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BSI PD CEN/TS 17710:2022

BSI PD CEN/TS 17710:2022

$142.49

Plant biostimulants. Detection of Listeria monocytogenes

Published By Publication Date Number of Pages
BSI 2022 28
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This document provides a method for the detection of Listeria monocytogenes in microbial plant biostimulants for verifying that the concentration of this human pathogen does not exceed the respective limits outlined in the EU Regulation on Fertilising Products [1].

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PDF Pages PDF Title
2 undefined
7 1 Scope
2 Normative references
3 Terms and definitions
4 Principle
4.1 General
4.2 Pre-enrichment in non-selective liquid medium
8 4.3 Enrichment in/on selective media
4.4 Plating out on selective solid media
4.5 Confirmation
5 Culture media, reagents, antisera
6 Equipment and consumables
9 7 Sampling
8 Preparation of test sample
9 Preparation procedure
9.1 Test portion and initial suspension
9.1.1 General
9.1.2 Liquid – water based– formulations
10 9.1.3 Liquid – oil based (emulsifiable concentrate – EC) formulations
9.1.4 Solid – Wettable Powder (WP) formulations
9.1.5 Solid – Water dispersible granules (WDG) formulations
9.1.6 Solid – Pellets, granules, microgranules (slow release) formulations
9.1.7 Solid substrates
9.2 Non-selective pre-enrichment
9.3 Selective enrichment
9.4 Plating out
9.4.1 General
11 9.4.2 Agar Listeria according to Ottaviani and Agosti (B.3)
9.4.3 Second selective medium
9.5 Confirmation of L. monocytogenes
9.5.1 General
9.5.2 Selection of colonies for confirmation
12 9.5.3 Confirmation tests for L. monocytogenes
9.5.3.1 General
9.5.3.2 Catalase reaction (optional)
9.5.3.3 Motility test (optional)
13 9.5.3.4 Microscopic aspect
9.5.3.5 Haemolysis tests
14 9.5.3.6 CAMP test (optional)
9.5.3.7 Carbohydrate utilization
9.6 Interpretation of morphological and physiological properties and of the biochemical reactions
9.7 Additional characterization of isolated strains (optional)
10 Expression of results
15 11 Performance characteristics of the method
11.1 Interlaboratory studies
11.2 Sensitivity
11.3 Specificity
11.4 LOD50
12 Test report
13 Quality assurance
16 Annex A (normative)Diagram of the procedures
17 Annex B (normative)Composition and preparation of culture media and reagents
B.1 Selective primary enrichment medium: half-Fraser broth
B.1.1 Base
B.1.1.1 Composition
B.1.1.2 Preparation
B.1.2 Lithium chloride solution
B.1.2.1 Composition
18 B.1.2.2 Preparation
B.1.3 Solution of sodium salt of nalidixic acid
B.1.3.1 Composition
B.1.3.2 Preparation
B.1.4 Acriflavine hydrochloride solution
B.1.4.1 Composition
B.1.4.2 Preparation
B.1.5 Ammonium iron(III) citrate solution
B.1.5.1 Composition
B.1.5.2 Preparation
19 B.1.6 Complete medium
B.1.6.1 Composition
B.1.6.2 Preparation
B.2 Selective secondary enrichment medium: Fraser broth
B.2.1 Base
B.2.1.1 Composition
B.2.1.2 Preparation
B.2.2 Acriflavine hydrochloride solution
20 B.2.3 Ammonium iron(III) citrate solution
B.2.4 Complete medium
B.3 Agar Listeria according to Ottaviani and Agosti (ALOA)
B.3.1 Base medium
B.3.1.1 Composition
B.3.1.2 Preparation
B.3.2 Nalidixic acid solution
B.3.2.1 Composition
21 B.3.2.2 Preparation
B.3.3 Ceftazidime solution
B.3.3.1 Composition
B.3.3.2 Preparation
B.3.4 Polymyxin B solution
B.3.4.1 Composition
B.3.4.2 Preparation
B.3.5 Antibiotic supplement
B.3.5.1 Cycloheximide solution
22 B.3.5.2 Amphotericin B solution (as an alternative to cycloheximide solution)
B.3.6 Supplement
B.3.7 Complete medium
B.3.7.1 Composition
B.3.7.2 Preparation
B.3.7.3 Preparation of agar plates
23 B.4 Second selective solid plating-out medium
B.5 Performance testing for the quality assurance of the culture media
B.6 Hydrogen peroxide solution
B.7 Motility agar
B.7.1 Composition
B.7.2 Preparation
B.8 Blood agar
B.8.1 Base
B.8.1.1 Composition
24 B.8.1.2 Preparation
B.8.2 Defibrinated blood (sheep, calf or bovine)
B.8.3 Complete medium
B.8.3.1 Composition
B.8.3.2 Preparation
B.9 Phosphate-buffered saline (PBS)
B.9.1 Composition
B.9.2 Preparation
B.10 Red blood corpuscle suspension
25 B.11 CAMP (Christie, Atkins, Munch-Petersen) medium and test strains
B.11.1 General
B.11.2 Base
B.11.3 Blood medium
B.11.4 Complete medium
B.12 Carbohydrate utilization broth (L-Rhamnose and D-Xylose)
B.12.1 Base
B.12.1.1 Composition
B.12.1.2 Preparation
26 B.12.2 Carbohydrate solutions
B.12.2.1 Composition
B.12.2.2 Preparation
B.12.3 Complete medium
B.13 Tryptone soya yeast extract agar (TSYEA)
B.13.1 Composition
B.13.2 Preparation
B.13.2.1 General
B.13.2.2 Preparation of agar plates

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BSI PD CEN/TS 17710:2022
$142.49