The authors would like to acknowledge the Infectious Disease Department at Brigham and Women&#8217;s Hospital especially Michael S. Calderwood, MD, MPH for helping us construct this protocol. We would also like to thank the Institution&#8217;s endoscopy center and it&#8217;s leadership team for their support.

注:在此稿件发布时的研究性新药（IND）申请不需要执行FMT在临床实践中经常CDI。但是，如果FMTS正在执行作为研究的一部分，或另一个迹象，一个IND或等效的应用程序可能需要18。监管机构可能会继续评估FMT的安全性和有效性，适用于CDI和其他医疗条件等，建议检查在各自的位置上监管要求的FMT程序的启动和每一至两个月一次前积极履行FMT。 注:考生必须愿意同意粪便移植和结肠镜检查。 1.确定合适的候选人FMT的<ol><li>患者谁都有复发的CDI或难治性疾病，没有响应传统抗生素都是适宜Ë候选人FMT。 注:复发CDI定义为CDI的三个或更多个情节由毒素检测或PCR，或两集即需要住院证实的历史。 </li><li>确保患者能够安全地接受结肠镜检查。这是在内镜的自由裁量权。 </li></ol> 2.粪便捐赠者的选择注:凳子施主选择适用于CDI的治疗，可能并不适用于其他疾病的治疗。 <ol><li>确保病人标识捐助。潜在的捐助者应该存在与收件人在初始消化内科就诊的时间。捐助者可能是配偶，家人或朋友。捐助者可以住在同一住所收件人只要其他条件都满足。 </li></ol> 3.捐助资格<ol><li>纳入标准:确保捐赠是一个健康的成年人（≥18岁），并提供每日排便形成米ovement。 </li><li>排除标准: <ol><li>确保捐助者未服用抗生素或没有采取抗生素的前3个月服用药物以减低感染。 </li><li>确保捐助者没有便秘或使用泻药史长期。此外，保证了供体不具有炎性肠病，慢性腹泻，显微镜性结肠炎，肠/结肠恶性肿瘤，或胃旁路/其他减肥手术史的个人历史的历史。 </li><li>确保捐助最近没有前往（最近三个月内）到一个领域，感染性腹泻是一个问题。如果他们旅行，出现腹泻应已排除。 </li><li>确保捐助者没有采取任何免疫抑制剂，是不是全身化疗。 </li><li>确保施主不具有代谢综合征，自体免疫疾病或过敏性疾病。 </li></ol></li><li>获得书面CON从筛查给出的筛查试验的敏感性之前，供体和受体发送。安抚双方测试结果将不会与对方分享。 </li></ol> 4.筛选注:此筛选协议被改编为患者布莱根妇女医院在马萨诸塞州，美国考虑咨询传染病部门和医院感染控制启动FMT协议，以确保一切适当的筛选试验被认为之前。 <ol><li>收件人: <ol><li>在最初就诊时，发送以下实验室检查收件人筛选（建立基准测试，这些感染性疾病，并确保没有在未来毫无疑问传输）。 </li><li>进行A型肝炎抗体（IgG和IgM），乙型肝炎（乙肝表面抗原/抗体和抗 - HBc），丙型肝炎抗体，HIV-1/2（Ab和病毒载量）和梅毒（TP-IgG）的实验室外周血TEST序列。 </li><li>获得食物过敏的彻底的历史，在这个初步评估。 </li></ol></li><li>捐赠: <ol><li>筛选，以确保成形粪便不丢弃之前与微生物实验室坐标。 </li><li>订购以下实验室检查献血者筛查，如果捐赠者符合资格按照第3条 。 </li><li>进行A型肝炎抗体（IgG和IgM），乙型肝炎（乙肝表面抗原/抗体和HBcAb阳性），丙型肝炎抗体，HIV-1/2（Ab和病毒载量）和梅毒（TP-IgG）的实验室验血。 </li><li>执行艰难梭菌 （由文化），常规粪便培养，贾第虫抗原，抗原隐孢子虫和虫卵和寄生虫实验室大便化验。 </li><li>屏幕捐助不超过30天，但最好是1 - 预定程序之前2周。 注意:如果捐赠者开发腹泻或大便液体他们不再符合捐赠。 </li></ol></li><li>律师对食品捐助，以避免基于收件人的食物过敏。捐赠者不应该摄取食物，收件人是过敏性的捐赠前的5天。 </li></ol> 5.预过程制备<ol><li>收件人: <ol><li>在最初就诊，如果收款人是不是已经对口腔万古霉素，万古霉素开始125毫克，每6小时以前发生的过程，至少七到十天。 </li><li>为受援人提供的内​​镜中心的指令前结肠镜检查肠道准备，以预定的程序前一天完成。