Laboratory Methods in Microbiology

PrefacePart 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 Sterilization 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 BacteriophagePart 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 StandardsAppendix 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 MoldsAppendix 2 Recipes for Stains, Reagents and Media Stains Reagents MediaAppendix 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 DilutionsSelected BibliographyReferencesAuthor IndexSubject Index