CHAPTER Ⅰ GENERAL BACTERIOLOGICAL APPARATUS AND PROCEDURES 1
Quality of Glass 1
Hard Glass 2
Soft Glass 2
Cleaning Glassware 2
Culture Tubes 3
Petri Dishes 4
Dilution Bottles and＂Water Blanks＂ 6
Fermentation Tubes 7
Baskets and Culture Tube Racks 10
Slides and Cover Glasses 10
Needles and Loops 13
Pipettes 14
Plugging Tubes，Flasks，etc 15
Incubators 16
Miscellaneous Apparatus 17
Sterilization and Disinfection 17
Dry Heat Methods 18
Incineration 19
Flaming 19
Hot Air Sterilization 19
Moist Heat 20
High Pressure Steam 22
Boiling 23
Free Flowing Steam 23
Sterilization by Filtration 24
Preparation and Storage of Sterile Apparatus 26
CHAPTER Ⅱ THE MICROSCOPE AND ITS USE 28
General Information 28
General Construction of the Microscope 28
Mechanical Parts 29
The Body with its Draw Tube 29
The Stage 30
The Pillar 31
The Coarse Adjustment 31
Care of the Coarse Adjustment 31
The Fine Adjustment 31
The Substage Diaphragm 32
Optical Parts 33
Magnification 33
Optical Glass 33
The Eyepieces 33
Which Eye to Use 33
The Objectives 34
Cover Glasses 36
To Attach the Objective 36
The Oil-immersion Objective 37
Care of Objectives and Eye Pieces 37
The Condenser 38
Purpose of the Condenser 38
Centering of Illumination 39
To Focus Condenser 39
The Mirror 41
Use of the Microscope 41
Finding an Object 42
Micrometry—Measurement of Microorganisms 46
CHAPTER Ⅲ CULTURE MEDIA 50
General Remarks 50
Standard Methods 50
Characteristics of a Good Medium 51
Constituents of Media 51
Water 51
Agar 52
Classification of Sugars 53
Gelatin 54
Peptones 54
Use of Indicators in Media 55
Classification of Bacteriological Culture Media 56
Liquid Media 56
Beef Extract Broth 56
Beef Infusion Broth 57
Carbohydrate Broths 57
Nitrate Broth 58
Stiritz＇s Malt Medium for Yeasts 58
Plain Milk 58
Litmus Milk 58
Brom-cresol-purple Milk 58
Whey Broth 59
Synthetic Media 59
Liquefiable Solid Media 59
Beef Extract Agar 59
Wort Agar for Yeasts 61
Lactic Acid Dextrose Agar for Yeasts 61
Gorodkowa＇s Agar Medium for Yeasts 62
Maneval＇s Modification of Gorodkowa＇s Agar Medium 62
Beef Extract Gelatin 62
Plain Gelatin 63
Carbohydrate Gelatin 63
Potato Slants 63
Carrot Slants 63
Dehydrated Media 64
Filling Tubes，etc 64
Reaction of Media and Cultures 64
Indicators 65
How is Reaction Expressed 66
Titration Methods 66
Measurement of Hydrogen Ion Concentration 67
The Electrometric Method 67
The Colorimetric Method 69
Action of Buffers，＂Buffer Index＂ 70
Method for Adjusting the Reaction of Culture Media，etc 71
Sterilization of Culture Media 73
Storage of Culture Media 74
References on Media and their Preparation 74
CHAPTER Ⅳ MICROSCOPIC EXAMINATION OF BACTERIA 75
Examination in Unstained Preparations 75
Hanging Drop Preparations 75
Lindner＇s Adhesion Culture 76
India Ink Preparations 78
Dark Field Illumination Examination 78
Examination in Stained Preparation 80
Stains and their Properties 80
Mordants 82
Theory of Staining 82
Preparation of Apparatus 83
Staining Solutions 85
Preparation of Film for Staining 85
General Microbiological Stains 86
Aqueous Alcoholic Stains 86
The Gram Stain 86
Hucker＇s Modification of the Gram Stain 89
Burke＇s Modification of the Gram Stain 89
Reaction of Some Common Bacteria to the Gram Stain 91
The Spore Stain 93
Neisser＇s Method for Staining Spores 93
Chromic Acid Method for Staining Spores 93
May＇s Spore Staining Procedure 93
Capsule Stains 94
Muir＇s Method 94
Welch＇s Method 94
Huntoon＇s Method 94
Hiss＇s Method 95
The Gins Method of Demonstrating Capsules 95
Acid Fast Staining 97
Flagella Stains 97
Loeffler＇s Flagella Stain 98
Plimmer and Paine＇s Modification of the Casares-Gil Flagella Staining Procedure 99
Van Ermengem＇s Flagella Stain 100
Staining of Granules 100
Preservation of Stained Preparations 101
Differential Counting of Living and Dead Bacteria 102
Bibliography 102
CHAPTER Ⅴ ISOLATION OF BACTERIA 103
Pure Cultures 103
Dilutions 103
Qualiative Method of Dilution 104
Dilution in Liquefiable Solid Media 105
Quantitative Method of Dilution 105
Preparation of Petri Dish Cultures 106
Method of Isolation of Pure Cultures 107
Inoculation of Liquid Media 109
Preparation of Stab Cultures 109
Preparation of Shake Cultures 110
Enumeration of Bacteria 110
Collection of Samples 111
Standard Agar Plate 111
Streak Plates 112
Disposal of Cultures 113
Anaerobic Methods 114
Fermentation Tube 114
Buchner＇s Method 114
Wright＇s Method 115
Replacement of Oxygen by Hydrogen 115
Dick＇s Method 115
Methods Using Ordinary Petri Dishes 116
Torrey＇s Method 116
Krumwiede and Pratt＇s Method 116
Frost＇s Method for the Study of Bacteria 117
