Name: a ligated intestinal loop model in anesthetized specific pathogen free chickens to study clostridium perfringens virulence
Uploaded: 2018-10-11T12:30:00
Description: Watch this Scientific Journal Video about A Ligated Intestinal Loop Model in Anesthetized Specific Pathogen Free Chickens to Study Clostridium Perfringens Virulence at JoVE.com

这项工作得到了蒙特利尔大学兽医学院家禽研究 (m. Boulianne) 主席的支持。Boulianne 和家长开发了模型。Boulianne 和家长进行了手术。伯恩斯博士协助制定了麻醉协议。家长博士编辑了这部影片并写了手稿。Drs 烧伤, Desrochers 和 Boulianne 编辑了手稿。

所有使用活体动物的程序均由蒙特利尔大学兽医学院道德委员会 (CÉUA, éthique ' Animaux ') 授权。
1. 手术前的注意事项
<ol><li>选择10周老特定病原体免费 (SPF) 里窝那鸡进行手术。 注: 重量必须介于1.0 到1.2 公斤之间。</li>
	<li>取出饲料12小时前的程序清空肠道。</li>
	<li>手术前, 按照麻醉师验证的协议将鸡置于全身麻醉下。Premedicate 每只鸡15分钟手术前与肌内 (IM) 注射咪唑安定1?...

