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Ahmed E. Yousef. Analytical Food Microbiology
Table of Contents
List of Tables
List of Illustrations
Guide
Pages
ANALYTICAL FOOD MICROBIOLOGY. A Laboratory Manual
PREFACE
PART I BASICS OF FOOD MICROBIOLOGY LABORATORY
CHAPTER 1 LABORATORY SAFETY
BACKGROUND
Laboratory Environment and Personal Safety
Biosafety Levels
Decontamination and Waste Disposal
PERSONAL SAFETY IN THE LABORATORY
Personal Protective Equipment
Observing Personal Safety
MATERIALS IN MICROBIOLOGY LABORATORY
PRACTICAL ASPECTS
Before the Laboratory Session
Immediately Before Entering the Laboratory
Immediately After Entering the Laboratory
While Executing the Exercise
Immediately After Completing the Laboratory Exercise
REFERENCE
CHAPTER 2 SAMPLING FOR MICROBIOLOGICAL ANALYSIS OF FOOD AND PROCESSING ENVIRONMENT
THEORETICAL ASPECTS
Sampling Principles. Introduction
Preparing A Sampling Plan
SAMPLING TECHNIQUES AND SAMPLE PREPARATION. Sampling Tips
Sample Preparation
Withdrawing the test sample
Homogenizing the test sample
Dilution of food homogenate
ENVIRONMENTAL SAMPLING
Surface Sampling
Swab method
Sponge method
Replicate organism direct agar contact (RODAC) method
Air Sampling
SELECTED REFERENCES
PRACTICING SAMPLING AND SAMPLE PREPARATION
OBJECTIVES
PROCEDURE OVERVIEW
MATERIALS. Per Group of Two
Group‐Shared
METHODS
QUESTIONS
CHAPTER 3 ENUMERATION OF MICROORGANISMS IN FOOD: DILUTION SERIES. COLONY COUNTING. MOST PROBABLE NUMBER. INTRODUCTION
PLATE COUNT METHOD
Dilution
Dilutions suitable for plating
Pipetting
Plating
Spread‐plating
Pour‐plating
Incubation
Colony Counting
Counting rules
Plates with colonies in the range of 20–200 (the best possible scenario)
Plates with < 20 colonies
Plates with no colonies
Plates with greater than 200 colonies
Plates with spreaders
Which counting rule to use
Population Counting
Important Considerations
MOST PROBABLE NUMBER METHOD. Principles
Scoring the Tubes and Calculating MPN
SELECTED REFERENCES
QUESTIONS
CHAPTER 4 PRACTICING BASIC TECHNIQUES: POUR‐PLATING. SPREAD‐PLATING. COLONY COUNTING. GRAM STAINING. MICROSCOPY. INTRODUCTION
Microbial Growth
Culture media
Sampling, Homogenization, Dilution, and Plating
Incubation, Colony Counting, and Population Determination
OBJECTIVES
MEDIA. Plate Count Agar (PCA)
Peptone Water
PROCEDURE OVERVIEW
SELECTED REFERENCES
SESSION 1: DILUTION AND PLATING
MATERIALS AND EQUIPMENT. Per Student
Class‐Shared
PROCEDURE
Dilution
Spread‐Plating
Pour‐Plating
SESSION 2: POPULATION COUNTING AND COLONY STREAKING
MATERIALS AND EQUIPMENT. Per Student
PROCEDURE. Determining Population Count. Spread plates
Pour plates
Streaking
SESSION 3: MORPHOLOGICAL EXAMINATION
MATERIALS
PROCEDURE. Examining Streak Plates
Examining Colony Morphology
