Microbiological Examination Methods of Food and Water: A Laboratory Manual

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Features

  • All methods and procedures are presented in a simple and didactic manner to facilitate understanding and are accompanied by figures that illustrate the sequence of each assay and give an overview of all the steps.
  • Every chapter begins with a comprehensive, in-depth and updated bibliographic reference on the microorganism(s) dealt with in that particular section of the book. Latest facts on the taxonomic position of each group, genus or species are given, as well as clear guidelines on how to update any changes in nomenclature on the internet.
  • All chapters provide schematic comparisons between the methods presented, highlighting the main differences and similarities. This allows the user to choose the method that best meets his needs. Moreover, each chapter lists validated alternative quick methods, which, though not described in the book, may and can be used for the analysis of the microorganism(s) dealt with in that particular chapter.
  • Support materials such as drawings, procedure schemes and laboratory sheets are available for downloading and customization.

Summary

Microbiological Examination Methods of Food and Water is an illustrated laboratory manual that provides an overview of current standard microbiological culture methods for the examination of food and water, adhered to by renowned international organizations, such as ISO, AOAC, APHA, FDA and FSIS/USDA. It includes methods for the enumeration of indicator microorganisms of general contamination, indicators of hygiene and sanitary conditions, sporeforming, spoilage fungi and pathogenic bacteria.

Every chapter begins with a comprehensive, in-depth and updated bibliographic reference on the microorganism(s) dealt with in that particular section of the book. The latest facts on the taxonomic position of each group, genus or species are given, as well as clear guidelines on how to deal with changes in nomenclature on the internet. All chapters provide schematic comparisons between the methods presented, highlighting the main differences and similarities. This allows the user to choose the method that best meets his/her needs. Moreover, each chapter lists validated alternative quick methods, which, though not described in the book, may and can be used for the analysis of the microorganism(s) dealt with in that particular chapter. The didactic setup and the visualization of procedures in step-by-step schemes allow the user to quickly perceive and execute the procedure intended. Support material such as drawings, procedure schemes and laboratory sheets are available for downloading and customization.

This compendium will serve as an up-to-date practical companion for laboratory professionals, technicians and research scientists, instructors, teachers and food and water analysts. Alimentary engineering, chemistry, biotechnology and biology (under)graduate students specializing in food sciences will also find the book beneficial. It is furthermore suited for use as a practical/laboratory manual for graduate courses in Food Engineering and Food Microbiology.

Table of Contents

Preface
About the authors
List of tables
List of figures

1 Sampling, transport and storage of samples for analysis
1.1 Introduction
1.1.1 Lot
1.1.2 Lot sample and sample unit
1.1.3 Lot sampling plans
1.1.3.1 The two-class sampling plan
1.1.3.2 The three-class sampling plan
1.1.4 Analytical unit
1.2 Collecting samples for analysis
1.2.1 Selection and preparation of containers for the sampling of foods contained in non-individual packages
1.2.2 Procedures for the sampling of foods contained in non-individual packages
1.2.3 Sampling of foods involved in foodborne diseases
1.2.4 Sampling of water
1.3 Transportation and storage of samples until analysis
1.3.1 Foods with low water activity
1.3.2 Frozen foods
1.3.3 Refrigerated foods
1.3.4 Commercially sterile foods in sealed packages
1.3.5 Water samples
1.4 References