收件人也应遵循内镜中心的标准膳食指令前程序的一周。 </li><li>指示接收者采取步骤之前，万古霉素的夜晚他们的最后一剂。 </li></ol></li><li>捐赠: <ol><li>提供多种粪便收集杯的捐助。标签马桶插入（帽）和杯子与收件人的标签，以及3个空SPecimen容器。向供体以开始粪便采集不超过6小时之前，处理（过程时间）。 注意:如果捐赠者提供了多种粪便样本，应标有日期和时间。使用最新鲜的样本（一个充满试样杯就足够了）。 </li><li>确保收件人带来了捐赠者的粪便与他们的程序。 </li><li>储存粪便试样在室温下。排除任何未成形大便。丢弃在下水道系统中任何未使用的凳子（除非储蓄用于研究目的）。 </li></ol></li></ol> 6.大便准备注:请确保此过程进行了讨论，并经医院感染控制部门。 <ol><li>准备大便时穿戴个人防护装备。 </li><li>大便准备在内镜室在指定的工作区域，其中有一个18英寸的飞溅区，防止了超视距污染呃材料。 </li><li>此前大便准备，清洁与医院批准消毒剂的指定区域。 </li><li>用一次性衬垫覆盖的区域，以避免污染。 </li><li>转移形成大便成14盎司单速搅拌器，加入500毫升的生理盐水。 </li><li>混合的内容为至少一分钟或直到它到达一个液体浆液的一致性。小心取出搅拌机的盖子时，以避免溢出。 </li><li>倒入经过过滤的溶液倒入一个塑料8夸脱盆地以消除任何固体废物。丢弃的过滤器。 </li><li>拟定紧张溶液倒入60毫升鲁尔锁注射器（约八九个月的标准FMT通过结肠镜检查）。制订一个额外的60毫升生理盐水冲洗的。 </li></ol>通过结肠镜检查7.粪便移植<ol><li>穿戴个人防护装备按照内窥镜设备的政策在执行或协助与infusioñ。 </li><li>考虑光镇静，如果可能的话，使病人可以是能够与在手术后保留的粪便进行合作。 </li><li>进行结肠镜检查，以回肠末端（或尽可能技术上可行）。上插入colonscope，灌洗和抽吸的任何残余液体粪便。 </li><li>检查粘膜壁的炎症，但粘膜活检的证据也没有惯常的FMT过程中进行的。 注:确定一个完整结肠镜检查是否是安全的，由于炎症的程度。如果没有，考虑更有限的乙状结肠镜输液（使用一半的大便浆）。 </li><li>通过使用具有延伸管活检通道帽结肠镜的活组织检查通道注入60立方厘米（一个针筒）一旦在回肠末端。 </li><li>大便注射器的应用程序之间，冲洗通道用生理盐水，避免堵塞。 </li><li>注入另外60毫升的粪便进入盲肠，然后重复正确的ç敖。 </li><li>继续注入30毫升每五to10厘米而均匀地分布在泥浆上的所有四个结肠壁象限直到中间横结肠为止。记得在大便注射器之间用生理盐水冲洗。 注意:如果该浆料是非常粘稠的，可能有必要推与注射器相当大的力。要小心，活检通道帽也不会从结肠镜分开并引起的粪便内容溢出。频繁冲洗将允许浆料更容易通过。如果存在持续的困难在按压浆料，考虑重新拉紧剩余施主粪便用于第二时间或用生理盐水进一步稀释。 </li><li>停药后更换针筒的时候，注意不要抽吸任何液体粪便吸出多余的空气。 注:如果患者已经知道大便失禁，最后注射器应注入不低于肝曲，以避免在泄漏后的程序。 </li><li>第一次Ë结肠镜检查与FMT完成后，建议患者保持大便只要他们能。他们可能恢复他们的正常饮食和药物治疗两小时后，结肠镜检查完成。 </li></ol> 8.结肠镜检查与FMT清理<ol><li>执行程序后的再加工设备，环境的清洁/消毒和废物处理按照医院的协议。 </li><li>在适当的生物危害容器处理所有粪便材料。 </li><li>把结肠镜的净化面积每标准协议常规处理。 </li><li>处置搅拌机和过滤与任何其他脏浪费。 注意:如果单用搅拌机和过滤器是不可行的，在一个单独的机构替代混合方法可以考虑。一个例子:使粪便通过不锈钢马铃薯ricer的，可以在高压釜中进行清洗。我们建议讨论与您设施的感染控制DEP任何潜在的方法artment。 </li><li>净化被用于使用该设施批准的产品的方法，所有的表面。 </li></ol> 9.跟进<ol><li>安排随访预约消化内科门诊的病人FMT后约30天。 </li><li>律师关于难辨梭状芽孢杆菌复发与抗生素的未来课程风险的患者。 </li></ol>不良事件报告10 <ol><li>虽然FMT被认为是一个安全的过程，强烈建议保持跟踪所有的不良事件认为将与过程相关联，并且将它们报告给适当的管理板。 </li><li>为了记录格式化的长期安全性，保留该可定期查询一个可靠的数据库。 </li></ol>

