Name: oral transmission of listeria monocytogenes in mice via ingestion of contaminated food
Uploaded: 2013-05-06T12:00:00
Description: Watch this Scientific Journal Video about Oral Transmission of Listeria monocytogenes in Mice via Ingestion of Contaminated Food at JoVE.com

这项工作是从美国国立卫生研究院（AI079442和AI091918）授予SEFD的补助支持

1。制备选择性琼脂培养基（BHI / L + G），以抑制肠道菌群<ol><li>称取26 g脑心浸液琼脂（Difco公司），7.5克氯化锂和5.0克甘氨酸，地点在1升烧瓶中。加入500毫升去离子水。 </li><li>热火在磁力搅拌器上直到琼脂沸腾，不断搅拌。烧瓶中，关火，让泡沫简要定居，然后再次带回烧开。 </li><li>高压灭菌30分钟，在121°C下16-19磅液体循环。 注意：离开搅拌棒的烧瓶中。 </li><li>平衡媒体55℃的?...

<ol>
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T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Probing+CD8+T+cell+responses+with+Listeria+monocytogenes+infection.">Probing CD8 T cell responses with Listeria monocytogenes infection.</a> Advances in Immunology. 113, 51-80 (2012).</li><li>Disson, O., 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=Modeling+human+listeriosis+in+natural+and+genetically+engineered+animals.">Modeling human listeriosis in natural and genetically engineered animals.</a> Nat Protoc. 4, 799-810 (2009).</li><li>Czuprynski, C. J., Faith, N. 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(Krakow). 50, 1-3 (2002).</li><li>Freitag, N. E., Port, G. C., Miner, M. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Listeria+monocytogenes+-+from+saprophyte+to+intracellular+pathogen.">Listeria monocytogenes - from saprophyte to intracellular pathogen.</a> Nature reviews. Microbiology. 7, 623-628 (2009).</li><li>Camejo, A., 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=The+arsenal+of+virulence+factors+deployed+by+Listeria+monocytogenes+to+promote+its+cell+infection+cycle.">The arsenal of virulence factors deployed by Listeria monocytogenes to promote its cell infection cycle.</a> Virulence. 2, 379-394 (2011).</li></ol>

近交系小鼠喂养一天的时间，统一接受，他们愿意吃了被污染的面包，将取决于应变类型和小鼠17岁。根据我们的经验，6-9周龄B6小鼠是在一天中的任何时间接受喂食，但BALB小鼠不会坚持吃面包片，除非它是在他们的黑暗周期提供。用于安置动物的房间，可以改变光周期，所以黑暗阶段恰逢实验室工作人员正常工作日。然而，应给予老鼠至少两个星期来适应环境的改变的光周期在感染前。...

单核细胞增生李斯特氏菌是兼性细胞内细菌病原体引起的食源性感染人类。细菌耐许多用来保护我们的食物供应，如干燥过程，腌制，或制冷1,2和感染通常与处理，“准备吃”的食物，食用前没有加热。在以往一些爆发的来源L.单核细胞增生李斯特菌污染的食物经鉴定，受限制的组暴露个体密切监测3-6。在这些例子中，其他健康人的变化从一个温和的，自限性胃肠炎的临床疾病，更严重的肠道感染和全身感染，需要住院治疗。L.已与菌感染在免疫系统受损的个人，包括新生儿和老年人，死亡率高（25-30％），即使使用抗生素治疗<s高达7,8。鲜为人知的是，感染剂量L.菌 ，或主机执政后对感染的易感性口头传播的因素，主要是由于缺乏一个小动物模型，概括这种大范围感染的结果。 李斯特菌病模型使用最广泛的是静脉注射（ⅳ）接种小鼠。 IV模型是高度重复性好，一直是非常有用的学习幼稚和记忆T细胞应答过程中感染9,10。静脉模型的缺点是，它完全绕过肠道感染期。食物传播后，肠道粘膜提供一个屏障，它可能减慢和限制的数量的细菌，可以不断地传播到外周组织。相比之下，整个接种可以发现在脾和肝静脉给药后几分钟内，这个大丸有机体可能压倒先天的免疫Ð在这些组织efenses。口腔感染小鼠灌胃不常用，因为通常需要大剂量（10 -10 11 CFU）实现肠道定植10。此外，胃内（ig）可以与馈电针接种不产生可再现的期间全身扩散前的胃肠道感染。 研究报告说，一些实验室单核细胞增生达到脾脏和肝脏感染后4-12小时内（HPI），而其他人没有表现出全身扩散，直到48 HPI 11-15。这个实验室到实验室的变异可能是灌胃接种侵入性的后果，这可能会导致轻微外伤的食道里，并直接促进血液入侵的细菌。 我们最近开发出一种新的小鼠模型口服L.单核细胞增生李斯特菌感染，密切模仿人类疾病的所有阶段16。感染发生时，老鼠摄取件续氨化食品，是一个过程，非创伤性，并不需要专门的技能实验室调查。对于一个离散的一段时间（36-48小时），L。菌只重复殖民胃肠道，从而使致病性李斯特菌所使用的机制，转移整个肠道黏膜和传播外围组织调查。重要的是，该模型可用于研究在感染宿主固有电阻的差异。在先前的研究中，我们发现，通过/ J小鼠BALB / C /极易受到食源性李斯特菌病，指数复制L.发生菌在肠道，脾，肝，胆16。相比之下，C57BL / 6小鼠耐食源性感染，只有短暂的殖民L.发生在每个这些组织中的单核细胞增生李斯特菌 。食源性，密切模仿人类疾病模型的另外一个特点是自然传播到大脑感染的后期阶段（5-7 dpi）的过程中发生。

