Laboratory Methods in Microbiology

Laboratory Methods in Microbiology

1st Edition - January 1, 1966

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  • Authors: W. F. Harrigan, Margaret E. McCance
  • eBook ISBN: 9781483274348

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Laboratory Methods in Microbiology is a laboratory manual based on the experience of the authors over several years in devising and organizing practical classes in microbiology to meet the requirements of students following courses in microbiology at the West of Scotland Agricultural College. The primary object of the manual is to provide a laboratory handbook for use by students following food science, dairying, agriculture and allied courses to degree and diploma level, in addition to being of value to students reading microbiology or general bacteriology. It is hoped that laboratory workers in the food manufacturing and dairying industries will find the book useful in the microbiological aspects of quality control and production development. The book is organized into two parts. Part I is concerned with basic methods in microbiology and would normally form the basis of a first year course. Abbreviated recipes and formulations for a number of typical media and reagents are included where appropriate, so that the principles involved are more readily apparent. Part II consists of an extension of these basic methods into microbiology as applied in the food manufacturing, dairying and allied industries. In this part, the methods in current use are given in addition to, or in place of, the ""classical"" or conventional techniques.

Table of Contents

  • Preface

    Part I Basic Methods

    General Notes on Laboratory Procedure

    Laboratory Reports

    Procedure for the Use of a Microscope with an Oil-Immersion Objective

    Examination of Cultures for Motility by "Hanging Drop" Preparations

    Staining Methods

    A. Preparation of Smears for Staining

    B. Simple Stains

    C. Gram'S Staining Method

    D. Ziehl-Neelsen Method for Staining Acid-Fast Bacteria

    E. Staining of Bacterial Flagella

    F. The Demonstration of Bacterial Capsules

    G. Negative Stain

    H. The Staining of Bacterial Spores

    Cultivation of Micro-Organisms

    A. Types of Culture

    B. Incubation of Cultures

    C. Method of Inoculation: Aseptic Technique

    D. Maintenance of Pure Cultures in the Laboratory

    The Description of the Morphological and Cultural Characteristics of an Organism

    A. Morphological Characters

    B. Cultural Characters

    Plate Cultures

    A. Preparation of Plates for Streaking

    B. Streak Plates: The Separation of Mixed Cultures

    Determination of the Number of Viable Organisms in a Sample

    A. Colony Count Methods

    B. Membrane Filtration

    C. Dilution Tube Count

    D. Dye Reduction Methods

    Determination of the Total Number of Organisms in a Sample

    A. the Breed's Smear Method for Direct Microscopic Counts

    B. Direct Microscopic Counts by Membrane Filtration

    C. Brown's Opacity Tubes

    Statistical Methods for the Selection and Examination of Microbial Colonies

    Composition of Culture Media

    A. Introduction

    B. Dehydrated Media

    C. The Determination of the pH of Culture Media

    D. Examples of Non-Selective Culture Media

    E. Separation of Mixed Cultures—Enrichment Procedures, Elective and Selective Media

    F. Examples of Selective Culture Media

    Methods of Anaerobic Culture

    A. Robertson's Cooked Meat Medium

    B. Shake Cultures

    C. Semi-Solid Media

    D. Vaseline, Paraffin Wax or Agar Seals

    E. The Anaerobic Jar

    Cultivation in a Carbon Dioxide-Enriched Atmosphere

    Biochemical Tests for Bacteria

    A. Reactions Involving Protein, Amino Acids and Other Nitrogen Compounds, Including Tests for Proteolytic Activity

    B. Reactions Involving Carbohydrates and Other Carbon Compounds, Including Tests for Saccharolytic Activity

    C. Reactions Involving Fats and Related Substances, Including Tests for Lipolytic Activity

    D. Miscellaneous Tests

    Cleaning of Glassware and Apparatus

    A. Treatment of New Glassware

    B. Treatment of Used Glassware and Other Apparatus

    C. Disposable Apparatus


    A. Sterilization by Dry Heat

    B. Sterilization by Moist Heat

    C. Sterilization by Filtration

    D. Chemical Disinfectants

    E. Preparation of Glassware and Materials for Sterilization

    F. To Test the Sterility of Laboratory Equipment

    Laboratory Evaluation of Disinfectants

    A. The Rideal-Walker Test

    B. The Suspension Test

    C. The Capacity Test

    Serological Methods

    A. Introduction

    B. The Agglutination Reaction

    C. The Precipitin Test

    Molds and Yeasts

    A. General Conditions for the Growth of Molds and Yeasts

    B. Media for the Growth of Molds and Yeasts

    C. Examination of Molds

    D. Examination of Yeasts

    E. Note on the Identification of Molds and Yeasts

    The Isolation of Bacteriophage

