Standard
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AS 4276.15:2014
[Current]Water microbiology, Method 15: Examination for Vibrio cholerae
This Standard sets out a method for detecting Vibrio cholerae in water using culture in enrichment broth and on selective agar media.
Published: 01/05/2014
Pages: 22
Table of contents
Cited references
Content history
Table of contents
Header
About this publication
Preface
1 Scope
2 Referenced documents
3 Definition
4 Principle
5 Culture media and reagents
5.1 Culture media
5.1.1 Alkaline peptone water (APW)
5.1.2 Thiosulphate citrate bile salts sucrose agar (TCBS)
5.1.3 Nutrient agar
5.1.4 8% NaCl tryptone water (8% NaCl TW)
5.1.5 3% NaCl tryptone water (3% NaCl TW)
5.1.6 0% NaCl tryptone water (0% NaCl TW)
5.1.7 Tryptone water
5.1.8 Buffered glucose broth
5.1.9 Arginine dihydrolase broth
5.1.10 Lysine decarboxylase broth
5.1.11 Ornithine decarboxylase broth
5.1.12 Control decarboxylase broth
5.1.13 Glucose fermentation broth
5.1.14 Arabinose fermentation broth
5.1.15 Cellobiose fermentation broth
5.1.16 Lactose fermentation broth
5.1.17 Maltose fermentation broth
5.1.18 Mannitol fermentation broth
5.1.19 Salicin fermentation broth
5.1.20 Sucrose fermentation broth
5.1.21 Nitrate reduction broth
5.1.22 O-nitrophenyl-β-D-galactopyranoside broth (ONPG)
5.1.23 Cellobiose-colistin (CC) Agar
5.1.24 Modified cellobiose polymyxin B-colistin (mCPC) Agar
5.1.25 Sodium dodecyl sulphate polymyxin sucrose (SDSPS) Agar
5.2 Reagents
5.2.1 Kovacs’ oxidase reagent
5.2.2 Kovacs’ indole reagent
5.2.3 5% α naphthol in ethanol
5.2.4 40% KOH
5.2.5 Nitrate reagent A (0.8% w/v sulphanilic acid in 5N acetic acid)
5.2.6 Nitrate reagent B (0.6% v/v N,N-Dimethly-α-naphthylamine in 5N acetic acid)
6 Apparatus
6.1 Apparatus and materials for membrane filtration
6.2 Incubators or water baths
7 Reference cultures
7.1 Positive culture
7.2 Use of reference culture
8 Samples
8.1 Laboratory sample
8.2 Sample volume
8.3 Concentration techniques
8.3.1 Moore swab
8.3.2 Membrane filtration
9 Procedure
9.1 Enrichment
9.2 Selective agar culture
9.3 Presumptive identification of Vibrio cholerae
9.4 Confirmation of Vibrio cholerae
9.4.1 Phenotypic identification and serological characterization
9.4.2 Genotypic tests
10 Test report
11 Bibliography
Appendix A
Appendix B
B1 Scope
B2 0%, 3%, 8% NaCl tryptone water
B3 Alkaline peptone water
B4 Buffered glucose broth
B5 Carbohydrate fermentation broths
B5.1 Preparation of complete medium
B5.2 Fermentation Broth base
B5.3 Andrade’s indicator
B6 Decarboxylase broth (Moeller’s)
B6.1 Preparation of complete medium
B6.2 Decarboxylase broth base (Moeller’s)
B7 Nitrate reduction broth
B8 Thiosulfate citrate bile salts sucrose agar (TCBS)
B9 Tryptone water
B10 ONPG broth
B10.1 Preparation of complete medium
B10.2 Preparation of ONPG solution
B11 Nutrient agar
B12 Kovacs’ oxidase reagent
B13 Kovacs’ indole reagent
B14 5% Alpha naphthol in ethanol
B15 40% KOH
B16 Nitrate reagent A
B17 Nitrate reagent B
B18 Cellobiose-colistin (CC) agar
B18.1 Preparation of complete medium
B18.2 Solution 1
B18.3 1000x dye stock solution
B18.4 Solution 2
B19 Modified cellobiose-polymyxin B-colistin (mCPC) agar
B19.1 Preparation of complete medium
B19.2 Solution 1
B19.3 1000x dye stock solution
B19.4 Solution 2
B20 Sodium dodecyl sulfate polymyxin sucrose (SDSPS) agar
B20.1 Preparation of complete medium
B20.2 Polymyxin B solution
Appendix C
Appendix D
D1 Indole test
D2 Voges-proskauer test (for the production of acetoin)
D3 Nitrate reduction test (Barritt)
Appendix E
E1 Purpose and scope
E2 Acknowledgements
E3 Principle
E4 Materials
E4.1 Primers and products
E4.2 PCR master mix recipe
E5 Method
E5.1 Isolate preparation
E5.2 PCR reaction
E5.3 Reaction volumes
E5.4 PCR thermocycler program
E5.5 PCR product analysis
E6 Quality control
E7 References
Cited references in this standard
[Current]
Water microbiology, Method 1: Water quality — General requirements and guidance for microbiological examinations by culture (ISO 8199:2018, MOD)
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