Examining Cell Morphology Under Bright‐Field Microscope
QUESTIONS
PART II FOOD MICROBIOTA
Transient and Resident Food Microbiota
Food Characteristics as Determinants of Its Microbiota
Temperature and Microbial Behavior in Food
Taxonomy of Food Microbiota
SELECTED REFERENCES
CHAPTER 5 AEROBIC MESOPHILIC PLATE COUNT: APPLICATION OF BASIC TECHNIQUES TO FOOD. INTRODUCTION. Measuring the Microbiological Quality of Food
Aerobic Mesophilic Count as a Measure of Food’sMicrobiological Quality
OBJECTIVES
MEDIA
Tryptic Soy Agar (TSA)
PROCEDURE OVERVIEW
Food Handling Precautions
Examination of Colonies
SELECTED REFERENCES
SESSION 1: SAMPLING, HOMOGENIZATION, DILUTION, PLATING, AND INCUBATION
MATERIALS AND EQUIPMENT. Per Two Students (i.e., one group)
Sample Preparation Utensils (as needed)
Class‐Shared
PROCEDURE. Labeling
Sampling
Homogenization
Preparing Dilutions
Spread‐Plating
Plate Incubation
SESSION 2: COLONY COUNTING AND ISOLATION
MATERIALS AND EQUIPMENT. Per Group of Two
Class‐Shared
PROCEDURE. Colony Counting
Streak Selected Colonies for Isolation
SESSION 3: EXAMINING COLONY AND CELL MORPHOLOGIES
MATERIALS AND EQUIPMENT. Per Group of Two
Class‐Shared
PROCEDURE. Colony Morphology
Cell Morphology
QUESTIONS
CHAPTER 6 MESOPHILIC SPOREFORMING BACTERIA: AEROBIC AND ANAEROBIC INCUBATION. DIFFERENTIAL STAINING. PHASE‐CONTRAST MICROSCOPY. INTRODUCTION
Thermal Requirements for Growth and Inactivation
Prevalence and Impact on Food
Life‐Cycle of a Sporeforming Bacterium
Spore formation
Spore structure and resistance
Spore‐to‐cell transition
Activation
Commitment
Germination step
Outgrowth
OBJECTIVES
MEDIA AND INCUBATION CONDITIONS. Thioglycolate Agar
Tryptone Glucose Extract (TGE) Agar
Anaerobic Incubation
MICROSCOPY METHODS
Spore Staining and Examination by Brightfield Microscope
Phase‐Contrast Microscope
PROCEDURE OVERVIEW
Organization
SELECTED REFERENCES
SESSION 1: SAMPLE PREPARATION, PLATING, AND INCUBATION
MATERIALS AND EQUIPMENT. Per Pair of Students
Class‐Shared
PROCEDURE
SESSION 2: SPORE ENUMERATION AND EXAMINING SPORULATION
MATERIALS AND EQUIPMENT. Per Pair of Students
Class‐Shared
PROCEDURE
SESSION 3: MICROSCOPIC EXAMINATION OF SPORES
MATERIALS AND EQUIPMENT. Per Student Pair
Class‐Shared
PROCEDURE
QUESTIONS
CHAPTER 7 Pseudomonas SPECIES AND OTHER SPOILAGE PSYCHROTROPHS: SELECTION BY INCUBATION TEMPERATURE; DETECTION OF PROTEOLYTIC AND LIPOLYTIC ACTIVITY. INTRODUCTION
Microbial Spoilage of Food
Psychrotrophic Bacteria in Food
Pseudomonas spp
OBJECTIVES
MEDIA AND TESTS. Milk Agar
Pseudomonas Isolation Agar (PIA)
Tributyrin Agar
Tryptic Soy Agar (TSA)
PROCEDURE OVERVIEW
Organization
SELECTED REFERENCES
SESSION 1: SAMPLING AND PLATING
MATERIALS. Per Student
Class‐Shared
PROCEDURE. Dilutions
Plating
SESSION 2: ENUMERATION OF Pseudomonas SPECIES AND TESTING FOR ENZYMATIC ACTIVITY
