SIST-TS CEN/TS 17712:2023
(Main)Plant biostimulants - Detection of Staphylococcus aureus
Plant biostimulants - Detection of Staphylococcus aureus
This document provides a method for verifying that the pathogen Staphylococcus aureus is present in microbial plant biostimulants according to the limits outlined in the EU Regulation on Fertilising Products [2].
Pflanzen-Biostimulanzien - Nachweis von Staphylococcus aureus
Dieses Dokument stellt ein Verfahren zur Verfügung, mit dem verifiziert werden kann, dass das Pathogen Staphylococcus aureus in Biostimulanzien für die pflanzliche Anwendung die Grenzwerte der EU Dünge-produkteverordnung [2] nicht überschreitet.
Biostimulants des végétaux - Détection de Staphylococcus aureus
Le présent document fournit une méthode permettant de vérifier que le pathogène Staphylococcus aureus est présent dans les biostimulants microbiens des végétaux dans le respect des limites énoncées dans le Règlement UE sur les fertilisants [2].
Rastlinski biostimulanti - Ugotavljanje prisotnosti Staphylococcus aureus
Ta dokument zagotavlja metodo za preverjanje, da je patogen Staphylococcus aureus prisoten v mikrobnih rastlinskih biostimulantih v skladu z omejitvami, določenimi v Uredbi EU o sredstvih za gnojenje [2].
General Information
Relations
Standards Content (Sample)
SLOVENSKI STANDARD
SIST-TS CEN/TS 17712:2023
01-februar-2023
Rastlinski biostimulanti - Ugotavljanje prisotnosti Staphylococcus aureus
Plant biostimulants - Detection of Staphylococcus aureus
Pflanzen-Biostimulanzien - Nachweis von Staphylococcus aureus
Biostimulants des végétaux - Détection de Staphylococcus aureus
Ta slovenski standard je istoveten z: CEN/TS 17712:2022
ICS:
65.080 Gnojila Fertilizers
SIST-TS CEN/TS 17712:2023 en,fr,de
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
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SIST-TS CEN/TS 17712:2023
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SIST-TS CEN/TS 17712:2023
CEN/TS 17712
TECHNICAL SPECIFICATION
SPÉCIFICATION TECHNIQUE
March 2022
TECHNISCHE SPEZIFIKATION
ICS 65.080
English Version
Plant biostimulants - Detection of Staphylococcus aureus
Biostimulants des végétaux - Détection de Pflanzen-Biostimulanzien - Nachweis von
Staphylococcus aureus Staphylococcus aureus
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 17712:2022 E
worldwide for CEN national Members.
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SIST-TS CEN/TS 17712:2023
CEN/TS 17712:2022 (E)
Contents Page
European foreword . 3
Introduction . 4
1 Scope . 6
2 Normative references . 6
3 Terms and definitions . 6
4 Principle . 7
5 Sampling . 7
6 Preparation of test sample . 7
7 Procedure . 7
7.1 General . 7
7.2 Diluent . 7
7.3 Modified Giolitti-Cantoni broth . 7
7.4 Baird-Parker agar medium . 7
8 Apparatus and glassware . 8
9 Procedure . 8
9.1 Test portion, initial suspension and dilutions . 8
9.2 Enrichment and isolation . 9
10 Precision . 10
11 Test report . 10
12 Performance characteristics of the method . 10
12.1 Interlaboratory studies . 10
12.2 Sensitivity . 10
12.3 Specificity . 10
Annex A (normative) Media composition and preparation . 11
Bibliography . 13
2
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SIST-TS CEN/TS 17712:2023
CEN/TS 17712:2022 (E)
European foreword
This document (CEN/TS 17712:2022) 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.
3
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SIST-TS CEN/TS 17712:2023
CEN/TS 17712:2022 (E)
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 fertilising 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.
Biostimulants used in agriculture can be applied in multiple ways to the soil, to 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 1 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. Mycorrhizae
- symbiotic exchanges i.e. Rhizobiaceae/Faba
- secretions mimicking plant hormones (i.e. Trichoderma)
By regulating plant physiological processes
- such as growth, metabolism or plant development…
Improve biogeochemical cycles
- improve absorption of nutritional elements
- improve bioavailability of nutritional elements in the soil
- stimulate degradation of organic matter
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.
4
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SIST-TS CEN/TS 17712:2023
CEN/TS 17712:2022 (E)
IMPORTANT — It is absolutely essential that tests conducted in accordance with this document
be carried out by suitably trained staff.