<table border="1">
	<tbody>
 <tr>
 <td>Name of Reagent/Material</td>
 <td>Company</td>
 <td>Catalog Number</td>
 <td>Comments</td>
 </tr>
 <tr>
 <td>Brain Heart Infusion Agar</td>
 <td>Difco</td>
 <td>BD-241830</td>
 <td></td>
 </tr>
 <tr>
 <td>Lithium chloride</td>
 <td>Sigma</td>
 <td>L9650</td>
 <td ></td>
 </tr>
 <tr>
 <td>Glycine</td>
 <td>Omnipur</td>
 <td>4840</td>
 <td ></td>
 </tr>
 <tr>
 <td>EDTA</td>
 <td>Gibco</td>
 <td>15575-038</td>
 <td ></td>
 </tr>
 <tr>
 <td>DTT</td>
 <td>Sigma</td>
 <td>D5545</td>
 <td ></td>
 </tr>
 <tr>
 <td>Collagenase, type IV</td>
 <td>Worthington</td>
 <td>LS004089</td>
 <td></td>
 </tr>
 <tr>
 <td>DNAse I</td>
 <td>Worthington</td>
 <td>LS002007</td>
 <td></td>
 </tr>
 <tr>
 <td>Diff-Quik</td>
 <td>Dade-Behring</td>
 <td>B4132-1A</td>
 <td></td>
 </tr>
 <tr>
 <td>PowerGen 1000 homogenizer</td>
 <td>Fisher</td>
 <td>14-261-06</td>
 <td></td>
 </tr>
 <tr>
 <td>stainless steel type 304 mesh #80</td>
 <td>Small Parts, Inc.</td>
 <td>CX-0080-C</td>
 <td></td>
 </tr>
 <tr>
 <td>Cytospin</td>
 <td>Statspin</td>
 <td>M801-22</td>
 <td></td>
 </tr>
	 </tbody>
 </table>

oral transmission of listeria monocytogenes in mice via ingestion of contaminated food

单核细胞增生李斯特菌是兼性细胞内细菌病原体引起的食源性感染人类。很少有人知道有关胃肠阶段的李斯特菌病，由于缺乏一个小的，密切模仿人类疾病的动物模型。本文介绍了一种新型的小鼠模型，口头传播研究菌等 。利用这个模型，小鼠灌胃L.单核细胞增生李斯特菌污染的面包有一个独立的阶段，胃肠道感染，易感（BALB / C / J /通过）或抗（C57BL / 6）小鼠品系不同程度的全身蔓延。在感染的后期阶段，胆和脑的传播观察。食源性李斯特菌病模型是高度可重复性，并不需要专门的技能，可以用各种各样的细菌菌株和实验室小鼠品系。因此，它是理想的模型来研究两个致病策略由L.单核细胞增生 </em&gt;的促进肠道定植，以及侵入性食源性细菌感染的宿主反应。

在37°C孵育36-48小时后， 单核细胞增生李斯特菌的菌落BHI / L + G板将是可见的的菌落有一个平滑的，圆顶形的奶白色的外观（ 图1A）。为广大的肠道菌群的生长受到抑制，但它是经常可以看到一些菌落，不L.菌 ，尤其是当镀小肠或结肠的情况下直接显着的稀释（ 图1B）。可疑菌落可以证实电镀在CHROMAGAR TM 李斯特板。L.菌出现蓝色菌落白色...

