ISO 11731:2017
Water quality — Enumeration of Legionella
ISO 11731:2017 specifies culture methods for the isolation of Legionella and estimation of their numbers in water samples.
These methods are applicable to all kinds of water samples including potable, industrial, waste and natural waters. These methods can be used for water related matrices, e.g. biofilms, sediments, etc.
Not all Legionella species are culturable; therefore, the methods described in this document do not recover all species of Legionella.
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Foreword
Introduction
1 Scope
2 Normative references
3 Terms and definitions
4 Principle
4.1 General
4.2 Examination
4.3 Confirmation
5 Apparatus and glassware
6 Culture media and reagents
7 Sampling
8 Procedure
8.1 Samples
8.2 Concentration of water samples
8.2.1 General
8.2.2 Membrane filtration and direct placing of the membrane filter on culture media
8.2.3 Membrane filtration followed by a washing procedure
8.3 Sample pre-treatment
8.3.1 Heat treatment
8.3.2 Acid treatment
8.4 Culture
8.4.1 General
8.4.2 Samples with a high concentration of Legionella species and a low concentration of interfering microorganisms
8.4.3 Samples with a low concentration of Legionella species and a low concentration of interfering microorganisms
8.4.4 Samples with a high concentration of interfering microorganisms
8.4.5 Samples with an extremely high concentration of interfering microorganisms
8.4.6 Incubation
8.4.7 Examination of the plates
8.5 Confirmation of presumptive Legionella colonies on culture media: BCYE agar and BCYE–cys agar
9 Expression of results
10 Test report
11 Quality assurance
11.1 General
11.2 Performance testing of Legionella culture media
11.3 Preparing working culture and test suspension for performance testing
Annex A (informative) Legionella species
Annex B (normative) Culture media
Annex C (normative) Diluents
Annex D (normative) Acid solution
Annex E (informative) Scraping or rubbing the bacteria from membrane filters
Annex F (informative) Centrifugation technique
Annex G (informative) Indirect immunofluorescent antibody assay for the identification of Legionella species
Annex H (informative) Performance data
Annex I (informative) Pre-treatment of water related matrices
Annex J (normative) Decision matrix
Bibliography
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Foreword
Introduction
1 Scope
2 Normative references
3 Terms and definitions
4 Principle
4.1 General
4.2 Examination
4.3 Confirmation
5 Apparatus and glassware
6 Culture media and reagents
7 Sampling
8 Procedure
8.1 Samples
8.2 Concentration of water samples
8.2.1 General
8.2.2 Membrane filtration and direct placing of the membrane filter on culture media
8.2.3 Membrane filtration followed by a washing procedure
8.3 Sample pre-treatment
8.3.1 Heat treatment
8.3.2 Acid treatment
8.4 Culture
8.4.1 General
8.4.2 Samples with a high concentration of Legionella species and a low concentration of interfering microorganisms
8.4.3 Samples with a low concentration of Legionella species and a low concentration of interfering microorganisms
8.4.4 Samples with a high concentration of interfering microorganisms
8.4.5 Samples with an extremely high concentration of interfering microorganisms
8.4.6 Incubation
8.4.7 Examination of the plates
8.5 Confirmation of presumptive Legionella colonies on culture media: BCYE agar and BCYE–cys agar
9 Expression of results
10 Test report
11 Quality assurance
11.1 General
11.2 Performance testing of Legionella culture media
11.3 Preparing working culture and test suspension for performance testing
Annex A (informative) Legionella species
Annex B (normative) Culture media
Annex C (normative) Diluents
Annex D (normative) Acid solution
Annex E (informative) Scraping or rubbing the bacteria from membrane filters
Annex F (informative) Centrifugation technique
Annex G (informative) Indirect immunofluorescent antibody assay for the identification of Legionella species
Annex H (informative) Performance data
Annex I (informative) Pre-treatment of water related matrices
Annex J (normative) Decision matrix
Bibliography