MATERIALS. Per Student
Class‐Shared
PROCEDURE. Inspection and Enumeration of Pseudomonas spp
Testing Enzymatic Activity of Isolates
SESSION 3: DETECTING ENZYMATIC ACTIVITY
MATERIALS. Per Pair of Students
PROCEDURE
SESSION 4: ENUMERATION OF AEROBIC PSYCHROTROPHS
MATERIALS. Per Student
Class‐Shared
PROCEDURE. Psychrotrophic Count
QUESTIONS
CHAPTER 8 DETECTION AND ENUMERATION OF Enterobacteriaceae IN FOOD: USE OF SELECTIVE‐DIFFERENTIAL MEDIA. MOST PROBABLE NUMBER TECHNIQUE. INTRODUCTION
Enterobacteriaceae in Food
OBJECTIVES
MEDIA AND TESTS. Enterobacteriaceae Enrichment Broth (EEB)
Glucose Agar
Tryptic Soy Agar (TSA)
Violet‐Red Bile Glucose (VRBG) Agar
Oxidase Test
PROCEDURE OVERVIEW
Organization
SELECTED REFERENCES
SESSION 1: PREPARATION OF MOST PROBABLE NUMBER TUBES
MATERIALS AND EQUIPMENT. Per Pair of Students
Class‐Shared
PROCEDURE. Dilutions
Inoculating EEB – MPN Method
SESSION 2: PRESUMPTIVE MPN DETERMINATION
MATERIALS AND EQUIPMENT. Per Pair of Students
Class‐Shared
PROCEDURE
SESSION 3: PRESUMPTIVE MPN DETERMINATION AND CONFIRMATION
MATERIALS AND EQUIPMENT. Per Pair of Students
Class‐Shared
PROCEDURE
SESSION 4: CONFIRMATION
MATERIALS AND EQUIPMENT. Per Pair of Students
Class‐Shared
PROCEDURE
DATA INTERPRETATION
QUESTIONS
CHAPTER 9 EXAMINATION AND ENUMERATION OF FOODBORNE FUNGI: FUNGI MORPHOLOGY AS IDENTIFICATION TOOL. SELECTIVE MEDIA COMPARISON. INTRODUCTION
Classification
Foodborne Fungi
Quantifying Foodborne Fungi
OBJECTIVES
MEDIA
Antibiotic Plate Count Agar (APCA)
Dichloran Rose Bengal Chloramphenicol (DRBC) Agar
Dichloran 18% Glycerol (DG18) Agar
Yeast and Mold Count Petrifilm™
PROCEDURE OVERVIEW
Foods Analyzed
Sample Preparation
SELECTED REFERENCES
SESSION 1: EXAMINATION OF FOOD FOR MOLDINESS AND PREPARING SAMPLES FOR FUNGI ENUMERATION
FOODS
ORGANIZATION
Session I. Visual inspection of food for moldiness. MATERIALS. Per Pair of Students
PROCEDURE
II. Preparation of slide culture
MATERIALS. Per Pair of Students
Class‐Shared
PROCEDURE. Microscope Examination of Mold on Food
Preparation of Slide Culture
III. Sample preparation for fungi enumeration
MATERIALS. Per Pair of Students
Class‐Shared
PROCEDURE. Sample Preparation. Strawberries (fresh and stored)
Cheddar cheese
Blue cheese
Dilution and Plating. Fresh strawberries
Stored strawberries
Cheddar cheese
Blue cheese
SESSION 2: EXAMINATION OF PRE‐MOUNTED FUNGAL SPECIMENS
MATERIALS AND EQUIPMENT. Class‐Shared
PROCEDURE. Demonstrated Yeast and Bacteria
Demonstrated Mold
SESSION 3: DETERMINING FUNGI POPULATION AND ISOLATE MORPHOLOGY
MATERIALS AND EQUIPMENT. Per Pair of Students
Class‐Shared
PROCEDURE. Colony Counting and Calculating Population Count
Microscopic Examination. Yeast isolates
Mold isolates
Slide culture
DATA ANALYSIS AND QUESTIONS. Group Data
Class Data
