GNOTOBIOTICS AND MICROBIOLOGY CORE

知生学和微生物学核心

• 批准号: 8565001
• 负责人: LYNN BRY
• 金额: $ 17.9万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-09-01 至 2015-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8565001
• 关键词: Aerobic; Animals; Area; Autopsy; Back; Bar Codes; Beds; Biochemical; Bioinformatics; Biology; Blood; Breeding; C57BL/6 Mouse; Charge; Cluster Analysis; Collaborations; Collection; Complex; Culture Media; DNA; Data; Derivation procedure; Dissection; Epithelial; Equipment; Evaluation; Fermentation; Fixatives; Frozen Sections; Gases; Germ-Free; Gnotobiotic; Gram' s stain; High Performance Computing; Histopathology; Human; Human Resources; Individual; Knock-out; Laboratories; Libraries; Liquid substance; Maintenance; Methods; Microbiology; Molecular; Mouse Strains; Mus; Nutrient; Organism; Paraffin Embedding; Polymerase; Preparation; Reagent; Recombinant DNA; Research Personnel; Resources; Ribosomal RNA; Rivers; Rodent; Sampling; Savings; Schedule; Services; Shipping; Ships; Site; Slide; Solid; Source; Sterility; System; Testing; Time; Tissue Harvesting; Tissues; Water; Wild Type Mouse; Work; animal breeding; base; cost; design; enzyme activity; investigator training; long chain fatty acid; member; microbial; microbiome; pathogen; pressure; rRNA Genes; research study; routine Bacterial stain; tool

SPECIFIC ACTIVITIES OF THE GNOTOBIOTICS and MICROBIOLOGY CORE A. Core Services Core services are provided on a charge-back basis. They are designed to achieve economies of scale for purchase of equipment, reagents/supplies, and labor so that the real costs of use are subsidized by up to 50% for individual investigators. 1) Maintenance of Germfree mice: The core maintains germfree mice for a number of investigators, including wild-type Swiss Webster and C57BL/6 strains, as well as knockout strains that have been derived for investigators by Taconic or other sites. Mr. Yeliseyev manages isolators for 4 investigators who use core isolators on a regular basis. Ms. Belavusava assists in microbiological sterility checks of mice, as performed by culture and 16S rRNA PCR. 2) Maintenance of Specifically-Associated Mice: The core has 4 small positive-pressure isolators for specific-association experiments with commensal organisms. The Core Director and Mr. Yeliseyev work with investigators to devise experimental plans for specific-association experiments and schedule use of isolators. 3) Maintenance of Specifically-Associated Mice with BL-2 pathogens: The core has 2 negative-pressure isolators for association of germfree mice with BL-2 pathogens alone or with commensal species. Mr. Yeliseyev, with assistance from Ms. Belavusava, maintains isolators with specifically associated mice while Ms. Belavusava assists with culturing of isolates for associations and quantitative culture of materials from colonized mice, as warranted. 4) On-site breeding and availability of wild-type germfree stocks: The core maintains 2 large breeding isolators, primarily to provide a local supply of germfree Swiss-Webster and C57BL/6 mice for experiments. Local breeding provides a significant savings in animal purchase costs to HDDC investigators (>70% savings) and often provides a more ready supply of wild-type mice than can be obtained from commercial sources. Per request, the core will also maintain small numbers of breeding pairs of other strains of germfree mice imported by HDDC investigators for designated experiments. Mr Yeliseyev maintains animal breeding colonies, including ordering commercially-maintained germfree mice to replenish breeding stocks as needed. Note, Core E does not currently undertaking germfree derivations given space constraints in available mouse facilities in the Longwood area. The core does provide information and options for deriving mouse lines to a germfree state through Taconic or Charles River. HDDC: Integrated Epithelial and Mucosal Biology Lencer, Wayne I. 5) Tissue harvest and histopathology: The core offers assistance with collection of mouse tissues and blood, and preparation of materials for frozen section or paraffin embedding. Mr. Yeliseyev is well-versed in animal dissection and is available to perform or otherwise assist investigators with tissue harvests. Dr. Bry is also able to assist and advise with optimal methods to perform mouse necropsies for downstream applications. 6) Aerobic and anaerobic culture: The core provides equipment, materials and expertise for the culture of aerobic, microaerophilic and anaerobic species. Resources are available in Dr. Bry and Dr. Onderdonk's labs and may be further supported through additional technical staff in their labs. 7) Microbial identification and analysis: The core has equipment, resources and expertise to speciate commensal isolates by microbiological, biochemical and molecular methods, including biochemical analyses of fermentation profiles and other enzyme activities, MIDI analysis of short- medium- or long-chain fatty acids, and comparison of 16S rRNA sequences against curated 16S libraries. 8) Analyses of the Microbiome. The core has the resources and expertise to assist investigators with the collection and preparation of human and rodent samples for deep sequencing of 16S rRNA gene signatures. HDDC investigators may access bioinformatics tools maintained on the BWH High Performance Computing Cluster for analysis of Roche 454 sequencing data of 16S rRNA gene signatures.

