Plant biostimulants - Detection of Shigella spp.

This document provides a method for verifying that the pathogen Shigella spp. is not present in microbial plant biostimulants.
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.
This document is applicable to the blends of fertilizing products where a blend is a mix of at least two of the following component EU fertilizing products categories: Fertilizers, Liming Materials, Soil Improvers, Growing Media, Plant Biostimulants and where the following category Plant Biostimulants is the highest percentage in the blend by mass or volume, or in the case of liquid form by dry mass. If Plant Biostimulants is not the highest percentage in the blend, the European Standard for the highest percentage of the blend applies. In case a blend of fertilizing products is composed of components in equal quantity or in case the component EU fertilising products used for the blend have identical formulations , the user decides which standard to apply.

Pflanzen-Biostimulanzien - Nachweis von Shigella spp.

Dieses Dokument liefert ein Verfahren zum Nachweis, dass das Pathogen Shigella spp. in mikrobiellen Pflanzen-Biostimulanzien nicht vorhanden ist.
Das Nachweisverfahren für Shigella-Pathogene ist nicht trennscharf, und eine Quantifizierung wird selten durchgeführt. Der Nachweis erfolgt in der Regel mit einem Anreicherungsmedium und anschließender Subkultivierung auf verschiedenen Selektivmedien.
Dieses Dokument ist für die Mischungen von Düngeprodukten anwendbar, in denen eine Mischung aus mindestens zwei der folgenden Kategorien von Komponenten-EU Düngeprodukten besteht: Düngemittel, Kalkdünger, Bodenverbesserungsmittel, Kultursubstrate, Pflanzen-Biostimulanzien und wenn die folgende Kategorie Pflanzen-Biostimulanzien prozentual den höchsten Massen- oder Volumenanteil in der Mischung bzw., bei flüssiger Form, den höchsten Trockenmasseanteil aufweist. Wenn Pflanzen-Biostimulanzien nicht den höchsten prozentualen Anteil in der Mischung ausmachen, ist die für den höchsten prozentualen Anteil geltende Europäische Norm anwendbar. Falls eine Düngemittelmischung aus Komponenten gleicher Menge besteht oder falls die für die Mischung verwendeten Komponenten-EU Düngeprodukte identische Formulierungen  haben, entscheidet der Anwender, welche Norm angewendet wird.

Biostimulants des végétaux - Recherche de Shigella spp.

Le présent document fournit une méthode permettant de vérifier que le pathogène Shigella spp. n’est pas présent dans les biostimulants microbiens des végétaux.
La méthode de recherche des pathogènes Shigella n’est pas sensible et la quantification est rarement réalisée. La recherche est généralement réalisée en utilisant un milieu d’enrichissement, puis en procédant à un repiquage sur une variété de milieux sélectifs.
Le présent document est applicable aux mélanges de fertilisants constitués d’au moins deux composants appartenant aux catégories de fertilisants UE suivantes : engrais, amendements minéraux basiques, amendements du sol, supports de culture, biostimulants des végétaux et, dans lesquels la catégorie Biostimulants des végétaux représente le pourcentage le plus élevé du mélange en masse ou en volume, ou dans le cas d’un liquide, en masse sèche. Si les biostimulants des végétaux ne représentent pas le pourcentage le plus élevé du mélange, la Norme européenne relative au composant représentant le pourcentage le plus élevé du mélange s’applique. Dans le cas où un mélange de fertilisants contient des composants en quantité égale ou dans le cas où les composants des fertilisants UE utilisés pour le mélange ont des formulations  identiques, l’utilisateur décide quelle norme appliquer.

Rastlinski biostimulanti - Ugotavljanje prisotnosti Shigella spp.

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Standards Content (Sample)

oSIST prEN 17715:2023
Rastlinski biostimulanti - Ugotavljanje prisotnosti Shigella spp.
Plant biostimulants - Detection of Shigella spp.
Pflanzen-Biostimulanzien - Nachweis von Shigella spp.
Biostimulants des végétaux - Détection de Shigella spp.
Ta slovenski standard je istoveten z: prEN 17715
65.080 Gnojila Fertilizers
oSIST prEN 17715:2023 en,fr,de
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

oSIST prEN 17715:2023
oSIST prEN 17715:2023
prEN 17715
April 2023
ICS 65.080 Will supersede CEN/TS 17715:2022
English Version
Plant biostimulants - Detection of Shigella spp.
Biostimulants des végétaux - Détection de Shigella spp. Pflanzen-Biostimulanzien - Nachweis von Shigella spp.
This draft European Standard is submitted to CEN members for enquiry. It has been drawn up by the Technical Committee
CEN/TC 455.
If this draft becomes a European Standard, CEN members are bound to comply with the CEN/CENELEC Internal Regulations
which stipulate the conditions for giving this European Standard the status of a national standard without any alteration.

This draft European Standard was established by CEN in three official versions (English, French, German). A version in any other
language made by translation under the responsibility of a CEN member into its own language and notified to the CEN-CENELEC
Management Centre has the same status as the official versions.

CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia,
Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway,
Poland, Portugal, Republic of North Macedonia, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Türkiye and
United Kingdom.
Recipients of this draft are invited to submit, with their comments, notification of any relevant patent rights of which they are
aware and to provide supporting documentation.

Warning : This document is not a European Standard. It is distributed for review and comments. It is subject to change without
notice and shall not be referred to as a European Standard.