Watch this Scientific Journal Video about Oral Transmission of Listeria monocytogenes in Mice via Ingestion of Contaminated Food at JoVE.com

Oral Transmission of Listeria monocytogenes in Mice via Ingestion of Contaminated Food

本文介绍了一种新方法，口腔感染的小鼠使用<em&gt;单核细胞增生李斯特氏菌</em&gt;受污染的食物。该协议可以很容易地适应使用其他食源性细菌病原体。

口头传播的<em&gt;单核细胞增生李斯特氏菌</em小鼠通过食入被污染的食物

L. monocytogenes are facultative intracellular bacterial pathogens that cause food borne infections in humans. Very little is known about the ...

Research

JoVE Journal

Immunology and Infection

Oral Transmission of Listeria monocytogenes in Mice via Ingestion of Contaminated Food

Measuring Bacterial Load and Immune Responses in Mice Infected with Listeria monocytogenes

Measuring Bacterial Load and Immune Responses in Mice Infected with Listeria monocytogenes

测量细菌感染小鼠和免疫反应李斯特菌</em

Listeria monocytogenes (Listeria) is a Gram-positive facultative intracellular pathogen1. Mouse studies typically employ intravenous injection of ...

科研

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Imaging InlC Secretion to Investigate Cellular Infection by the Bacterial Pathogen Listeria monocytogenes

Imaging InlC Secretion to Investigate Cellular Infection by the Bacterial Pathogen Listeria monocytogenes

成像InlC的分泌由细菌病原体调查感染蜂窝<em&gt;单核细胞增生李斯特氏菌</em

Bacterial intracellular  pathogens can be conceived as molecular tools to dissect cellular signaling  cascades due to their capacity to exquisitely ...

Assessing Bacterial Invasion of Cardiac Cells in Culture and Heart Colonization in Infected Mice Using Listeria monocytogenes

Assessing Bacterial Invasion of Cardiac Cells in Culture and Heart Colonization in Infected Mice Using Listeria monocytogenes

使用李斯特菌单细胞基因评估受感染小鼠培养和心脏殖民化中心脏细胞的细菌入侵

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Transformation of Probiotic Yeast and Their Recovery from Gastrointestinal Immune Tissues Following Oral Gavage in Mice

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Development of recombinant oral therapy would allow for more direct targeting of the mucosal immune system and improve the ability to combat ...

RNA Purification from Intracellularly Grown Listeria monocytogenes in Macrophage Cells

RNA Purification from Intracellularly Grown Listeria monocytogenes in Macrophage Cells

RNA纯化在细胞内生长<em&gt;李斯特菌</em&gt;巨噬细胞

Analysis of the transcriptome of bacterial pathogens during mammalian infection is a valuable tool for studying genes and factors that mediate infection. ...

Experimental Infection with Listeria monocytogenes as a Model for Studying Host Interferon-&#947; Responses

实验性感染<em&gt;李斯特菌</em&gt;作为研究主机干扰素γ响应模型

L. monocytogenes is a gram-positive bacterium that is a cause of food borne disease in humans. Experimental infection of mice with this pathogen has been ...

An In Vitro Caseum Binding Assay that Predicts Drug Penetration in Tuberculosis Lesions

An In Vitro Caseum Binding Assay that Predicts Drug Penetration in Tuberculosis Lesions

一个<em&gt;体外</em&gt; Caseum结合测定，预测药物在渗透结核病变

The eradication of tuberculosis disease requires drug regimens that can penetrate the multiple layers of complex pulmonary lesions. Drug distribution in ...

Listeria monocytogenes Infection of the Brain

Listeria monocytogenes Infection of the Brain

李斯特菌脑部感染

Listeria monocytogenes is an intracellular bacterial pathogen that is frequently associated with food-borne infection. The ability of&#160;L. ...

Imaging of In Situ Interferon Gamma Production in the Mouse Spleen following Listeria monocytogenes Infection

Imaging of In Situ Interferon Gamma Production in the Mouse Spleen following Listeria monocytogenes Infection

李斯特菌单细胞基因感染后小鼠脾脏原位干扰素伽马生产的成像

Cytokines are small proteins secreted by cells, mediating cell-cell communications that are crucial for effective immune responses. One characteristic of ...

Inducing Meningococcal Meningitis Serogroup C in Mice via Intracisternal Delivery

通过细胞内分娩诱导小鼠脑膜炎球菌性脑膜炎血清群C

Neisseria meningitidis (meningococcus) is a narrow-host-range microorganism, globally recognized as the leading cause of bacterial meningitis. ...