Microbiology: A Laboratory Manual

Title Page......Page 6
Copyright Page......Page 7
Contents......Page 8
Preface......Page 11
Laboratory Safety......Page 14
Laboratory Protocol......Page 16
 Introduction......Page 18
 Experiment 1: Culture Transfer Techniques......Page 24
  Part A: Isolation of Discrete Colonies from a Mixed Culture......Page 32
  Part B: Isolation of Pure Cultures from a Spread-Plate or Streak-Plate Preparation......Page 35
 Experiment 3: Cultural Characteristics of Microorganisms......Page 42
 Introduction......Page 48
 Experiment 4: Microscopic Examination of Stained Cell Preparations......Page 50
 Experiment 5: Microscopic Examination of Living Microorganisms Using a Hanging-Drop Preparation or a Wet Mount......Page 58
 Introduction......Page 64
 Introduction 6: Preparation of Bacterial Smears......Page 68
 Introduction 7: Simple Staining......Page 74
 Introduction 8: Negative Staining......Page 80
 Introduction 9: Gram Stain......Page 84
 Introduction 10: Acid-Fast Stain......Page 92
  Part A: Spore Stain (Schaeffer-Fulton Method)......Page 98
  Part B: Capsule Stain (Anthony Method)......Page 100
Part 4: Cultivation of Microorganisms: Nutritional and Physical Requirements, and Enumeration of Microbial Populations......Page 108
 Experiment 12: Nutritional Requirements: Media for the Routine Cultivation of Bacteria......Page 110
 Experiment 13: Use of Differential, Selective, and Enriched Media......Page 116
 Experiment 14: Physical Factors: Temperature......Page 126
 Experiment 15: Physical Factors: pH of the Extracellular Environment......Page 132
 Experiment 16: Physical Factors: Atmospheric Oxygen Requirements......Page 136
 Experiment 17: Techniques for the Cultivation of Anaerobic Microorganisms......Page 142
 Experiment 18: Serial Dilution—Agar Plate Procedure to Quantitate Viable Cells......Page 148
 Experiment 19: The Bacterial Growth Curve......Page 156
 Introduction......Page 164
 Experiment 20: Extracellular Enzymatic Activities of Microorganisms......Page 166
 Experiment 21: Carbohydrate Fermentation......Page 172
 Experiment 22: Triple Sugar–Iron Agar Test......Page 178
 Experiment 23: Imvic Test......Page 184
  Part A: Indole Production Test......Page 185
  Part B: Methyl Red Test......Page 186
  Part C: Voges-Proskauer Test......Page 187
  Part D: Citrate Utilization Test......Page 188
 Experiment 24: Hydrogen Sulfide Test......Page 196
 Experiment 25: Urease Test......Page 200
 Experiment 26: Litmus–Milk Reactions......Page 204
 Experiment 27: Nitrate Reduction Test......Page 210
 Experiment 28: Catalase Test......Page 214
 Experiment 29: Oxidase Test......Page 218
  Part A: Decarboxylase Test......Page 222
  Part B: Phenylalanine Deaminase Test......Page 224
 Experiment 31: Genus Identification of Unknown Bacterial Cultures......Page 228
 Introduction......Page 234
 Experiment 32: Free-Living Protozoa......Page 236
 Experiment 33: Parasitic Protozoa......Page 242
 Introduction......Page 250
  Part A: Slide Culture Technique......Page 252
  Part B: Mold Cultivation on Solid Surfaces......Page 254
 Experiment 35: Yeast Morphology, Cultural Characteristics, and Reproduction......Page 260
 Experiment 36: Identification of Unknown Fungi......Page 268
 Introduction......Page 274
 Experiment 37: Cultivation and Enumeration of Bacteriophages......Page 278
 Experiment 38: Isolation of Coliphages from Raw Sewage......Page 284
 Experiment 39: Propagation of Isolated Bacteriophage Cultures......Page 290
 Introduction......Page 294
 Experiment 40: Physical Agents of Control: Moist Heat......Page 296
 Experiment 41: Physical Agents of Control: Electromagnetic Radiations......Page 302
 Experiment 42: Chemical Agents of Control: Chemotherapeutic Agents......Page 306
  Part A: The Kirby-Bauer Antibiotic Sensitivity Test Procedure......Page 307
