SIST-TS CEN/TS 17719:2023
(Main)Plant biostimulants - Determination of the anaerobic plate count
Plant biostimulants - Determination of the anaerobic plate count
This document provides a horizontal method for enumeration of microorganisms that are able to grow and form colonies in a solid medium after anaerobic incubation at 30 °C.
The method is applicable to microbial plant biostimulants for verifying that the concentration of anaerobes does not exceed the respective limits outlined in the EU Regulation on Fertilising Products [1].
This method does not apply to the microbiological monitoring of the environment in which microbial plant biostimulants are manufactured.
No information about potential human pathogens can be inferred from anaerobic plate counts.
Pflanzen-Biostimulanzien - Bestimmung der anaeroben Keimzahl
Dieses Dokument stellt ein horizontales Verfahren zur Zählung von nach anaerober Bebrütung bei 30 °C in einem festen Medium wachsenden und Kolonien bildenden Mikroorganismen zur Verfügung.
Das Verfahren gilt für mikrobielle Biostimulanzien für die pflanzliche Anwendung, um zu überprüfen, ob die Konzentration der Anaerobier die entsprechenden Grenzwerte der EU-Düngeprodukteverordnung [1] nicht überschreitet.
Dieses Verfahren gilt nicht für die mikrobiologische Überwachung der Umgebung, in der die mikrobiellen Biostimulanzien für die pflanzliche Anwendung hergestellt werden.
Es können keine Informationen über potentielle humanpathogene Keime aus der Auszählung der anaeroben koloniebildenden Einheiten abgeleitet werden.
Biostimulants des végétaux - Détermination du dénombrement sur plaque des germes anaérobies
Le présent document fournit une méthode horizontale pour le dénombrement des microorganismes qui sont capables de croître et de former des colonies dans un milieu solide après une incubation anaérobie à 30 °C.
La méthode est applicable aux biostimulants microbiens des végétaux pour vérifier que la concentration en anaérobies ne dépasse pas les limites correspondantes indiquées dans le Règlement UE sur les fertilisants [1].
La présente méthode ne s’applique pas à la surveillance microbiologique de l’environnement dans lequel les biostimulants microbiens des végétaux sont fabriqués.
Aucune information sur les pathogènes humains potentiels ne peut être déduite des dénombrements sur plaque des germes anaérobies.
Rastlinski biostimulanti - Določanje števila na anaerobnih mikrotitrskih ploščah
Dokument določa vodoravno metodo za ugotavljanje števila mikroorganizmov, ki so sposobni rasti in tvoriti kolonije v trdnem gojišču po anaerobni inkubaciji pri 30 °C.
Metoda se uporablja za mikrobne rastlinske biostimulante za preverjanje, da koncentracija anaerobov ne presega ustreznih mejnih vrednosti, navedenih v Uredbi EU o sredstvih za gnojenje [1].
Metoda se ne uporablja pri mikrobiološkem nadzoru okolja, v katerem se proizvajajo mikrobni rastlinski biostimulanti.
Na podlagi števila anaerobnih mikrotitrskih plošč ni mogoče sklepati na nobene informacije o potencialnih človeških patogenih.
General Information
Relations
Standards Content (Sample)
SLOVENSKI STANDARD
SIST-TS CEN/TS 17719:2023
01-februar-2023
Rastlinski biostimulanti - Določanje števila na anaerobnih mikrotitrskih ploščah
Plant biostimulants - Determination of the anaerobic plate count
Pflanzen-Biostimulanzien - Bestimmung der anaeroben Keimzahl
Biostimulants des végétaux - Détermination du dénombrement sur plaque des germes
anaérobies
Ta slovenski standard je istoveten z: CEN/TS 17719:2022
ICS:
65.080 Gnojila Fertilizers
SIST-TS CEN/TS 17719:2023 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 17719:2023
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SIST-TS CEN/TS 17719:2023
CEN/TS 17719
TECHNICAL SPECIFICATION
SPÉCIFICATION TECHNIQUE
March 2022
TECHNISCHE SPEZIFIKATION
ICS 65.080
English Version
Plant biostimulants - Determination of the anaerobic plate
count
Biostimulanzien für die pflanzliche Anwendung -
Bestimmung der anaeroben Keimzahl
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 NORMALISATION
EUROPÄISCHES KOMITEE FÜR NORMUNG
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 17719:2022 E
worldwide for CEN national Members.
