Laboratory Exercises in Microbiology – 10th Edition

PART ONE Microscopic Techniques

۱. Bright-Field Light Microscope (Dark Image, Brighter Background) and Microscopic Measurement of Organisms

۲. The Hanging Drop Slide and Bacterial Motility

۳. Dark-Field Light Microscope: Bright Image, Dark Background

۴. Phase-Contrast Light Microscope

۵. Fluorescence Microscope

PART TWO Bacterial Morphology and Staining

۶. Negative Staining

۷. Smear Preparation and Simple Staining

۸. Gram Stain

۹. Acid-Fast Staining (Ziehl-Neelsen and Kinyoun) Procedures

۱۰. Endospore Staining (Schaeffer-Fulton or Wirtz-Conklin)

۱۱. Capsule Staining

۱۲. Flagella Staining: West and Difco’s SpotTest Methods

PART THREE Basic Laboratory and Culture Techniques

۱۳. Sterilization and Microbiological Culture Media Preparation

۱۴. Culture Transfer Instruments, Techniques, and Isolation and Maintenance of Pure Cultures

۱۵. Spread-Plate Technique

۱۶. The Streak-Plate Technique: Selective and Differential Media

۱۷. Pour-Plate Technique

۱۸. Cultivation of Anaerobic Bacteria

۱۹. Determination of Bacterial Numbers

PART FOUR Biochemical Activities of Bacteria

۲۰. Carbohydrates I: Fermentation and β-Galactosidase Activity

۲۱. Carbohydrates II: Triple Sugar Iron Agar Test

۲۲. Carbohydrates III: Starch Hydrolysis

۲۳. Lipids: Lipid Hydrolysis

۲۴. Proteins, Amino Acids, and Enzymes I: Hydrogen Sulfide Production and Motility

۲۵. Proteins, Amino Acids, and Enzymes II: The IMViC Tests

۲۶. Proteins, Amino Acids, and Enzymes III: Casein Hydrolysis

۲۷. Proteins, Amino Acids, and Enzymes IV: Gelatin Hydrolysis

۲۸. Proteins, Amino Acids, and Enzymes V: Catalase Activity

۲۹. Proteins, Amino Acids, and Enzymes VI: Coagulase and DNase Activity

۳۰. Proteins, Amino Acids, and Enzymes VII: Oxidase Test and Bioluminescence

۳۱. Proteins, Amino Acids, and Enzymes VIII: Urease Activity

۳۲. Proteins, Amino Acids, and Enzymes IX: Lysine and Ornithine Decarboxylase Tests

۳۳. Proteins, Amino Acids, and Enzymes X: Phenylalanine Deamination

۳۴. Proteins, Amino Acids, and Enzymes XI: Nitrate Reduction

PART FIVE Rapid Multitest Systems

۳۵. The API® 20E System

۳۶. The EnteroPluri-Test System

PART SIX Unknown Identification

۳۷. Using the First Edition of Bergey’s Manual of Systematic Bacteriology to Identify Bacteria

۳۸. General Unknown

PART SEVEN Environmental Factors Affecting Growth of Microorganisms

۳۹. Temperature

۴۰. pH

۴۱. Osmotic Pressure

۴۲. The Effects of Chemical Agents on Bacteria I: Disinfectants and Other Antimicrobial Products

۴۳. The Effects of Chemical Agents on Bacteria II: Antimicrobial Agents (Kirby-Bauer Method) and the Etest®

۴۴. Hand Washing, Environmental Sampling, and Microbiological Monitoring

۴۵. Determination of a Bacterial Growth Curve: Classical and Two-Hour Methods

PART EIGHT Environmental and Food Microbiology

۴۶. Standard Coliform Most Probable Number (MPN) Test and Colilert®-۱۸ Coliform Test

۴۷. Membrane Filter Technique for Coliforms and Fecal Streptococci; KONFIRMTM Test for Fecal Coliforms

۴۸. Isolation of Escherichia coli Bacteriophages from Sewage and Determining Bacteriophage Titers

۴۹. Enumeration of Soil Microorganisms

۵۰. Bacterial Count of a Food Product

۵۱. Examination of Milk for Bacteria

PART NINE Medical Microbiology

۵۲. Hemagglutination Reactions: Blood Groups

۵۳. Isolation of Normal Microbiota from the Human Body

۵۴. Staphylococci

۵۵. Pneumococci

۵۶. Streptococci

۵۷. Neisseriae

۵۸. Aerobic and Anaerobic Endospore-Forming Bacteria

PART TEN Survey of Selected Eucaryotic Microorganisms

۵۹. Fungi (Ascomycota; also Ascomycetes) I: Yeasts

۶۰. Fungi (Eumycota) II: Zygomycota (Rhizopus), Ascomycota (Penicillium), and Basidiomycota(Agaricus)

PART ELEVEN Microbial Genetics and Genomics

۶۱. Bacterial Mutation

۶۲. Bacterial Transformation

۶۳. Bacterial Conjugation: The Transfer of Antibiotic-Resistant Plasmids

۶۴. Isolation of Genomic DNA from Baker’s Yeast (Saccharomyces cerevisiae)

۶۵. Isolation and Purifi cation of Genomic DNA from Escherichia coli

PART TWELVE Scientific Inquiry

۶۶. Independent Group Projects

۶۷. Case Studies

Appendix A Dilutions with Sample Problems

Appendix B Transmission-Absorbance Table for Spectrophotometry

Appendix C Logarithms; pH and pH Indicators

Appendix D Scientifi c Notation

Appendix E Identifi cation Charts

Appendix F Reagents, Solutions, Stains, and Tests

Appendix G Culture Media Appendix H Sources and Maintenance of Microbiological Stock Cultures

Appendix I Laboratory Skills and Laboratory Thinking Skills

Appendix J Orientation to the Laboratory: Rules of Conduct and General Safety

Appendix K Summary of Universal Precautions and Laboratory Safety Procedures

Appendix L Biosafety Level (BSL)

Index