2 Preparation of sample for analysis
2.1 Introduction
2.2 Homogenization of samples and withdrawal of the analytical unit
2.2.1 Procedure for homogenization and withdrawal of analytical units from liquid products
2.2.2 Procedure for homogenization and withdrawal of analytical units from solid or concentrated liquid products
2.2.3 Procedure for withdrawing the analytical unit using the surface swabbing technique
2.2.3.1 Swab sampling
2.2.3.2 Sponge sampling
2.2.4 Procedure for withdrawing the analytical unit using the surface washing technique
2.2.4.1 Procedure for washing poultry carcasses
2.2.4.2 Procedure for washing other foods
2.2.4.3 Procedure for washing packages
2.2.5 Keeping of counter-samples
2.3 Preparation of the first dilution of the analytical unit
2.3.1 Diluents for presence/absence tests
2.3.2 Diluents for tests requiring differentiated handling of the sample
2.3.3 Diluents for general quantification tests
2.3.4 How to prepare an initial 1:10 (10–1) dilution
2.3.5 How to prepare an initial dilution different from 1:10
2.3.6 Procedure for the preparation of the first dilution of liquid samples
2.3.7 Procedure for the preparation of the first dilution of solid or concentrated liquid samples
2.3.8 Procedure for the preparation of the first dilution of samples obtained by surface swabbing or surface washing
2.4 Serial decimal dilution of the sample
2.5 References
Annex 2.1 – Procedures for the homogenization of the content and withdrawal of the analytical unit of different types of foods
Annex 2.2 – Special cases in which there are variations in the analytical unit and/or dilution and/or diluents recommended for the preparation of the first dilution of samples of different types of foods

3 Basic plate count techniques for the enumeration of microorganisms
3.1 Introduction
3.2 Pour plate technique
3.2.1 Material required for the analyses
3.2.2 Procedure
3.3 Spread plate technique
3.3.1 Material required for the analyses
3.3.2 Procedure
3.4 Drop plate technique
3.4.1 Material required for the analyses
3.4.2 Procedure
3.5 Membrane filtration
3.5.1 Material required for the analyses
3.5.2 Procedure
3.6 Counting colonies and calculating results
3.6.1 Pour plate calculations
3.6.1.1 Calculating the pour plate results in the standard situation
3.6.1.2 Calculating the pour plates results for samples prepared by the surface swabbing technique (swabs or sponges)
3.6.1.3 Calculating the pour plate results for samples prepared by the surface washing technique
3.6.2 Spread plate calculations
3.6.3 Drop plate calculations
3.6.4 Membrane filtration calculations
3.7 Counting colonies and calculating results according to ISO 7218:2007
3.8 References

4 Basic techniques for microbial enumeration by the most probable number method (MPN)
4.1 Introduction
4.2 Multiple dilution test
4.2.1 Material required for the analyses
4.2.2 Procedure
4.3 Single dilution test
4.3.1 Material required for the analyses
4.3.2 Procedure
4.4 Calculation of the results
4.4.1 Calculating the results of the multiple dilution test
4.4.1.1 Calculation using the MPN tables (for decimal dilutions)
4.4.1.2 Calculating using the Thomas formula (for non-decimal dilutions)
4.4.1.3 Calculating the results for samples prepared by the surface swabbing or surface washing techniques
4.4.2 Calculating the results of the single dilution test
4.4.2.1 Rules for calculations performed using the MPN-3 Table
4.4.2.2 Calculation for samples prepared by the surface swabbing or surface washing techniques
4.5 References
Annex 4.1 – MPN tables

5 Basic techniques for the detection of the presence/absence of microorganisms
5.1 Introduction
5.1.1 Enrichment
5.1.1.1 Pre-enrichment
5.1.1.2 Selective enrichment
5.1.2 Isolation in solid media (selective differential plating)
5.1.3 Confirmation
5.1.3.1 Catalase test
5.1.3.2 Citrate test
5.1.3.3 Amino acid decarboxylation tests
5.1.3.4 Phenylalanine deaminase test
5.1.3.5 Carbohydrate fermentation tests
5.1.3.6 Indole test
5.1.3.7 Malonate test
5.1.3.8 Oxidation/Fermentation test (O/F)
5.1.3.9 Oxidase test
5.1.3.10 Nitrate reduction test
5.1.3.11 Urease test
5.1.3.12 Methyl Red test (MR)
5.1.3.13 Voges-Proskauer test (VP)
5.2 Material required for the analyses
5.3 Procedure
5.3.1 Pre-enrichment
5.3.2 Selective enrichment
5.3.3 Selective differential plating
5.3.3.1 Streak plating technique for obtaining pure cultures
5.3.4 Selection of colonies and subculturing of cultures for confirmation
5.3.4.1 Technique for the subculturing of pure cultures starting from colonies isolated from plates
5.3.5 Confirmation tests
5.3.5.1 Gram-staining (Hucker’s method)
5.3.5.2 Spore-staining (Schaeffer-Fulton’s method)
5.3.5.3 Spore-staining (Ashby’s method)
5.3.5.4 Wet mounts for direct (fresh) microscopic observation
5.4 References 56