<ol>
	<li>Vitale, A., Chiarotti, F., Alleva, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+use+of+animal+models+in+disease+research.">The use of animal models in disease research.</a> Rare diseases and orphan drugs. 2 (1), 1-4 (2015).</li><li>Uzal, F. 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=Animal+models+to+study+the+pathogenesis+of+human+and+animal+Clostridium+perfringens+infections.">Animal models to study the pathogenesis of human and animal Clostridium perfringens infections.</a> Veterinary Microbiology. , (2015).</li><li>Bernier, G., Phaneuf, J. B., Filion, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Necrotic+enteritis+in+broiler+chickens.+III.+Study+of+the+factors+favoring+the+multiplication+of+Clostridium+perfringens+and+the+experimental+transmission+of+the+disease.">Necrotic enteritis in broiler chickens. III. Study of the factors favoring the multiplication of Clostridium perfringens and the experimental transmission of the disease.</a> Canadian Journal of Comparative Medicine. 41 (1), 112-116 (1977).</li><li>Al-Sheikhly, F., Truscott, R. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+pathology+of+necrotic+enteritis+of+chickens+following+infusion+of+broth+cultures+of+Clostridium+perfringens+into+the+duodenum.">The pathology of necrotic enteritis of chickens following infusion of broth cultures of Clostridium perfringens into the duodenum.</a> Avian Diseases. 21 (2), 230-240 (1977).</li><li>Wicker, D. L., Iscrigg, W. N., Trammell, J. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+control+and+prevention+of+necrotic+enteritis+in+broilers+with+zinc+bacitracin.">The control and prevention of necrotic enteritis in broilers with zinc bacitracin.</a> Poultry Science. 56 (4), 1229-1231 (1977).</li><li>Keyburn, A. 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=NetB,+a+new+toxin+that+is+associated+with+avian+necrotic+enteritis+caused+by+Clostridium+perfringens.">NetB, a new toxin that is associated with avian necrotic enteritis caused by Clostridium perfringens.</a> PLoS Pathog. 4 (2), e26 (2008).</li><li>Timbermont, 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=Origin+of+Clostridium+perfringens+isolates+determines+the+ability+to+induce+necrotic+enteritis+in+broilers.">Origin of Clostridium perfringens isolates determines the ability to induce necrotic enteritis in broilers.</a> Comp Immunol Microbiol Infect Dis. 32 (6), 503-512 (2009).</li><li>Paradis, M. 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=Efficacy+of+avilamycin+for+the+prevention+of+necrotic+enteritis+caused+by+a+pathogenic+strain+of+Clostridium+perfringens+in+broiler+chickens.">Efficacy of avilamycin for the prevention of necrotic enteritis caused by a pathogenic strain of Clostridium perfringens in broiler chickens.</a> Avian Pathol. 45 (3), 365-369 (2016).</li><li>Valgaeren, B., 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=Lesion+development+in+a+new+intestinal+loop+model+indicates+the+involvement+of+a+shared+Clostridium+perfringens+virulence+factor+in+haemorrhagic+enteritis+in+calves.">Lesion development in a new intestinal loop model indicates the involvement of a shared Clostridium perfringens virulence factor in haemorrhagic enteritis in calves.</a> Journal of Comparative Pathology. 149 (1), 103-112 (2013).</li><li>Goossens, E., 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+C-terminal+domain+of+Clostridium+perfringens+alpha+toxin+as+a+vaccine+candidate+against+bovine+necrohemorrhagic+enteritis.">The C-terminal domain of Clostridium perfringens alpha toxin as a vaccine candidate against bovine necrohemorrhagic enteritis.</a> Vet Res. 47 (1), 52 (2016).</li><li>Goossens, E., 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=Toxin-neutralizing+antibodies+protect+against+Clostridium+perfringens-induced+necrosis+in+an+intestinal+loop+model+for+bovine+necrohemorrhagic+enteritis.">Toxin-neutralizing antibodies protect against Clostridium perfringens-induced necrosis in an intestinal loop model for bovine necrohemorrhagic enteritis.</a> BMC Vet Res. 12 (1), 101 (2016).</li><li>Parent, E., Archambault, M., Charlebois, A., Bernier-Lachance, J., Boulianne, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+chicken+intestinal+ligated+loop+model+to+study+the+virulence+of+Clostridium+perfringens+isolates+recovered+from+antibiotic-free+chicken+flocks.">A chicken intestinal ligated loop model to study the virulence of Clostridium perfringens isolates recovered from antibiotic-free chicken flocks.</a> Avian Pathol. 46 (2), 138-149 (2017).</li><li>Janvilisri, T., 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=Temporal+differential+proteomes+of+Clostridium+difficile+in+the+pig+ileal-ligated+loop+model.">Temporal differential proteomes of Clostridium difficile in the pig ileal-ligated loop model.</a> PLoS One. 7 (9), e45608 (2012).</li><li>Aabo, 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=Development+of+an+in+vivo+model+for+study+of+intestinal+invasion+by+Salmonella+enterica+in+chickens.">Development of an in vivo model for study of intestinal invasion by Salmonella enterica in chickens.</a> Infection and Immunity. 68 (12), 7122-7125 (2000).</li><li>Meurens, F., 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=Early+immune+response+following+Salmonella+enterica+subspecies+enterica+serovar+Typhimurium+infection+in+porcine+jejunal+gut+loops.">Early immune response following Salmonella enterica subspecies enterica serovar Typhimurium infection in porcine jejunal gut loops.</a> Vet Res. 40 (1), 5 (2009).</li><li>Gerdts, V., 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=Multiple+intestinal+'loops'+provide+an+in+vivo+model+to+analyse+multiple+mucosal+immune+responses.">Multiple intestinal 'loops' provide an in vivo model to analyse multiple mucosal immune responses.</a> J Immunol Methods. 256 (1-2), 19-33 (2001).</li><li>Wade, B., 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+adherent+abilities+of+Clostridium+perfringens+strains+are+critical+for+the+pathogenesis+of+avian+necrotic+enteritis.">The adherent abilities of Clostridium perfringens strains are critical for the pathogenesis of avian necrotic enteritis.</a> Vet Microbiol. 197, 53-61 (2016).</li><li>Cooper, K. 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=Virulence+for+chickens+of+Clostridium+perfringens+isolated+from+poultry+and+other+sources.">Virulence for chickens of Clostridium perfringens isolated from poultry and other sources.</a> Anaerobe. 16 (3), 289-292 (2010).</li><li>Cooper, K. K., Songer, J. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Virulence+of+Clostridium+perfringens+in+an+experimental+model+of+poultry+necrotic+enteritis.">Virulence of Clostridium perfringens in an experimental model of poultry necrotic enteritis.</a> Veterinary Microbiology. 142 (3-4), 323-328 (2010).</li><li>Chalmers, G., Bruce, H. L., Toole, D. L., Barnum, D. A., Boerlin, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Necrotic+enteritis+potential+in+a+model+system+using+Clostridium+perfringens+isolated+from+field+outbreaks.">Necrotic enteritis potential in a model system using Clostridium perfringens isolated from field outbreaks.</a> Avian Diseases. 51 (4), 834-839 (2007).</li></ol>

