Benson's Microbiological Applications Laboratory Manual in General Microbiology

Preface......Page 2
Laboratory Protocol......Page 3
PART 1 Microscopy......Page 6
1 Brightfield Microscopy......Page 7
2 Darkfield Microscopy......Page 13
3 Phase-Contrast Microscopy......Page 15
4 Fluorescence Microscopy......Page 21
5 Microscopic Measurements......Page 26
PART 2 Survey of Microrganisms......Page 29
6 Protozoa, Algae, and Cyanobacteria......Page 30
7 Microscopic Invertebrates......Page 39
8 Aseptic Technique......Page 42
9 The Bacteria......Page 49
10 The Fungi: Yeasts and Molds......Page 51
PART 3 Microscope Slide Techniques (Bacterial Morphology)......Page 57
11 Negative Staining......Page 58
12 Smear Preparation......Page 60
13 Simple Staining......Page 64
14 Capsular Staining......Page 65
15 Gram Staining......Page 66
16 Spore Staining: Two Methods......Page 69
17 Acid-Fast Staining: Ziehl-Neelsen Method......Page 71
18 Acid-Fast Staining: Fluorescence Method......Page 72
19 Motility Determination......Page 74
PART 4 Culture Methods......Page 77
20 Culture Media Preparation......Page 78
21 Pure Culture Techniques......Page 84
22 Cultivation of Anaerobes......Page 91
23 Bacterial Population Counts......Page 95
24 Slide Culture: Autotrophs......Page 101
25 Slime Mold Culture......Page 102
26 Slide Culture: Molds......Page 105
27 Isolation of Anaerobic Phototrophic Bacteria: Using the Winogradsky Column......Page 108
PART 5 Bacterial Viruses: Isolation and Propagation......Page 112
28 Isolation of Phage from Sewage......Page 114
29 Isolation of Phage from Flies......Page 117
30 Burst Size Determination: A One-Step Growth Curve......Page 121
PART 6 Microbila Interrelationships......Page 126
31 Bacterial Commensalism......Page 127
32 Bacterial Synergism......Page 128
33 Microbial Antagonism......Page 129
PART 7 Environmental Influences and Control of Microbial Growth......Page 130
34 Temperature: Effects on Growth......Page 131
35 Temperature: Lethal Effects......Page 133
36 pH and Microbial Growth......Page 135
37 Osmotic Pressure and Bacterial Growth......Page 136
38 Oligodynamic Action......Page 137
39 Ultraviolet Light: Lethal Effects......Page 138
40 Evaluation of Disinfectants: The Use-Dilution Method......Page 140
41 Evaluation of Alcohol: Its Effectiveness as a Skin Degerming Agent......Page 142
42 Evaluation of Antiseptics: The Filter Paper Disk Method......Page 144
43 Antimicrobic Sensitivity Testing: The Kirby-Bauer Method......Page 146
44 Effectiveness of Hand Scrubbing......Page 149
PART 8 Identification of Unknown Bacteria......Page 152
45 Preparation and Care of Stock Cultures......Page 153
46 Morphological Study of Unknown......Page 155
47 Cultural Characteristics......Page 158
48 Physiological Characteristics: Oxidation and Fermentation Tests......Page 162
49 Physiological Characteristics: Hydrolytic Reactions......Page 171
50 Physiological Characteristics: Miscellaneous Tests......Page 175
51 Use of Bergey’s Manual and Identibacter Interactus......Page 178
PART 9 Miniaturized Multitest Systems......Page 183
52 Enterobacteriaceae Identification: The API 20E System......Page 184
53 Enterobacteriaceae Identification: The Enterotube II System......Page 188
54 O/F Gram-Negative Rods Identification: The Oxi/Ferm Tube II System......Page 193
55 Staphylococcus Identification: The API Staph-Ident System......Page 197
PART 10 Microbiology of Soil......Page 200
56 Microbial Population Counts of Soil......Page 201
57 Isolation of an Antibiotic Producer from Soil......Page 202
58 The Nitrogen Cycle......Page 205
59 Nitrogen-Fixing Bacteria......Page 207
60 Ammonification in Soil......Page 211
61 Isolation of a Denitrifier from Soil: Using Nitrate Agar......Page 212
62 Isolation of a Denitrifier from Soil: Using an Enrichment Medium......Page 215
PART 11 Microbiology of Water......Page 219
63 Bacteriological Examination of Water: Qualitative Tests......Page 220
64 The Membrane Filter Method......Page 224
