute who performed this work are: J.H.K., an employee of Tunnell Government Services, Inc.; and M.G.L., an employee of Lovelace Respiratory Research Institute.

该协议符合下列动物护理指南。动物被安置在由该协会的评估和实验动物国际评审认可的设施。所有的实验过程由国家过敏和传染病研究所，临床研究部，动物护理和使用委员会批准，均符合动物福利法的规定，公共卫生服务政策和指南的保养和使用实验动物的建议。 1.大气生物学：动物生物安全4级（ABSL-4）套装实验室<ol><li> 实验室制 <ol><li>完成ABSL-4实...

<ol>
	<li>Alibek, K., Handelman, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The chilling true story of the largest covert biological weapons program in the world-told from inside by the man who ran it. , (1999).</li><li>Roy, C. J., Pitt, L. M., Swearingen, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Infectious+disease+aerobiology:+aerosol+challenge+methods.">Infectious disease aerobiology: aerosol challenge methods.</a> Biodefense: research methodology and animal models. , 61-76 (2006).</li><li>Lackemeyer, M. G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=ABSL-4+aerobiology+biosafety+and+technology+at+the+NIH/NIAID+integrated+research+facility+at+Fort+Detrick.">ABSL-4 aerobiology biosafety and technology at the NIH/NIAID integrated research facility at Fort Detrick.</a> Viruses. 6 (1), 137-150 (2014).</li><li>Bohannon, J. K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Generation+and+characterization+of+large-particle+aerosols+using+a+center+flow+tangential+aerosol+generator+with+a+non-human-primate,+head-only+aerosol+chamber.">Generation and characterization of large-particle aerosols using a center flow tangential aerosol generator with a non-human-primate, head-only aerosol chamber.</a> Inhal Toxicol. , (2015).</li><li>Chosewood, L. C., Wilson, D. E., eds, <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Biosafety in Microbiological and Biomedical Laboratories. , (2009).</li><li>Janosko, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Safety+Precautions+and+Operating+Procedures+in+an+(A)BSL4+Laboratory:+1.+Biosafety+level+4+suit+laboratory+suite+entry+and+exit+procedures.">Safety Precautions and Operating Procedures in an (A)BSL4 Laboratory: 1. Biosafety level 4 suit laboratory suite entry and exit procedures.</a> J Vis Exp. , (2015).</li><li>Mazur, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Safety+Precautions+and+Operating+Procedures+in+an+(A)BSL4+Laboratory:+2.+General+Practices.">Safety Precautions and Operating Procedures in an (A)BSL4 Laboratory: 2. General Practices.</a> J Vis Exp. , (2015).</li><li>Mortola, J. P., Frappell, P. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=On+the+barometric+method+for+measurements+of+ventilation,+and+its+use+in+small+animals.">On the barometric method for measurements of ventilation, and its use in small animals.</a> Can J Physiol Pharmacol. 76 (10-11), 937-944 (1998).</li><li>Zhang, Z., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+a+respiratory+inductive+plethysmography+module+supporting+multiple+sensors+for+wearable+systems.">Development of a respiratory inductive plethysmography module supporting multiple sensors for wearable systems.</a> Sensors (Basel). 12 (10), 13167-13184 (2012).</li><li>Ingram-Ross, J. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cardiorespiratory+safety+evaluation+in+non-human+primates.">Cardiorespiratory safety evaluation in non-human primates.</a> J Pharmacol Toxicol Meth. 66 (2), 114-124 (2012).</li><li>Besch, T. K., Ruble, D. L., Gibbs, P. H., Pitt, M. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Steady-state+minute+volume+determination+by+body-only+plethysmography+in+juvenile+rhesus+monkeys.">Steady-state minute volume determination by body-only plethysmography in juvenile rhesus monkeys.</a> Lab Anim Sci. 46 (5), 539-544 (1996).</li></ol>

我们概述在IRF-弗雷德里克用于高危险（风险组4）病原体的工作程序，大气生物学。可视化生物气溶胶程序的目的之一就是与这些病原体在实验过程中使用的是III类BSC时，为了避免实验室感染强调工作人员的安全。第三类的BSC保持一个向内定向的气流即排出到双HEPA过滤器，以确保病原体包含在实验室（ 图1）内。 作为Ⅲ类BSC是在生物气溶胶研究防止潜在的病原体?...