肠道循环模型已被描述在许多物种, 以研究寄主病原体相互作用和疾病的发病机制, 如梭菌荚, 梭杆菌和沙门氏菌起因7,9,13,14,15。它也被用来分析黏膜免疫反应16, 抗体的效果, 以消除肠道病原体排泄的细菌毒素11和评估疫...

使用动物模型来研究影响人类和动物的传染性疾病, 是评估控制特定疾病发病机制的毒力因素以及制定预防或治疗疾病的策略的中心工具1。尽管使用动物模型来研究各种疾病有许多优点, 但对这种做法的伦理关注却是产生的。研究人员需要尽量减少动物的使用, 同时获得显著和有效的结果。制定了3《 Rs 原则 (替换、减少和改进) 的概念, 以确保在这种审判中处理福利问题。在鸡坏死性肠炎研究中,体内鸡模型用于调查这种情况的病因、预防和治疗2、3、4、5。荚携带特定毒力因子, 如 NetB 毒素6的致病菌株, 被管理导致鸡坏死性肠炎7。因此, 替换原则在这种情况下很难实现, 因为这些毒力因子在其他动物物种中可能不那么重要。大多数鸡坏死性肠炎模型使用的危险因素的组合, 如球虫病和改变的饮食诱导坏死肠炎后, 灌胃的肉汤培养含有大量的C. 荚2.这些模型将导致大量的鸡的疾病, 死亡率可以是重要的在这种试验8, 从而引起关注的减少和细化原则在动物研究。
肠道结扎循环模型是研究肠道病原体引起的疾病的一种可取的替代方法, 以减少其原理和细化。在这些模型中, 被称为 ' 循环 ' 的肠道片段通过在肠道上放置连字来形成独立和密闭的隔间, 在这种情况下, 病原体可以单独注射9或与其他分子, 如疫苗候选10,11. 有兴趣的病原体与肠道细胞密切接触, 经过几个小时的感染时间, 可以恢复肠道样本以作进一步分析。这允许使用在同一动物的多个治疗和控制组。统计分析可以通过重复测量模型进行, 这增加了群体之间的歧视的力量, 减少了必要的鸡的数量相比口服灌胃试验。此外, 手术过程和随后的感染时间在持续全身麻醉和镇痛下进行, 从而尽量减少动物疼痛。闭环结扎是减少主机数量和创建更人性化的动物研究系统的理想模板。
肠道结扎循环模型在不同的物种, 如小牛, 兔子和小鼠2,9, 但在鸡7描述很差的描述。为了最有效地使用这个手术模型, 正确的技术和执行对于建立结扎的肠道循环, 以避免损害肠道完整性是必不可少的。本手稿的目的是描述一步一步的方法在创建在鸡模型的多个肠道循环。这种技术受外科医生的技能和经验的限制, 因为准确的程序对于项目的成功至关重要。