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The authors have nothing to disclose relevant to the material presented in this manuscript.

粪便移植已被证明是一种安全有效的治疗复发性CDI。我们提出的标准协议，通过 FMT结肠镜检查，其中包括我们的研究结果为22例患者谁接受的程序反复CDI。我们的研究结果迄今媲美的病情反复发作公布的数据显示，只有一名患者复发后一个月FMT。该过程的一个重要步骤是将预筛选访问，以确保供体是合适的，并且该患者不具有禁忌接受结肠镜检查。筛查一般需要三到四天才能完成的过程可?...

粪便移植的pseumembranous结肠炎的治疗于1958年首次描述了1，但摄入粪便食物中毒和严重腹泻日期的治疗回来，早在四世纪中国2的报道。粪便菌群移植（FMT）也被称为在文献"粪bacteriotherapy"，"人类的益生菌输液"，"粪便移植"，"肠菌群恢复"和"粪便转移"。它包括来自健康，预筛选供体采集粪 ​​便和经由鼻胃管，esophagogastricduodenoscopy，结肠镜检查，或灌肠剂3递送准备好的浆体到受体的胃肠道。很多情况下，系列报道，通过NGT和灌肠给药的药效，但是最近的研究表明，捐助粪便的管理，通过结肠镜检查是安全，高效4。而其它的方法，如鼻胃管给药已经报道，通过结肠镜检查输送给体大便具有较少即时副作用（ 例如腹胀和恶心）4和我们已发现，患者更接受FMT的概念时，它是通过执行结肠镜检查。一个FMT 通过结肠镜检查的治疗优点是在结肠生物量的减少是由于被该过程4之前进行肠灌洗。 多项研究已经调查了FMT的作用，用于治疗结肠炎和腹泻导致的机会性病原体C.难辨 （CDI）。 CDI的发病率继续上升，并且是发病和死亡的一个主要原因与在世界各地的大的经济负担。一线治疗CDI包括抗生素治疗，但复发率已经报道的15-35％5。一些病例和报告都做cumented FMT的CDI难治抗生素4,6-16标准的医疗治疗的安全性和有效性。一项研究观察长期随访的患者FMT后经肠镜检查的CDI报告了91％，主要的治愈率在77例17。 在我们中心（布里格姆与消化内科，肝病及妇女医院内镜科），我们发起的患者复发或难治性CDI粪便移植方案。合适的候选人被定义为谁有反复CDI（3个或更多的情节，或2次发作史是需要住院），或难治性疾病，没有响应传统抗生素的病人患者。我们相信，一个系统的方法来此过程的所有阶段最大限度地提高疗效。在这个手稿和影随行，我们详细介绍我们使用了我们的中心，其中包括患者和供者的筛选，大便准备了协议，和大便交付吨结肠镜检查的时间。这种方法已经产生了相媲美的已发表的文献阳性结果。 病例报告: 这名患者代表提到了粪便移植一个典型的病人。 该患者是一名69岁的女子与慢性淋巴细胞白血病病史谁曾病需要化疗进一步治疗的复发。她患有CDI的三集，在过去的一年中，并因此被称为我们的诊所审议FMT之前重新开始化疗。发生CDI她的第一集于2012年10月，她没有上过前面的抗生素。她病得很重，在ICU脓毒性休克提出后进行分流循环回肠造口顺行灌肠万古霉素为6周的课程，结合给她病情的严重程度万古霉素口服。她的表现非常好，她腹泻分辨率和她能来关闭抗生素。她开发了她周围的造口疝因此它被推翻。不久后造瘘逆转，她再次出现腹泻，被发现再次是C.艰难梭菌毒素阳性。她完成了万古霉素口服另外6周的课程，并能够它逐渐停止成功。然而几个月后，她又开始腹泻，这是C.艰难梭菌毒素阳性，并开始在她的第三万古霉素口服疗程。她对抗生素表现不错时，有人指出，她的白细胞计数增加到&gt; 100，她发现有复发她的骨髓活检CLL的。她的肿瘤学家担心化疗开始没有她CDI完全解决，所以我们被要求进行格式化。