病毒的传播一般通过直接或身体接触，但许多重要的病毒性疾病（ 如麻疹，水痘，流感）通过由气雾剂或呼吸道飞沫传播的病原体引起的有发生。这样的病原体有可能导致其后果从与工作相关的损失普遍轻微的疾病大流行的可能性（ 如普通感冒），以罕见的高致命性（ 如天花）严重的疾病。后果严重的病原体，通过气溶胶或故意释放烟雾自然传播（生物武器）特别感兴趣大气生物学1。人类可以通过大的呼吸道飞沫或小颗粒核迅速成为感染了一些这些病原体，容易通过唾液分泌物传播这些病原体对他人，咳嗽，打喷嚏2。在美国生物防御社区，后果严重的病原体（ 如丝状病毒或其他NIAID的Category AC优先病原体和CDC生物恐怖代理）是由于相关感染3,4的高致命性的气溶胶研究项目的重点。在空气生物学领域内的科学显著进步已经在过去的十年内提出由于气溶胶设备和高封闭设施5,6的技术进步。研究在美国国立卫生研究院国家过敏和传染病研究所（NIH / NIAID），在位于弗雷德里克，MD，USA（IRF-弗雷德里克）德特里克堡综合研究机构的重点需要动物生物安全后果严重的新兴病原体4级（ABSL-4）遏制。在IRF-弗雷德里克的总体任务是评估和推动候选疫苗和疗法（医疗措施）的发展。 研究与在IRF-弗雷德里克后果严重的病原体是通过严格的生物安全和动物护理和使用要求的约束。这些requir对此语句在微生物和生物医学实验室 （BMBL）手册7和联邦动物福利法规的生物安全概述。这些必要的要求可能会限制可以进行的研究类型，并影响整体的研究设计。正如我们在本杂志如前所述，在ABSL-4环境中进行的所有研究需要特别谨慎，高度专业化的培训，以及强大的和冗余的设备基础设施8,9。 进入IRF-弗雷德里克ABSL-4实验室的西装穿上需要正压密封服8。不需要用于输入ABSL -4-柜实验室正压包封西装。穿上擦洗服，橡胶或丁腈手套，并靠近趾鞋在ABSL-4箱体实验室7认证的III级生物安全柜（BSC）内操作风险组4传染性物质时是合适的。 在IRF-弗雷德里克，气溶胶设备被设计，组装，并保持两个密封，不锈钢，气密，负压III类的BSC， 图1. IRF-弗雷德里克大气生物学核心采用一种自动化气雾剂管理平台（ AAMP）来控制和监视这些生物安全柜中气溶胶的实验， 如图2所示 。先前公布所述的IRF-腓特烈III级生物安全柜，并通过直通端口5连接到西装实验室的特定功能。之前的实验制备III类BSC的方法是特定于在IRF中。在其他机构使用的其他III类生物安全柜的功能类似于III类BSC在IRF的使用，但可能有交通工具，访问或对接不同的机制。 为了进一步了解高后果的病原体如何保持传染性，通过空气传播蔓延，安全AErobiological实验必须在这些III类的BSC根据一个特定的工作流程程序来进行。研究人员已经过仔细而彻底培训，确保此工作流之后在安全和一致的方式。到非人灵长类（NHP）气溶胶攻击，一些气溶胶表征或假气雾剂运行之前被执行，以测试的试剂的气雾剂形式的稳定性和生存能力时。气溶胶表征过程模拟实际的气溶胶挑战，研究人员评估与气溶胶研究相关的变量。 工作流的另一部分是记录于图表为每个NHP物理操作，管理或麻醉剂或其它试剂，或例行程序。这些主题图表深入分析，以确保程序的一致性和标准化。科目气溶胶曝光前麻醉。例如麻醉剂包括替来他明/唑拉西泮，氯胺酮/乙酰丙嗪，和ketami东北。麻醉药是基于减少呼吸抑制和促进控制，稳态呼吸选择。附加麻醉用品保持在动物操作室和输送在与NHP转印车到大气生物学ABSL -4-柜实验室。 内ABSL-4实验室的西装，NHPS通过两种方法（ 即头出体积描记法，呼吸感应体积描记[RIP]）来确定吸气潮气卷一接受体积描记法和呼吸率变化10-12。这些衍生的参数到或气溶胶曝光期间用于估计吸入剂量的病原体的精确计算之前立即。头出体积描记法使用长，圆柱形腔，里面的NHP 13。当动物是在气缸内产生的压力降由一个呼吸速度捕获，中继到放大器，由交流电流/直接柯伦处理T转换器，以及集成到软件来导出上述肺参数。 RIP使用由嵌入在围着人的胸部和腹部11,12松紧带感性盘绕铜线传感器。的感应电容的传感器产生的磁场。呼吸改变磁场，将所得的电压变化是由一个发射机通过短波长的超高频无线电波中继弹性带旁向接收机在计算机中。专用软件决定呼吸频率和总胸排量潮气量。 通过体积描记法得到的分钟量（MV）中的估计吸入剂量（D）的计算被使用。在产生和采样气雾剂，气溶胶浓度（AC）是由媒体（V）的该卷的biosampler浓度（BC）相乘并通过由所述biosampler（佛罗里达州）的流速乘以结果除以计算曝光时间（T）。简化式表示为AC = BC x垂直÷FL点¯xT.反过来，对于在NHPS实际气溶胶攻击时，D由MV和曝光时间（时间= T），乘以交流计算。该简化公式表示为D = AC点¯xMV点¯xT. 本文的目的是为了在视觉上演示使用NHPS从两个观点出发，ABSL -4-西装实验室侧和ABSL -4-柜实验室侧的整个气溶胶攻击过程。虽然这些程序可以针对提到的若干做法是一般性的，它们是专用于在IRF-弗雷德里克大气生物学Core和表示在该机构中使用的实际做法。本文着重讨论安全执行气溶胶的挑战，而不是实际的气溶胶攻击本身所需要的生物安全程序。在这些过程中，我们使用一个虚拟的主题展现生物安全的做法，由于与麻醉的NHP相关的风险。然而，PERFO的过程中最差气溶胶的挑战是写在一般的方式，因为程序是相同的，不管使用的高后果的病原体。我们的目标是提高有关最高封闭条件下进行严重后果的病原体气溶胶研究的严酷科学家的认识和了解。