<table>
	<tbody>
		<tr>
			<td>EZ-Scrub 747</td>
			<td>Becton Dickinson and Company</td>
			<td></td>
			<td>4% chlorhexidine gluconate detergent impregnated sterile brush. No catalog number available. Web address: https://www.bd.com/en-us/offerings/capabilities/infection-prevention/surgical-hand-scrubs/ez-scrub-preoperative-surgical-scrub-brushes&#160;</td>
		</tr>
		<tr>
			<td>Isopropyl alcohol 70% USP 4 L</td>
			<td>Commercial isopropyl alcohol</td>
			<td>P016IP70</td>
			<td>Web address: http://www.comalc.com/products/</td>
		</tr>
		<tr>
			<td>Chlorexidine</td>
			<td>Sigma-Aldrich</td>
			<td>282227-1G</td>
			<td>Chlorhexidine gluconate solution, must be diluted to 4%. Web address: http://www.sigmaaldrich.com/catalog/product/aldrich/282227?lang=fr&#38;region=CA&#38;gclid=EAIaIQobChMI8vHr4_OY1wIV3rrACh2ZWQKuEAAYAiAAEgLKx_D_BwE</td>
		</tr>
		<tr>
			<td>Surgical Drape Small (27 inch x 24 inch) with 3 x 6 inch Fenestration</td>
			<td>Veterinary Specialty Products</td>
			<td>32724</td>
			<td>Web address: http://www.vetspecialtyproducts.com/index.cfm?fuseaction=ecommercecatalog.detail&#38;productgroup_id=15</td>
		</tr>
		<tr>
			<td>Scalpel blades #3</td>
			<td>Swann-Morton</td>
			<td>301</td>
			<td>Web address: https://www.swann-morton.com/product/16.php</td>
		</tr>
		<tr>
			<td>Sterile saline NaCl 0.9%</td>
			<td>Sigma-Aldrich</td>
			<td>S8776-100ML</td>
			<td>Web address: http://www.sigmaaldrich.com/catalog/product/sigma/s8776?lang=fr&#38;region=CA</td>
		</tr>
		<tr>
			<td>Vicryl 3-0</td>
			<td>Ethicon</td>
			<td>D9003</td>
			<td>Polyglactin multifilament absorbable suture material. Web address: http://www.ethicon.com/healthcare-professionals/products/wound-closure/absorbable-sutures/coated-vicryl-polyglactin-910-suture</td>
		</tr>
		<tr>
			<td>Syringe 1 ml with 26G needle</td>
			<td>Becton Dickinson and Company</td>
			<td>329652</td>
			<td>Web address: https://www.bd.com/en-us/offerings/capabilities/diabetes-care/insulin-syringes/bd-1-ml-conventional-insulin-syringes</td>
		</tr>
		<tr>
			<td>Brain Heart Infusion</td>
			<td>Sigma-Aldrich</td>
			<td>1104930500</td>
			<td>Web address: http://www.sigmaaldrich.com/catalog/product/mm/110493?lang=fr&#38;region=CA&#38;cm_sp=Insite-_-prodRecCold_xorders-_-prodRecCold2-1</td>
		</tr>
		<tr>
			<td>Eppendorf tube</td>
			<td>Sigma-Aldrich</td>
			<td>T9661-500EA</td>
			<td>Microcentrifuge tube. Web address: http://www.sigmaaldrich.com/catalog/product/sigma/t9661?lang=fr&#38;region=CA&#38;gclid=EAIaIQobChMI-YizzfiY1wIVRkCGCh30SQjuEAAYAiAAEgLK5fD_BwE</td>
		</tr>
		<tr>
			<td>Formalin solution, buffered neutral, 10%</td>
			<td>Sigma-Aldrich</td>
			<td>HT501128-4L</td>
			<td>Ratio tissue : formalin of 1: 10 for adequate fixation. Web address: http://www.sigmaaldrich.com/catalog/product/sigma/ht501128?lang=fr&#38;region=CA</td>
		</tr>
		<tr>
			<td>Clostridium perfringens strains</td>
			<td>Universit&#233; de Montr&#233;al, Chaire en recherche avicole</td>
			<td>N/A</td>
			<td>Specific to each laboratory, available upon request to correspondant author</td>
		</tr>
	</tbody>
</table>

a ligated intestinal loop model in anesthetized specific pathogen free chickens to study clostridium perfringens virulence

