CEN/TS 17711:2022

SLOVENSKI STANDARD
01-februar-2023
Rastlinski biostimulanti - Ugotavljanje prisotnosti Vibrio spp.
Plant biostimulants - Detection of Vibrio spp
Pflanzen-Biostimulanzien - Nachweis von Vibrio spp.
Biostimulants des végétaux - Recherche des espèces de Vibrio
Ta slovenski standard je istoveten z: CEN/TS 17711:2022
ICS:
65.080 Gnojila Fertilizers
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

CEN/TS 17711
TECHNICAL SPECIFICATION
SPÉCIFICATION TECHNIQUE
March 2022
TECHNISCHE SPEZIFIKATION
ICS 65.080
English Version
Plant biostimulants - Detection of Vibrio spp.
Biostimulants des végétaux - Recherche des espèces de Pflanzen-Biostimulanzien - Nachweis von Vibrio spp.
Vibrio
This Technical Specification (CEN/TS) was approved by CEN on 3 January 2022 for provisional application.

The period of validity of this CEN/TS is limited initially to three years. After two years the members of CEN will be requested to
submit their comments, particularly on the question whether the CEN/TS can be converted into a European Standard.

CEN members are required to announce the existence of this CEN/TS in the same way as for an EN and to make the CEN/TS
available promptly at national level in an appropriate form. It is permissible to keep conflicting national standards in force (in
parallel to the CEN/TS) until the final decision about the possible conversion of the CEN/TS into an EN is reached.

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, Turkey and
United Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATIO N

EUROPÄISCHES KOMITEE FÜR NORMUN G

CEN-CENELEC Management Centre: Rue de la Science 23, B-1040 Brussels
© 2022 CEN All rights of exploitation in any form and by any means reserved Ref. No. CEN/TS 17711:2022 E
worldwide for CEN national Members.

Contents Page
European foreword . 3
Introduction . 4
1 Scope . 5
2 Normative references . 5
3 Terms and definitions . 5
4 Principle. 6
4.1 General . 6
4.2 Primary enrichment in a liquid selective medium. 6
4.3 Secondary enrichment in a liquid selective medium . 6
4.4 Isolation and identification . 6
4.5 Confirmation . 7
5 Culture media and reagents . 7
5.1 Enrichment medium: alkaline saline peptone water (ASPW) . 7
5.2 Solid selective isolation media . 7
5.3 Saline nutrient agar (SNA) . 8
5.4 Reagent for detection of oxidase . 8
5.5 Biochemical tests . 8
6 Equipment and consumables . 8
7 Sampling . 9
8 Preparation of the test sample . 9
9 Procedure (see Figure A.1) . 9
9.1 Test portion and initial suspension . 9
9.2 Primary selective enrichment . 10
9.3 Secondary selective enrichment . 10
9.4 Isolation and identification . 11
9.5 Confirmation . 11
10 Expression of results . 14
11 Performance characteristics of the method . 14
11.1 Sensitivity . 14
11.2 Specificity . 15
11.3 LOD50 . 15
12 Test report . 15
Annex A (normative) Diagram of procedure . 16
Annex B (normative) Composition and preparation of the culture media and reagents . 17
Annex C (informative) Conventional PCR for the detection of Vibrio parahaemolyticus,
thermostable direct haemolysin (tdh) and thermostable direct related haemolysin
(trh) genes, Vibrio cholerae and Vibrio vulnificus . 23
Annex D (informative) Real-time PCR for the detection of Vibrio parahaemolyticus,
thermostable direct haemolysin gene (tdh) and Vibrio vulnificus . 29
Bibliography . 32

European foreword
This document (CEN/TS 17711:2022) has been prepared by Technical Committee CEN/TC 455
“Plant biostimulants”, the secretariat of which is held by AFNOR.
Attention is drawn to the possibility that some of the elements of this document may be the subject
of patent rights. CEN shall not be held responsible for identifying any or all such patent rights.
This document has been prepared under a Standardization Request given to CEN by the European
Commission and the European Free Trade Association.
Any feedback and questions on this document should be directed to the users’ national standards
body. A complete listing of these bodies can be found on the CEN website.
According to the CEN/CENELEC Internal Regulations, the national standards organisations of the
following countries are bound to announce this Technical Specification: 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, Turkey and the United Kingdom.
Introduction
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 M/564, also contributes to the Communication on
“Innovating for Sustainable Growth: A Bio economy for Europe”. The Working Group 5 “Labelling
and denominations”, was created to develop a work program as part of this Request. The 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 commercialisation of the
European biostimulant industry.
Because of the large variety of Plant Biostimulant products, the horizontal method described in
this document may not be appropriate in every detail for certain products. In this case, different
methods, which are specific to these products may be used if absolutely necessary for justified
technical reasons. Nevertheless, every attempt will be made to apply this horizontal method as far
as possible.
The harmonization of test methods cannot be immediate and, for certain groups of products,
International Standards and/or national standards may already exist that do not comply with this
horizontal method. It is hoped that when such standards are reviewed they will be changed to
comply with this document so that eventually the only remaining departures from this horizontal
method will be those necessary for well-established technical reasons.
WARNING — Persons using this document should be familiar with normal laboratory practice.
This document does not purport to address all of the safety problems, if any, associated with its
use. It is the responsibility of the user to establish appropriate safety and health practices and to
ensure compliance with any national regulatory conditions.
IMPORTANT — It is absolutely essential that tests conducted in accordance with this document
be carried out by suitably trained staff.

1 Scope
This document specifies a horizontal method for the detection of enteropathogenic Vibrio spp.,
which causes human illness in or via the intestinal tract [1]. The species detectable by the methods
specified include Vibrio parahaemolyticus, Vibrio cholerae and Vibrio vulnificus.
It is applicable to the following:
— microbial plant biostimulants.
NOTE 1 The World Health Organization (WHO) has identified that V. parahaemolyticus, V. cholerae and
V. vulnificus are the major contaminants of Vibrio spp. [1].
NOTE 2 For confirmation, it is possible to use PCR tests; in this case the laboratory validates the
procedure and data generated.
2 Normative references
The following documents are referred to in the text in such a way that some or all of their content
constitutes requirements of this document. For dated references, only the edition cited applies.
For undated references, the latest edition of the referenced document (including any
amendments) applies.
CEN/TS 17702-1, Plant biostimulants — Sampling and sample preparation — Part 1: Sampling
CEN/TS 17724, Plant biostimulants — Terminology
EN ISO 7218:2007, Microbiology of food and animal feeding stuffs — General requirements and
guidance for microbiological examinations (ISO 7218:2007)
EN ISO 11133:2014, Microbiology of food, animal feed and water — Preparation, production,
storage and performance testing of culture media (ISO 11133:2014)
EN ISO 3696:1995, Water for analytical laboratory use — Specification and test methods (ISO
3696:1987)
3 Terms and definitions
For the purposes of this document, the terms and definitions given in CEN/TS 17724 and the
following apply.
3.1
potentially enteropathogenic Vibrio spp
microorganism which forms typical colonies on solid selective media and which possesses the
described biochemical or molecular characteristics when the test is performed in accordance with
this document
Note 1 to entry: This document describes specific procedures for V. parahaemolyticus, V. cholerae and V.
vulnificus.
As amended by EN ISO 7218:2007/A1:2013.
As amended by EN ISO 11133:2014/A1:2018 and EN ISO 11133:2014/A2:2020.
3.2
detection of potentially enteropathogenic Vibrio spp
determination of the presence or absence o
...