Steve K. Alexander Dennis Strete VW ioaiytl lo Pa Te Re eI Ya) DYContents 1 Microscopes and Their Use in the Study of Cells and Viruses 1 Lab Procedure: Bright-Field Microscope 1 Fluorescent Microscope 4 Electron Microscope 5 Scanning Electron Microscope (SEM) Transmission Electron Microscope (TEM) 5 2 Microscopic Observation of Bacteria, Fungi, and Protozoa 11 Bacteria 12 Size 12 Cell Shapes 12 Cell Arrangements 14 ‘Actinomycetes 17 Lab Procedure: Using Wet Mounts to Observe Bacteria 18 Fungi 19 Growth Forms and Reproduction 19 Divisions of Fungi 19 Using Wer Mounts to Observe Fungi 25 Protozoa 28 Protozoan Groups 28 3 Staining Bacteria 33 Lab Procedure: Preparing Cells for Staining 33 Morphological Stains 34 Lab Procedure: Simple Stain 34 Lab Procedure: Negative Stain 35 Differential Stains 35 Lab Procedure: Gram Stain 35 Lab Procedure: Acid-Fast Stain 40 Structural Stains 41 Lab Procedure: Capsule Stain 42 Lab Procedure: Endospore Stain 42 Lab Procedure: Flagella Stain 45 4 Cultivation of Bacteria and Fungi 47 Sample Collection 47 Environmental Samples 47 Clinical Samples 47 Incubation of Inoculated Media 49 ‘Temperature Considerations 49 [Atmospheric Considerations 50 Examination of Growth on Agar Media 51 Growth of Organisms from the Enviconment $1 Growth of Organisms from Clinical Samples 53 Examination of Growth in Broth 61 Colony Selection 62 Lab Procedure: Isolation by the Streak-Plate Method 63 Maintenance of Bacterial Cultures 68 Biochemical Tests for the Identification of Bacteria 69 Lab Procedure: Carbohydrate Utilization 69. Lab Procedure: Catalase Test 71 Lab Procedure: Citrate Utilization 72 Lab Procedure: Decarboxylase Tests 73 Lab Procedure: Denitrification 74 Lab Procedure: Gelatin Utilization 75 Lab Procedure: Hydrogen Sulfide (HS) Production 76 Indole Production 77 Litmus Milk Reactions 78 Aethyl Red Test 80 : Motility Test 81 xidase Test 82 Lab Procedure: Oxidation-Fermentation {O-F) Test 83 Lab Procedure: Phenylalanine Deaminase Test 85 (Contents continued) ixChapter 5, continued Lab Procedure: Skim Milk Utilization 86 Lab Procedure: Starch Hydrolysis Test 87 Lab Procedure: Triple Sugar Iron (TSI) Agar and Kligler Iron Agar (KIA) Tests 88 Lab Procedure: Urea Utilization 90 Lab Procedure: Voges-Proskauer Test 91 Bacterial Identification Methods 93 Identification Using Conventional Methods 93 Identification Using Multitest Systems 96 Identification Using Automated Methods 98 Evaluating Antibacterial Chemical Agents 101 Lab Procedure: Evaluating Handwashing Effectiveness 101 Lab Procedure: Evaluating Antisepties and Disinfectants: The Filter Paper Method 102 Antibiotic Effectiveness 104 Lab Procedure: Evaluating Antibioti The Kirby-Bauer Method 105 Lab Procedure: Evaluating Antibiotics: Etest® 108 Antibiotic Resistance 109 Soil and Water Microbiology 111 Lab Procedure: Microbial Enumeration By Plate Count 111 Lab Procedure: Most Probable Number (MPN) Method 116 Membrane Filter Method 120 Contents 9 Survey of Microbial Pathogens 121 Bacterial Pathogens 121 Fungal Pathogens 134 Protozoan Pathogens 138 10 Arthropod Vectors of Human Diseases 145 Vectors of Human Diseases 146 Acarines 146 Insects 148 11 Multicellular Parasites 151 Basie Structural Characteristics of Multicellular Parasites 151 Life Cycles of Multicellular Parasites Phylum Platyhelminthes (Flatworms) 155 Phylum Nematoda (Roundworms) 166 12. Clinical Applications of Host Responses to Infection 171 Identifying White Blood Cell Types 171 Lab Procedure: White Blood Cell Counts 174 ‘Organs and Tissues of the Immune System 176 Immunization 179 Lab Procedure: Antigen-Antibody Precipitation Reactions in Solution 179 Immunodiffusion Methods 181 Lab Procedure: Agglutination Reactions 183 Enzyme-Linked Immunosorbent Assay (ELISA) 184 SDS-Polyacrylamide Gel Electrophoresis and Western Blot 185 Index 187