    Part II Techniques in Applied Microbiology

    The Bacteriological Examination of Water

    A. Introduction

    B. Sampling Procedure

    C. Plate Counts

    D. Presumptive Coliform Counts and the Eijkman Test

    E. Enumeration of Fecal Streptococci

    F. Detection of Clostridium Welchii

    G. Membrane Filtration Methods

    The Microbiological Examination of Soil

    A. Introduction

    B. General Viable Counts of Bacteria by the Pour Plate Method

    C. Examination for Molds and Yeasts

    D. Buried Slide Technique (Rossi-Riccardo-Cholodny Technique)

    E. Isolation of Nitrogen-Fixing Bacteria

    F. Demonstration of the Presence of Nitrifying Bacteria

    G. Demonstration of the Presence of Denitrifying Bacteria

    H. Demonstration of Cellulose-Decomposing Bacteria

    The Bacteriological Examination of Silage

    A. The Preparation of Experimental Silage

    B. The Quantitative Bacteriological Examination of Grass or Silage

    C. Qualitative Bacteriological Examination of Grass or Silage

    D. Identification of Isolates

    The Microbiological Examination of Foods

    A. Introduction

    B. Sampling Methods

    C. Preparation of Dilutions

    D. The Isolation of Pathogenic and Indicator Organisms

    E. Meat and Meat Products

    F. Fish, Shellfish and Other Seafoods

    G. Eggs

    H. Fruit Juices and Squashes

    I. Sugar Syrups

    J. Fresh Fruit and Vegetables

    K. Salted, Pickled and Fermented Vegetables

    L. Bread, Cakes etc

    M. Frozen Foods

    N. Canned Foods

    0. Note on the Rationale of the Microbiological Analysis of Foods

    The Examination of Milk

    A. Determination of Total Numbers

    B. Determination of General Viable Numbers

    C. Dye Reduction Methods

    D. Presumptive Coliform Test

    E. The Laboratory Pasteurization Test

    F. Enumeration and Isolation of Thermophiles in Milk

    G. Enumeration and Isolation of Aerobic Spore-Formers in Milk

    H. Enumeration and Isolation of Psychrophiles

    I. Investigation of Faults and Taints in Milk and Isolation of Causative Organisms

    J. Examination of Mastitis Milk

    K. the Detection of Antibiotics in Milk

    The Examination of Dairy Utensils and Equipment

    A. Examination of Washed Milk Cans

    B. Examination of Washed Milk Bottles—Rinse Method

    C. Swab Method for the Examination of Milk Plant and Equipment

    D. Examination of Farm Dairy Equipment by the Rinse Method

    The Examination of Fermented Milks and Isolation of Constituent Flora

    A. Microscopic Examination

    B. Isolation of Constituent Flora

    The Examination of Dairy Starter Cultures

    A. Biochemical Tests

    B. Microscopic Examination

    C. Cultural Examination

    D. Demonstration of the Effect of Phage on Starter Bacteria

    E. Demonstration of the Effect of Penicillin on Starter Bacteria and Test Organisms

    The Microbiological Examination of Cheese

    A. Sampling

    B. Microscopic Examination

    C. Cultural Examination

    D. Isolation of Propionobacteria from Swiss Cheese

    E. Isolation of Moulds from Mold Ripened Cheese

    F. Isolation of Brevibacterium Linens from Bacterial-Ripened Soft Cheese

    The Microbiological Examination of Butter

    A. Sampling

    B. Cultural Examination

    C. The Isolation and Further Study of Lipolytic Micro-Organisms

    The Microbiological Examination of Milk Powder

    A. Sampling

    B. Microscopic Examination

    C. Cultural Examination

    D. Results and Suggested Standards

    The Microbiological Examination of Canned Concentrated Milk

    A. Evaporated Milk (Unsweetened Condensed Milk)

    B. Sweetened Condensed Milk

    The Microbiological Examination of Ice-Cream

    A. Collection of Samples

    B. Treatment of Samples

    C. Cultural Examination

    D. Methylene Blue Dye Reduction Test

    E. Results and Recommended Standards

    The Microbiological Examination of Cream

    A. Sampling

    B. Methylene Blue Dye Reduction Test

    C. Crossley's Bromcresol Purple Milk Test

    D. Cultural Examination

    E. Results and Recommended Standards

    Appendix 1 Schemes for the Identification of Micro-Organisms

    1. Diagnostic Tables for Gram-Negative Bacteria Which Can Grow on Nutrient Agar or Plate Count Agar

    2. Simple Key for the Identification of Gram-Positive Bacteria

    3. Identification of Yeasts and Molds

    Appendix 2 Recipes for Stains, Reagents and Media




    Appendix 3 Probability Tables for the Estimation of Bacterial Numbers by the Dilution Tube Technique

    Table 1

    Table 2

    Table 3

    Determination of MPNS from Series of More Than Three Dilutions

    Selected Bibliography


    Author Index

    Subject Index

Product details

  • No. of pages: 374
  • Language: English
  • Copyright: © Academic Press 1966
  • Published: January 1, 1966
  • Imprint: Academic Press
  • eBook ISBN: 9781483274348

About the Authors

W. F. Harrigan

Margaret E. McCance

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