6 Aerobic plate count
6.1 Introduction
6.1.1 The importance and significance of the total aerobic mesophilic count
6.1.2 Definition of psychrotrophics
6.1.3 Methods of analysis
6.2 Plate count method APHA 2001 for aerobic mesophilic bacteria in foods and water
6.2.1 Material required for analysis
6.2.2 Procedure
6.2.2.1 Pour plate technique
6.2.2.2 Spread plate technique
6.2.2.3 Membrane filtration technique
6.3 Petrifilm™ AOAC official methods 990.12 - 989.10 - 986.33 for aerobic mesophilic bacteria in foods
6.3.1 Material required for analysis
6.3.2 Procedure
6.4 Plate count method APHA 2001 for aerobic psychrotrophic bacteria in foods
6.4.1 Material required for analysis
6.4.2 Procedure
6.5 References

7 Yeasts and molds
7.1 Introduction
7.1.1 Yeasts and molds in foods
7.1.2 Methods of analysis for total yeast and mold counts
7.1.3 Psychrotrophic fungi
7.1.4 Heat-resistant molds
7.1.5 Preservative-resistant yeasts (PRY)
7.1.5.1 Zigosaccharomyces bailii (Lindner) Guilliermond 1912
7.1.5.2 Zygosaccharomyces bisporus (Naganishi) Lodder and Kreger 1952
7.1.5.3 Schizosaccharomyces pombe Lindner 1893
7.1.5.4 Candida krusei (Castellani) Berkhout 1923
7.1.5.5 Pichia membranaefaciens Hansen 1904
7.1.6 Osmophilic yeasts
7.1.6.1 Zygosaccharomyces rouxii (Boutroux) Yarrow
7.2 Plate count method APHA 2001 for yeasts and molds in foods
7.2.1 Material required for analysis
7.2.2 Procedure
7.3 Plate count method APHA 2001 for psychrotrophic fungi in foods
7.3.1 Material required for analysis
7.3.2 Procedure
7.4 Plate count method APHA 2001 for heat-resistant molds in foods
7.4.1 Material required for analysis
7.4.2 Procedure
7.5 Presence/absence method Pitt and Hocking 2009 and Plate count method Pitt and Hocking 2009 for preservative-resistant yeasts in foods
7.5.1 Material required for analysis
7.5.2 Procedure
7.6 Membrane filtration method APHA 2001 and Plate count method APHA 2001 for osmophilic yeasts in foods
7.6.1 Material required for analysis
7.6.2 Procedure
7.7 References

8 Enterobacteriaceae
8.1 Introduction
8.1.1 Taxonomy
8.1.2 Methods of analysis
8.2 Plate count method APHA 2001 for Enterobacteriaceae in foods
8.2.1 Material required for analysis
8.2.2 Procedure
8.3 Most probable number (MPN) method APHA 2001 for Enterobacteriaceae in foods
8.3.1 Material required for analysis
8.3.2 Procedure
8.4 Petrifilm™ AOAC official method 2003.1 for Enterobacteriaceae in selected foods
8.4.1 Material required for analysis
8.4.2 Procedure
8.5 References

9 Total and thermotolerant coliforms and Escherichia coli
9.1 Introduction
9.1.1 Definition of total coliforms
9.1.2 Definition of thermotolerant coliforms
9.1.3 Escherichia coli
9.1.4 Use as indicators
9.1.5 Methods of analysis
9.2 Most probable number (MPN) method APHA 2001 for total coliforms, thermotolerant coliforms and E. coli in foods
9.2.1 Material required for analysis
9.2.2 Procedure
9.3 Most probable number (MPN) methods ISO 4831:2006 and ISO 7251:2005 for total coliforms and presumptive E. coli in foods
9.3.1 Material required for analysis
9.3.2 Procedure
9.4 Most probable number (MPN) method APHA/AWWA/WEF 2005 for total and thermotolerant coliforms and E. coli in water
9.4.1 Material required for analysis
9.4.2 Procedure 1
9.5 Plate count method APHA 2001 for total coliforms in foods
9.5.1 Material required for analysis
9.5.2 Procedure
9.6 References

