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BSI PD CEN/TS 17715:2022

BSI PD CEN/TS 17715:2022

$142.49

Plant biostimulants. Detection of Shigella spp.

Published By Publication Date Number of Pages
BSI 2022 30
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This document provides a method for verifying that the pathogen Shigella spp. is not present in microbial plant biostimulants in a concentration that exceeds the respective limits outlined in the EU Regulation on Fertilising Products. The detection method for Shigella pathogens is not sensitive and quantification is rarely performed. Detection is usually performed using an enrichment medium followed by subculturing onto a variety of selective media.

PDF Catalog

PDF Pages PDF Title
2 undefined
7 1 Scope
2 Normative references
3 Terms and definitions
4 Principle
4.1 General
4.2 Enrichment in selective liquid medium
4.3 Plating out and identification of colonies
8 4.4 Biochemical and serological confirmation
5 Culture media, reagents and antisera
6 Apparatus and glassware
7 Sampling
8 Procedure
8.1 General
8.2 Test portion
8.3 Enrichment
8.3.1 General
8.3.2 Liquid formulations
9 8.3.3 Solid formulations
8.4 Plating out and colony selection
8.5 Confirmation of colonies
8.5.1 General
8.5.2 Purification of colonies
10 8.5.3 Biochemical confirmation
8.5.3.1 General
8.5.3.2 Triple sugar iron agar (TSI slopes) (B.5)
8.5.3.3 Semi-solid nutrient agar for motility tests (B.6)
8.5.3.4 Urea agar (B.7)
8.5.3.5 L-Lysine decarboxylase medium (B.8)
11 8.5.3.6 L-Ornithine decarboxylase medium (B.9)
8.5.3.7 Detection of indole formation (B.10)
8.5.3.8 Detection of β-galactosidase (B.11)
8.5.3.9 Utilization of carbohydrates (B.12)
8.5.3.10 Interpretation of biochemical results
13 8.5.4 Additional biochemical differentiation
8.5.4.1 General
8.5.4.2 Sodium acetate
8.5.4.3 Christensen’s citrate
8.5.4.4 Sodium mucate
14 8.6 Serological confirmation
8.6.1 Antigenic differentiation
15 8.6.2 Agglutination tests
8.6.3 Definitive confirmation
9 Expression of results
10 Test report
16 Annex A (normative)Diagram of test procedure
17 Annex B (normative)Composition and preparation of culture media and reagents
B.1 General
B.2 Selective enrichment broth
B.2.1 Shigella broth
B.2.1.1 Composition
B.2.1.2 Preparation
B.2.2 Novobiocin solution
B.2.2.1 Composition
B.2.2.2 Preparation
18 B.2.3 Complete medium
B.3 Selective differential agars
B.3.1 MacConkey agar
B.3.1.1 Composition
B.3.1.2 Preparation
B.3.2 Xylose lysine desoxycholate (XLD) agar
B.3.2.1 Composition
19 B.3.2.2 Preparation
B.3.3 Hektoen enteric (HE) agar
B.3.3.1 Composition
B.3.3.2 Preparation
20 B.4 Nutrient agar
B.4.1 Composition
B.4.2 Preparation
B.4.3 Preparation of nutrient agar plates
B.5 Triple iron sugar (TSI) agar
B.5.1 Composition
21 B.5.2 Preparation
B.6 Semi-solid nutrient agar
B.6.1 Composition
B.6.2 Preparation
B.7 Urea agar (Christensen’s medium)
B.7.1 Base
B.7.1.1 Composition
22 B.7.1.2 Preparation
B.7.2 Urea solution
B.7.2.1 Composition
B.7.2.2 Preparation
B.7.3 Complete medium
B.7.3.1 Composition
B.7.3.2 Preparation
B.8 L-Lysine decarboxylase medium
B.8.1 Composition
B.8.2 Preparation
23 B.9 L-Ornithine decarboxylase medium
B.10 Reagents for indole reaction
B.10.1 Tryptone/DL-tryptophan medium
B.10.1.1 Composition
B.10.1.2 Preparation
B.10.2 Kovac’s indole reagent
B.10.2.1 Composition
B.10.2.2 Preparation
B.11 β-Galactosidase reagent
B.11.1 Buffer solution
B.11.1.1 Composition
24 B.11.1.2 Preparation
B.11.2 ONPG solution
B.11.2.1 Composition
B.11.2.2 Preparation
B.11.3 Complete medium
B.11.3.1 Composition
B.11.3.2 Preparation
B.12 Bromocresol purple broth
B.12.1 Composition
B.12.2 Preparation
25 B.13 Saline solution
B.13.1 Composition
B.13.2 Preparation
B.14 Sodium acetate agar
B.14.1 Composition
B.14.2 Composition
B.15 Christensen’s citrate agar
B.15.1 Composition
26 B.15.2 Preparation
B.16 Mucate broth
B.16.1 Test broth
B.16.1.1 Composition
B.16.1.2 Preparation
B.16.2 Control broth
B.16.2.1 Composition
27 B.16.2.2 Preparation
B.17 Antisera of Shigella species
28 Annex C (normative) Description of Shigella colony morphology and colour on selective agars, for both identification and quality control purposes

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BSI PD CEN/TS 17715:2022
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