<table border="0" cellpadding="0" cellspacing="0" width="917">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:340px;">Micro-Chem Plus</td>
			<td style="width:239px;">National Chemical Laboratories</td>
			<td style="width:139px;">255</td>
			<td style="width:200px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Ethanol&#160;</td>
			<td>Fisher&#160;</td>
			<td>BP2818500</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Paraformaldehyde</td>
			<td>Sigma-Aldrich</td>
			<td>441244</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Class III BSC</td>
			<td>Germfree</td>
			<td>DGB-10</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Integrated BSC gloves</td>
			<td>Piercan</td>
			<td>10UY2032-9</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Aerosol Management Platform (AeroMP)</td>
			<td>Biaera Technologies</td>
			<td>NA</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Head-out plethysmography</td>
			<td>Buxco/Data Sciences International</td>
			<td>NA</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Respriatory inductive plethysmography</td>
			<td>Data Sciences International</td>
			<td>NA</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Centered flow tangential aerosol generator (CenTAG)</td>
			<td>CH Technologies</td>
			<td>NA</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Collison nebulizer</td>
			<td>BGI Inc.&#160;</td>
			<td>CN25</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Autoclave</td>
			<td>Getinge</td>
			<td>GEB 2404 AMB-2</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sperian positive-pressure suit</td>
			<td>Honeywell Safety Products</td>
			<td>BSL 4-2</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Outer suit gloves (latex, Ansell Canners and Handlers)</td>
			<td>Fisher</td>
			<td style="width:139px;">19-019-601</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Outer suit gloves (nitrile/rubber, MAPA)</td>
			<td>Fisher</td>
			<td>2MYU1</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Scrubs</td>
			<td>Cintas</td>
			<td>60975/60976</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Socks</td>
			<td>Cintas</td>
			<td>944</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Duct tape</td>
			<td>Pack-N-Tape</td>
			<td>51131069695</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Towels</td>
			<td>Cintas</td>
			<td>2720</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">O-rings</td>
			<td>O-ring warehouse</td>
			<td>AS568-343</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Overshoes</td>
			<td>Amazon</td>
			<td>B0034KZE22</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Zip lube</td>
			<td>Amazon</td>
			<td>B000GKBEJA</td>
			<td></td>
		</tr>
	</tbody>
</table>

safety precautions and operating procedures in an (a)bsl-4 laboratory: 3. aerobiology

Aerosol or inhalational studies of high-consequence pathogens have recently been increasing in number due to the perceived threat of intentional aerosol releases or unexpected natural aerosol transmission. Specific laboratories designed to perform these experiments require tremendous engineering controls to provide a safe and secure working environment and constant systems maintenance to sustain functionality. Class III biosafety cabinets, also referred to as gloveboxes, are gas-tight enclosures with non-opening windows. These cabinets are maintained under negative pressure by double high-efficiency-particulate-air (HEPA)-filtered exhaust systems and are the ideal primary containment for housing aerosolization equipment. A well planned workflow between staff members within high containment from, for instance, an animal biosafety level-4 (ABSL-4) suit laboratory to the ABSL-4 cabinet laboratory is a crucial component for successful experimentation. For smooth study execution, establishing a communication network, moving equipment and subjects, and setting up and placing equipment, requires staff members to meticulously plan procedures prior to study initiation. Here, we provide an overview and a visual representation of how aerobiology research is conducted at the National Institutes of Health, National Institute of Allergy and Infectious Diseases Integrated Research Facility at Fort Detrick, Maryland, USA, within an ABSL-4 environment.

