Table of Contents # of Pages Select Introduction: Safety and Laboratory Guidelines 14

Section 1: Fundamental Skills for the Microbiology Laboratory 40

Section 1 Introductory Pages 2

Exercise 1-1: A Comparison of Hand-Cleansing Agents 4

Exercise 1-2: Nutrient Broth and Nutrient Agar Preparation 6

Exercise 1-3: Common Aseptic Transfers and Inoculation Methods (includes pg. 27) 14

Exercise 1-4: Streak Plate Methods of Isolation 8

Exercise 1-5: Spread Plate Method of Isolation 6

Section 2: Microbial Growth 86

Section 2 Introductory Pages 2

Exercise 2-1: Ubiquity of Microorganisms 6

Exercise 2-2: Colony Morphology 12

Exercise 2-3: Growth Patterns on Slants 4

Exercise 2-4: Growth Patterns in Broth 4

Exercise 2-5: Evaluation of Media 8

Exercise 2-6: Agar Deep Stabs 4

Exercise 2-7: Fluid Thioglycollate Broth 4

Exercise 2-8: Anaerobic Jar 4

Exercise 2-9: The Effect of Temperature on Microbial Growth 6

Exercise 2-10: The Effect of pH on Microbial Growth 4

Exercise 2-11: The Effect of Osmotic Pressure on Microbial Growth 8

Exercise 2-12: Steam Sterilization (includes pg. 121) 8

Exercise 2-13: The Lethal Effect of Ultraviolet Radiation on Microbial Growth 6

Exercise 2-14: Chemical Germicides: Disinfectants and Antiseptics 6

Section 3: Microscopy and Staining 94

Section 3 Introductory Pages 1

Exercise 3-1: Introduction to the Light Microscope (includes pg. 142) 13

Exercise 3-2: Calibration of the Ocular Micrometer 6

Exercise 3-3: Microscopic Examination of Eukaryotic Microbes 8

Exercise 3-4: Microscopic Examination of Freshwater Microbes 12

Exercise 3-5: Simple Stains (includes pgs. 181-184) 10

Exercise 3-6: Negative Stains 4

Exercise 3-7: Gram Stain 8

Exercise 3-8: Acid-Fast Stains 8

Exercise 3-9: Capsule Stain 4

Exercise 3-10: Endospore Stain 6

Exercise 3-11: Wet Mount and Hanging Drop Preparations 4

Exercise 3-12: Flagella Stain 4

Exercise 3-13: Morphological Unknown 6

Section 4: Selective Media 50

Section 4 Introductory Pages 3

Exercise 4-1: Phenylethyl Alcohol Agar 6

Exercise 4-2: Columbia CNA With 5% Sheep Blood Agar 6

Exercise 4-3: Bile Esculin Agar 4

Exercise 4-4: Mannitol Salt Agar 6

Exercise 4-5: MacConkey Agar 8

Exercise 4-6: Eosin Methylene Blue Agar (Levine) 6

Exercise 4-7: Hektoen Enteric Agar 6

Exercise 4-8: Xylose Lysine Desoxycholate Agar 6

Section 5: Differential Tests 182

Section 5 Introductory Pages 4

Exercise 5-1: Reduction Potential 6

Exercise 5-2: Oxidation-Fermentation Test 8

Exercise 5-3: Phenol Red Broth 8

Exercise 5-4: Methyl Red and Voges-Proskauer Tests 6

Exercise 5-5: ONPG Test 4

Exercise 5-6: Catalase Test 6

Exercise 5-7: Oxidase Test 6

Exercise 5-8: Nitrate Reduction Test 6

Exercise 5-9: Citrate Utilization Test 6

Exercise 5-10: Malonate Utilization Test 4

Exercise 5-11: Decarboxylation Tests 8

Exercise 5-12: Phenylalanine Deaminase test 4

Exercise 5-13: Starch Hydrolysis (Amylase Test) 6

Exercise 5-14: DNA Hydrolysis (DNase Test) 4

Exercise 5-15: Lipid Hydrolysis (Lipase Test) 4

Exercise 5-16: Casein Hydrolysis (Casease Test) 4

Exercise 5-17: Gelatin Hydrolysis (Gelatinase Test) 4

Exercise 5-18: Urea Hydrolysis (Urease Test) 6

Exercise 5-19: PYR Test 4

Exercise 5-20: SIM Medium (Sulfur Reduction, Indole Production, Motility) 8

Exercise 5-21: Triple Sugar Iron Agar / Kligler Iron Agar 6

Exercise 5-22: Lysine Iron Agar 4

Exercise 5-23: Litmus Milk Medium 6

Exercise 5-24: Bacitracin, Novobiocin, and Optochin Susceptibility Tests 6

Exercise 5-25: Blood Agar 6

Exercise 5-26: Camp Test 4

Exercise 5-27: Coagulase and Clumping Factor Tests 4

