Laboratory Exercises in Microbiology

Häftad, Engelska, 2021

Av Robert A. Pollack, Lorraine Findlay, Walter Mondschein, R. Ronald Modesto

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The Laboratory Exercises in Microbiology, 5e by Pollack, et al. presents exercises and experiments covered in a 1 or 2-semester undergraduate microbiology laboratory course for allied health students. The labs are introduced in a clear and concise manner, while maintaining a student-friendly tone. The manual contains a variety of interactive activities and experiments that teach students the basic concepts of microbiology. The 5th edition contains new and updated labs that cover a wide array of topics, including identification of microbes, microbial biochemistry, medical microbiology, food microbiology, and environmental microbiology.

Produktinformation

Utgivningsdatum2021-03-02
Mått201 x 252 x 15 mm
Vikt567 g
FormatHäftad
SpråkEngelska
Antal sidor320
Upplaga5
FörlagJohn Wiley & Sons Inc
ISBN9781119462668

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Biologi inom Naturvetenskap och teknik

Preface iiiIntroduction: Laboratory Operations and Safety xiiiBasic Rules for Microbiology Laboratory Safety xvPart 1 General Microscopy and Aseptic Technique1 Laboratory Safety: Introduction to the Microscope 3Laboratory Safety and Procedures 4Laboratory Procedures 4Simple Stain Technique 5The Microscope 6Microscope Components (Plate 1) 7Eukaryotic Versus Prokaryotic Cells (Plates 2A, 2B) 10Laboratory Cleanup 10Microscope Cleanup 10Discards 11General Cleanup 11Speaking of Safety 112 Transfer and Isolation Techniques, Microbes in the Environment 17Transfer Technique 17Tube-to-Tube Transfers 18Basic Aseptic Technique Procedures 18Isolation Techniques: Streak Plate and Pour Plate (Optional) 22Streak Plate (Plate 3) 22Prepare the Following Streak Plates Using the Cultures Provided 25To Lap or Not to Lap— That is the Question 25Pour Plate (Plate 4) Optional, May be Done as a Demonstration 25Microbes in the Environment 26Human Environment: Procedure 27Classroom/School Environment: Procedure 27Incubation 27Results 27Laboratory Cleanup 28Discards 28General Cleanup 28Part 2 Microbial Morphology, Differential Stains3 Cultural and Cellular Morphology 33Cultural Characteristics of Bacteria 33Procedure: Plates (Plate 5) 34Procedure: Agar Slants 35Procedure: Broth (Plate 6) 35Microbial Cellular Morphology 35Smear Preparation 36Broth Preparation 37Agar Slant and Plate Preparation 37Simple Stain Procedure 37Results from the Simple Stain Procedure (Plates 7–16) 38Results from Prepared Slide 38Laboratory Cleanup 384 Bacterial Growth 43Factors Needed for Bacterial Growth 44Measuring Bacterial Growth 45Procedure for Preparation of Spread Plate 45Procedure for Demonstration of a Bacterial Growth Curve 475 Gram Stain and Acid-Fast Stain 53The Gram Stain 53The Gram Stain Technique (Traditional Method) (Plate 17A, B, C) 54Gram Stain: (Alternative Method) 55The Acid-Fast Stain Technique (Plate 18) 56Acid-Fast Technique 56Laboratory Cleanup 58Incubation 58Discards 58General Cleanup 58Slides and Microscopes 586 Endospore Stain, Capsule Stain, and the Hanging Drop Technique 61The Endospore Stain 61The Spore Stain Technique (Plates 19, 20) 62Results 63The Capsule Stain 63The Capsule Stain Technique (Plate 21) 63The Hanging Drop Technique 64Results 647 Fungi 69The Fungi 69Results 728 Viruses—Visualization and Enumeration 77Viruses 77Bacteriophage Enumeration 78Results 809 Parasitology 85Part 3 Microbial Control and Biochemistry10 Microbial Sensitivity Testing 97Part 1: Physical Methods Ultraviolet Light Sensitivity 98Sterilization by the Use of UV Light 98The Penetrating Power of UV Light (Plate 34) 99Heat Sensitivity (may be done as an alternative to the UV light procedure) 101Effect of Cold Temperature and Slow Freezing (Optional) 102Part 2: Chemical Methods Chemical Sensitivity 102Chemotherapeutic Agent Testing: The Kirby–Bauer Plate 103The Kirby–Bauer Technique 104Results 10711 Bacterial Biochemistry 113Carbohydrate Metabolizing Enzymes 114Results 115Carbohydrate Fermentation (Phenol Red Broth) Tubes (Plate 37) 115Amino Acid and Nitrogen Metabolism 116Results 117Decarboxylase Tubes (Plate 42) 11712 Gas Requirements of Microorganisms 123Growth of Anaerobes (may be done as a demonstration) 124Demonstration of Catalase 12513 Specialized Media 129Examples 130Inoculation of Blood Agar Plates for Demonstration of Alpha Hemolysis and Transport Medium 131Inoculation of Blood, Phenylethyl Alcohol, Mannitol Salt, Macconkey, and Eosin–Methylene Blue Agar Plates 132Results 133Inoculation of Triple Sugar Iron Agar and Sulfide–Indole Motility Medium 133Results 134Results of Transport