SIST EN 17711:2025
(Main)Plant biostimulants - Detection of Vibrio spp.
Plant biostimulants - Detection of Vibrio spp.
This document specifies a horizontal method for the detection of enteropathogenic Vibrio species (spp.),
which causes human illness in or via the intestinal tract. The species detectable by the methods specified
include Vibrio parahaemolyticus, Vibrio cholerae and Vibrio vulnificus.
It is applicable to the 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. [2].
NOTE 2 For confirmation, it is possible to use PCR (Polymerase Chain Reaction) tests; in this case a validation is
carried out by the laboratory for the procedure and data generated.
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 fertilising products categories: Fertilizers, Liming Materials, Soil Improvers,
Growing Media, Plant Biostimulants, and where the following category Plant Biostimulants is the highest
% in the blend by mass or volume, or in the case of liquid form by dry mass. If Plant Biostimulants is not
the highest % in the blend, the European Standard for the highest % 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 formulations1, the user decides which standard to
apply.
Pflanzen-Biostimulanzien - Nachweis von Vibrio spp.
Dieses Dokument legt ein horizontales Verfahren zum Nachweis von enteropathogenen Vibrio spp. fest, die Erkrankungen des Menschen am oder über den Darmtrakt verursachen [1]. Die Spezies, deren Nachweis die hier aufgeführten Verfahren einbeziehen, sind Vibrio parahaemolyticus, Vibrio cholerae und Vibrio vulnificus.
Das Dokument gilt für
- mikrobielle Pflanzen-Biostimulanzien.
ANMERKUNG 1 Die Weltgesundheitsorganisation (WHO; en: World Health Organization) hat V. parahaemolyticus, V. cholerae und V. vulnificus als die wichtigsten Kontaminanten von Vibrio spp. identifiziert [1].
ANMERKUNG 2 Eine Bestätigung durch PCR Tests ist möglich; in diesem Fall muss das Labor das Verfahren und die erzeugten Daten validieren.
Biostimulants des végétaux - Détection de Vibrio spp.
Le présent document spécifie une méthode horizontale pour la recherche des espèces entéropathogènes de Vibrio provoquant des maladies dans ou via le tractus intestinal chez l’homme [1]. Les espèces détectables par les méthodes spécifiées incluent Vibrio parahaemolyticus, Vibrio cholerae et Vibrio vulnificus.
Il s’applique :
- aux biostimulants microbiens des végétaux.
NOTE 1 L’Organisation Mondiale de la Santé (OMS) a identifié V. parahaemolyticus, V. cholerae et V. vulnificus comme les principales espèces de Vibrio contaminantes [1].
NOTE 2 Pour la confirmation, il est possible de recourir aux essais de PCR ; dans ce cas, le laboratoire doit valider le mode opératoire et les données générées.
Rastlinski biostimulanti - Ugotavljanje prisotnosti Vibrio spp.
Ta dokument določa horizontalno metodo za ugotavljanje prisotnosti enteropatogenih bakterij vrste Vibrio (spp.), ki povzročajo bolezni v prebavilih ali prek prebavil ljudi. Vrste bakterij, ki jih je mogoče zaznati z navedenimi metodami, vključujejo Vibrio parahaemolyticus, Vibrio cholerae in Vibrio vulnificus.
Dokument se uporablja za mikrobne rastlinske biostimulante.
OPOMBA 1: Svetovna zdravstvena organizacija (WHO) je ugotovila, da so V. parahaemolyticus, V. cholerae in V. vulnificus glavni onesnaževalci Vibrio spp. [2].
OPOMBA 2: Za potrditev je možna uporaba preskusov s polimerazno verižno reakcijo (PCR); v tem primeru laboratorij validira postopek in pridobljene podatke.
Ta dokument se uporablja za mešanice sredstev za gnojenje, ki vsebujejo najmanj dve od naslednjih kategorij sredstev za gnojenje EU: gnojila, sredstva za apnenje, izboljševalci tal, rastni substrati in rastlinski biostimulanti, pri čemer rastlinski biostimulanti predstavljajo največji masni ali prostorninski odstotek mešanice ali, v primeru tekoče oblike, odstotek suhe mase. Če rastlinski biostimulanti ne predstavljajo največjega odstotka v mešanici, se uporablja evropski standard za največji odstotek mešanice. Če vsebuje mešanica sredstev za gnojenje sestavine v enakih količinah ali če imajo posamezna sredstva za gnojenje EU, uporabljena za mešanico, enake formulacije1, standard za uporabo izbere uporabnik.
General Information
Relations
Standards Content (Sample)
SLOVENSKI STANDARD
01-februar-2025
Nadomešča:
SIST-TS CEN/TS 17711:2023
Rastlinski biostimulanti - Ugotavljanje prisotnosti Vibrio spp.