采用各种体内感染模型对鸡进行坏死肠炎的研究。大多数这些使用的诱发因素的组合, 如球虫病和饮食, 与灌胃或管理通过饲料使用荚梭菌。在这些模型中, 多荚菌株对毒力研究的比较需要大量的主机来获得显著的结果。在研究过程中, 死亡率可能会很高, 这取决于实验模型, 因此在研究中提高了对动物福利的伦理关注。新的感染模型的开发需要较少的动物来研究发病机制, 但提供统计学上的显著和有效的结果, 在减少动物在研究中的使用是重要的。肠道结扎环模型已被用于研究梭菌感染的各种物种, 如小鼠, 兔和小牛。在手术过程中创建结扎环段, C. 荚菌株直接注入循环中, 在细菌和肠道粘膜之间建立密切接触。小肠和管腔的样本在程序终止后几个小时后采取。可以在每种动物中接种多个细菌菌株, 从而减少实验中所需的受试者数量。同时, 在全身麻醉下进行手术以减少动物疼痛。在鸡中, 这种模式比口服施用更适合荚菌株致病性, 因为需要较少的动物, 不需要诱发因素诱发疾病, 疼痛由止痛药控制。.鸡小肠结扎环模型的描述不充分, 标准化是其最佳使用的必要条件。这份手稿提供了所有必要的步骤, 以创建大量的肠道结扎循环在鸡和带来的关键点信息, 以获得有效的结果。

图 1显示了9肠道循环和 8 interloops 的示意图。在此模型中, 使用简单连字创建总共9个循环和 8 interloops。一个循环由近端和远端连字间隔2厘米测量从近距离结扎。两个相邻的环路由1厘米的 interloop 隔开。为了减少交叉污染循环之间的可能的风险从一个连字的泄漏, interloop 连字被放置在中间的方式从远端结扎和近端结扎2相邻环路。循环的数量可以根据?...

Watch this Scientific Journal Video about A Ligated Intestinal Loop Model in Anesthetized Specific Pathogen Free Chickens to Study Clostridium Perfringens Virulence at JoVE.com

A Ligated Intestinal Loop Model in Anesthetized Specific Pathogen Free Chickens to Study Clostridium Perfringens Virulence

在这里, 我们提出了一个在鸡小肠中手术创建 ' 肠结扎循环 ' 的协议。本程序允许在单个宿主中对多个荚菌株的毒力进行原位比较。这种方法明显地减少了通常需要的鸡的数量在类似的体内实验。

麻醉特异性病原体游离鸡结扎肠环模型研究荚梭菌毒力

Necrotic enteritis was studied in chickens using various in vivo infection models. Most of these use a combination of predisposing factors, such as ...

