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BSI PD CEN/TS 17716:2022

BSI PD CEN/TS 17716:2022

$142.49

Plant biostimulants. Determination of Escherichia coli

Published By Publication Date Number of Pages
BSI 2022 24
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This document gives general guidelines for the detection and identification of the specified microorganism Escherichia coli in technical and formulated biostimulant products, both in liquid and solid state, and also the horizontal method for the enumeration of ß-glucuronidase-positive Escherichia coli in plant biostimulants products (both in liquid and solid state). The qualitative method described in this document is based on the detection of Escherichia coli in a non-selective liquid medium (enrichment broth), followed by isolation on a selective agar medium. Other methods can be appropriate, depending on the level of detection required. NOTE For the detection of Escherichia coli, subcultures can be performed on non-selective culture media followed by suitable identification steps (e.g. using identification kits). The quantitative method described in this document uses a colony-count technique at 44 °C on a solid medium containing a chromogenic ingredient for detection of the enzyme ß-glucuronidase. WARNING — Strains of Escherichia coli which do not grow at 44 °C and, in particular, those that are ß glucuronidase negative, such as Escherichia coli O157, will not be detected.

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PDF Pages PDF Title
2 undefined
7 1 Scope
2 Normative references
3 Terms and definitions
8 4 Principle
4.1 Qualitative method
4.2 Quantitative method
5 Diluent and culture media
5.1 General
5.2 Broth and culture media in the qualitative method
5.2.1 Enrichment broth
9 5.2.2 Selective culture media: Tryptone-bile-glucuronic medium (TBX) for isolation of Escherichia coli
5.3 Diluent and culture media in the quantitative method
5.3.1 Diluent
5.3.2 Culture media: Tryptone-bile-glucuronic medium (TBX)
6 Apparatus and glassware
7 Handling of plant biostimulants products and sampling
8 Procedure
8.1 General
8.2 Qualitative method
8.2.1 General
10 8.2.2 Solid formulations: Wettable Powder (WP), Water dispersible granules (WDG), Pellets, granules, microgranules (slow release) formulation
8.2.3 Liquid formulations: water based formulations and oil based (emulsifiable concentrate – EC) formulations
8.2.4 Incubation of the inoculated enrichment broth
8.2.5 Detection and identification of Escherichia coli
8.2.5.1 Isolation
8.2.5.2 Identification of Escherichia coli
11 8.3 Quantitative method
8.3.1 Test portion and initial suspension
8.3.1.1 General
8.3.1.2 Liquid – water based – formulations
8.3.1.3 Liquid – oil based (emulsifiable concentrate – EC) formulations
8.3.1.4 Solid – Wettable Powder (WP) formulations
8.3.1.5 Solid – Water dispersible granules (WDG) formulations
8.3.1.6 Solid – Pellets, granules, microgranules (slow release) formulations
8.3.1.7 Solid – substrate
8.3.2 Serial dilutions
12 8.3.3 Inoculation (surface plate method) and incubation
8.3.4 Counting the colony-forming units
9 Expression of results
9.1 Expression of results in the qualitative test
9.2 Expression of results in the quantitative test
13 10 Test report
14 Annex A (informative)Enrichment broth in the qualitative method
A.1 General
A.2 Enrichment broth
A.2.1 Eugon LT 100 broth
A.2.1.1 General
A.2.1.2 Composition
A.2.1.3 Preparation
15 A.3 Other enrichment broth
A.3.1 General
A.3.2 Fluid lactose medium
A.3.2.1 Composition
A.3.2.2 Preparation
16 Annex B (informative)Selective agar medium in the qualitative and quantitative method
B.1 General
B.2 Selective agar medium for isolation
B.2.1 Tryptone-bile-glucuronic medium (TBX)
B.2.1.1 Composition
B.2.1.2 Preparation
17 B.3 Other selective agar medium for isolation
B.3.1 MacConkey agar medium
B.3.1.1 Composition
B.3.1.2 Preparation
B.3.1.3 Morphological characteristics of Escherichia coli on MacConkey agar medium
B.4 Selective agar medium for confirmation
B.4.1 Levine eosin-methylene blue agar medium (EMB agar medium)
B.4.1.1 Composition
18 B.4.1.2 Preparation
B.4.1.3 Morphological characteristics of Escherichia coli on EMB agar medium
19 Annex C (informative)Diluent in the quantitative method
C.1 General
C.2 Phosphate Buffered Saline (PBS)
C.2.1 Composition
C.2.2 Preparation
20 Annex D (informative)Neutralization of the antimicrobial properties of the product
D.1 General
D.2 Suitability of the detection method
21 D.3 Interpretation of suitability test results

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BSI PD CEN/TS 17716:2022
$142.49