Questions
PART III FOODBORNE PATHOGENS
Foodborne Disease Hazards
Detection vs. Enumeration of Pathogens in Food
PATHOGEN DETECTION METHODS
Sampling and Sample Preparation
Enrichment
Sample Screening
Pathogen Isolation
Identification of Pathogen Isolate
Biochemical tests
Immunological assays
Genetic approach
Typing
IMPORTANT CONSIDERATIONS
Controls and Surrogates
Method Sensitivity and Specificity
SELECTED REFERENCES
CHAPTER 10 Staphylococcus aureus. INTRODUCTION
The Disease
The Toxins
Incidence of The Pathogen in Food
Analyzing Food for S. aureus and Its Toxins
SELECTED REFERENCES
EXERCISE I: ENUMERATION, ISOLATION, AND IDENTIFICATION OF Staphylococcus aureus IN FOOD. PATHOGEN ENUMERATION. PATHOGEN IDENTIFICATION. MOST‐PROBABLE NUMBER TECHNIQUE. INTRODUCTION
OBJECTIVES
MEDIA AND TESTS. Baird‐Parker Agar
Blood Agar
Brain‐Heart Infusion Broth Supplemented with Yeast Extract
Tryptic Soy Agar
Tryptic Soy Broth Supplemented with Salt and Pyruvate (TSB‐SP)
Catalase Test
Gram Reaction
Coagulase Test
PROCEDURE OVERVIEW
Organization
Personal Safety
SESSION 1: SAMPLING, HOMOGENIZATION, DILUTION, AND INOCULATION OF MPN TUBES
MATERIALS AND EQUIPMENT. Per Pair of Students
Class‐Shared
PROCEDURE
SESSION 2: SCREENING MPN TUBES USING Staphylococcus SELECTIVE‐DIFFERENTIAL MEDIUM
MATERIALS AND EQUIPMENT. Per Pair of Students
Class‐Shared
PROCEDURE
SESSION 3: INSPECTING AND ISOLATING PRESUMPTIVE Staphylococcus aureus COLONIES
MATERIALS. Per Pair of Students
Class‐Shared
PROCEDURE
SESSION 4: CONFIRMING Staphylococcus aureus IDENTITY AND CALCULATING S. AUREUS MPN
MATERIALS AND EQUIPMENT. Per Pair of Students
Class‐Shared
PROCEDURE
DATA INTERPRETATION
QUESTIONS
EXERCISE 2: EXPRESSION OF ENTEROTOXIN GENES BY Staphylococcus aureus FOOD ISOLATES. POLYMERASE CHAIN REACTION. ENZYME‐LINKED IMMUNOSORBENT ASSAY. GENE EXPRESSION. INTRODUCTION
OBJECTIVES
PROCEDURE OVERVIEW
Organization
Personal Safety
SESSION 1: DETECTION OF SEA GENE: DNA EXTRACTION AND PREPARING FOR PCR
MATERIALS. Per Pair of Students
Class‐Shared
PROCEDURE
SESSION 2: DETECTION OF SEA GENE: GEL ELECTROPHORESIS
MATERIALS AND EQUIPMENT. Per Pair of Students
Class‐Shared
PROCEDURE
SESSION 3: DETECTION OF SEA GENE: ELECTROPHORESIS RESULTS OBSERVATION
MATERIALS AND EQUIPMENT. Per Pair of Students
PROCEDURE
SESSION 4: DETECTING THE EXPRESSION OF STAPHYLOCOCCAL ENTEROTOXINS USING ELISA
MATERIALS AND EQUIPMENT
Per Pair of Students
Class‐Shared
PROCEDURE
QUESTIONS
CHAPTER 11 Listeria monocytogenes: SAMPLING PROCESSING ENVIRONMENT. CULTURE‐BASED DETECTION METHOD. PCR‐BASED DETECTION METHOD. INTRODUCTION
Characteristics
The Disease
Foods Implicated
Detection
PROCEDURE OVERVIEW. Organization
Personal Safety
SELECTED REFERENCES
EXERCISE I: DETECTION OF Listeria SPECIES USING CULTURE‐BASED METHOD. INTRODUCTION