<table><tbody><tr><td>Blender</td><td>Hamilton Beach</td><td>51101</td><td>&lt;a href=&quot;http://www.amazon.com/Hamilton-Beach-51101-Personal-Blender/dp/B00065L6CU/ref=pd_sim_k_2?ie=UTF8&amp;amp;refRID=1Q06MXAE9XKTB4QG0AEN&quot; target=&quot;_blank&quot;&gt;Link to Amazon.com&lt;/a&gt;</td></tr><tr><td>Strainer</td><td>Winco</td><td>MS3A-8S</td><td>&lt;a href=&quot;http://www.amazon.com/Winco-MS3A-8S-Strainer-Single-Wooden/dp/B001VZ5E40/ref=sr_1_3?s=home-garden&amp;amp;ie=UTF8&amp;amp;qid=1395001295&amp;amp;sr=1-3&amp;amp;keywords=kitchen+strainer+wooden+handle&quot; target=&quot;_blank&quot;&gt;Link to Amazon.com&lt;/a&gt;</td></tr><tr><td>Disposable pads (chux), 17 x 24 in.</td><td>hospital standard</td><td></td><td>one package</td></tr><tr><td>Pink rectangular hospital standard wash basin, 8 quart</td><td>hospital standard</td><td></td><td>1 or 2</td></tr><tr><td>500 cc bottle of sterile normal saline</td><td>hospital standard</td><td></td><td>2</td></tr><tr><td>60 cc luer lock syringes</td><td>hospital standard</td><td></td><td>8 or 9</td></tr><tr><td>Biopsy channel cap with extension</td><td>hospital standard</td><td></td><td></td></tr></tbody></table>

fecal microbiota transplantation via colonoscopy for recurrent c. difficile infection

粪便菌群移植（FMT）是一种安全，高效的治疗复发和难治C.难辨感染（CDI）。 FMT管理的各种方法已经有文献报道，包括鼻胃管，上消化道内镜检查，灌肠和结肠镜检查。 通过肠镜产量出色的治愈率FMT，也很好的耐受性。我们已发现，患者发现这是一个可以接受的和可耐受的递送模式。在我们中心，我们已经开始为患者复发或难治性CDI粪便移植方案。我们已经制定了使用协议修订的迭代过程，并在22例成功率堪比目前公开发表的文献进行24粪便移植。系统的方法对病人和捐赠者的筛选，准备大便，和交付的凳子最大限度地提高治疗成功。下面我们详细的FMT协议，可以进行任何胃镜的每一步中心与安全和成功的程度高。

我们已经完成对22例24 FMTS在我们使用上述协议中心（ 表1）。 9例患者并发IBD（6克罗恩病和三个溃疡性结肠炎）。所有9例CDI的分辨率。其中一名患者与克罗恩病是状态发布全结肠切除克罗恩结肠炎。他最初不过在经历了复发后两周FMT他通过导致CDI分辨率上消化道内镜检查进行了第二次FMT通过结肠镜检查进行格式化。十一出的非IBD患者的13具有相关的为C.症状完全分辨率艰难</e...