CHAPTER Ⅵ PROCEDURES FOR STUDYING THE CHARACTERISTICS OF BACTERIA 121
Invigoration of Cultures 121
Sketches 122
Study of Morphological Characteristics 122
Vegetative Cells 122
Abnormal Forms 123
Motility 124
Endospores 125
Capsules 127
Study of Physiological Characteristics 128
Cultural Characteristics 128
Oxygen Relations 128
Staining Reactions 131
Nutrient Broth Cultures 132
Agar and Gelatin Colonies 134
Gelatin Stab 135
Growth on Potato and the Hydrolysis of Starch 136
Temperature Relations 139
Thermal Death Point 139
Action of Bacteria on Milk 140
Litmus Milk，Purple Milk，etc 141
Production of Hydrogen Sulfide 143
Nitrate Reduction 144
Detection of Indol Formation 145
Salkowski Test 145
Ehrlich＇s Test 145
Gore Modification of Ehrlich＇s Test 145
The Oxalic Acid Test for Indol 146
Fermentation Reactions 146
Detection of Gas Formation 146
Acid Formation 147
Pathogenicity to Animals 148
The Index Number 148
Agglutination Technic 148
The Microscopic Agglutination Reaction 149
The Macroscopic Agglutination Reaction 150
Study of an Unknown Organism 150
CHAPTER Ⅶ STUDY OF YEASTS 152
Outline of Procedures for Studying Yeasts 152
Detection of Ascospore Formation by Yeasts 153
Hansen＇s Gypsum Block Method 153
Maneval＇s Method 153
Staining of the Nucleus in Yeasts 154
Wager＇s Method 154
Demonstration of Glycogen in Yeasts 155
Demonstration of Fat in Yeasts 155
Exercise：Study of Compressed Yeast 155
Exercise：Preparation of Pure Culture of Yeast by Lindner＇s Method 156
Exercise：Demonstration of Budding of Yeasts 156
Exercise：Determination of the Fermenting Power of Yeasts 156
Exercise：Determination of the Fermenting Activity of Yeasts 157
Exercise：The Effect of Freezing on Yeasts 158
CHAPTER Ⅷ STUDY OF MOLDS 159
Outline of Observations to be Made on Molds 159
Himmel＇s Method for Studying Mold Mycelium 159
Exercise：Isolation of Oidium Lactis 160
Exercise：Isolation of Rhizopus nigricans 160
Exercise：Observations on Germination of Mold Spores 160
Exercise：Effect of Heat on Mold Spores 160
Exercise：The Effect of Freezing upon Mold Spores 161
CHAPTER Ⅸ PLANS FOR WORKING 163
Outlines for Studying Microorganisms in Classes 163
CHAPTER Ⅹ SPECIAL EXERCISES 196
Exercise：Characteristics of Unknown Organisms 196
Exercise：Bacteriological Examination of Milk 196
Exercise：Effect of Drying on Bacteria 197
Exercise：Effect of Sunlight on Bacteria 197
Exercise：Action of Disinfectants on Bacteria 198
Exercise：Bacteriological Examination of Water 199
Exercise：Determination of the Bacterial Content of Rain Water and Snow 200
Exercise：Determination of the Number and Types of Bacteria in Ice 200
Exercise：The Isolation and Study of Thermophilic Bacteria 201
Exercise：Molds on Bread，Fruits，etc 201
Exercise：Biological Test for Arsenic 201
Exercise：Bacteriological Examination of Dish Water 202
Exercise：To Determine the Types of Bacteria in Milk 202
Exercise：Study of the Influence of Freezing on Bacteria 202
Exercise：Isolation of Rhizobium leguminosarum(Bacillus radicicola) 203
Exercise：Study of the Types of Curd Formed in Milk 203
Exercise：Study of Bacteria in the Mouth 203
Exercise：Effect of Dyes on Bacteria 204
Exercise：Study of Bacteria in Air，Dust，etc 204
Exercise：Bacteria on the Hands 205
Exercise：Isolation of Bacteria from Boils or Pimples 205
Exercise：Bacteria on Coins and the Oligodynamic Action of Metals 205
Exercise：Examination of Candy 206
APPENDIX 211
Preparation of Reagents 211
Normal Sodium Hydroxide 211
Normal Hydrochloric Acid 211
Preparation of Staining Solutions 212
Methylene Blue—Saturated Aqueous Solution 212
Methylene Blue—Saturated Alcoholic Solution 212
Loeffler＇s Methylene Blue 212
Gentian Violet—Saturated Aqueous Solution 212
Gentian Violet—Stabilized(Stovall and Nichols，1916) 213
Gentian Violet—Stabilized(Kilduffe，1909) 213
Gentian Violet—Saturated Alcoholic Solution 213
Carbol Gentian Violet 213
Aniline Water Gentian Violet 213
Fuchsin Basic—Saturated Aqueous Solution 214
Fuchsin Basic—Saturated Alcoholic Solution 214
Carbol Fuchsin 214
Eosin—Aqueous Solution 214
Eosin—Alcoholic Solution 214
Safranine—Aqueous Solution 214
Neutral Red—Aqueous Solution 214
Bismarck Brown—Aqueous Solution 215
Haematoxylin 215
Lugol＇s Iodine Solution(Gram＇s) 215
Core Test for Indol，Reagents 215
Metric Equivalents 216
Azolitmin Solution 217
Andrade＇s Indicator 217
Solutions for La Motte Comparator Sets 217
Solutions for La Motte Hydrogen-ion Testing Sets 218
Preparation of Indicator Solutions 219
Indicators for the Determination of Hydrogen-ion Concentration 219
Comparison of Thermometer Scales 222
Index 231