Microbiology: Laboratory Theory and Application Michael J. Leboffe, Burton E Pierce

Cover
title
copyright
Preface
Acknowledgments
INTRODUCTION: Safety and Laboratory Guidelines
	Student Conduct
	Basic Laboratory Safety
	Reducing Contamination of Self, Others, Cultures, and the Environment
	Guidelines Governing Handling of BSL-2 Organisms
	Disposing of Contaminated Materials
	Student Compliance with Laboratory Safety Regulations
	A Word About Experimental Design
	Data Presentation: Tables and Graphs
	Data Presentation: Be Creative, But Complete
	Student Safety Contract
SECTION 1: Fundamental Skills for the Microbiology Laboratory
	Exercise 1-1: A Comparison of Hand-Cleansing Agents
	Basic Growth Media
	Exercise 1-2: Nutrient Broth and Nutrient Agar Preparation
	Aseptic Transfers and Inoculation Methods
	Exercise 1-3: Common Aseptic Transfers and Inoculation Methods
	Exercise 1-4: Streak Plate Methods of Isolation
	Exercise 1-5: Spread Plate Method of Isolation
SECTION 2: Microbial Growth
	Diversity and Ubiquity of Microorganisms
	Exercise 2-1: Ubiquity of Microorganisms
	Exercise 2-2: Colony Morphology
	Exercise 2-3: Growth Patterns on Slants
	Exercise 2-4: Growth Patterns in Broth
	Environmental Factors Affecting Microbial Growth
	Exercise 2-5: Evaluation of Media
	Aerotolerance
	Exercise 2-6: Agar Deep Stabs
	Exercise 2-7: Fluid Thioglycollate Broth
	Exercise 2-8: Anaerobic Jar
	Effect of Environmental Factors on Bacterial Growth Rate
	Exercise 2-9: The Effect of Temperature on Microbial Growth
	Exercise 2-10: The Effect of pH on Microbial Growth
	Exercise 2-11: The Effect of Osmotic Pressure on Microbial Growth
	Control of Pathogens: Physical and Chemical Methods
	Exercise 2-12: Steam Sterilization
	Exercise 2-13: The Lethal Effect of UltravioletRadiation on Microbial Growth
	Exercise 2-14: Chemical Germicides:Disinfectants and Antiseptics
SECTION 3: Microscopy and Staining
	Exercise 3-1: Introduction tothe Light Microscope
	Exercise 3-2: Calibration of the Ocular Micrometer
	Exercise 3-3: Microscopic Examination of Eukaryotic Microbes
	Exercise 3-4: Microscopic Examination of Freshwater Microbes
	Bacterial Structure and Simple Stains
	Exercise 3-5: Simple Stains
	Exercise 3-6: Negative Stains
	Differential and Structural Stains
	Exercise 3-7: Gram Stain
	Exercise 3-8: Acid-Fast Stains
	Exercise 3-9: Capsule Stain
	Exercise 3-10: Endospore Stain
	Exercise 3-11: Wet Mount and Hanging Drop Preparation
	Exercise 3-12: Flagella Stain
	Exercise 3-13: Morphological Unknown
SECTION 4: Selective Media
	A Word about Selective Media
	Selective Media for Isolation of Gram-Positive Cocci
	Exercise 4-1: Phenylethyl Alcohol Agar
	Exercise 4-2: Columbia CNA with 5% Sheep Blood Agar
	Exercise 4-3: Bile Esculin Agar
	Exercise 4-4: Mannitol Salt Agar
	Selective Media for Isolation of Gram-negative Rods
	Exercise 4-5: MacConkey Agar
	Exercise 4-6: Eosin Methylene Blue Agar (Levine)
	Exercise 4-7: Hektoen Enteric Agar
	Exercise 4-8: Xylose Lysine Desoxycholate Agar
SECTION 5: Differential Tests
	A Word about Biochemical Tests and Acid-Base Reactions
	Introduction to Energy Metabolism Tests
	Exercise 5-1: Reduction Potential
	Exercise 5-2: Oxidation–Fermentation Test
	Fermentation Tests
	Exercise 5-3: Phenol Red Broth
	Exercise 5-4: Methyl Red and Voges–Proskauer Tests
	Exercise 5-5: ONPG Test
	Exercise 5-6: Catalase Test
	Exercise 5-7: Oxidase Test
	Exercise 5-8: Nitrate Reduction Test
	Nutrient Utilization Tests
	Exercise 5-9: Citrate Utilization Test
	Exercise 5-10: Malonate Utilization Test
	Decarboxylation and Deamination Tests
	Exercise 5-11: Decarboxylation Tests
	Exercise 5-12: Phenylalanine Deaminase Test
	Tests Detecting Hydrolytic Enzymes
	Exercise 5-13: Starch Hydrolysis (Amylase Test)
	Exercise 5-14: DNA Hydrolysis (DNase Test)
	Exercise 5-15: Lipid Hydrolysis (Lipase Test)
	Exercise 5-16: Casein Hydrolysis (Casease Test)
	Exercise 5-17: Gelatin Hydrolysis (Gelatinase Test)
	Exercise 5-18: Urea Hydrolysis (Urease Test)
	Exercise 5-19: PYR Test
	Combination Differential Media
	Exercise 5-20: SIM Medium Sulfur Reduction, Indole Production, Motility
	Exercise 5-21: Triple Sugar Iron Agar / Kligler Iron Agar
	Exercise 5-22: Lysine Iron Agar