Watch this Scientific Journal Video about Fecal Microbiota Transplantation via Colonoscopy for Recurrent C. difficile Infection at JoVE.com

Fecal Microbiota Transplantation via Colonoscopy for Recurrent C. difficile Infection

通过结肠镜检查粪便菌群移植是一种安全，有效的治疗复发和难治艰难梭菌感染。结肠镜检查过程中的系统方法对病人和捐赠者的筛选，准备大便，和交付的凳子将最大限度地提高治疗成功。

通过结肠镜检查粪便菌群移植治疗复发性<em&gt;温度。难辨</em&gt;感染

Fecal Microbiota Transplantation (FMT) is a safe and highly effective treatment for recurrent and refractory C. difficile infection (CDI). Various methods ...

fecal-microbiota-transplantation-via-colonoscopy-for-recurrent-c

Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

Fecal Microbiota Transplantation via Colonoscopy for Recurrent C. difficile Infection

27.0K 次观看.  Brigham and Women‘s Hospital.  通过结肠镜检查粪便菌群移植是一种安全，有效的治疗复发和难治艰难梭菌感染。结肠镜检查过程中的系统方法对病人和捐赠者的筛选，准备大便，和交付的凳子将最大限度地提高治疗成功。 

视频: Fecal Microbiota Transplantation via Colonoscopy for Recurrent C. difficile Infection

A Genetically Engineered Mouse Model of Sporadic Colorectal Cancer

Despite the advantages of easy applicability and cost-effectiveness, colorectal cancer mouse models based on tumor cell injection have severe limitations and do not accurately simulate tumor biology and tumor cell dissemination. Genetically engineered mouse models have been introduced to overcome these limitations; however, such models are technically demanding, especially in large organs such as the colon in which only a single tumor is desired.
As a result, an immunocompetent, genetically engineered mouse model of colorectal cancer was developed which develops highly uniform tumors and can be used for tumor biology studies as well as therapeutic trials. Tumor development is initiated by surgical, segmental infection of the distal colon with adeno-cre virus in compound conditionally mutant mice. The tumors can be easily detected and monitored via colonoscopy. We here describe the surgical technique of segmental adeno-cre infection of the colon, the surveillance of the tumor via high-resolution colonoscopy and present the resulting colorectal tumors.

A protocol for the establishment of a genetically engineered mouse model of colorectal cancer by segmental adeno-cre infection and its surveillance via high-resolution colonoscopy is presented.

遗传工程小鼠模型的散发性结肠直肠癌

Despite the advantages of easy applicability and cost-effectiveness, colorectal cancer mouse models based on tumor cell injection have severe limitations ...

科研

癌症研究

Culturing and Maintaining Clostridium difficile in an Anaerobic Environment

Clostridium difficile is a Gram-positive, anaerobic, sporogenic bacterium that is primarily responsible for antibiotic associated diarrhea (AAD) and is a significant nosocomial pathogen. C. difficile is notoriously difficult to isolate and cultivate and is extremely sensitive to even low levels of oxygen in the environment. Here, methods for isolating C. difficile from fecal samples and subsequently culturing C. difficile for preparation of glycerol stocks for long-term storage are presented. Techniques for preparing and enumerating spore stocks in the laboratory for a variety of downstream applications including microscopy and animal studies are also described. These techniques necessitate an anaerobic chamber, which maintains a consistent anaerobic environment to ensure proper conditions for optimal C. difficile growth. We provide protocols for transferring materials in and out of the chamber without causing significant oxygen contamination along with suggestions for regular maintenance required to sustain the appropriate anaerobic environment for efficient and consistent C. difficile cultivation.

Culturing and Maintaining Clostridium difficile in an Anaerobic Environment

Clostridium difficile is a pathogenic bacterium that is a strict anaerobe and causes antibiotic associated diarrhea (AAD). Here, methods for isolating, culturing and maintaining C. difficile vegetative cells and spores are described. These techniques necessitate an anaerobic chamber, which requires regular maintenance to ensure proper conditions for optimal C. difficile cultivation.

培养和维护<em&gt;难辨梭状芽孢杆菌</em&gt;在厌氧环境

Clostridium  difficile is a Gram-positive, anaerobic, sporogenic bacterium  that is primarily responsible for antibiotic associated diarrhea (AAD) and is ...