Enrichment
Isolation
Identification
OBJECTIVES
MEDIA. Brilliance Listeria Agar (BLA)
Half‐Fraser Broth
Modified Oxford (MOX) Agar
Motility Medium
Tryptic Soy Agar with Blood (TSA‐Blood)
Tryptic Soy Agar with Yeast Extract (TSA‐YE)
TESTS. Catalase Test
Christie‐Atkins‐Munch‐Peterson (CAMP) Test
Gram Reaction
Oxidase Test
PROCEDURE OVERVIEW
Organization
SESSION 1: SAMPLE PREPARATION AND ENRICHMENT
MATERIALS AND EQUIPMENT. Per Group of Two
Class‐Shared
PROCEDURE
SESSION 2: ISOLATION
MATERIALS AND EQUIPMENT. Per Group of Two
Class‐Shared
PROCEDURE
SESSION 3: IDENTIFICATION
MATERIALS AND EQUIPMENT. Per Group of Two
Class‐Shared
PROCEDURE
SESSION 4: MORPHOLOGICAL AND BIOCHEMICAL CHARACTERIZATION
MATERIALS AND EQUIPMENT. Per Group of Two
Class‐Shared
PROCEDURE
EXERCISE II: DETECTION OF Listeria monocytogenes USING PCR‐BASED METHOD. INTRODUCTION
OBJECTIVES
PROCEDURE OVERVIEW
DNA Extraction
DNA Amplification (PCR Technique)
DNA Detection (Gel Electrophoresis)
Gel Photographing
Results Explanation
Data Interpretation
Organization
SESSION 1: SAMPLE PREPARATION AND ENRICHMENT
SESSION 2: DNA EXTRACTION AND PCR. MATERIALS AND EQUIPMENT
PROCEDURE
SESSION 3: GEL ELECTROPHORESIS
MATERIALS AND EQUIPMENT. Per Group of Two
Class‐Shared
Gel Electrophoresis
SESSION 4: ELECTROPHORESIS RESULTS
MATERIALS AND EQUIPMENT
PROCEDURE
QUESTIONS
CHAPTER 12 Salmonella enterica: CULTURE‐BASED DETECTION. SELECTIVE ENRICHMENT. BIOCHEMICAL IDENTIFICATION. IMMUNOASSAY. INTRODUCTION
Classification and Nomenclature
Somatic (O) antigens
Capsular (Vi) antigens
Flagellar (H) antigens
Diseases
Foods Implicated in Salmonellosis Outbreaks
Detection of Salmonella in Food
Enrichment
Screening
Isolation
Biochemical identification
Confirmation and typing
OBJECTIVES
MEDIA. Buffered Peptone Water (BPW)
Rappaport‐Vassiliadis (RV) + Soya Broth
Tetrathionate (TT) Broth
Bismuth Sulfite (BS) Agar
Xylose Lysine Desoxycholate (XLD) Agar
PROCEDURE OVERVIEW
Organization
Personal Safety
SELECTED REFERENCES
SESSION 1: PRE‐ENRICHMENT
MATERIALS AND EQUIPMENT. Per Pair of Students
Class‐Shared
PROCEDURE. Pre‐enrichment
SESSION 2: SELECTIVE ENRICHMENT
MATERIALS AND EQUIPMENT. Per Pair of Students
Class‐Shared
PROCEDURE
SESSION 3: SAMPLE SCREENING AND Salmonella ISOLATION
MATERIALS AND EQUIPMENT. Per Pair of Students
Class‐Shared
PROCEDURE
SESSION 4: ISOLATION
MATERIALS AND EQUIPMENT. Per Pair of Students
Class‐Shared
PROCEDURE
SESSION 5: IDENTIFICATION
MATERIALS AND EQUIPMENT. Per Pair of Students
Class‐Shared
PROCEDURE
SESSION 6: SCORING API STRIPS
MATERIALS AND EQUIPMENT. Per Pair of Students
Class‐Shared
PROCEDURE
DATA INTERPRETATION
QUESTIONS
CHAPTER 13 SHIGA TOXIN‐PRODUCING Escherichia coli : RAPID MOLECULAR SCREENING. MULTIPLEX PCR. PCR‐ASSISTED ISOLATION OF STEC STRAINS. INTRODUCTION