5
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SIST-TS CEN/TS 17712:2023
CEN/TS 17712:2022 (E)
1 Scope
This document provides a method for verifying that the pathogen Staphylococcus aureus is present
in microbial plant biostimulants according to the limits outlined in the EU Regulation on
Fertilising Products [2].
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.
EN ISO 7218, Microbiology of food and animal feeding stuffs — General requirements and guidance
for microbiological examinations (ISO 7218)
CEN/TS 17708, Plant biostimulants — Preparation of sample for microbial analysis
CEN/TS 17724, Plant biostimulants — Terminology
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
Staphylococcus aureus
bacterium which forms colonies fitting the description of the species on the specified selective
medium after incubation of 24 h at a temperature of 37 °C under aerobic conditions
Note 1 to entry: S.aureus colony description:
— circular;
— convex;
— entire margin;
— grey to black (due to the reduction of potassium tellurite to telluride).
Note 2 to entry: Colony size varies between 1 mm and 2 mm in diameter.
Note 3 to entry: S. aureus is a facultatively anaerobic, Gram-positive coccus, which appears as grape-like
clusters when visualized under a microscope, and has a round, usually golden-yellow colonies, often with
haemolysis, when grown on selective blood agar plates.
Note 4 to entry: The term ‘Coagulase-positive staphylococci’ refers to bacteria that form typical and/or
atypical colonies on the surface of a selective culture medium and show a positive coagulase reaction when
the test is performed following the method specified in this document.
3.2
detection of the coagulase-positive staphylococci
determination of the detection or non detection of Staphylococcus aureus (3.2), in 25 g or 25 ml of
product, when tests are carried out in accordance with this document
6
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SIST-TS CEN/TS 17712:2023
CEN/TS 17712:2022 (E)
4 Principle
After sample preparation that is specific to microbial biostimulants, for detection of
Staphylococcus aureus, refer to the sections of EN ISO 6888-3:2003 that are pertinent to the
physical/chemical nature of the test sample for the detection of Staphylococcus aureus. If the
product is liquid, a specified volume of the test sample will be inoculated onto a liquid selective
culture medium. For products formulat
...
SLOVENSKI STANDARD
kSIST-TS FprCEN/TS 17712:2021
01-november-2021
[Not translated]
Plant biostimulants - Detection of Staphylococcus aureus
Biostimulanzien für die pflanzliche Anwendung - Nachweis von Staphylococcus aureus
Ta slovenski standard je istoveten z: FprCEN/TS 17712
ICS:
65.080 Gnojila Fertilizers
kSIST-TS FprCEN/TS 17712:2021 en,fr,de
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
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kSIST-TS FprCEN/TS 17712:2021
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kSIST-TS FprCEN/TS 17712:2021
FINAL DRAFT
TECHNICAL SPECIFICATION
FprCEN/TS 17712
SPÉCIFICATION TECHNIQUE
TECHNISCHE SPEZIFIKATION
September 2021
ICS 65.080
English Version
Plant biostimulants - Detection of Staphylococcus aureus
Biostimulanzien für die pflanzliche Anwendung -
Nachweis von Staphylococcus aureus
This draft Technical Specification is submitted to CEN members for Vote. It has been drawn up by the Technical Committee
CEN/TC 455.
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.
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 Technical Specification. It is distributed for review and comments. It is subject to change
without notice and shall not be referred to as a Technical Specification.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION
EUROPÄISCHES KOMITEE FÜR NORMUNG
CEN-CENELEC Management Centre: Rue de la Science 23, B-1040 Brussels
© 2021 CEN All rights of exploitation in any form and by any means reserved Ref. No. FprCEN/TS 17712:2021 E
worldwide for CEN national Members.
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kSIST-TS FprCEN/TS 17712:2021
FprCEN/TS 17712:2021 (E)
Contents Page
European foreword . 3
Introduction . 4
1 Scope . 6
2 Normative references . 6
3 Terms and definitions . 6
4 Principle . 7
5 Sampling . 7
6 Preparation of test sample . 7
7 Procedure . 7
7.1 Diluent . 7
7.2 Modified Giolitti-Cantoni broth . 7
7.3 Baird-Parker agar medium . 8
8 Apparatus and glassware . 8
9 Procedure . 9
9.1 Test portion, initial suspension and dilutions . 9
9.1.1 Solid formulations . 9
9.1.2 Liquid formulations . 9
9.2 Enrichment and isolation . 9
10 Precision . 10
11 Test report . 10
12 Performance characteristics of the method . 10
12.1 Interlaboratory studies . 10
12.2 Sensitivity . 11
12.3 Specificity . 11
Annex A (normative) (Media) . 12
A.1 Media composition and preparation . 12
A.1.1 Baird Parker Rabbit Plasma Fibrinogen (RFP) agar [4], [5], [6] . 12
A.1.2 Buffered peptone water (BPW) . 12
A.1.3 Modified Giolitti-Cantoni broth . 12
Bibliography . 14
2
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kSIST-TS FprCEN/TS 17712:2021
FprCEN/TS 17712:2021 (E)
European foreword
This document (FprCEN/TS 17712:2021) 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 Vote on TS.