10 Staphylococcus aureus
10.1 Introduction
10.1.1 Taxonomy
10.1.1.1 The genus Staphylococcus
10.1.1.2 The coagulase positive staphylococci
10.1.1.3 Staphylococcus aureus
10.1.2 Pathogenicity
10.1.2.1 Staphylococcus aureus enterotoxins
10.1.2.2 Staphylococcal food poisoning
10.1.3 Methods of analysis
10.2 Plate count method APHA 2001 for coagulase positive staphylococci and S. aureus in foods
10.2.1 Material required for analysis
10.2.2 Procedure
10.3 Most probable number (MPN) method APHA 2001 for coagulase positive staphylococci and S. aureus in foods
10.3.1 Material required for analysis
10.3.2 Procedure 1
10.4 Presence/absence method APHA 2001 for coagulase positive staphylococci and S. aureus in foods
10.4.1 Material required for analysis
10.4.2 Procedure
10.5. References

11 Bacillus cereus
11.1 Introduction
11.1.1 B. cereus Group
11.1.2 Main characteristics of B. cereus
11.1.3 Methods of analysis
11.2 Plate count method APHA 2001 for Bacillus cereus in foods
11.2.1 Material required for analysis
11.2.2 Procedure
11.3 Most probable number (MPN) method APHA 2001 for Bacillus cereus in foods
11.3.1 Material required for analysis
11.3.2 Procedure
11.4 References

12 Clostridium perfringens
12.1 Introduction
12.1.1 Main characteristics of C. perfringens
12.1.2 Epidemiology
12.1.2.1 C. perfringens type A food poisoning
12.1.2.2 C. perfringens type C necrotic enteritis
12.1.3 Methods of analysis
12.2 Plate count method APHA 2001 for Clostridium perfringens in foods
12.2.1 Material required for analysis
12.2.2 Procedure
12.3 Presence/absence method APHA 2001 for Clostridium perfringens in foods
12.3.1 Material required for analysis
12.3.2 Procedure
12.4 References

13 Enterococci
13.1 Introduction
13.1.1 Enterococci
13.1.1.1 Species of intestinal origin
13.1.1.2 Species found in plants, soil and water
13.1.1.3 Species found in foods
13.1.1.4 Biochemical characteristics of the genus Enterococcus
13.1.2 Fecal streptococci
13.1.2.1 Biochemical characteristics of the genus Streptococcus
13.1.3 Diferentiation of enterococci from fecal streptococci
13.1.4 Methods of analysis
13.2 Plate count method APHA 2001 for enterococci and fecal streptococci in foods
13.2.1 Material required for analysis
13.2.2 Procedure
13.3 Most probable number (MPN) method APHA 2001 for enterococci and fecal streptococci in foods
13.3.1 Material required for analysis
13.3.2 Procedure
13.4 Membrane filtration method APHA/AWWA/WEF 2005 for enterococci and fecal streptococci in water
13.4.1 Material required for analysis
13.4.2 Procedure
13.5 Membrane filtration method ISO 7899-2:2000 for intestinal enterococci in water
13.5.1 Material required for analysis
13.5.2 Procedure
13.6 References