该方法有助于回答鸡坏死性肠炎研究领域的一些关键问题。如比较引起这种疾病的特定梭菌菌株的毒性。这种技术的主要优点是，它显著减少了通常为类似的比约实验而需要的鸡的数量。虽然这种方法可以提供洞察梭菌的毒性潜力的孢子素菌株，它也可以适用于其他肠道病原体，如梭菌，或沙门氏菌肠。一般来说，由于外科手术需要特定的培训，而只有那些已经获得技术的人才能进行手术，例如在实验前获得技术的技术。麻醉10周大的特异性无病原体无腿角鸡后，设置重要测量的监测器，并开始通过温和的牵引力手动去除腹部的羽毛。接下来，用软刷刷用氯己丁葡萄糖酸盐擦洗手术现场。轻轻将皮肤从中心擦洗到外围五分钟，无需反转路线。然后，使用氯己定葡萄糖酸溶液和使用无菌纱布在手术现场进行三个交替通道。再次，从中心到外围，然后，披上鸡。首先打开手术部位上方的窗帘，然后用三号手术刀做一个L形低中线皮肤切口，开始一厘米拥抱胸骨，最后以一厘米的颅面结束。接下来，继续 L 形切口垂直于第一个切口，从第一个切口的拥抱端开始做第二个切口，然后按照骨盆线继续到腹部左侧五厘米。这个开口将允许容易提取的肠道。接下来，打开腹腔和腹部肌肉使用相同的L形图案进入腹腔。气囊现在将暴露，必须保持湿润使用盐水浸泡纱布。接下来，按照左腹壁和在腹囊周围工作钩来提取肠道，将斯诺克钩插入腹腔。然后，使用干纱布，继续轻轻地外化肠道，以暴露的Jejunum。分散肠道，用盐水频繁喷洒，用盐水浸泡纱布覆盖肠道，保持肠道湿润。取出肠子时保持非常温和很重要，因为过度紧张会破裂肠内血管，这可能导致肠道缺血性病变，并大大损害手术的结果。外化肠道后，在肠道中进行一系列循环，从Jejunum开始。每个循环可用于不同的处理。在近体Jejunum中，用聚葡萄糖胺多丝合成可吸收材料放置一个简单的连结，同时避免主要肠血管的连体。接下来，将远近连字放在离近线连字两厘米远的地方，以完成第一个循环。所有近向和大肠连字应按大约两厘米的空格。每对连字都成一个环，这是一个闭闭段。接下来，在循环之间创建一个间环或一段肠。将一个简单的连字放在第一个环的直结字上半厘米。此连结位于循环间的中间，将减少环路之间可能的交叉污染。现在，继续从环间连字中创建另一个半厘米的循环的模式。在这里，9个环和8个环创建覆盖26厘米的肠道。可以根据需要调整循环数。接下来，继续将梭菌素的菌株注射到循环中。进入环的抗肠侧，使用26测量针在45度角注射0.2毫升的BHI溶液，含有100万菌落形成单位的细菌在中生生阶段。在这种情况下，使用九个回路时，将五个不同的菌株注入交替循环中，其中介入回路单独注入车辆作为负控制。注射病原体后，轻轻将肠道更换到气囊下方腹腔的背侧区域。然后，关闭腹腔。使用聚葡萄糖胺多丝合成可吸收材料，用简单的连续缝合图案缝合骨盆沿岸的腹腔和腹部肌肉。继续用简单的连续缝合线将腹膜和腹部肌肉从胸骨缝合到胸骨。接下来，使用聚葡萄糖胺多丝合成材料和简单的连续图案缝合皮肤。首先，沿着骨盆缝合，然后缝合从胸骨到丁香。关闭切口后，请适当使用镇痛药，将鸡放在全身麻醉下，以尽量减少感染期间的动物疼痛。继续麻醉监测，以确保麻醉水平充足。在所需的感染时间（如本实验中的7小时）后，对鸟实施安乐死并收集肠道组织。使用三号手术刀，切出半到一厘米的环段。然后，将节转移到 10% formalin 进行隔夜固定和进一步组织病理学分析。然后，在同一循环上使用相同的手术刀刀片切出循环的其余部分，并转移该组织到微熔管隔离细菌。每次使用新的手术刀刀片对每个循环重复此过程。注射7小时后，注射控制无菌介质的肠道循环有一个完好无损的粘膜刷边框，无坏死。段肠道层的轻度充血并不罕见。注射病原体7小时后，在尖端周围用坏死的进入部位侵蚀了梭菌，维利尖端被侵蚀。然后坏死物质围绕维利，有大棒状细菌的集群。有时坏死是由于程序执行不力，可能被误解为细菌产生的病变。一个明显的迹象是组织学部分没有细菌。此外，肌肉层的血管因许多退化的红细胞的积累而严重扩张，这些红细胞表明血管充血。这是由连接性肠内血管引起的，这些血管可能连接，对肠道造成过度的身体压力。在外科手术过程中，可以很容易从微观上识别缺血回路。在连结几分钟后严重血管充血后，塞罗萨将是深蓝色，而不是其正常的粉红色红色。看完此视频后，您应该对如何在鸡模型中通过在肠道上放置连字来创建多个肠道循环有一个很好的理解。一旦掌握了外科手术，不包括感染时间，可以在不到三个小时，如果它执行得当。在尝试此程序时，重要的是要记住保持鸡在深度麻醉下，以控制疼痛和监测所有程序中的生命体征，包括手术部分和感染时间。按照这个程序或他们的方法，如细菌分离，和直接识别，如PCR的基因检测，以回答外部问题，如恢复和识别初始注射的病原体。经过这一开发，这项技术为梭菌病原体领域的研究人员探索鸡的坏死性肠炎铺平了道路。不要忘记，与活的动物一起工作可能是极其多变的。执行此程序时，应始终采取预防措施，如对鸡肉进行连续麻醉监测。

