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AS 5013.10:2022
[Current]Food microbiology, Method 10: Microbiology of the food chain — Horizontal method for the detection, enumeration and serotyping of Salmonella — Detection of Salmonella spp. (ISO 6579-1:2017, MOD)
Adopts and modifies ISO 6579 1:2017 and its Amendment No. 1 (2020), for Australia, specifying a horizontal method for the detection of Salmonella. Applicable to: products intended for human consumption and the feeding of animals; environmental samples in the area of food production and food handling; and samples from the primary production stage such as animal faeces, dust, and swabs.
Published: 21/01/2022
Pages: 55
Table of contents
Cited references
Content history
Table of contents
Header
About this publication
Preface
Foreword
Introduction
1 Scope
2 Normative references
3 Terms and definitions
4 Principle
4.1 General
4.2 Pre-enrichment in non-selective liquid medium
4.3 Enrichment in/on selective media
4.4 Plating out on selective solid media
4.5 Confirmation
5 Culture media, reagents, and antisera
6 Equipment and consumables
6.1
6.2
6.3
6.4
6.5
6.6
6.7
6.8
6.9
6.10
6.11
6.12
6.13
6.14
7 Sampling
8 Preparation of test sample
9 Procedure (see diagrams in Annex A)
9.1 Test portion and initial suspension
9.2 Non-selective pre-enrichment
9.3 Selective enrichment
9.3.1 General
9.3.2 Procedure for food, animal feed samples, and environmental samples from the food production area
9.3.3 Procedure for samples from the primary production stage
9.4 Plating out
9.4.1 General
9.4.2 Procedure for food, animal feed samples, and environmental samples from the food production area
9.4.3 Procedure for samples from the primary production stage
9.5 Confirmation
9.5.1 General
9.5.2 Selection of colonies for confirmation
9.5.3 Biochemical testing
9.5.3.1 General
9.5.3.2 TSI agar(B.8)
9.5.3.3 Urea agar (B.9)
9.5.3.4 L-Lysine decarboxylation medium(LDC, B.10)
9.5.3.5 Detection of β -galactosidase (B.11)(optional)
9.5.3.6 Medium for indole reaction (B.12)(optional)
9.5.4 Serological testing
9.5.4.1 General
9.5.4.2 Elimination of auto-agglutinable strains
9.5.4.3 Examination for O-antigens
9.5.4.4 Examination for Vi-antigens (optional)
9.5.4.5 Examination for H-antigens
9.5.5 Interpretation of biochemical and serological reactions
9.5.6 Serotyping
10 Expression of results
11 Performance characteristics of the method
11.1 Interlaboratory studies
11.2 Sensitivity
11.3 Specificity
11.4 LOD50
12 Test report
Annex A
Annex B
B.1 General
B.2 Buffered peptone water (BPW)
B.2.1 Composition
B.2.2 Preparation
B.3 Rappaport-Vassiliadis medium with soya (RVS broth)
B.3.1 Solution A
B.3.1.1 Composition
B.3.1.2 Preparation
B.3.2 Solution B
B.3.2.1 Composition
B.3.2.2 Preparation
B.3.3 Solution C
B.3.3.1 Composition
B.3.3.2 Preparation
B.3.4 Complete medium
B.3.4.1 Composition
B.3.4.2 Preparation
B.4 Modified semi-solid Rappaport-Vassiliadis (MSRV) agar
B.4.1 Solution A
B.4.1.1 Composition
B.4.1.2 Preparation
B.4.2 Solution B
B.4.2.1 Composition
B.4.2.2 Preparation
B.4.3 Solution C
B.4.3.1 Composition
B.4.3.2 Preparation
B.4.4 Base medium
B.4.4.1 Composition
B.4.4.2 Preparation
B.4.5 Novobiocin solution
B.4.5.1 Composition
B.4.5.2 Preparation
B.4.6 Complete medium
B.4.6.1 Composition
B.4.6.2 Preparation
B.5 Muller-Kauffmann tetrathionate-novobiocin (MKTTn) broth
