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Overview

This laboratory manual for allied health or general microbiology has been written with the student in mind. The authors have used their years of teaching microbiology and microbiology laboratory at all levels to identify and relate the fundamental concepts that are important to the understanding of the science and students’ success in their future field. They have included case studies to exemplify the relevance of the science and extensive visual imagery to help students understand and learn the content. Most importantly, the authors hope this manual will help students experience the thrill of bench science and share some of the enthusiasm they have for microbiology, a field of science that is dynamic, exciting and touches every aspect of your life. The third edition lab manual compliments content covered in Cowan’s Microbiology Fundamentals: A Clinical Approach, 3/e

Table of Contents

Laboratory Overview: Introduction

Laboratory Safety, Equipment, and Materials

Tools of the Laboratory: Basic Techniques

Ex 1 Use of the Microscope

Ex 2 Aseptic Techniques

Ex 3 Isolation Streak Plate

Eukaryotic Cells and Microorganisms

Ex 4 Microbial Phototrophs: Algae and Cyanobacteria

Ex 5 Protozoans

Ex 6 The Fungi

Ex 7 Bacterial Motility: Hanging Drop Method

Methods for Microscopic Analysis: Staining

Ex 8 Simple Staining and Smear Preparation

Ex 9 Negative Stain

Ex 10 Capsule Stain

Ex 11 Gram Stain

Ex 12 Acid-Fast Staining (Ziehl-Neelsen and Kinyoun Methods)

Ex 13 Endospore Stain (Schaeffer-Fulton Method)

Methods for Culturing and Diagnosing Microbes: Media

Ex 14 Blood Agar

Ex 15 Mannitol Salt Agar

Ex 16 Eosin Methylene Blue Agar

Ex 17 MacConkey Agar

Microbial Nutrition, Ecology, and Growth

Ex 18 Chemical Content of Media

Ex 19 Osmotic Pressure and Growth

Ex 20 Oxygen and Growth: Aerotolerance

Ex 21 Oxygen and Growth: Catalase

Ex 22 Oxidase

Food and Water Safety

Ex 23 Plate Count

Treatment and Control of Microbial Growth

Ex 24 Killing by Ultraviolet Light

Ex 25 Antimicrobial Susceptibility Testing (Kirby-Bauer Method)

Microbial Genetics and Genetic Engineering

Ex 26 ELISA and Using Nucleotide BLAST (Dry Lab)

Ex 27 Analysis of DNA: Electrophoresis

Ex 28 Gene Transfer: Transformation

Infectious Diseases

Ex 29 Parasitic Protozoa

Ex 30 Parasitic Worms

Ex 31 Transmission: Vectors of Disease

Diagnosing Infections

Ex 32 Gel Immunoprecipitation (Immunodiffusion)

Ex 33 Passive (Indirect) Agglutination

Ex 34 Rapid Immunoassays

Ex 35 Hemagglutination Inhibition Assay

Ex 36 EnteroPluri-Test

Environmental Microbiology

Ex 37 Soil Microbiology

Hydrolytic Enzymes and Disease Mechanisms

Ex 38 DNase Test

Ex 39 Gelatin Hydrolysis

Ex 40 Starch Hydrolysis

Ex 41 Casein Hydrolysis

Biochemical Testing and Diagnosis of Disease

Ex 42 Phenol Red Broth

Ex 43 Triple Sugar Iron Agar

Ex 44 Litmus Milk

Ex 45 IMViC (Indole, Methyl Red, Voges-Proskauer, Citrate Reactions)

Ex 46 Urease Test

Ex 47 Nitrate Reduction

Putting It All Together

Ex 48 Known Bacteria for Biochemical Testing

Ex 49 Identification of Unknown Bacteria

Index

Author Bios

About the Author

Steven Obenauf

Steven Obenauf, Ph.D., has been teaching microbiology lectures and laboratories at Broward College since 1993. For six years before moving to the Fort Lauderdale area, he taught medical immunology and medical virology to first- and second-year medical students as well as first-year pharmacy students. His dedication to his students has led him to be a recipient of both Professor of the Year and Endowed Teaching Chair awards for undergraduate teaching and the “Golden Apple” teaching award bestowed by the graduating medical class. Dr. Obenauf is a registered microbiologist and belongs to both the American Society for Microbiology (ASM) and National Association of Biology Teachers. For 10 years, he served as the secretary and newsletter editor for the Florida branch of ASM. He loves using various forms of technology as part of his teaching and has given workshops and mentored other faculty at his college on in-class and online technology tools. Dr. Obenauf is currently Professor of Biological Sciences at Broward College. This manual brings together two of his passions: microbiology and photography

Susan Finazzo

Susan Finazzo, M.S., Ph.D., has a diverse academic background with degrees in biology, geology, microbiology, and horticulture. Her background, clinical work experience, and innate interest in the living world have allowed her to teach a broad variety of subjects. Her first academic love has always been microbiology. Throughout the past 20 years, Dr. Finazzo has taught students at both the 2-year college level and university level. She is a member of the National Association of Biology Teachers and has held several leadership positions including serving as president in 2017. She has been nominated for several teaching and service awards, and has twice been the recipient of an Endowed Teaching Award. Dr. Finazzo is very interested in new pedagogical approaches and tools to address the learning styles of the millennial student.

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