	Exercise 5-23: Litmus Milk Medium
	Antimicrobial Susceptibility Testing
	Exercise 5-24: Bacitracin, Novobiocin, and Optochin Susceptibility Tests
	Other Differential Tests
	Exercise 5-25: Blood Agar
	Exercise 5-26: CAMP Test
	Exercise 5-27: Coagulase and Clumping Factor Tests
	Exercise 5-28: Motility Test
	Multiple Test Systems
	Exercise 5-29: api 20 E Identification System for Enterobacteriaceae and Other Gram-negative Rods
	Exercise 5-30: EnteroPluri–Test (Enterotube® II)
	Bacterial Unknowns Project
	Exercise 5-31: Bacterial Unknowns Project
SECTION 6: Quantitative Techniques
	Exercise 6-1: Standard Plate Count (Viable Count)
	Exercise 6-2: Urine Culture
	Exercise 6-3: Direct Count (Petroff–Hausser)
	Exercise 6-4: Closed-System Growth
	Exercise 6-5: Plaque Assay of Virus Titer
	Exercise 6-6: Thermal Death Time Versus Decimal Reduction Value
SECTION 7: Medical Microbiology
	Exercise 7-1: Snyder Test
	Exercise 7-2: Lysozyme Assay
	Exercise 7-3: Antimicrobial Susceptibility Test: Disk Diffusion (Kirby-Bauer) Method
	Exercise 7-4: Clinical Biofilms
	Exercise 7-5: Morbidity and Mortality Weekly Report (MMWR) Assignment
	Exercise 7-6: Epidemic Simulation
	Exercise 7-7: Identification of Enterobacteriaceae
	Exercise 7-8: Identification of Gram-positive Cocci
	Exercise 7-9: Identification of Gram-positive Rods
SECTION 8: Environmental Microbiology
	A Word about Trophic Groups
	Exercise 8-1: Winogradsky Column
	Introduction to the Nitrogen Cycle
	Exercise 8-2: Nitrogen Fixation
	Exercise 8-3: Nitrification: The Production of Nitrate
	Exercise 8-4: Ammonification
	Exercise 8-5: Denitrification: Nitrate Reduction
	Sulfur Cycle—Introduction
	Exercise 8-6: Photosynthetic Sulfur Bacteria
	Exercise 8-7: Chemolithotrophic Sulfur-Oxidizing Bacteria
	Exercise 8-8: Sulfur-Reducing Bacteria
	Exercise 8-9: Bioluminescence
	Exercise 8-10: Soil Slide Culture
	Exercise 8-11: Soil Microbial Count
	Water Quality Testing
	Exercise 8-12: Membrane Filter Technique
	Exercise 8-13: Multiple Tube Fermentation Methodfor Determining Water Quality
SECTION 9: Food Microbiology
	Exercise 9-1: Methylene Blue Reductase Test
	Exercise 9-2: Making Yogurt
	Exercise 9-3: Yogurt CFU Density of an Active and Live Culture and Identification of a Yogurt Unknown
SECTION 10: Microbial Genetics
	Exercise 10-1: Extraction of DNA from Bacterial Cells
	Exercise 10-2: Restriction Digest
	Exercise 10-3: Bacterial Transformation: the pGLO™ System
	Exercise 10-4: Polymerase Chain Reaction
	Exercise 10-5: Ultraviolet Radiation Damage and Repair
	Exercise 10-6: Ames Test
	Exercise 10-7: Phage Typing of E. Coli Strains
SECTION 11: Hematology and Serology
	Exercise 11-1: Differential Blood Cell Count
	Introduction to Antigens and Antibodies
	Simple Serological Reactions
	Exercise 11-2: Precipitin Ring Test
	Exercise 11-3: Double Gel Immunodiffusion
	Exercise 11-4: Slide Agglutination
	Exercise 11-5: Blood Typing
	Exercise 11-6: ELISA for Detecting Antibodies in a Patient’s Sample (The Antibody Capture Method)
SECTION 12: Eukaryotic Microbes
	Eukaryotic Supergroups—A Refresher
	Exercise 12-1: Representative Fungi of Medical and Economic Importance
	Exercise 12-2: Fungal Slide Culture
	Exercise 12-3: Examination of Simple Eukaryotic Pathogens
	Exercise 12-4: Parasitic Helminths
APPENDICES
	APPENDIX A: Biochemical Pathways
	APPENDIX B: Miscellaneous Transfer Methods
	APPENDIX C: Transfer from a Broth Culture Using a Glass Pipette
	APPENDIX D: Transfer from a Broth Culture Using a Digital Pipette
	APPENDIX E: The Spectrophotometer
	APPENDIX F: Alternative Procedures for Section 6
	APPENDIX G: Agarose Gel Electrophoresis
	APPENDIX H: Medium, Reagent, and Stain Recipes
Glossary
Index
Copyright
Title Page
Dedication
Contents
Chapter 1: ‘I’m thinking’ – Oh, but are you?
Chapter 2: Renegade perception
Chapter 3: The Pushbacker sting
Chapter 4: ‘Covid’: The calculated catastrophe
Chapter 5: There is no ‘virus’
Chapter 6: Sequence of deceit
Chapter 7: War on your mind
Chapter 8: ‘Reframing’ insanity
Chapter 9: We must have it? So what is it?
Chapter 10: Human 2.0
Chapter 11: Who controls the Cult?
Chapter 12: Escaping Wetiko
Postscript
Appendix: Cowan-Kaufman-Morell Statement on Virus Isolation
Bibliography
Index