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E-raamat: Gnotobiotics

Edited by (Department of Genetics (Genomics Division) and Animal Resources Program Comparative Pathology Laboratory, University of Alabama at Birmingham), Edited by (Department of Microbiology and Immunology, University of Michigan Medical School)
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Gnotobiotics summarizes and analyzes the research conducted on the use of gnotobiotes, providing detailed information regarding actual facility operation and derivation of gnotobiotic animals.

In response to the development of new tools for microbiota and microbiome analysis, the increasing recognition of the various roles of microbiota in health and disease, and the consequent expanding demand for gnotobiotic animals for microbiota/microbiome related research, this volume collates the research of this expanding field into one definitive resource.

  • Reviews and defines gnotobiotic animal species
  • Analyzes microbiota in numerous contexts
  • Presents detailed coverage of the protocols and operation of a gnotobiotic facility

Muu info

Comprehensively covers the expanding field of gnotobiotic animals, the implications of microbiota research and how to run a gnotobiotic facility
List of Contributors
ix
Foreword xi
1 Historical Overview
Trenton R. Schoeb
Roger P. Orcutt
References
14(8)
2 Principles of Establishing and Operating a Gnotobiotic Facility
Trenton R. Schoeb
Richard J. Rahija
Cassie Boyd
Roger P. Orcutt
Kathryn A. Eaton
1 Introduction
22(1)
2 Institutional Factors
22(5)
2.1 Financial Support
22(1)
2.2 Space Requirements
22(3)
2.3 Autoclave
25(1)
2.4 Personnel
25(1)
2.5 Microbiology Laboratory
25(1)
2.6 Facility Productivity and Experimental Utilization
25(1)
2.7 Facility Operation and Compliance
26(1)
3 Isolator Selection
27(5)
3.1 Isolator Types and Construction
27(5)
3.2 Individual Positive Pressure Cage Systems
32(1)
4 Sterilization
32(6)
4.1 Steam
33(1)
4.2 Ionizing Radiation
34(1)
4.3 Liquid Chemical
34(3)
4.4 Gas and Vapor Chemical
37(1)
4.5 Dry Heat
38(1)
4.6 Filtration
38(1)
4.7 Further Information
38(1)
5 Operation
38(14)
5.1 Standard Operating Procedures
38(1)
5.2 Isolator Setup
38(2)
5.3 Sterilization and Entry of Supplies
40(5)
5.4 Introduction and Transfer of Animals
45(1)
5.5 Working in Isolators
45(1)
5.6 Monitoring for Contamination
46(4)
5.7 Derivation of Germfree Mice
50(1)
5.8 Establishing Defined Microbiota
51(1)
6 Resources
52(14)
6.1 Recommended Reading
52(2)
6.2 Online
54(1)
6.3 Manufacturers
54(1)
6.4 Laboratories
55(1)
Glossary
56(2)
References
58(8)
3 Procedures for Rodents
Cassie Boyd
Kathryn A. Eaton
Trenton R. Schoeb
Alton G. Swennes
Hayden Bickerton
Billie J. Parsons
Stacey Sinclair
Chriss Vowles
1 Introduction
66(1)
2 Background and Acknowledgments
67(1)
2.1 University of Texas Southwestern Medical Center (UT Southwestern)
67(1)
2.2 University of Michigan (Michigan)
67(1)
2.3 University of Alabama at Birmingham (UAB)
67(1)
