1st Edition

Gnotobiotic Mouse Technology An Illustrated Guide

    256 Pages 275 Color Illustrations
    by CRC Press

    256 Pages
    by CRC Press

    The popularity of germ-free animal models, particularly mice, for investigation of human physiology and disease has recently exploded. Gnotobiotic Mouse Technology: An Illustrated Guide provides the first manual for the maintenance, husbandry, and experimental manipulation of germ-free and gnotobiotic mice. It includes information on all aspects of establishing and operating a germ-free mouse research facility, from basic principles and equipment to detailed instructions for assembling and maintaining isolators, sterilizing supplies, handling animals, and monitoring sterility.

    The book breaks down techniques and procedures into modules, each of which contains a step-by-step practical and visual guide to a set of related procedures. Each description consists of a materials list, introduction, and general overview, followed by a thoroughly illustrated walkthrough of the steps for each procedure. The descriptions end with troubleshooting tips. Technical chapters are heavily illustrated and include notations of potential pitfalls and alternatives. Other chapters discuss management procedures and practices such as the hiring and training of personnel, setting fee schedules, and record keeping.

    This book is a landmark resource for establishing and maintaining a facility for germ-free mouse research. Useful to both technicians and investigators, it presents every step necessary to establish a successful facility. It also gives direction in expanding multi-user facilities and applying new technologies to your current practices.

    A Brief History of Germ-Free Life

    Overview of Gnotobiotic Technology
    How Do We Get Germ-Free Mice?
    Germ-Free Housing and Barrier Types

    Equipment and Terminology

    Personal Protective Equipment
    Types of PPE
    Applying Sterile PPE

    Sterilants and Sterilization
    Sterilization Methods
    Cold Sterilization
    Wrapping Materials for Autoclaving
    Gas Sterilization (Ethylene Oxide)

    Isolator Setup
    Part 1: Small (5.5 x 3 feet) Isolator Assembly
    Part 2: Large Isolator Assembly

    Port Entry and Exit
    Preparing, Opening, and Closing the Port
    Proper Placement of Materials

    Sterilizing Food and Supplies
    Biological Indicators
    Assembling Supply Cylinders: Installing HEPA Filtration Media
    Prepare the Cylinder
    Fill the Cylinder
    Water Sterilization

    Supply Cylinder Entry

    Isolator Maintenance
    Part 1: Cleaning and Maintenance
    Part 2: Replacing Gloves on an Active Isolator
    Part 3: Repairing Holes in Isolators
    Part 4: Working in the Isolator

    Aseptic Mouse Transfer
    Part 1: Internal Transfer between Isolators
    Part 2: External Transfer of Mice

    Working with Germ-Free or Gnotobiotic Mice in the Class II Biosafety Cabinet
    Experimental Manipulations in the Biosafety Cabinet

    Shipping Mice

    Derivation by Cesarean Section
    Derivation by Embryo Transfer

    Microbiological Testing
    Serologic Screening
    Part 1: Bacterial Culture
    Part 2: Examination of Gram-Stained Slides for the Presence of Bacteria

    Genetic Testing

    Record Keeping
    Colony Management Documents
    Project-Planning Documents
    Other Documents

    Facility Setup and Management
    Establishing a New Facility

    Appendix: Sources for Equipment and Supplies


    Chriss J. Vowles co-manages a multi-investigator germ-free research laboratory at the University of Michigan in Ann Arbor. He began his career at the University of Michigan in 2003, working full-time as a husbandry technician in the Unit for Laboratory Animal Medicine. In 2006, he discovered gnotobiotic technology. At that time, the Germ Free Laboratory was just starting. Chriss joined the research group on the ground floor and has been growing with it ever since. The idea of maintaining a germ-free, complex organism fascinated him then and still captivates him today. After eight years of maintaining a germ-free colony of mice, the laboratory is still evolving, he is still learning, and his trials, tribulations, and rewards are constant. Natalie E. Anderson is a research technician lead at the University of Michigan's Germ Free Core. In 2009, she graduated from Michigan State University with a bachelor's degree in animal science and accepted a position with SoBran's contract for the National Institute of Allergy and Infectious Diseases (NIAID). In December 2010, she returned to Michigan to take a position with the University of Michigan's Unit for Lab Animal Medicine. Shortly thereafter, she started working part-time in the Germ Free Core and quickly learned to love the daily challenges involved in maintaining a germ-free colony. Natalie joined the core full time in July 2013. Kathryn A. Eaton has been working with germ-free and gnotobiotic animal models for 30 years. She started her gnotobiology career at Ohio State University, where she did her PhD research on Helicobacter pylori in gnotobiotic piglets. She went on to study mice several years later, and in 2004 she established the University of Michigan Germ Free Mouse Laboratory, which she now directs. Dr. Eaton is a board-certified veterinary pathologist with research interests in bacterial enteric disease and immunology. In addition to H. pylori ,

    "This book introduces and describes techniques utilized for the production, maintenance, and experimental manipulation of germ-free and gnotobiotic mice in a research support facility. The introductory chapter provides history and identifies animals that were successfully derived and maintained in a germ-free setting. …

    The middle portion of the book provides detailed information regarding isolator setup. … provides step-by-step descriptions and illustrations for assembly, inflation, sterilization, and decontamination….

    The next portion of the book describes the proper technique for port entry and exit, isolator maintenance, and the use of supply cylinders and supply isolators.

    The final chapters of the book discuss methods of microbiological testing such as the Gram stain procedure and quantitative PCR. … Genetic testing and record keeping are discussed. … Chapter 18 describes facility setup and management of maintaining a gnotobiotic laboratory. Gnotobiotic laboratories are labor intensive and extremely expensive. Funding options and space requirements when establishing and/or maintaining an existing gnotobiotic laboratory must be discussed with institutional directors.

    Overall, this book is reasonably priced and available in electronic and paperback versions. The combination of procedural descriptions and detailed illustrations are an advantage of this book. The chapter describing isolator setup is very descriptive with complementary illustrations. This book serves as a good reference for new and/or existing gnotobiotic facilities."

    Dondrae Coble, DVM, DACLAM in Laboratory Animal Practitione