14 Lactic acid bacteria
14.1 Introduction
14.1.1 Carnobacterium Collins et al. 1987
14.1.2 Enterococcus (ex Thiercelin & Jouhaud 1903) Schleifer & Kilpper-Bälz 1984
14.1.3 Fructobacillus Endo and Okada 2008
14.1.4 Lactobacillus Beijerinck 1901 emend. Haakensen et al. 2009
14.1.5 Lactococcus Schleifer et al. 1986
14.1.6 Leuconostoc van Tieghem 1878
14.1.7 Oenococcus Dicks et al. 1995 emend. Endo and Okada 2006
14.1.8 Pediococcus Balcke 1884
14.1.9 Streptococcus Rosenbach 1884
14.1.10 Tetragenococcus Collins et al. 1993
14.1.11 Weissella Collins et al. 1994
14.1.12 Methods of analysis
14.2 Plate count method APHA 2001 for lactic acid bacteria in foods
14.2.1 Material required for analysis
14.2.2 Procedure
14.3 Most probable number (MPN) methods APHA 2001 for lactic acid bacteria in foods
14.3.1 Material required for analysis
14.3.2 Procedure using the MRS broth
14.3.3 Procedure using the Rogosa SL Broth
14.4 References

15 Campylobacter
15.1 Introduction
15.1.1 Taxonomy
15.1.2 Epidemiology
15.2 Presence/absence method ISO 10272-1:2006 for thermotolerant Campylobacter in foods
15.2.1 Material required for analysis
15.2.2 Procedure
15.3 References

16 Cronobacter
16.1 Introduction
16.1.1 Taxonomy
16.1.1.1 Cronobacter Iversen et al. 2008, gen. nov.
16.1.2 Epidemiology
16.1.3 Codex Alimentarius microbiological criteria for Cronobacter spp. in powdered infant formulae
16.2 Presence/absence method ISO 22964:2006 for Cronobacter [Enterobacter sakazakii] in milk powder and powdered infant formula
16.2.1 Material required for analysis
16.2.2 Procedure
16.3 References

17 Pseudomonas
17.1 Introduction
17.1.1 Taxonomy
17.1.1.1 Pseudomonas Migula 1894
17.1.1.2 Shewanella MacDonell & Colwell 1986
17.1.1.3 Janthinobacterium De Ley et al. 1978 emend. Lincoln et al. 1999
17.1.1.4 Stenotrophomonas Palleroni & Bradbury 1993
17.2 Most probable number (MPN) method APHA/AWWA/WEF 2005 for Pseudomonas aeruginosa in water
17.2.1 Material required for analysis
17.2.2 Procedure
17.3 Membrane filtration method ISO 16266:2006 for Pseudomonas aeruginosa in water
17.3.1 Material required for analysis
17.3.2 Procedure
17.4 Plate count method ISO 13720:2010 for presumptive Pseudomonas spp. in meat and meat products
17.4.1 Material required for analysis
17.4.2 Procedure
17.5 Plate count method ISO 11059:2009 IDF/RM 225:2009 for Pseudomonas spp. in milk and milk products
17.5.1 Material required for analysis
17.5.2 Procedure
17.6 References

18 Listeria monocytogenes
18.1 Introduction
18.1.1 Taxonomy
18.1.2 Epidemiology
18.1.3 Methods of analysis
18.2 Presence/absence method BAM/FDA 2011 for Listeria monocytogenes in foods
18.2.1 Material required for analysis
18.2.2 Procedure
18.3 Presence/absence method MLG/FSIS/USDA 2009 for Listeria monocytogenes in foods
18.3.1 Material required for analysis
18.3.2 Procedure
18.4 Plate count method ISO 11290-2:1998 Amendment 1:2004 for Listeria monocytogenes in foods
18.4.1 Material required for analysis
18.4.2 Procedure
18.5 Presence/absence method ISO 11290-1:1996 Amendment 1:2004 for Listeria monocytogenes in foods
18.5.1 Material required for analysis
18.5.2 Procedure
18.6 References

19 Salmonella
19.1 Introduction
19.1.1 Taxonomic classification of Salmonella
19.1.2 Serological classification of Salmonella
19.1.3 Biochemical characteristics of Salmonella
19.1.4 Epidemiology
19.1.5 Traditional methods used for the examination of Salmonella
19.1.6 Alternative methods for the analysis of Salmonella
19.1.7 Composite samples for analysis
19.2 Presence/absence method ISO 6579:2002 Amendment 1:2007 for Salmonella in foods
19.2.1 Material required for analysis
19.2.2 Procedure
19.3 Presence/absence method BAM/FDA 2011 for Salmonella in foods
19.3.1 Material required for analysis
19.3.2 Procedure
19.4 Presence/absence method MLG/FSIS/USDA 2011 for Salmonella in foods
19.4.1 Material required for analysis
19.4.2 Procedure
19.5 References