B.5.1 Base medium
B.5.1.1 Composition
B.5.1.2 Preparation
B.5.2 Iodine-iodide solution
B.5.2.1 Composition
B.5.2.2 Preparation
B.5.3 Novobiocin solution
B.5.3.1 Composition
B.5.3.2 Preparation
B.5.4 Complete medium
B.5.4.1 Composition
B.5.4.2 Preparation
B.6 Xylose Lysine Deoxycholate agar (XLD agar)
B.6.1 Composition
B.6.2 Preparation
B.6.3 Preparation of the agar plates
B.7 Nutrient agar (example of non-selective medium)
B.7.1 Composition
B.7.2 Preparation
B.7.3 Preparation of nutrient agar plates
B.8 Triple sugar/iron agar (TSI agar)
B.8.1 Composition
B.8.2 Preparation
B.9 Urea agar (Christensen)
B.9.1 Base medium
B.9.1.1 Composition
B.9.1.2 Preparation
B.9.2 Urea solution
B.9.2.1 Composition
B.9.2.2 Preparation
B.9.3 Complete medium
B.9.3.1 Composition
B.9.3.2 Preparation
B.10 L-Lysine decarboxylation medium (LDC)
B.10.1 Composition
B.10.2 Preparation
B.11 β-Galactosidase reagent (optional)
B.11.1 Buffer solution
B.11.1.1 Composition
B.11.1.2 Preparation
B.11.2 ONPG solution
B.11.2.1 Composition
B.11.2.2 Preparation
B.11.3 Complete reagent
B.11.3.1 Composition
B.11.3.2 Preparation
B.12 Medium and reagent for indole reaction (optional)
B.12.1 Tryptone/tryptophan medium
B.12.1.1 Composition
B.12.1.2 Preparation
B.12.2 Kovacs reagent
B.12.2.1 Composition
B.12.2.2 Preparation
B.13 Saline solution
B.13.1 Composition
B.13.2 Preparation
B.14 Antisera
B.15 Performance testing for the quality assurance of the culture media
Annex C
C.1 Performance characteristics RVS broth and MKTTn broth
C.2 Performance characteristics of MSRV agar for detection of Salmonella spp. in food and animal feed
C.3 Performance characteristics of MSRV agar for detection of Salmonella spp. in animal faeces and in environmental samples from the primary production stage
Annex D
D.1 General
D.2 Detection of Salmonella Typhi and Salmonella Paratyphi
D.2.1 Principle
D.2.1.1 General
D.2.1.2 Pre-enrichment in non-selective liquid medium
D.2.1.3 Enrichment in selective liquid media
D.2.1.4 Plating out and identification
D.2.1.5 Confirmation of identity
D.3 Culture media
D.3.1 Selenite cystine medium (SC)
D.3.1.1 Base medium
D.3.1.1.1 Composition
D.3.1.1.2 Preparation
D.3.1.2 L-Cystine solution
D.3.1.2.1 Composition
D.3.1.2.2 Preparation
D.3.1.3 Complete medium
D.3.1.3.1 Composition
D.3.1.3.2 Preparation
D.3.2 Bismuth sulfite agar (BS)
D.3.2.1 Composition
D.3.2.2 Preparation
D.3.3 Performance testing
D.4 Equipment and consumables
D.4.1 General
D.4.2 Volumetric flasks, sterile, 100 ml nominal capacity.
D.5 Procedure
D.5.1 General
D.5.2 Enrichment in selective liquid media
D.5.3 Plating out
D.5.3.1
D.5.3.2
D.5.3.3
D.5.3.4
D.5.3.5
D.6 Confirmation
D.6.1 General
D.6.2 Interpretation of the biochemical tests
Annex E
Bibliography
Amendment 1
Appendix ZZ
ZZ.1 Scope
ZZ.2 Variations
Cited references in this standard
[Current]
Microbiology of food, animal feed and water - Preparation, production, storage and performance testing of culture media
[Current]
Microbiology of the food chain - General requirements and guidance for microbiological examinations
ISO 6887 (all parts)
Microbiology of food and animal feed — Preparation of test samples, initial suspension and decimal dilutions for microbiological examination
Content history
[Current]
[Current]
[Superseded]
DR AS 5013.10:2021
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