Standard
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AS 4276.17.2:2016
[Current]Water microbiology, Method 17.2: Spores of Clostridium perfringens — Estimation of most probable number (MPN) using the multiple tube dilution technique
Sets out a method for determining the most probable number (MPN) of spores of C. perfringens in water, using a multiple tube dilution technique, with a confirmation procedure for identifying C. perfringens.
Published: 12/04/2016
Pages: 10
Table of contents
Cited references
Content history
Table of contents
Header
About this publication
Preface
Foreword
1 Scope
2 Referenced documents
3 Definitions
4 Principle
5 Media, reagents, and reference cultures
5.1 Culture media (see Appendix B)
5.1.1 Differential reinforced clostridial medium (DRCM)
5.1.2 Tryptose sulfite cycloserine (TSC) agar
5.1.3 Oleandomycin polymyxin sulphadiazine perfringens (OPSP) agar
5.1.4 Columbia blood agar or other suitable nutrient-rich agar
5.1.5 Columbia blood agar with neomycin (0.01%)
5.1.6 Acid phosphatase reagent
5.2 Reference cultures
5.2.1 Positive culture
5.2.2 Negative culture
5.2.3 Use of reference cultures
6 Apparatus
6.1 Screw cap containers
6.2 Pipettes or equivalent measuring devices
6.3 Anaerobic incubators/jar
6.4 Incubators or water bath
7 Samples
8 Test procedures
9 Confirmation of C. Perfringens
9.1 Waters with low levels of competing spore forming bacteria
9.2 Waters with high levels of competing spore forming bacteria
9.3 Selection of colonies
9.4 Biochemical confirmation
9.4.1 Acid phosphatase test
9.4.2 Alternative biochemical tests
9.5 Confirmed C. perfringens
10 Calculations
11 Report
12 Bibliography
Appendix A
Appendix B
B1 Scope
B2 Differential reinforced clostridial medium (single strength)
B3 Tryptose sulfite cycloserine (TSC) agar
B3.1 TSC base agar
B3.2 TSC selective agent (per 1 L TSC base)
B3.3 Preparation of complete TSC agar
B4 Oleandomycin polymyxin sulphadiazine perfringens (OPSP) agar
B4.1 OPSP base agar
B4.2 OPSP selective agents (per 1 L OPSP base)
B4.3 Preparation of complete OPSP agar
B5 Columbia blood agar
B6 Columbia blood agar with neomycin (0.01%)
B7 Acid phosphatase reagent
Cited references in this standard
[Pending Revision]
Water quality — Sampling for microbiological analysis (ISO 19458:2006, MOD)
[Current]
Food microbiology, Method 16: Microbiology of food and animal feeding stuffs — Horizontal method for the enumeration of Clostridium perfringens — Colony-count technique
[Superseded]
Water microbiology, Method 1: General information and procedures (ISO 8199:2005, MOD)
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