Diseases
Pathogen Characteristics
OBJECTIVES
DETECTION OF STEC IN FOOD
Enrichment
Detection of Virulence Genes
Isolation and Identification
MEDIA. E. coli Broth with Antibiotics (EC Broth+VCC)
Eosin Methylene Blue (EMB) Agar
Tryptic Soy Agar + Yeast Extract (TSA‐YE)
PROCEDURE OVERVIEW
Organization
Control Strains
Personal Safety
SELECTED REFERENCES
SESSION 1: ENRICHMENT
MATERIALS AND EQUIPMENT. Per Group of Two
Class‐Shared
PROCEDURE
SESSION 2: RAPID SCREENING BY MULTIPLEX PCR
MATERIALS AND EQUIPMENT. Per Group of Four
Class‐Shared
PROCEDURE
SESSION 3: GEL ELECTROPHORESIS
MATERIALS AND EQUIPMENT. Per Group of Four
Class‐Shared
PROCEDURE
SESSION 4: ISOLATION
MATERIALS AND EQUIPMENT. Per Group of Two
Class‐Shared
PROCEDURE
SESSION 5: ISOLATION (CONTINUED)
MATERIALS AND EQUIPMENT. Per Group of Four
Class‐Shared
PROCEDURE
SESSION 6: SCREENING ISOLATES FOR VIRULENCE GENES: PCR. MATERIALS AND EQUIPMENT. Per Group of Four
Class‐Shared
PROCEDURE
SESSION 7: SCREENING OF ISOLATES FOR VIRULENCE GENES: GEL ELECTROPHORESIS AND ISOLATE SELECTION. MATERIALS AND EQUIPMENT. Per Group of Four
Class‐Shared
PROCEDURE
SESSION 8: ISOLATE IDENTITY CONFIRMATION: BIOCHEMICAL TESTING
MATERIALS AND EQUIPMENT. Per Group of Four
PROCEDURE
DATA INTERPRETATION
QUESTIONS
PART IV CONTROL OF FOODBORNE MICROORGANISMS
REFERENCES
CHAPTER 14 THERMAL RESISTANCE OF MICROORGANISMS IN FOOD: MICROBIAL INACTIVATION KINETICS. INTRODUCING THERMAL PROCESSING CONCEPT. INTRODUCTION
Determining Thermal Resistance of Targeted Microorganisms
Methods for Determining Thermal Resistance
Calculating Thermal Resistance
D‐value
z‐Value
Challenges in Determining Microbial Thermal Resistance. Linearity of inactivation kinetics
Food type affects thermal resistance
Physiological status affects thermal resistance of the test microorganism
Difficulty in determining temperature come‐up time
OBJECTIVES
MEDIA. Tryptic Soy Agar + Yeast Extract (TSA+YE)
Tryptic Soy Broth (TSB)
PROCEDURE OVERVIEW
Culture Preparation for Consistent Thermal Resistance
SELECTED REFERENCES
SESSION 1: INOCULATION AND THERMAL TREATMENTS OF TWO MICROORGANISMS
MATERIALS AND EQUIPMENT. Per Student Pair
Class‐Shared
PROCEDURE
SESSION 2: HEAT TREATMENTS OF Enterococcus sp. AT 60 AND 65°C
MATERIALS AND EQUIPMENT. Per Student Pair
Class‐Shared
PROCEDURE. Part 1: Colony Counting
Part 2: Heat Treatments
SESSION 3: COUNTING ENTEROCOCCUS SP. SURVIVORS
MATERIALS AND EQUIPMENT. Per Student Pair
Class‐Shared
PROCEDURE
SESSION 4: DETERMINATION OF D‐VALUES AND Z‐VALUE
Determining D‐valuves at 55°C
Determining D‐values at 60°C and 65°C
Determining z‐value for Enterococcus sp
QUESTIONS
CHAPTER 15 PRODUCTION OF BACTERIOCINS IN MILK: BACTERIOCIN BIOASSAY. MICRO‐DILUTION SERIES FOR POPULATION ENUMERATION. INTRODUCTION. What are Bacteriocins?