20 Vibrio cholerae and Vibrio parahaemolyticus
20.1 Introduction
20.1.1 Taxonomy
20.1.2 Epidemiology
20.1.2.1 V. cholerae
20.1.2.2 V. parahaemolyticus
20.1.2.3 V. vulnificus
20.1.3 Methods of analysis
20.2 Presence/absence method APHA 2001 and BAM/FDA 2004 for Vibrio cholerae in foods
20.2.1 Material required for analysis
20.2.2 Procedure
20.3 Most probable number (MPN) method APHA 2001 and BAM/FDA 2004 for Vibrio parahaemolyticus in foods
20.3.1 Material required for analysis
20.3.2 Procedure
20.4 Presence/absence method ISO 21872-1:2007 for presumptive enteropathogenic Vibrio cholerae and Vibrio parahaemolyticus in foods
20.4.1 Material required for analysis
20.4.2 Procedure
20.5 References

21 Yersinia enterocolitica
21.1 Introduction
21.1.1 Taxonomy
21.1.2 Epidemiology
21.2 Presence/absence method ISO 10273:2003 for presumptive pathogenic Yersinia enterocolitica in foods
21.2.1 Material required for analysis
21.2.2 Procedure
21.3 References

22 Bacterial spore count
22.1 Introduction
22.1.1 The bacterial spore
22.1.1.1 Sequence of spore formation
22.1.1.2 Spore ultrastructure
22.1.1.3 Mechanisms of spore resistance
22.1.2 Taxonomy of sporeforming bacteria important in foods
22.1.2.1 Aeribacillus Miñana-Galbis et al. 2010
22.1.2.2 Alicyclobacillus Wisotzkey et al. 1992 emend. Goto et al. 2003 emend. Karavaiko et al. 2005
22.1.2.3 Aneurinibacillus Shida et al. 1996 emend. Heyndrickx et al. 1997
22.1.2.4 Anoxybacillus Pikuta et al. 2000 emend. Pikuta et al. 2003
22.1.2.5 Bacillus Cohn 1872
22.1.2.6 Brevibacillus Shida et al. 1996
22.1.2.7 Clostridium Prazmowski 1880
22.1.2.8 Cohnella Kämpfer et al. 2006
22.1.2.9 Desulfotomaculum Campbell and Postgate 1965
22.1.2.10 Geobacillus Nazina et al. 2001
22.1.2.11 Jeotgalibacillus Yoon et al. 2001 emend. Chen et al. 2010
22.1.2.12 Lentibacillus Yoon et al. 2002 emend. Jeon et al. 2005
22.1.2.13 Lysinibacillus Ahmed et al. 2007
22.1.2.14 Moorella Collins et al. 1994
22.1.2.15 Oceanobacillus Lu et al. 2002 emend. Lee et al. 2006
22.1.2.16 Paenibacillus Ash et al. 1994 emend. Shida et al. 1997
22.1.2.17 Sporolactobacillus Kitahara and Suzuki 1963
22.1.2.18 Thermoanaerobacter Wiegel and Ljungdahl 1982 emend. Lee et al. 2007
22.1.2.19 Thermoanaerobacterium Lee et al. 1993
22.1.2.20 Virgibacillus Heyndrickx et al. 1998 emend. Wainø et al. 1999 emend. Heyrman et al. 2003
22.2 Methods APHA 2001 for spores of total and “flat sour” thermophilic aerobic sporeformers in foods
22.2.1 Material required for analysis
22.2.2 Procedure for the analysis of sugar
22.2.3 Procedure for the analysis of starch
22.2.4 Procedure for the analysis of whole tomatoes, tomato pulp, tomato puree and concentrated milk
22.2.5 Procedure for the analysis of nonfat dry milk
22.2.6 Procedure for the analysis of milk cream
22.2.7 Procedure for the analysis of other foods and ingredients (general)
22.3 Methods APHA 2001 for spores of thermophilic anaerobic sporeformers in foods
22.3.1 Material required for analysis
22.3.2 Procedure for the analysis of sugar and powdered milk
22.3.3 Procedure for the analysis of starches and flours
22.3.4 Procedure for the analysis of cereals and alimentary pastes
22.3.5 Procedure for the analysis of fresh mushrooms
22.3.6 Procedure for the analysis of “in-process” products
22.4 Methods APHA 2001 for spores of sulfide spoilage anaerobic sporeformers in foods
22.4.1 Material required for analysis
22.4.2 Procedure for the analysis of sugar
22.4.3 Procedure for the analysis of starch and flour
22.4.4 Procedure for the analysis of skim milk powder
22.4.5 Procedure for the analysis of soy protein isolates
22.5 Methods APHA 2001 for spores of mesophilic aerobic sporeformers in foods
22.5.1 Material required for analysis
22.5.2 Procedure for foods in general
22.5.3 Procedure for the analysis of milk and dairy products
22.5.4 Procedure for the analysis of water
22.6 Methods APHA 2001 for spores of mesophilic anaerobic sporeformers in foods
22.6.1 Material required for analysis
22.6.2 Procedure for the analysis of sugar
22.6.3 Procedure for the analysis of starch, flours and other cereal products
22.6.4 Procedure for the analysis of dehydrated vegetables
22.6.5 Procedure for the analysis of seasonings and spices
22.6.6 Procedure for the analysis of egg powder, milk powder and other powdered dairy products
22.6.7 Procedure for the analysis of fluid milk and cheeses
22.6.8 Other procedures for mesophilic anaerobic sporeformers
22.7 Methods IFU 12:2007 for Alicyclobacillus in foods
22.7.1 Material required for analysis
22.7.2 Procedure for the analysis of raw material
22.7.3 Procedure for analysis of the finished product
22.7.4 Interpretation and calculation of the results
22.8 References