Exercise 5-28: Motility Test 4

Exercise 5-29: api 20 E Identification System for Enterobacteriaceae 8

Exercise 5-30: EnteroPluri-Test (Enterotube® II) 8

Exercise 5-31: Bacterial Unknowns Project 10

Section 6: Quantitative Techniques 50

Section 6 Introductory Pages 2

Exercise 6-1: Standard Plate Count (Viable Count) 10

Exercise 6-2: Urine Culture 4

Exercise 6-3: Direct Count (Petroff-Hausser) 4

Exercise 6-4: Closed-System Growth 10

Exercise 6-5: Plaque Assay of Virus Titre 10

Exercise 6-6: Thermal Death Time Versus Decimal Reduction Value 10

Section 7: Medical Microbiology 64

Section 7 Introductory Pages 2

Exercise 7-1: Snyder Test 4

Exercise 7-2: Lysozyme Assay 6

Exercise 7-3: Antimicrobial Susceptibility Test: Disk Diffusion (Kirby-Bauer) Method 8

Exercise 7-4: Clinical Biofilms 4

Exercise 7-5: Morbidity and Mortality Weekly Report (MMWR) Assignment 10

Exercise 7-6: Epidemic Simulation 6

Exercise 7-7: Identification of Enterobacteriaceae 8

Exercise 7-8: Identification of Gram-positive Cocci 8

Exercise 7-9: Identification of Gram-positive Rods 8

Section 8: Environmental Microbiology 84

Section 8 Introductory Pages 2

Exercise 8-1: Winogradsky Column 6

Exercise 8-2: Nitrogen Fixation (includes pgs. 589-590) 10

Exercise 8-3: Nitrification: The Production of Nitrate 6

Exercise 8-4: Ammonification 4

Exercise 8-5: Denitrification: Nitrate Reduction 4

Exercise 8-6: Photosynthetic Sulfur Bacteria (includes pgs. 613-614) 8

Exercise 8-7: Chemolithotrophic Sulfur-Oxidizing Bacteria 6

Exercise 8-8: Sulfur-Reducing Bacteria 6

Exercise 8-9: Bioluminescence 4

Exercise 8-10: Soil Slide Culture 4

Exercise 8-11: Soil Microbial Count 6

Exercise 8-12: Membrane Filter Technique 8

Exercise 8-13: Multiple Tube Fermentation Method for Determining Water Quality 10

Section 9: Food Microbiology 20

Section 9 Introductory Pages 2

Exercise 9-1: Methylene Blue Reductase Test 4

Exercise 9-2: Making Yogurt 4

Exercise 9-3: Yogurt CFU Density of an Active and Live Culture and Identification of a Yogurt Unknown 10

Section 10: Microbial Genetics 58

Section 10 Introductory Pages 2

Exercise 10-1: Extraction of DNA from Bacterial Cells 6

Exercise 10-2: Restriction Digest 12

Exercise 10-3: Bacterial Transformation: The pGLO^{TM} System 8

Exercise 10-4: Polymerase Chain Reaction 14

Exercise 10-5: Ultraviolet Radiation Damage and Repair 6

Exercise 10-6: Ames Test 6

Exercise 10-7: Phage Typing of E. coli Strains 4

Section 11: Hematology and Serology 38

Section 11 Introductory Pages 2

Exercise 11-1: Differential Blood Cell Count 6

Exercise 11-2: Precipitin Ring Test (includes pgs. 751-752) 6

Exercise 11-3: Double Gel Immunodiffusion 4

Exercise 11-4: Slide Agglutination 4

Exercise 11-5: Blood Typing 6

Exercise 11-6: ELISA for Detecting Antibodies in a Patient's Sample (The Antibody Capture Method) 10

Section 12: Eukaryotic Microbes 46

Section 12 Introductory Pages 2

Exercise 12-1: Representative Fungi of Medical and Economic Importance 12

Exercise 12-2: Fungal Slide Culture 6

Exercise 12-3: Examination of Simple Eukaryotic Pathogens 12

Exercise 12-4: Parasitic Helminths 14

Appendix A: Biochemical Pathways 8

Appendix B: Miscellaneous Transfer Methods 4

Appendix C: Transfer from a Broth Culture Using a Glass Pipette 4

Appendix D: Transfer from a Broth Culture Using a Digital Pipette 4

Appendix E: The Spectrophotometer 4

Appendix F: Alternative Procedures for Section 6 6

Appendix G: Agarose Gel Electrophoresis 6

Appendix H: Medium, Reagent, and Stain Recipes 18

Glossary 8

Index 8