Laboratory Exercises in Microbiology

John P. Harley, Lansing M. Prescott0072333456, 9780072333459

Like the text, the laboratory manual provides a balanced introduction to laboratory techniques and principles that are important in each area of microbiology.

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Table of contents :
Cover Page……Page 1
P R E F A C E……Page 2
Acknowledgments……Page 7
Orientation to the Laboratory: Rules of Conduct and General Safety……Page 8
Summary of Universal Precautions and Laboratory Safety Procedures……Page 10
Part I -Microscopic Techniques……Page 12
1. Bright-Field Light Microscope and Microscopic Measurement of Organisms……Page 13
2. The Hanging Drop Slide and Bacterial Motility……Page 24
3. Dark-Field Light Microscope……Page 28
4. Phase-Contrast Light Microscope……Page 32
5. Fluorescence Microscope……Page 37
Part II – Bacterial Morphology and Staining……Page 41
6. Negative Staining……Page 42
7. Smear Preparation and Simple Staining……Page 47
8. Gram Stain……Page 53
9. Acid-Fast Staining (Ziehl-Neelsen and Kinyoun) Procedures……Page 60
10. Endospore Staining (Schaeffer-Fulton or Wirtz-Conklin)……Page 65
11. Capsule Staining……Page 70
12. Flagella Staining:West and Difco’s SpotTest Methods……Page 75
Part III – Basic Laboratory and Culture Techniques……Page 81
13. Microbiological Culture Media Preparation and Sterilization……Page 82
14. Culture Transfer Instruments, Techniques, and Isolation of Pure Cultures and Their Maintenance……Page 89
15. Spread-Plate Technique……Page 98
16. The Streak-Plate Technique and Differential Media……Page 103
17. Pour-Plate Technique……Page 109
18. Cultivation of Anaerobic Bacteria……Page 113
19. Determination of Bacterial Numbers……Page 121
Part IV – Biochemical Activities of Bacteria……Page 129
20. Carbohydrates I: Fermentation and beta-Galactosidase Activity……Page 130
21. Carbohydrates II:Triple Sugar Iron Agar Test……Page 137
22. Carbohydrates III: Starch Hydrolysis……Page 143
23. Lipids: Lipid Hydrolysis……Page 147
24. Proteins, Amino Acids, and Enzymes I: Hydrogen Sulfide Production and Motility……Page 151
25. Proteins, Amino Acids, and Enzymes II: The IMViC Tests……Page 156
26. Proteins,Amino Acids, and Enzymes III: Casein Hydrolysis……Page 164
27. Proteins, Amino Acids, and Enzymes IV: Gelatin Hydrolysis……Page 168
28. Proteins, Amino Acids, and Enzymes V: Catalase Activity……Page 172
29. Proteins, Amino Acids, and Enzymes VI: Coagulase and DNase Activity……Page 176
30. Proteins,Amino Acids, and Enzymes VII: Oxidase Test……Page 181
31. Proteins, Amino Acids, and Enzymes VIII: Urease Activity……Page 186
32. Proteins, Amino Acids, and Enzymes IX: Lysine and Ornithine Decarboxylase Test……Page 190
33. Proteins, Amino Acids, and Enzymes X: Phenylalanine Deamination……Page 196
34. Proteins, Amino Acids, and Enzymes XI: Nitrate Reduction……Page 201
Part V – Rapid Multitest Systems……Page 206
35. The API 20E System……Page 207
36. The Enterotube II System……Page 214
Part VI – Unknown Identification……Page 221
37. Using Bergey’s Manual of Systematic Bacteriology to Identify Bacteria……Page 222
38. General Unknown……Page 227
Part VII – Environmental Factors Affecting Growth of Microorganisms……Page 234
39. Temperature……Page 235
40. pH……Page 239
41. Osmotic Pressure……Page 243
42. The Effects of Chemical Agents on Bacteria I: Disinfectants……Page 247
43. The Effects of Chemical Agents on Bacteria II: Antimicrobial Agents (Kirby-Bauer Method)……Page 252
44. Hand Washing, Environmental Sampling, and Microbiological Monitoring……Page 258
45. Determination of a Bacterial Growth Curve: Classical and Two-Hour Methods……Page 265
Part VIII – Environmental and Food Microbiology……Page 276
46. Standard Coliform Most Probable Number (MPN) Test and Presence-Absence Coliform Test……Page 278
47. Membrane Filter Technique for Coliforms and Fecal Streptococci; KONFIRM Test for Fecal Coliforms……Page 284
48. Isolation of Escherichia coli Bacteriophages from Sewage and Determining Bacteriophage Titers……Page 291
49. Enumeration of Soil Microorganisms……Page 297
50. Bacterial Count of a Food Product……Page 302
51. Examination of Milk for Bacteria……Page 306
Part IX – Medical Microbiology……Page 312
52. Agglutination Reactions: Blood Groups……Page 313
53. Isolation of Normal Microbiota from the Human Body……Page 318
54. Staphylococci……Page 324
55. Pneumococci……Page 332
56. Streptococci……Page 337
57. Neisseriae……Page 344
58. Aerobic and Anaerobic Endospore-Forming Bacteria……Page 349
Part X – Survey of Selected Eucaryotic Microorganisms……Page 354
59. Fungi I:Yeasts……Page 355
60. Fungi II: Phycomycetes,Ascomycetes, and Basidiomycetes……Page 359
Part XI – Microbial Genetics and Genomics……Page 366
61. Bacterial Mutation……Page 367
62. Bacterial Transformation……Page 371
63. Bacterial Conjugation: The Transfer of Antibiotic-Resistant Plasmids……Page 376
64. Isolation of Genomic DNA from Saccharomyces cerevisiae……Page 382
65. Isolation and Purification of Genomic DNA from Escherichia coli……Page 386
66. Identifying Archaea and Bacteria Using the Internet and Computer-Assisted Gene Analysis……Page 392
Appendix A – Dilutions with Sample Problems……Page 399
Appendix B – Metric and English Measurement Equivalents……Page 409
Appendix C – Transmission-Absorbance Table for Spectrophotometry……Page 411
Appendix D – Logarithms……Page 412
Appendix E – pH and pH Indicators……Page 413
Appendix F – Scientific Notation……Page 414
Appendix G – Identification Charts……Page 416
Appendix H – Reagents, Solutions, Stains, and Tests……Page 421
Appendix I – Culture Media……Page 425
Appendix J – Sources and Maintenance of Microbiological Stock Cultures……Page 432
SPECIFIC GUIDELINES FOR LAB SAFETY……Page 436
CREDITS……Page 437
INDEX……Page 438