Bacteriocins and Food Preservation
Bacteriocin Bioassay
Bioassay without bacteriocin standard
Bioassay with bacteriocin reference standard
OBJECTIVES
MEDIA. Lactobacilli de Man, Rogosa, & Sharpe (MRS) Media. MRS broth
MRS agar
MRS soft agar
PROCEDURE OVERVIEW
Organization
SELECTED REFERENCES
SESSION 1: FERMENTATION
MATERIALS AND EQUIPMENT. Per Group of Two
Class‐Shared
PROCEDURE
SESSION 2: ANALYSIS OF FERMENTED MILK
MATERIALS AND EQUIPMENT. Per Group of Two
Class‐Shared
PROCEDURE
SESSION 3: LACTIC ACID BACTERIA COUNT (CONTINUED) AND BACTERIOCIN BIOASSAY
MATERIALS AND EQUIPMENT. Per Group of Two
Class‐Shared
PROCEDURE
SESSION 4: BACTERIOCIN BIOASSAY (CONTINUED) MATERIALS. Per Group of Two
Class‐Shared
PROCEDURE
QUESTIONS
APPENDIX I LABORATORY EXERCISE REPORT
OUTLINE
Cover Page
Introduction
Objectives
Methods
Results
Discussion
References
DATA PRESENTATION
Tables
Figures
Scientific Notation and Significant Digits
MISCELLANOUS WRITING ISSUES. Microorganisms’ Nomenclature
Grammar and Style
Plagiarism
APPENDIX II BACTERIAL AND FUNGAL STRAINS RECOMMENDED FOR USE AS CONTROL ORGANISMS
General Cultural Handling Methods
Specific Strains
APPENDIX III MICROBIOLOGICAL MEDIA
MEDIA PREPARATION GUIDELINES
Media Reconstitution and Autoclaving
MEDIA RECIPES. Antibiotic Plate Count Agar (APCA)
Baird‐Parker Agar
Bismuth Sulfite (BS) Agar (Wilson and Blair)
Brain‐Heart Infusion Agar with Yeast Extract (BHI‐YE)
Brilliance Listeria Agar (BLA) (Oxoid)
Buffered Peptone Water (BPW)
Dichloran Rose Bengal Chloramphenicol (DRBC) Agar
Dichloran 18% Glycerol (DG18) Agar
E. coli Broth with Antibiotics (EC‐broth+VCC)
Enterobacteriaceae Enrichment Broth, Mossel (Neogen)
Eosin Methylene Blue (EMB) Agar, Levine
Glucose Agar (also called Glucose OF Medium) (Neogen)
Half‐Fraser Broth (also called Demi‐Fraser Broth)
Lactobacilli deMan, Rogosa, & Sharpe (MRS) Agar (Neogen, Oxoid)
Lactobacilli deMan, Rogosa, & Sharpe (MRS) Soft Agar
Lactobacilli deMan, Rogosa, & Sharpe (MRS) Broth
Modified Oxford (MOX) Agar
Motility Medium
Peptone Water (1%)
Phosphate Buffered Saline
Pseudomonas Isolation Agar (PIA)
Rappaport‐Vassiliadis (RV) Broth (Soya Peptone Formula)
Tetrathionate (TT) Broth
FDA formulation
FSIS formulation (Hajna – Hajna & Damon, 1956)
Thioglycolate Agar
Tributyrin Agar (Sigma Aldrich)
Tryptic Soy Agar (TSA)
Tryptic Soy Agar + Blood (TSA‐Blood) (Also Called Blood Agar)
Tryptic Soy Agar with Yeast Extract (TSA‐YE)
Tryptic Soy Broth (TSB)
Tryptic Soy Broth with Salt and Pyruvate (TSB‐SP)
Tryptone Glucose Extract (TGE) Agar
Violet Red Bile Glucose (VRBG) Agar
Xylose Lysine Desoxycholate (XLD) Agar
APPENDIX IV SUPPLIES AND EQUIPMENT AVAILABILITY
EQUIPMENT. Balance, top loading
Bunsen burner
Calculator
Colony counter
Computer or computing tablet
Homogenizer or blender
Incubator
Micropipette
Microscope
Microtiter plate reader
Molecular analysis equipment
pH meter
Refrigerator
Spectrophotometer
Vortex
Water bath or heating block (dry bath)
SUPPLIES. Bacteriological needles and loops
Biohazard boxes and containers
Disposable gloves, all sizes
Markers
Micropipette tips (sterile)
Paper towels
Rulers
Safety goggles
Sample preparation supplies
Sanitizer
Serological pipettes
Spread‐plating supplies
Staining supplies
Stomacher bags and holders
Striker or lighter
Tape
Tube racks
INDEX
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