Medium 134Part 4 Medical Microbiology14 Genetics 143Ames Test 144UV Light Procedure 145Kirby–Bauer Procedure 145Results: Ames Test (Plate 55) 145Results: UV Light Procedure (Plate 35) 146Results: Kirby–Bauer Test (Plates 36, 55) 14615 Epidemiology 149Handwashing Procedure (Done with Three Students) 150Results: Handwashing (Plate 56) 151Fomite and Direct Transmission of Microbes 151Results: Fomite and Direct Transmission 152Airborne Infections: Cough and Sneeze Plates 152Results: Airborne and Cough/Sneeze Plates 153Microbes in Makeup (Optional) 153Results: Microbes in Makeup 15316 Specimen-Handling Protocols 159Quantitative Urinalysis 160Results: Urine Samples 162Specimens from the Gastrointestinal Tract 163Results: GI Tract Isolation Technique 16317 Specific Laboratory Tests 169Catalase Test 171Alternative Method (to be done at the end of this exercise) 171Results: Catalase Test (Plate 46) 171Bacitracin Sensitivity—Demonstration 171Results: Bacitracin Sensitivity (Plate 57) 172Oxidase Test 172Results: Oxidase Test (Plate 59) 172Coagulase Test 172Rapid Slide Test 172Tube Test 172Results: Coagulase Tube Test (Plate 60) 172Novobiocin Sensitivity 172Results: Novobiocin Sensitivity 173Camp Test (Plate 58) 173Results: Camp 17318 Serology 179Elisa Test: Test for Toxins A and B from Clostridium Difficile 180Results 182Rapid Identification of Group A Antigen 182Results 184Differentiation of Streptococci Using Latex Agglutination 184Slidex Strepto-Kit® 184Results 186Procedure Controls 186Part 5 Identification of a Bacterial Unknown19 Identification of Enteric Pathogens: Traditional Methods 191Results 194IMV IC Test (See Plate 61) 194Results of Gelatin Hydrolysis 194Results of the Oxidase Test (See Plate 59) 19420 Identification of Enteric Pathogens: Rapid Identification Methods 201EnteroPluri Test (Plate 62) 201Compartment 1: Glucose Fermentation and Gas Production 202Compartments 2 and 3: Lysine and Ornithine Decarboxylation 202Compartment 4: Sulfide and Indole Production 202Compartments 5–8: Adonitol, Lactose, Arabinose, and Sorbitol Fermentation 202Compartment 9: Voges–Proskauer 202Compartment 10: Phenylalanine Deaminase and Dulcitol 202Compartment 11: Urea 202Compartment 12: Citrate 202Api® 20 E System (Plate 63) 203Microtube 1: Ortho-Nitrophenyl-Beta-D-Ortho-Nitrophenyl-Beta-D-Galactopyranoside (Onpg) 203Microtubes 2–4: Arginine Dihydrolase (Decarboxylase), Lysine Decarboxylase, and Ornithine Decarboxylase (Adh, Ldc, Odc) 203Microtube 5: Citrate (Cit) 203Microtube 6: Sulfide (H2s) 203Microtube 7: Urea (Ure) 203Microtube 8: Tryptophan Deaminase (Tda) 203Microtube 9: Indole (Ind) 203Microtube 10: Voges–Proskauer (Vp) 203Microtube 11: Gelatin Hydrolysis (Gel) 203Microtubes 12–20: Utilization of the Carbohydrates Glucose, Mannitol, Inositol, Sorbitol, Rhamnose, Saccharose (Sucrose), Melibiose, Amygdalin, and Arabinose (Glu, Man, Ino, Sor, Rha, Sac, Mel, Amy, And Ara) 203Inoculation of Rapid Identification Media 204Inoculation of EnteroPluri Test 204Inoculation of the Api 20 E System 204Results 20521 Identification of a Bacterial Unknown: The Gram-Negative Unknown 209Review 209Comparison Chart 210Flowchart 210Computer Identification 210ID Value Worksheet for EnteroPluri Test II 212Results 21322 Identification of a Bacterial Unknown: The Gram-Positive Cocci 217Part 6 Food and Environmental Microbiology23 Identification and Quantitation of Microbial Numbers in a Water Sample 225Presumptive Test: Analysis of a Contaminated Water Sample for the Presence of Coliforms 226Results 227Quantitation of Microbial Number in a Water Sample 227Results (Plate 64) 22824 Identification of Microbes in Beef and Poultry and the Quantitation of Microbial Numbers 231Microbial Presence in a Food Product and Colony Counts 232Results 233Recognition of Organism Genera 23325 Soil Microbiology 237Part 1: The Recovery of Microorganisms from Soil 238Results 239Isolation Of Soil Bacteria 239Results 239Part 2: Isolation of Soil Bacteriophages of Bacillus Subtilis 239Results 24026 Microbial Ecology 245Competition Between Bacteria 246Results 246Bacteriocin Production 246Results 247Bacterial–Fungal Interaction (Plate 65) 247Results 24727 Biofilms 251Growth of Dental Plaque in situ 252Results 253Soil Biofilm 253Results 253Growth of Bacterial Biofilms in vitro 253Results 254Bibliography 261Answer Key for the Exercise Pre-Tests 263Appendix 1 Flow Charts and Comparison Charts for the Identification of Enteric Gram Negative Rods Using Traditional Testing Procedures 265Appendix 2 Flow Chart for the Identification of Enterobacteriaceae based on the API 20 E Rapid Identification System 269Appendix 3 Gram-Positive Flow Chart 275Photo Credit List 277Index 279Photographic Atlas PA-1