This document has been prepared under a mandate given to CEN by the European Commission
and the European Free Trade Association.
3
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kSIST-TS FprCEN/TS 17712:2021
FprCEN/TS 17712:2021 (E)
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 fertilising products (“FPR” or “Fertilising Products Regulation”). This request,
presented as SR 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.
Biostimulants used in agriculture can be applied in multiple ways to the soil, to 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 Fertilizing
Products Regulation.
This document is applicable to all microbial biostimulants in agriculture.
The Table 1 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
- such as growth, metabolism or plant development…
Improve biogeochemical cycles
- improve absorption of nutritional elements
- improve bioavailability of nutritional elements in the soil
- stimulate degradation of organic matter
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.
4
---------------------- Page: 6 ----------------------
kSIST-TS FprCEN/TS 17712:2021
FprCEN/TS 17712:2021 (E)
IMPORTANT — It is absolutely essential that tests conducted in accordance with this document
be carried out by suitably trained staff.
5
---------------------- Page: 7 ----------------------
kSIST-TS FprCEN/TS 17712:2021
FprCEN/TS 17712:2021 (E)
1 Scope
This document provides a method for verifying that the pathogen Staphylococcus aureus is present
in microbial plant biostimulants according to the limits outlined in the EU Regulation on
Fertilising Products [2].
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.
EN ISO 7218:2007, Microbiology of food and animal feeding stuffs — General requirements and
guidance for microbiological examinations (ISO 7218:2007)
FprCEN/TS 17724, Plant biostimulants — Terminology
3 Terms and definitions
For the purposes of this document, the terms and definitions given in FprCEN/TS 17724 and the
following apply.
ISO and IEC maintain terminological databases for use in standardization at the following
addresses:
• IEC Electropedia: available at https://www.electropedia.org/
• ISO Online browsing platform: available at https://www.iso.org/obp
3.1
staphylococcus aureus
bacterium which forms colonies fitting the description of the species on the specified selective
medium after incubation of 24 h at a temperature of 37 °C under aerobic conditions
Note 1 to entry: S.aureus colony description:
— circular;
— convex;
— entire margin;
— grey to black (due to the reduction of potassium tellurite to telluride).
Note 2 to entry: Colony size varies between 1 mm and 2 mm in diameter.
Note 3 to entry: S. aureus is a facultatively anaerobic, Gram-positive coccus, which appears as grape-like
clusters when visualized under a microscope, and has a round, usually golden-yellow colonies, often with
haemolysis, when grown on selective blood agar plates.
Note 4 to entry: The term ‘Coagulase-positive staphylococci’ refers to bacteria that form typical and/or
atypical colonies on the surface of a selective culture medium and show a positive coagulase reaction when
the test is performed following the method specified in this document.
6
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kSIST-TS FprCEN/TS 17712:2021
FprCEN/TS 17712:2021 (E)
3.2
detection of the coagulase-positive staphylococci
determination of the detection or non detection of Staphylococcus aureus (3.2), in 25 g or 25 ml of
product, when tests are carried out in accordance with this document
4 Principle
After sample preparation that is specific to microbial biostimulants, for detection of
Staphylococcus aureus, we refer to the sections of EN ISO 6888-3:2003 that are pertinent to the
physical/chemical nature of the test sample for the detection of Staphylococcus aureus. If the
product is liquid, a specified volume of the test sample will be inoculated onto a liquid selective
culture medium. .For products formulated differently, a specified volume of an initial suspension
will be inoculated onto the selective medium. Incubation under anaerobic conditions at 37 °C for
24 and 48 h.
Pour plate and spread plate techniques could be used. Spread plate is considered enough for
detecting the presence of this species. In this case, the inoculation could be done with a sterile
loop (containing around 10 μl of product, dilution o
...
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