Research

JoVE Journal

Medicine

A Ligated Intestinal Loop Model in Anesthetized Specific Pathogen Free Chickens to Study Clostridium Perfringens Virulence

Biomarkers in an Animal Model for Revealing Neural, Hematologic, and Behavioral Correlates of PTSD

在动物模型揭示神经，血液，创伤后应激障碍和行为相关因素的生物标志物

Identification of biomarkers representing the evolution of the pathophysiology of Post Traumatic Stress Disorder (PTSD) is vitally important, not only for ...

科研

医学

Functional Assessment of Intestinal Motility and Gut Wall Inflammation in Rodents: Analyses in a Standardized Model of Intestinal Manipulation

在啮齿类动物的肠动力及胃肠壁的炎症分析功能评价一个标准化的模式肠道操作

Inflammation of the gastrointestinal tract is a common reason for a variety of human diseases. Animal research models are critical in investigating the ...

A Human Ex Vivo Atherosclerotic Plaque Model to Study Lesion Biology

A Human Ex Vivo Atherosclerotic Plaque Model to Study Lesion Biology

一个人力<em&gt;前体内</em&gt;动脉粥样硬化斑块模型研究病变的生物学

Atherosclerosis is a chronic inflammatory disease of the vasculature. There are various methods to study the inflammatory compound in atherosclerotic ...

Facial Nerve Axotomy in Mice: A Model to Study Motoneuron Response to Injury

面神经干切断小鼠：模型，研究运动神经元损伤反应

The goal of this surgical protocol is to expose the facial nerve, which innervates the facial musculature, at its exit from the stylomastoid foramen and ...

Murine Model of Intestinal Ischemia-reperfusion Injury

肠缺血再灌注损伤的小鼠模型

Intestinal ischemia is a life-threatening condition associated with a broad range of clinical conditions including atherosclerosis, thrombosis, ...

Vein Interposition Model: A Suitable Model to Study Bypass Graft Patency

静脉介入型号：研究旁路移植通畅一个合适的模型

Bypass grafting is an established treatment method for coronary artery disease. Graft patency continues to be the Achilles heel of saphenous vein grafts. ...

A Model of Free Tissue Transfer: The Rat Epigastric Free Flap

游离组织移植的模型：鼠上腹部游离皮瓣

Free tissue transfer has been increasingly used in clinical practice since the 1970s, allowing reconstruction of complex and otherwise untreatable defects ...

Intestinal Stem Cell Isolation and Culture in a Porcine Model of Segmental Small Intestinal Ischemia

猪节段性小肠缺血模型中肠道干细胞的分离与培养

Intestinal ischemia remains a major cause of morbidity and mortality in human and veterinary patients. Many disease processes result in intestinal ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

用线 Myography 测定小鼠肠系膜动脉的等轴收缩力

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...

A Cryoinjury Model to Study Myocardial Infarction in the Mouse

研究小鼠心肌梗死的低温损伤模型

The use of animal models is essential for developing new therapeutic strategies for acute coronary syndrome and its complications. In this article, we ...