2.4 Baylor College of Medicine (BCM)
68(1)
3 General Information
68(3)
3.1 UT Southwestern
68(1)
3.2 UAB
68(1)
3.3 BCM
69(2)
4 Sterilization and Sterilization Monitoring
71(15)
4.1 UT Southwestern
74(1)
4.2 UAB
75(8)
4.3 BCM
83(3)
5 Sterilizing Cylinders
86(50)
5.1 UT Southwestern
86(28)
5.2 UAB
114(11)
5.3 BCM
125(11)
6 Water
136(5)
6.1 UT Southwestern
136(2)
6.2 UAB
138(3)
6.3 BCM
141(1)
7 Transferring Supplies Into Isolators
141(42)
7.1 UT Southwestern
142(19)
7.2 UAB
161(12)
7.3 BCM
173(10)
8 Isolator Setup
183(107)
8.1 UT Southwestern
184(39)
8.2 Michigan
223(1)
8.3 UAB
224(44)
8.4 BCM
268(22)
9 Husbandry
290(8)
9.1 UAB
293(3)
9.2 BCM
296(2)
10 Microbiological Monitoring
298(12)
10.1 UT Southwestern
298(6)
10.2 Michigan
304(1)
10.3 UAB
305(5)
11 Records and Maintenance
310(2)
11.1 UAB
311(1)
11.2 BCM
312(1)
12 Derivation of Germfree Mice
312(5)
12.1 Michigan
313(1)
12.2 UAB
313(4)
13 Establishing Microbiota in Germfree Mice
317(1)
13.1 UAB
317(1)
14 Alternatives to Isolators
318(2)
14.1 UAB
319(1)
15 Shipping and Receiving Mice
320(3)
15.1 UAB
321(2)
16 Materials Lists and Fabrication Diagrams
323(12)
16.1 UAB
323(10)
References
333(2)
4 Developing a Business Plan for Your Gnotobiotics Program
George Langan
Betty Theriault
1 Introduction
335(1)
2 Operational Management Considerations in Establishing a Gnotobiotics Program
335(2)
3 Developing a Business Model
337(5)
3.1 Identifying the Users and Volume of Work
337(2)
3.2 Identifying Space and Equipment Requirements
339(2)
3.3 Labor---What Products and Services Will Be provided and Who Will Provide Them
341(1)
4 Identifying Expenses
342(4)
4.1 Consumable Supplies
342(2)
4.2 Labor
344(1)
4.3 Support Costs
345(1)
4.4 Quality Control Costs
346(1)
5 Establishing Fees
346(1)
6 Subsidization
347(1)
7 Financial Reviews
347(1)
8 Summary
347(2)
References
348(1)
5 Gnotobiotic Pigs: From Establishing Facility to Modeling Human Infectious Diseases
Lijuan Yuan
Peter M. Jobst
Mariah Weiss
1 History of Gnotobiotic Pigs
349(1)
2 Establishing and Operating a Gnotobiotic Pig Facility
350(10)
2.1 Facilities
350(1)
2.2 Specialized Equipment
351(2)
2.3 Isolation Systems
353(1)
2.4 Space Requirements
354(1)
2.5 Preparation of Isolators
354(1)
2.6 Sow Hysterectomy
355(3)
2.7 Feeding
358(1)
2.8 Spraying Materials Prior to Entry Into an Isolator
359(1)
2.9 Sample Collection and Administration of Inocula
359(1)
2.10 Cleaning a Dirty Isolator
360(1)
3 Gnotobiotic Pig Models for Studies of Infectious Diseases
360(9)
3.1 Gnotobiotic Pig Models for Human Enteric Viruses
360(2)
3.2 Gnotobiotic Pig Models for Bacterial and Fungal Infections
362(1)
3.3 Studies of Probiotics and Prebiotics Using Gnotobiotic Pigs
362(1)
3.4 Gnotobiotic Pigs Colonized With Human Gut Microbiota
363(1)
3.5 Concluding Remarks
363(1)
References
364(5)
6 Gnotobiotic Fish as Models to Study Host--Microbe Interactions
Torunn Forberg
Kathryn Milligan-Myhre
1 Introduction
369(1)
2 Brief Review of Fish Gnotobiology
370(1)
3 Protocol for Derivation and Husbandry of Gnotobiotic Fish