65 Standard Plate Count: A Quantitative Test......Page 226
PART 12 Microbiology of Milk and Food Products......Page 227
66 Standard Plate Count of Milk......Page 228
67 Direct Microscopic Count of Organisms in Milk: The Breed Count......Page 229
68 Reductase Test......Page 232
69 Bacterial Counts of Foods......Page 234
70 Microbial Spoilage of Canned Food......Page 235
71 Microbial Spoilage of Refrigerated Meat......Page 237
72 Microbiology of Alchohol Fermentation......Page 239
73 Microbiology of Yogurt Production......Page 241
PART 13 Bacterial Genetic Variations......Page 243
74 Mutant Isolation by Gradient Plate Method......Page 244
75 Mutant Isolation by Replica Plating......Page 246
76 Bacterial Mutagenicity and Carcinogenesis: The Ames Test......Page 248
PART 14 Medical Microbiology and Immunology......Page 251
77 A Synthetic Epidemic......Page 252
78 The Staphylococci: Isolation and Identification......Page 255
79 The Streptococci: Isolation and Identification......Page 260
80 Gram-Negative Intestinal Pathogens......Page 268
81 Urinary Tract Pathogens......Page 272
82 Slide Agglutination Test: Serological Typing......Page 277
83 Slide Agglutination (Latex) Test: For S. aureus Identification......Page 279
84 Tube Agglutination Test: The Heterophile Antibody Test......Page 281
85 Tube Agglutination Test: The Widal Test......Page 283
86 Phage Typing......Page 285
87 White Blood Cell Study: The Differential WBC Count......Page 286
88 Total WBC Count......Page 290
89 Blood Grouping......Page 294
90 The Snyder Caries Susceptibility Test......Page 297
1, 2......Page 299
3-5......Page 301
6......Page 303
8, 9......Page 307
10......Page 309
11–14......Page 311
15–18......Page 313
19......Page 315
20......Page 317
21......Page 319
22......Page 321
23......Page 323
24, 25......Page 325
27......Page 327
28, 29......Page 329
30......Page 331
31–33......Page 333
34......Page 335
35......Page 337
36......Page 339
37, 38......Page 341
39......Page 343
40......Page 345
41......Page 347
42......Page 349
43......Page 351
44......Page 353
45......Page 355
48–50......Page 357
52......Page 359
53......Page 361
54......Page 363
55......Page 365
56, 57......Page 367
59......Page 369
60......Page 371
61, 62......Page 373
63......Page 375
64, 65......Page 377
66, 67......Page 379
68......Page 381
69......Page 383
70, 71......Page 385
72, 73......Page 387
74, 75......Page 389
76......Page 391
77......Page 393
78......Page 395
79......Page 397
80......Page 401
81......Page 403
82, 83......Page 405
84, 85......Page 407
86......Page 409
87–89......Page 411
90......Page 413
Descriptive Charts......Page 415
Table I International Atomic Weights......Page 420
Table II Four-Place Logarithms......Page 421
Table III Temperature Conversion Table Centigrade to Fahrenheit......Page 423
Table IV Autoclave Steam Pressures and Corresponding Temperatures......Page 424
Table V Autoclave Temperatures as Related to the Presence of Air......Page 425
Table VI MPN Determination from Multiple Tube Test......Page 426
Table VII Significance of zones of inhibition in Kirby-Bauer Method of antimicrobic sensitivity testing (1995)......Page 427
Table VIII Indicators of Hydrogen Ion Concentration......Page 428
STAINS AND REAGENTS......Page 429
Appendix C - Media......Page 432
Appendix D - Identification Charts......Page 438
Chart I Interpretation of Test Results of API 20E System......Page 439
Chart II Symbol Interpretation of API 20E System......Page 440
Chart III Characterization of Gram-Negative Rods—The API 20E System......Page 441
Chart IV Characterization of Enterobacteriaceae—The Enterotube II System......Page 443
Chart V Reaction Interpretations for API Staph-Ident......Page 444
Chart VI Biochemistry of API Staph-Ident Tests......Page 445
Chart VII API Staph-Ident Profile Register......Page 446
Appendix E - The Streptococci: Classification, Habitat, Pathology, and Biochemical Characteristics......Page 447
Appendix F - Identibacter Interactus......Page 451
Index......Page 452