免疫与感染

Murine Ileocolic Bowel Resection with Primary Anastomosis

Intestinal resections are frequently required for treatment of diseases involving the gastrointestinal tract, with Crohn&#8217;s disease and colon cancer being two common examples. Despite the frequency of these procedures, a significant knowledge gap remains in describing the inherent effects of intestinal resection on host physiology and disease pathophysiology. This article provides detailed instructions for an ileocolic resection with primary end-to-end anastomosis in mice, as well as essential aspects of peri-operative care to maximize post-operative success. When followed closely, this procedure yields a 95% long-term survival rate, no failure to thrive, and minimizes post-operative complications of bowel obstruction and anastomotic leak. The technical challenges of performing the procedure in mice are a barrier to its wide spread use in research. The skills described in this article can be acquired without previous surgical experience. Once mastered, the murine ileocolic resection procedure will provide a reproducible tool for studying the effects of intestinal resection in models of human disease.

Ileocolic resection is commonly performed in several human diseases; however, little is known regarding the impact of intestinal resection on surgical illnesses. This article provides instruction on executing the procedure in mice with high success, providing a means to study the effects of ileocolic resection in models of disease.

鼠回结肠肠管切除原发性吻合

Intestinal resections are frequently required for treatment of diseases involving the gastrointestinal tract, with Crohn&#8217;s disease and colon cancer ...

医学

Cefoperazone-treated Mouse Model of Clinically-relevant Clostridium difficile Strain R20291

Clostridium difficile is an anaerobic, gram-positive, spore-forming enteric pathogen that is associated with increasing morbidity and mortality and consequently poses an urgent threat to public health. Recurrence of a C. difficile infection (CDI) after successful treatment with antibiotics is high, occurring in 20-30% of patients, thus necessitating the discovery of novel therapeutics against this pathogen. Current animal models of CDI result in high mortality rates and thus do not approximate the chronic, insidious disease manifestations seen in humans with CDI. To evaluate therapeutics against C. difficile, a mouse model approximating human disease utilizing a clinically-relevant strain is needed. This protocol outlines the cefoperazone mouse model of CDI using a clinically-relevant and genetically-tractable strain, R20291. Techniques for clinical disease monitoring, C. difficile bacterial enumeration, toxin cytotoxicity, and histopathological changes throughout CDI in a mouse model are detailed in the protocol. Compared to other mouse models of CDI, this model is not uniformly lethal at the dose administered, allowing for the observation of a prolonged clinical course of infection concordant with the human disease. Therefore, this cefoperazone mouse model of CDI proves a valuable experimental platform to assess the effects of novel therapeutics on the amelioration of clinical disease and on the restoration of colonization resistance against C. difficile.

Cefoperazone-treated Mouse Model of Clinically-relevant Clostridium difficile Strain R20291

This protocol outlines the cefoperazone mouse model of Clostridium difficile infection (CDI) using a clinically-relevant and genetically-tractable strain, R20291. Emphasis on clinical disease monitoring, C. difficile bacterial enumeration, toxin cytotoxicity, and histopathological changes throughout CDI in a mouse model are detailed in the protocol.

头孢哌酮治疗小鼠模型的临床相关的<em&gt;艰难梭菌</em&gt;应变R20291

Clostridium difficile is an anaerobic, gram-positive, spore-forming enteric pathogen that is associated with increasing morbidity and mortality and ...

Therapeutic Evaluation of Fecal Microbiota Transplantation in an Interleukin 10-Deficient Mouse Model

With the development of microecology in recent years, the relationship between intestinal bacteria and inflammatory bowel disease (IBD) has attracted considerable attention. Accumulating evidence suggests that dysbiotic microbiota plays an active role in triggering or worsening the inflammatory process in IBD and that fecal microbiota transplantation (FMT) is an attractive therapeutic strategy since transferring a healthy microbiota to IBD patient could restore the appropriate host-microbiota communication. However, the molecular mechanisms are unclear, and the efficacy of FMT has not been very well established. Thus, further studies in animal models of IBD are necessary. In this method, we applied FMT from wild-type C57BL/6J mice to IL-10 deficient mice, a widely used mouse model of colitis. The study elaborates on collecting fecal pellets from the donor mice, making the fecal solution/suspension, administering the fecal solution, and monitoring the disease. We found that FMT significantly mitigated the cardiac impairment in IL-10 knockout mice, underlining its therapeutic potential for IBD management.

The interaction of genetic susceptibility, mucosal immunity, and intestinal microecological environment is involved in the pathogenesis of inflammatory bowel disease (IBD). In this study, we applied fecal microbiota transplantation to IL-10 deficient mice and investigated its impact on colonic inflammation and heart function.