所述III类生物安全柜（BSC）为含有ABSL -4-柜实验室内负压的ABSL-4环境密封的不锈钢箱体（ 图1）。材料可以被引入到由工作人员通过一个橱柜下式不锈钢罐中的ABSL -4-柜实验室工作在BSC（通常被称为在ABSL-4或BSL-4设置的“扣篮罐”）含有5 ％的双季铵盐（正烷基二甲基苄基氯化铵，正 - 烷基二甲基乙基苄基氯化铵）消毒剂溶液。因为在BSC内置于壁分离从ABSL -4-西?...

Watch this Scientific Journal Video about Safety Precautions and Operating Procedures in an ABSL-4 Laboratory: 3. Aerobiology at JoVE.com

Safety Precautions and Operating Procedures in an ABSL-4 Laboratory: 3. Aerobiology

As high-consequence pathogens can potentially infect subjects through airborne particles, aerobiology has been increasingly applied in pathogenesis research and medical countermeasure development. We present a detailed visual demonstration of aerobiology procedures during an aerosol challenge in nonhuman primates in an animal biosafety level 4 maximum containment environment.

安全注意事项和操作步骤在（A）BSL-4实验室：3，大气生物学

Aerosol or inhalational studies of high-consequence pathogens have recently been increasing in number due to the perceived threat of intentional aerosol ...

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Safety Precautions and Operating Procedures in an (A)BSL-4 Laboratory: 3. Aerobiology

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This protocol describes a simple approach for adhesive-tape-based sampling of tomato and other fresh produce surfaces, followed by on-tape fluorescence in ...

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We present the video about assisting anti-retroviral therapy (ART) by an apt laboratory service - representing a South-African role model for economical ...

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建立的液体覆盖的文化偏光人类呼吸道上皮的CALU-3细胞研究呼吸系统病原宿主细胞的反应<em在体外</em

The  apical and basolateral surfaces of airway epithelial cells demonstrate  directional responses to pathogen exposure in  vivo. Thus, ideal in vitro ...

Production and Purification of Non Replicative Canine Adenovirus Type 2 Derived Vectors

生产的非复制型犬2型腺病毒衍生载体与纯化

Adenovirus (Ad) derived vectors have been widely used for short or long-term gene transfer, both for gene therapy and vaccine applications. Because of the ...

Procedures for Identifying Infectious Prions After Passage Through the Digestive System of an Avian Species

用于识别传染性朊病毒通过后通过一个鸟类的消化系统程序

Infectious prion (PrPRes) material is likely the cause of fatal, neurodegenerative transmissible spongiform encephalopathy (TSE) diseases1. Transmission ...

A Method for Generating Pulmonary Neutrophilia Using Aerosolized Lipopolysaccharide

采用雾化脂多糖一种产生肺中性粒细胞增多

Acute lung injury (ALI) is a severe disease characterized by alveolar neutrophilia, with limited treatment options and high mortality. Experimental models ...

Safety Precautions and Operating Procedures in an (A)BSL-4 Laboratory: 1. Biosafety Level 4 Suit Laboratory Suite Entry and Exit Procedures

Safety Precautions and Operating Procedures in an ABSL-4 Laboratory: 1. Biosafety Level 4 Suit Laboratory Suite Entry and Exit Procedures

安全注意事项和操作步骤在（A）BSL-4实验室：1.生物安全4级实验室西服套房出入境手续

Biosafety level 4 (BSL-4) suit laboratories are specifically designed to study high-consequence pathogens for which neither infection prophylaxes nor ...

Safety Precautions and Operating Procedures in an (A)BSL-4 Laboratory: 2. General Practices

Safety Precautions and Operating Procedures in an ABSL-4 Laboratory: 2. General Practices

安全注意事项和操作步骤在（A）BSL-4实验室：2.常规做法

Work in a biosafety level 4 (BSL-4) containment laboratory requires time and great attention to detail. The same work that is done in a BSL-2 laboratory ...

Safety Precautions and Operating Procedures in an (A)BSL-4 Laboratory: 4. Medical Imaging Procedures

Safety Precautions and Operating Procedures in an ABSL-4 Laboratory: 4. Medical Imaging Procedures

安全注意事项和操作步骤在（A）BSL-4实验室：4.医疗成像过程

Medical imaging using animal models for human diseases has been utilized for decades; however, until recently, medical imaging of diseases induced by ...

Establishment of a High-throughput Setup for Screening Small Molecules That Modulate c-di-GMP Signaling in Pseudomonas aeruginosa

Establishment of a High-throughput Setup for Screening Small Molecules That Modulate c-di-GMP Signaling in Pseudomonas aeruginosa

高通量设置的建立筛选小分子能调节C-二GMP信令<em&gt;绿脓杆菌</em

Bacterial resistance to traditional antibiotics has driven research attempts to identify new drug targets in recently discovered regulatory pathways. ...