Plant biostimulants - Detection of Vibrio spp.
Pflanzen-Biostimulanzien - Nachweis von Vibrio spp.
Biostimulants des végétaux - Détection de Vibrio spp.
Ta slovenski standard je istoveten z: EN 17711:2024
ICS:
65.080 Gnojila Fertilizers
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
EN 17711
EUROPEAN STANDARD
NORME EUROPÉENNE
November 2024
EUROPÄISCHE NORM
ICS 65.080 Supersedes CEN/TS 17711:2022
English Version
Plant biostimulants - Detection of Vibrio spp.
Biostimulants des végétaux - Détection de Vibrio spp. Pflanzen-Biostimulanzien - Nachweis von Vibrio spp.
This European Standard was approved by CEN on 26 August 2024.
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. Up-to-date lists and bibliographical references
concerning such national standards may be obtained on application to the CEN-CENELEC Management Centre or to any CEN
member.
This European Standard exists 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.
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
© 2024 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN 17711:2024 E
worldwide for CEN national Members.
Contents Page
European foreword . 5
Introduction . 6
1 Scope . 7
2 Normative references . 7
3 Terms and definitions . 8
4 Principle . 8
4.1 General . 8
4.2 Primary enrichment in a liquid selective medium . 8
4.3 Secondary enrichment in a liquid selective medium . 9
4.4 Isolation and identification . 9
4.5 Confirmation . 9
5 Culture media and reagents . 9
5.1 Enrichment medium: alkaline saline peptone water (ASPW) . 9
5.2 Solid selective isolation media . 9
5.2.1 First medium: thiosulphate citrate bile and sucrose agar (TCBS) medium . 9
5.2.2 Second medium .10
5.3 Saline nutrient agar (SNA) .10
5.4 Reagent for detection of oxidase .10
5.5 Reagent for biochemical tests.10
5.5.1 L-lysine decarboxylase saline medium (LDC) .10
5.5.2 Arginine dihydrolase saline medium (ADH) .10
5.5.3 Reagent for detection of β-galactosidase .10
5.5.4 Saline medium for detection of indole .10
5.5.5 Saline peptone water .10
5.5.6 Sodium chloride solution .10
5.5.7 Toluene, p.a. .11
5.5.8 Sterile mineral oil, p.a. .11
6 Equipment and consumables .11
7 Sampling .11
8 Preparation of the test sample .11
9 Procedure (see Figure A.1) .12
9.1 Test portion and initial suspension .12
9.2 Primary selective enrichment .12
9.3 Secondary selective enrichment .13
9.4 Isolation and identification .13
9.5 Confirmation .14
9.5.1 General .14
9.5.2 Selection of colonies for confirmation and preparation of pure cultures .14
9.5.3 Tests for presumptive identification .14
9.5.4 Biochemical confirmation .15
10 Expression of results .17
11 Performance characteristics of the method . 17
11.1 Sensitivity . 17
11.2 Specificity . 17
11.3 LOD50 . 17
11.4 Precision . 17
12 Test report . 17
Annex A (normative) Diagram of procedure . 19
Annex B (normative) Composition and preparation of culture media and reagents . 20
B.1 General . 20
B.2 Water . 20
B.3 Alkaline saline peptone water (ASPW) . 20
B.4 Thiosulfate citrate bile and sucrose agar (TCBS) . 21
B.5 Saline nutrient agar (SNA) . 21
B.6 Reagent for detection of oxidase . 22
B.7 L-lysine decarboxylase saline medium (LDC) . 22
B.8 Arginine dihydrolase saline medium (ADH) . 23
B.9 Detection of β-galactosidase . 23
B.10 Saline medium for detection of indole . 24
B.11 Saline peptone water . 25
B.12 Sodium chloride solution . 25
B.13 Tris acetate EDTA (TAE) buffer . 25
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 . 26
C.1 General . 26
C.2 Equipment . 26
C.3 DNA extraction . 27
C.4 Procedure conventional PCR . 27
C.5 Primers and probes . 28
C.6 Control material — conventional PCR . 30
Annex D (informative) Real-time PCR for the detection of Vibrio parahaemolyticus,
thermostable direct haemolysin gene (tdh) and Vibrio vulnificus . 32
D.1 General . 32
D.2 Equipment . 32
D.3 DNA extraction . 32
D.4 Real-time PCR. 33
D.5 Vibrio parahaemolyticus — primers and hydrolysis probes . 33
D.6 Thermostable direct haemolysin (tdh) gene Vibrio parahaemolyticus — primers and
hydrolysis probes . 33
D.7 Vibrio vulnificus — primers and hydrolysis probes .34
D.8 Cycling parameters .34
D.9 Control material — real-time PCR .34
Annex E (informative) Repeatability and reproducibility data .35
E.1 General .35
E.2 Samples .35
E.3 Procedure.
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