23 Commercial sterility
23.1 Introduction
23.1.1 Parameters for evaluating the heat resistance of microorganisms
23.1.1.1 Survival curve and decimal reduction time (D value)
23.1.1.2 Number of decimal reductions
23.1.1.3 Thermal destruction curve and temperature coefficient (z value)
23.1.2 D and z values of microorganisms of importance in foods
23.1.3 Dimensioning heat treatments and thermal processing
23.1.4 Microbial spoilage of canned foods 317
23.2 Method APHA 2001 for commercial sterility or cause of spoilage of low acid canned foods
23.2.1 Material required for analysis
23.2.2 Procedure
23.2.3 Interpretation of the results
23.3 Method APHA 2001 for commercial sterility or cause of spoilage of acid canned foods
23.3.1 Material required for analysis
23.3.2 Procedure
23.3.3 Interpretation of the results
23.4 References

24 Guidelines on preparation of culture media
24.1 Introduction
24.1.1 Ingredients used in the formulation of culture media
24.1.1.1 Water for preparing media and reagents
24.1.1.2 Nutrient sources for culture media
24.1.1.3 Selective agents
24.1.1.4 Differential agents
24.1.1.5 Reducing agents
24.1.1.6 Buffering agents
24.1.1.7 Chromogenic and fluorogenic substrates
24.1.1.8 Agar
24.1.2 Types of culture media
24.2 Procedure for the preparation of culture media
24.2.1 Storing supplies and ingredients for preparation of culture media
24.2.2 Weighing and rehydration
24.2.3 Dissolution and dispersion
24.2.4 Verification and adjustment of the pH before sterilization
24.2.5 Distribution
24.2.6 Sterilization by moist heat
24.2.7 Sterilization by filtration
24.2.8 Verification after sterilization
24.2.9 Preparation of supplements for culture media
24.2.10 Storage of sterilized media until the moment of use
24.2.11 Preparation of the media at the moment of use
24.3 References

Annex 1 – Preparation of media and reagents

Annex 2 – Sampling plans and microbiological limits recommended by ICMSF for foods

Subject index