Standard
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AS 4276.19:2014
[Current]Water microbiology, Method 19: Examination for thermophilic Campylobacter spp. — Membrane filtration
This Standard sets out a method for detecting thermophilic Campylobacters in water using culture in enrichment broth and on selective agar media.
Published: 01/05/2014
Pages: 15
Table of contents
Cited references
Content history
Table of contents
Header
About this publication
Preface
1 Scope
2 Referenced documents
3 Principle
4 Culture media and reagents
4.1 Culture media
4.1.1 Modified exeter broth (mExeter)
4.1.2 Bolton broth
4.1.3 Modified charcoal cefoperazone dexoxycholate agar (mCCD)
4.1.4 Blood agar
4.2 Reagents
4.2.1 Kovacs’oxidase reagent
4.2.2 Dilute carbol fuschin
4.2.3 Physiological saline solution, 0.85% aqueous solution
4.2.4 Sodium hippurate, 10 g/L aqueous solution
4.2.5 Ninhydrin solution, 3.5% in 1:1 mixture of acetone and butanol
4.2.6 Catalase reagent, 3% hydrogen peroxide (H2O2)
4.2.7 Indoxyl acetate discs
5 Reference cultures
5.1 Campylobacter jejuni, WDCM 00156 or WDCM 00005 or NCTC 11351 or a culture traceable to any of these strains.
5.2 Campylobacter coli, WDCM 00072 or WDCM 00004 or a culture traceable to any of these strains.
5.3 Use of reference cultures
6 Apparatus
6.1 Gas cylinders or kits
6.2 Filtration apparatus
6.3 Filter membranes 0.2 or 0.22 μm pore size
6.4 Sterile diatomaceous earth (‘filter-aid’) plus sterile absorbent pads (optional)
6.5 Incubators
7 Sampling
8 Procedures
8.1 Microaerobic incubation
8.2 Membrane filtration
8.3 Enrichment
8.3.1 General
8.3.2 Waters with low background counts
8.3.3 Waters with high background counts
8.4 Subculture onto selective medium
8.5 Examination of selective agar plates
9 Confirmation tests
9.1 General
9.2 Screening of colonies
9.3 Oxidase test
9.4 Gram stain
9.5 Motility
9.6 Confirmed thermophilic Campylobacter spp.
9.7 Genotypic tests
10 Speciation tests (Optional)
10.1 General
10.2 Hippurate hydrolysis
10.3 Catalase test
10.4 Indoxyl acetate
11 Test report
12 Bibliography
Appendix A
Appendix B
B1 Scope
B2 Complete modified exeter (mExeter) broth
B2.1 Composition
B2.2 mExeter basal broth
B2.3 Solution of mExeter broth growth supplement
B2.4 Preparation of lysed blood
B2.5 Antibiotic supplement
B2.6 Preparation of complete mExeter broth
B3 Complete bolton broth
B3.1 Composition
B3.2 Bolton basal broth
B3.3 Solution of bolton broth selective supplement
B3.4 Preparation of lysed blood
B3.5 Preparation of complete bolton broth
B4 Complete modified charcoal cefoperazone desoxycholate agar (mCCD)
B4.1 Composition
B4.2 Basal medium
B4.3 mCCD agar selective supplement
B4.4 Preparation of complete mCCD agar
B5 Complete blood agar
B5.1 General
B5.2 Nutrient agar
B6 Oxidase reagent (Kovacs’)
B7 Carbol fuchsin
B7.1 Strong
B7.2 Dilute
B8 Physiological saline solution
B9 Sodium hippurate, 10 g/L aqueous solution
B10 Ninhydrin solution, 3.5% in 1:1 mixture of acetone and butanol
B11 Catalase reagent, 3% solution of hydrogen peroxide, H2O2
B12 Indoxyl acetate discs
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)
[Pending Revision]
Water quality — Sampling for microbiological analysis (ISO 19458:2006, MOD)
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