لایب سان

Introduction 1 A Tribute to the Past, a Moment in the Present, and an Openness to the Future 1 SECTION 2 Isolation Techniques and Selective Media 5 Streak Plate Methods of Isolation 5 Bacteroides Bile Esculin (BBE) Agar 7 Bismuth Sulfite Glucose Glycine Yeast (BiGGY) Agar 7 Chocolate II Agar 8 Columbia CNA With 5% Sheep Blood Agar 9 Desoxycholate Agar 9 Endo Agar 11 Eosin Methylene Blue Agar 11 Hektoen Enteric Agar 12 MacConkey Agar 13 Mannitol Salt Agar 14 Phenylethyl Alcohol Agar 15 Pseudomonas Isolation Agar 15 Salmonella–Shigella Agar 16 Tellurite Glycine Agar 16 TCBS Agar 17 Xylose Lysine Desoxycholate Agar 18 SECTION 3 Bacterial Growth 19 Growth Patterns on Agar 19 Growth Patterns in Broth 27 Aerotolerance 28 Cultivation of Anaerobes—Anaerobic Jar 29 Cultivation of Anaerobes—Thioglycollate Broth 30 Cultivation of Anaerobes—Cooked Meat Broth 30 SECTION 4 Microscopy 31 Types of Microscopy 31 SECTION 5 Bacterial Cellular Morphology and Simple Stains 37 Simple Stains 37 Negative Stain 38 A Gallery of Bacterial Cell Diversity 39 v Contents SECTION 6 Bacterial Cell Structures and Differential Stains 45 Gram Stain 45 Acid-fast Stain 49 Capsule Stain 51 Endospore Stain 51 Flagella Stain 54 Wet Mount and Hanging Drop Preparations 55 Miscellaneous Structures 56 SECTION 7 Differential Media 57 API 20 E Identification System for Enterobacteriaceae and Other Gram-negative Rods 57 Bacitracin Susceptibility Test 58 􀀀-Lactamase Test 59 Bile Esculin Test 60 Blood Agar 61 CAMP Test 62 Casein Hydrolysis Test 62 Catalase Test 63 Citrate Utilization Test 64 Coagulase Tests 65 BBL CRYSTAL™ Identification System— Enteric/Nonfermenter (E/NF) 66 Decarboxylation Test 67 DNase Test Agars 68 Enterotube® II 70 Fermentation Tests (Purple Broth and Phenol Red Broth) 71 Gelatinase Hydrolysis Test 73 Indole Test (SIM Medium) 74 Kligler’s Iron Agar 75 Lipase Test 77 Litmus Milk Medium 78 Lysine Iron Agar 80 Malonate Test 81 Methyl Red Test 82 Motility Test 82 Nitrate Reduction Test 83 Novobiocin Susceptibility Test 85 ONPG Test 86 Optochin Susceptibility Test 87 Oxidase Test 87 Oxidation–Fermentation Test 89 Phenylalanine Deaminase Test 91 PYR Test 91 Starch Hydrolysis 92 Sulfur Reduction (SIM Medium) 93 SXT Susceptibility Test 94 Triple Sugar Iron Agar 95 Urease Tests 96 Voges-Proskauer Test 98 SECTION 8 Serology 99 Precipitation Reactions 99 Agglutination Reactions 101 Enzyme-Linked Immunosorbent Assay (ELISA) 103 Fluorescent Antibody (FA) Technique 105 Western Blot Technique 106 SECTION 9 Molecular Techniques 109 DNA Extraction 109 Electrophoresis 110 Polymerase Chain Reaction 111 DNA Sequencing 112 Ultraviolet Radiation Damage and Repair 114 Ames Test 115 SECTION 10 Viruses 117 Introduction to Viruses 117 Human Immunodeficiency Virus (HIV) 119 Viral Cytopathic Effects in Cell Culture 120 Hemadsorption in Cell Culture 122 vi 􀀀 A Photographic Atlas for the Microbiology Laboratory SECTION 11 Domain Bacteria 123 Phylum Aquificae 123 Phylum