白细胞介素10缺陷小鼠模型中粪便微生物群移植的治疗评估

With the development of microecology in recent years, the relationship between intestinal bacteria and inflammatory bowel disease (IBD) has attracted ...

Murine Fecal Isolation and Microbiota Transplantation

Gut microbiota dysbiosis plays a role in the pathophysiology of cardiovascular and metabolic disorders, but the mechanisms are not well understood. Fecal microbiota transplantation (FMT) is a valuable approach to delineating a direct role of the total microbiota or isolated species in disease pathophysiology. It is a safe treatment option for patients with recurrent Clostridium difficile infection. Preclinical studies demonstrate that manipulating the gut microbiota is a useful tool to study the mechanistic link between dysbiosis and disease. Fecal microbiota transplantation may help elucidate novel gut microbiota-targeted therapeutics for the management and treatment of cardiometabolic disease. Despite a high success rate in rodents, there remains translational changes associated with the transplantation. The goal here is to provide guidance in studying the effects of gut microbiome in experimental cardiovascular disease. In this study, a detailed protocol for the collection, handling, processing, and transplantation of fecal microbiota in murine studies is described. The collection and processing steps are described for both human and rodent donors. Lastly, we describe using a combination of the Swiss-rolling and immunostaining techniques to assess gut-specific morphology and integrity changes in cardiovascular disease and related gut microbiota mechanisms.

The goal here is to outline a protocol to investigate the mechanisms of dysbiosis in cardiovascular disease. This paper discusses how to aseptically collect and transplant murine fecal samples, isolate intestines, and use the "Swiss-roll" method, followed by immunostaining techniques to interrogate changes in the gastrointestinal tract.

小鼠粪便分离和微生物群移植

Gut microbiota dysbiosis plays a role in the pathophysiology of cardiovascular and metabolic disorders, but the mechanisms are not well understood. Fecal ...

Preparation and Maintenance of Bioexclusion IsoPositive Cage Experiment for Human Fecal Transplantation into Germ-Free Mice

Germ-free mice are an important investigation tool for understanding the contribution of microorganisms in host health and disease, enabling assessment of the specific role of individuals, defined or complex groups of microorganisms in host response. Traditionally bred and reared in flexible-film or semi-rigid isolators, germ-free mouse husbandry and experimental manipulation are costly and require numerous trained staff and a large space footprint in animal housing facilities. The IsoPositive caging system allows for experimental manipulation of germ-free mice in individual, hermetically-sealed, positive-pressure isolator cages (isocages), reducing cost and enabling greater flexibility in experimental manipulations. 
Here, a protocol is described for transferring germ-free mice from breeding isolators to isocages and subsequent fecal transfer from human donor stool into mice to create stable long-term gut "humanized" mice for future studies. The materials and preparation needed for the utilization of the isocage system are described, including the use of chlorine-dioxide sterilant chemical sterilant to clean cages, supplies, equipment, and personal protective equipment. The methods for confirming the germ-free status of transferred mice and how to determine contamination in the caging system are discussed. The procedure for husbandry, including bedding, food, and water supply, is further discussed. The protocol for human fecal slurry preparation and gavage into germ-free mice to create gut "humanized" mice, along with stool collection to monitor the microbial community composition of these mice, are described. An experiment illustrates that two weeks post-human fecal transplant allows for stable colonization of donor microbiota in the murine hosts, enabling subsequent experimental usage. Furthermore, the collection of humanized mouse feces in viability preservation media, enabling use in further functional experiments, is described. Overall, these methods allow for the safe and effective establishment of humanized mouse communities in experimental gnotobiotic cages for further manipulation.

This protocol describes the best practices for germ-free mouse transfer to and housing in experimental single-cage isolators (isocages) while maintaining sterile conditions. Methods for fecal transplant into germ-free mice and the collection of viable bacteria from these gut "humanized" mice for further applications are discussed.

用于人粪便移植到无菌小鼠体内的生物排斥 IsoPositive 笼实验的制备和维持

Germ-free mice are an important investigation tool for understanding the contribution of microorganisms in host health and disease, enabling assessment of ...