特定活动的GNOTOBIOTICS和微生物学核心 A.核心服务 核心服务按收费方式提供。它们旨在实现设备、试剂/供应品和劳动力采购的规模经济，从而使个体研究者的真实的使用成本得到高达50%的补贴。 1)无菌小鼠的维持：该中心为许多研究者维持无菌小鼠，包括野生型Swiss韦伯斯特和C57 BL/6品系，以及由Taconic或其他研究中心为研究者衍生的敲除品系。Yeliseyev先生为4名定期使用核心隔离器的研究者管理隔离器。Belavusava女士协助通过培养和16 S rRNA PCR对小鼠进行微生物无菌检查。 2)特定关联小鼠的维护：核心有4个小型正压隔离器，用于与微生物的特定关联实验。核心主任和Yeliseyev先生与研究人员合作，为特定关联实验制定实验计划，并安排隔离器的使用。 3)保持与BL-2病原体特异性相关的小鼠：核心有2个负压隔离器，用于将无菌小鼠与单独的BL-2病原体或与其他物种相关联。在Belavusava女士的协助下，Yeliseyev先生负责维护与特定关联小鼠的隔离器，而Belavusava女士则协助培养用于关联的隔离物，并对来自 殖民小鼠，如有必要。 4)现场饲养和野生型无菌种群的可用性：核心保持2个大型饲养隔离器，主要是为实验提供无菌Swiss-Webster和C57 BL/6小鼠的本地供应。 本地育种为HDDC研究者提供了动物购买成本的显著节省（>70%的节省），并且通常提供比从商业来源获得的野生型小鼠更容易的供应。根据要求，该中心还将保留少量进口的其他无菌小鼠品系的繁殖对 由HDDC研究人员进行指定实验。Yeliseyev先生维护动物繁殖群，包括订购商业化的无菌小鼠，以根据需要补充繁殖种群。 请注意，鉴于Longwood地区可用小鼠设施的空间限制，核心E目前不进行无菌衍生。核心确实提供了通过Taconic或Charles River将小鼠品系衍生到无菌状态的信息和选项。 HDDC：综合上皮和粘液生物学Lencer，韦恩I。 5)组织采集和组织病理学：核心可帮助采集小鼠组织和血液，并准备用于冷冻切片或石蜡包埋的材料。Yeliseyev先生精通动物解剖，可以进行或以其他方式协助研究人员进行组织采集。Bry博士还能够为下游应用提供最佳方法的协助和建议，以进行小鼠尸检。 6)好氧和厌氧培养：核心为好氧、微好氧和厌氧物种的培养提供设备、材料和专业知识。Bry博士和Onderdonk博士的实验室提供了资源，并可能通过他们实验室的额外技术人员提供进一步支持。 7)微生物鉴定和分析：该中心拥有通过微生物、生物化学和分子方法鉴定微生物分离株的设备、资源和专业知识，包括发酵概况和其他酶活性的生物化学分析、短链、中链或长链脂肪酸的微生物分析，以及16 S rRNA序列与精选的16 S文库的比较。 8)微生物组的分析。该中心拥有资源和专业知识，可协助研究人员收集和制备人类和啮齿动物样本，以进行16 S rRNA基因特征的深度测序。 HDDC研究人员可以访问BWH高性能计算集群上维护的生物信息学工具，以分析16 S rRNA基因签名的Roche 454测序数据。