CEN-CENELEC Management Centre: Rue de la Science 23, B-1040 Brussels
© 2023 CEN All rights of exploitation in any form and by any means reserved Ref. No. prEN 17715:2023 E
worldwide for CEN national Members.

oSIST prEN 17715:2023
prEN 17715:2023 (E)
Contents Page
European foreword . 4
Introduction . 5
1 Scope . 6
2 Normative references . 6
3 Terms and definitions . 6
4 Principle . 7
4.1 General . 7
4.2 Enrichment in selective liquid medium . 7
4.3 Plating out and identification of colonies. 7
4.4 Biochemical and serological confirmation . 7
5 Culture media, reagents and antisera . 7
6 Apparatus and glassware . 8
7 Sampling . 8
8 Procedure. 8
8.1 General . 8
8.2 Test portion . 8
8.3 Enrichment . 8
8.3.1 General . 8
8.3.2 Liquid formulations. 8
8.3.3 Solid formulations . 8
8.4 Plating out and colony selection . 9
8.5 Confirmation of colonies . 9
8.5.1 General . 9
8.5.2 Purification of colonies . 9
8.5.3 Biochemical confirmation . 9
8.5.4 Additional biochemical differentiation . 13
8.6 Serological confirmation . 14
8.6.1 Antigenic differentiation . 14
8.6.2 Agglutination tests . 15
8.6.3 Definitive confirmation . 15
9 Expression of results . 15
10 Test report . 15
Annex A (normative) Diagram of test procedure . 16
Annex B (normative) Composition and preparation of culture media and reagents . 17
B.1 General . 17
B.2 Selective enrichment broth . 17
B.3 Selective differential agars . 18
B.4 Nutrient agar . 20
oSIST prEN 17715:2023
prEN 17715:2023 (E)
B.5 Triple iron sugar (TSI) agar . 20
B.6 Semi-solid nutrient agar . 21
B.7 Urea agar (Christensen’s medium) . 21
B.8 L-Lysine decarboxylase medium . 22
B.9 L-Ornithine decarboxylase medium . 22
B.10 Reagents for indole reaction . 23
B.11 β-Galactosidase reagent . 23
B.12 Bromocresol purple broth . 24
B.13 Saline solution . 25
B.14 Sodium acetate agar . 25
B.15 Christensen's citrate agar . 25
B.16 Mucate broth . 26
B.17 Antisera of Shigella species . 27
Annex C (normative) Description of Shigella colony morphology and colour on selective
agars, for both identification and quality control purposes . 28
Annex D (informative) Interlaboratory study . 29
D.1 Materials used in the interlaboratory study . 29
D.2 Results of the interlaboratory study . 30
Annex ZA (informative) Relationship of this European Standard and the essential
requirements of Regulation (EU) 2019/1009 making available on the market of EU
fertilising products aimed to be covered . 31
Bibliography . 32
oSIST prEN 17715:2023
prEN 17715:2023 (E)
European foreword
This document (prEN 17715:2023) has been prepared by Technical Committee CEN/TC 455 “Plant
Biostimulants”, the secretariat of which is held by AFNOR.
This document is currently submitted to the CEN enquiry.
This document will supersede CEN/TS 17715:2022.
This document has been prepared under a Standardization Request given to CEN by the European
Commission and the European Free Trade Association, and supports essential requirements of EU
Directive(s) / Regulation(s).
For relationship with EU Directive(s) / Regulation(s), see informative Annex ZA, which is an integral part
of this document.
oSIST prEN 17715:2023
prEN 17715:2023 (E)
This document was prepared by the experts of CEN/TC 455 “Plant Biostimulants”. The European
Committee for Standardization (CEN) was requested by the European Commission (EC) to draft
European standards or European standardization deliverables to support the implementation of
Regulation (EU) 2019/1009 of 5 June 2019 laying down rules on the making available on the market of
EU fertilizing products (“FPR” or “Fertilising Products Regulation”). This Standardization Request,
presented as SR M/564 and M/564 Amd1, also contributes to the Communication on “Innovating for
Sustainable Growth: A Bio economy for Europe”. Working Group 5 “Labelling and denominations”, was
created to develop a work program as part of this request.
Technical committee CEN/TC 455 “Plant Biostimulants” was established to carry out the work program
that will prepare a series of standards. The interest in biostimulants has increased significantly in Europe
as a valuable tool to use in agriculture. Standardization was identified as having an important role in
order to promote the use of biostimulants. The work of CEN/TC 455 seeks to improve the reliability of
the supply chain, thereby improving the confidence of farmers, industry, and consumers in biostimulants,
and will promote and support commercialization of the European biostimulant industry.
Biostimulants used in agriculture can be applied in multiple ways: on soil, on plants, as seed treatment,
etc. A microbial plant biostimulant consists of a microorganism or a consortium of microorganisms, as
referred to in Component Material Category 7 of Annex II of the EU Fertilising Products Regulation.
This document is applicable to all microbial biostimulants in agriculture.
The table below summarizes many of the agro-ecological principles and the role played by biostimulants.
Table 1 — Agro-ecological principles and the role played by biostimulants [1]
Increase biodiversity
By improving soil microorganism quality/quantity
Reinforce biological regulation and interactions
By reinforcing plant-microorganism interactions
— symbiotic exchanges i.e. mycorrhize
— symbiotic exchanges i.e. rhizobiaciae/fava
— secretions mimicking plant hormones (i.e. trichoderma)
By regulating plant physiological processes
— e.g. growth, metabolism, plant development
Improve biogeochemical cycles
— improve absorption of nutritional elements
— impro

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