Fecal Microbiota Transplantation and Detection of Prevalence of IgA-Coated Bacteria in the Gut

Gut microbiota exert pleiotropic roles in human health and disease. Fecal microbiota transplantation (FMT) is an effective method to investigate the biological function of intestinal bacteria as a whole or at the species level. Several different FMT methods have been published. Here, we present an FMT protocol that successfully depletes gut microbiota in a matter of days, followed by transplantation of fecal microbiota from fresh or frozen donor intestinal contents to conventional mice. Real time-PCR is applied to test the efficacy of bacterial depletion. Sequencing of the 16S ribosomal RNA (rRNA) is then applied to test the relative abundance and identity of gut microbiota in recipient mice. We also present a flow cytometry-based detection method of immunoglobulin A (IgA)-coated bacteria in the gut.

We established an efficient way to deplete intestinal bacteria in three days, and subsequently transplant fecal microbiota via gavage of fecal fluid prepared from fresh or frozen intestinal contents in mice. We also present an optimized method to detect the frequency of IgA-coated bacteria in the gut.

Gut microbiota exert pleiotropic roles in human health and disease. Fecal microbiota transplantation (FMT) is an effective method to investigate the ...

生物学

Investigating the Alleviating Effects of Bacillus cereus Administration on Colitis through Gut Microbiota Modulation

Gut microbiota dysbiosis is thought to exert a role in the progression of colitis. However, the precise standards for probiotic administration in alleviating colitis remain undefined. Most analysis methods rely on limited diversity and abundance of gut microorganisms. Therefore, observational studies cannot establish causation. In this study, we applied antibiotic-induced pseudo-germ-free mice to investigate the role of gut microbiota in regulating the probiotic effects of Bacillus&#160;cereus (B. cereus)&#160;on dextran sulfate sodium (DSS)-induced colitis in mice. This process allows for evaluating the bidirectional regulating effect of B. cereus&#160;supplementation on health and provides stable and reproducible results. Here, the detailed protocols for B. cereus cultivation, gavage operation, stool collection, and antibiotic clearance treatment on colitis mice are provided. The optimization methods are also applicable for other chronic inflammatory-associated disorders. The results showed that B. cereus administration decreased body weight loss, colon length shortening, disease activity index, and histopathological scores. However, treatment with antibiotics suppressed the positive effect of B. cereus on colitis. These results indicate that gut microbiota are needed for the alleviating effects of B. cereus on colitis. Therefore, exploring the beneficial effects of probiotics in this research is a promising approach for developing novel treatment strategies for alleviating the symptoms of chronic inflammatory-associated disorders.

Investigating the Alleviating Effects of Bacillus cereus Administration on Colitis through Gut Microbiota Modulation

Here, we present a protocol to establish the antibiotic-induced pseudo-germ-free dextran sulfate sodium-induced colitis mouse model to investigate the role of gut microbiota in regulating the positive effects of Bacillus cereus on colitis.

Gut microbiota dysbiosis is thought to exert a role in the progression of colitis. However, the precise standards for probiotic administration in ...

Generating a Double Humanized BLT (Bone-Marrow, Liver, Thymus) Mouse Model

Source: Daharsh, L. et al., <a href="https://appjove.unicartagenaproxy.elogim.com/v/59773">A Double Humanized BLT-mice Model Featuring a Stable Human-Like Gut Microbiome and Human Immune System.</a>&#160;J. Vis. Exp. (2019)
This video demonstrates the method for generating a double humanized bone BLT (marrow, liver, and thymus) mouse with a functional human immune system and a stable engrafted human-like gut microbiome.

生成双人源化 BLT（骨髓、肝脏、胸腺）小鼠模型

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. Mice ...

通过结肠镜检查粪便菌群移植治疗复发性

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此视频中的章节

遗传工程小鼠模型的散发性结肠直肠癌

培养和维护

鼠回结肠肠管切除原发性吻合

头孢哌酮治疗小鼠模型的临床相关的

白细胞介素10缺陷小鼠模型中粪便微生物群移植的治疗评估

小鼠粪便分离和微生物群移植

用于人粪便移植到无菌小鼠体内的生物排斥 IsoPositive 笼实验的制备和维持

Fecal Microbiota Transplantation and Detection of Prevalence of IgA-Coated Bacteria in the Gut

Investigating the Alleviating Effects of Bacillus cereus Administration on Colitis through Gut Microbiota Modulation

生成双人源化 BLT（骨髓、肝脏、胸腺）小鼠模型