371(1)
4 Facilities
371(1)
5 Considerations for Disinfection of Eggs
371(3)
5.1 Polyvinylpyrrolidone--lodine
372(1)
5.2 Sodium Hypochlorite Solution
373(1)
5.3 Glutaraldehyde
373(1)
5.4 Hydrogen Peroxide
373(1)
5.5 Fish Husbandry in Water Containing Antimicrobial Agents
373(1)
6 Confirmation of Sterility
374(1)
6.1 Microscopic Visualization of Flask
374(1)
6.2 Culture-Based Detection
374(1)
6.3 PCR Amplification of the Bacteria-Specific 16S Ribosomal RNA Gene
374(1)
6.4 In Situ Hybridization
374(1)
6.5 Flow Cytometry
375(1)
7 Long-Term Husbandry
375(2)
7.1 Tissue Culture Flasks
375(1)
7.2 Aerated Bottles
376(1)
7.3 Beakers/Tanks in Gnotobiotic Bubble
376(1)
8 Food Considerations for Long-Term Experiments
377(1)
8.1 Formulated Food
377(1)
8.2 Rotifers
377(1)
8.3 Paramecium
377(1)
8.4 Artemia
377(1)
8.5 Copepods
378(1)
8.6 Tetrahymena
378(1)
8.7 Algae
378(1)
9 Other Considerations for Gnotobiotic Fish Husbandry
378(1)
10 Derivation of Germfree Stickleback Embryos
379(6)
10.1 Solutions
380(1)
References
380(5)
7 Use of Gnotobiotic Mice in the Study of Metabolic Syndrome
Benoit Chassaing
Andrew T. Gewirtz
1 Introduction
385(4)
1.1 To What Extent Is a Microbiota Required for Metabolic Syndrome?
386(1)
1.2 Is an Altered Microbiota Sufficient to Drive Metabolic Syndrome?
387(2)
2 Conclusion and Future Directions
389(2)
Acknowledgments
389(1)
References
389(2)
8 Gnotobiotics and Inflammatory Bowel Disease
Trenton R. Schoeb
1 Introduction
391(3)
2 Cytokines and Cytokine Signaling
394(2)
2.1 Interleukin-10
394(1)
2.2 Interleukin-2
395(1)
2.3 TGF-β1
396(1)
2.4 TNF-α
396(1)
2.5 Other Cytokines
396(1)
3 T-Cell Receptors and MHC
396(1)
3.1 T-Cell Receptor and MHC II Deficient Mice
396(1)
3.2 HLA B27 β2m Transgenic Rats
397(1)
4 Other Mutants
397(2)
4.1 Core 1-Derived O-Glycan Deficiency
397(1)
4.2 Fas-Associated via Death Domain Protein
397(1)
4.3 Glutathione Peroxidase Deficiency
397(1)
4.4 Nfil3 Deficiency
397(1)
4.5 PI3 Kinase p110δ Deficiency
398(1)
4.6 Telomerase Deficiency
398(1)
4.7 T-Bet Deficiency
398(1)
4.8 TLR5 Deficiency
398(1)
5 T-Cell Transfer
399(2)
6 Inbred Strains
401(1)
6.1 SAMP1/Yit
401(1)
6.2 C3H/HeJBir
401(1)
7 Conclusion
401(1)
References 401(10)
Index 411
Dr Trenton Schoebs research focuses on gnotobiotic and specific pathogen-free (SPF) rodents and the phenotyping of mutant mice. He has participated in a wide variety of projects involving evaluation of disease expression in rodents and other species. His current interests are in development of gnotobiotic mouse models of inflammatory bowel disease and identification of specific intestinal bacteria involved in initiation and/or perpetuation of intestinal inflammation in these models. Dr. Kathryn Eaton is a board-certified veterinary pathologist (diplomate of the American College of Veterinary Pathologists) with research interests in bacterial pathogens of the gastrointestinal tract, host-pathogen interactions, and intestinal immunity.
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