  Part B: Synergistic Effect of Drug Combinations......Page 309
 Experiment 43: Determination of Penicillin Activity in The Presence and Absence of Penicillinase......Page 316
  Part A: Mic Determination Using a Spectrophotometer......Page 317
  Part B: MIC Determination Using a Plate Reader......Page 318
 Chemical Agents of Control: Disinfectants and Antiseptics......Page 322
  Part A: Disc Diffusion Testing of Disinfectants and Antiseptics......Page 325
  Part B: Modified Use Dilution Testing of Disinfectants and Antiseptics......Page 326
 Introduction......Page 332
 Experiment 45: Microbiological Analysis of Food Products: Bacterial Count......Page 334
  Part A: Alcohol Fermentation......Page 338
  Part B: Lactic Acid Fermentation......Page 340
Part 11: Microbiology of Water......Page 344
 Experiment 47: Standard Qualitative Analysis of Water......Page 346
 Experiment 48: Quantitative Analysis of Water: Membrane Filter Method......Page 354
 Introduction......Page 360
 Experiment 49: Microbial Populations in Soil: Enumeration......Page 362
 Experiment 50: Isolation of Antibiotic-Producing Microorganisms and Determination of Antimicrobial Spectrum of Isolates......Page 368
  Part A: Isolation of Antibiotic-Producing Microorganisms......Page 369
  Part B: Determination of Antimicrobial Spectrum of Isolates......Page 370
 Experiment 51: Isolation of Pseudomonas Species by Means of the Enrichment Culture Technique......Page 374
 Introduction......Page 380
 Experiment 52: Enzyme Induction......Page 382
 Experiment 54: Bacterial Conjugation......Page 388
 Experiment 55: Isolation of a Streptomycinresistant Mutant......Page 394
 Experiment 56: The Ames Test: A Bacterial Test System for Chemical Carcinogenicity......Page 398
Part 14: Biotechnology......Page 404
 Experiment 56: Bacterial Transformation......Page 406
 Experiment 57: Isolation of Bacterial Plasmids......Page 414
 Experiment 58: Restriction Analysis and Electrophoretic Separation of Bacteriophage Lambda DNA......Page 424
Part 15: Medical Microbiology......Page 434
 Experiment 59: Microbial Flora of the Mouth: Determination of Susceptibility to Dental Caries......Page 436
  Part A: Isolation of Microbial Flora......Page 440
  Part B: Effectiveness of Handwashing......Page 444
 Experiment 61: Identification of Human Staphylococcal Pathogens......Page 452
 Experiment 62: Identification of Human Streptococcal Pathogens......Page 460
 Experiment 63: Identification of Streptococcus pneumoniae......Page 468
 Experiment 64: Identification of Enteric Microorganisms Using Computerassisted Multitest Microsystems......Page 474
 Experiment 65: Isolation and Presumptive Identification of Campylobacter......Page 484
 Experiment 66: Microbiological Analysis of Urine Specimens......Page 488
 Experiment 67: Microbiological Analysis of Blood Specimens......Page 494
 Experiment 68: Species Identification of Unknown Bacterial Cultures......Page 500
Part 16: Immunology......Page 508
 Experiment 69: Precipitin Reaction: The Ring Test......Page 510
 Experiment 70: Agglutination Reaction: The Febrile Antibody Test......Page 514
 Experiment 71: Enzyme-Linked Immunosorbent Assay......Page 520
  Part A: Rapid Plasma Reagin Test for Syphilis......Page 524
  Part B: Genital Herpes: Isolation and Identification of Herpes Simplex Virus......Page 526
  Part C: Detection of Sexually Transmitted Chlamydial Diseases......Page 527
Appendix 1: Scientific Notation......Page 532
Appendix 2: Methods for the Preparation of Dilutions......Page 534
Appendix 3: MicrobiologicalMedia......Page 536
Appendix 4: Biochemical TestReagents......Page 542
Appendix 5: Staining Reagents......Page 545
Appendix 6: ExperimentalMicroorganisms......Page 546
Art & Photo Credits......Page 548
Index......Page 550