Thermotogae 123 Phylum “Deinococcus-Thermus” 124 Phylum Chloroflexi 125 Phylum Spirochaetes 125 Phylum Cyanobacteria 125 Phylum Proteobacteria 127 Phylum Firmicutes 130 Phylum Actinobacteria 131 SECTION 12 Bacterial Pathogens 133 Aeromonas hydrophila 133 Bacillus anthracis 134 Bacillus cereus 135 Bacteroides fragilis 136 Bordetella pertussis 136 Borrelia burgdorferi 137 Brucella melitensis 138 Campylobacter jejuni 139 Citrobacter spp. 139 Clostridium botulinum 140 Clostridium difficile 141 Clostridium perfringens 142 Clostridium septicum 142 Clostridium tetani 143 Corynebacterium diphtheriae 143 Escherichia coli 144 Fusobacterium spp. 145 Haemophilus influenzae 146 Helicobacter pylori 147 Klebsiella pneumoniae 147 Legionella pneumophila 148 Listeria monocytogenes 149 Mycobacterium leprae 150 Mycobacterium tuberculosis 150 Neisseria gonorrhoeae 151 Neisseria meningitidis 152 Nocardia asteroides 152 Proteus mirabilis 153 Pseudomonas aeruginosa 154 Salmonella enteritidis 155 Salmonella typhi 156 Shigella dysenteriae 156 Staphylococcus aureus 157 Staphylococcus epidermidis 158 Streptococcus agalactiae 159 Streptococcus mutans 159 Streptococcus pneumoniae 160 Streptococcus pyogenes 161 Treponema pallidum 162 Vibrio cholerae 162 Yersinia pestis 163 SECTION 13 Domain Archaea 165 Crenarchaeota 165 Euryarchaeota 166 SECTION 14 Domain Eukarya: Simple Eukaryotes 169 Group Excavata 169 Group Chromalveolata 170 Group Rhizaria 174 Group Archaeplastida 174 Group Unikonta 176 SECTION 15 Fungi of Clinical Importance 183 Yeasts of Medical Importance 183 Monomorphic Molds 185 Dimorphic Molds 190 SECTION 16 Protozoans of Clinical Importance 193 Group Excavata 193 Group Chromalveolata 196 Group Unikonta 200 SECTION 17 Parasitic Helminths 203 Trematode Parasites Found in Clinical Specimens 203 Cestode Parasites Found in Clinical Specimens 207 Nematode Parasites Found in Clinical Specimens 211 Contents 􀀀 vii SECTION 18 Quantitative Microbiology 217 Standard Plate Count (Viable Count) 217 Direct Count (Petroff-Hausser) 219 Plaque Assay for Determination of Phage Titer 220 Urine Streak—Semiquantitative Method 222 SECTION 19 Medical, Environmental, and Food Microbiology 223 Antimicrobial Susceptibility Test (Kirby-Bauer Method and E Test) 223 Sample Collection and Transport 225 Environmental Sampling: The RODAC™ Plate 226 Colilert® Method for Testing Drinking Water 227 Membrane Filter Technique 228 Multiple Tube Fermentation Method for Total Coliform Determination 229 Bioluminescence 231 Winogradsky Column 231 Nitrogen Cycle 232 Sulfur Cycle—Introduction 235 Methylene Blue Reductase Test 237 Snyder Test 237 SECTION 20 Host Defenses 239 Differential Blood Cell Count 239 Other Immune Cells and Organs 241 APPENDIX A Biochemical Pathways 243 Oxidation of Glucose: Glycolysis, Entner-Doudoroff, and Pentose-Phosphase Pathways 243 Oxidation of Pyruvate: The Krebs Cycle and Fermentation 246 References 251 Index 253

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