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SIST-TS CEN/TS 17719:2023
CEN/TS 17719:2022 (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 Brief description . 7
5 Culture media and reagents . 7
6 Equipment and consumables . 7
7 Preparation of test sample . 8
7.1 General. 8
7.2 Liquid – water-based formulations . 8
7.3 Liquid – oil-based (emulsifiable concentrate - EC) formulations . 8
7.4 Solid – wettable powder (WP) formulations . 8
7.5 Solid – water dispersible granules (WDG) formulations . 8
7.6 Solid – pellets, granules, microgranules (slow release) formulations . 9
7.7 Solid substrates . 9
8 Procedure . 9
8.1 Test portion, initial suspension and dilutions . 9
8.2 Inoculation and incubation . 9
8.3 Critical Control Point . 9
9 Calculation . 9
10 Expression of results . 10
11 Performance characteristics of the method . 10
11.1 Interlaboratory studies . 10
11.2 Sensitivity . 10
11.3 Specificity . 10
12 Test report . 10
13 Quality assurance . 10
Annex A (informative) Guidelines for calculating and reporting the Anaerobic Plate Counts
(ANPC) . 11
A.1 General. 11
A.2 Example . 11
Annex B (normative) Composition and preparation of culture media and reagents . 12
B.1 Anaerobe Agar (ANA) . 12
B.1.1 Base . 12
B.1.2 Hemin solution . 13
B.1.3 Vitamin K1 solution . 13
2
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SIST-TS CEN/TS 17719:2023
CEN/TS 17719:2022 (E)
B.1.4 Buffered peptone water . 13
Bibliography . 14
3
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SIST-TS CEN/TS 17719:2023
CEN/TS 17719:2022 (E)
European foreword
This document (CEN/TS 17719:2022) 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.
4
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SIST-TS CEN/TS 17719:2023
CEN/TS 17719:2022 (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 the European Parliament and of the Council 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”. Working Group 5 “Labelling and
denominations” was created to develop a work program as part of this standardization 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 commercialisation 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
2019/1009 [1].
Table 1 summarizes many of the agro-ecological principles and the role played by biostimulants.
Table 1 — Agro-ecological principles and the role played by biostimulants
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/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
— 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.
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 17719:2023
CEN/TS 17719:2022 (E)
1 Scope
This document provides a horizontal method for enumeration of microorganisms that are able to grow
and form colonies in a solid medium after anaerobic incubation at 30 °C.
The method is applicable to microbial plant biostimulants for verifying that the concentration of
anaerobes does not exceed the respective limits outlined in the EU Regulation on Fertilising Products [1].
This method does not apply to the microbiological monitoring of the environment in which microbial
plant biostimulants are manufactured.
No information about potential human pathogens can be inferred from anaerobic plate counts.
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 17708, Plant biostimulants - Preparation of sample for microbial analysis
CEN/TS 17724, Plant biostimulants - Terminology
1
EN ISO 7218:2007, Microbiology of food and animal feeding stuffs - General requirements and guidance
for microbiological examinations (ISO 7218:2007)
2
EN ISO 11133:2014, Microbiology of food, animal feed and water - Preparation, production, storage and
performance testing of culture media (ISO 11133:2014)
3 Terms and definitions
For the purposes of this document, the terms and definitions given in CEN/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
microorganism
any microbiological entity, including lower fungi, bacteria and viruses, cellular or non-cellular, capable of
replication or of transferring genetic material
[SOURCE: Regulation (EC) No 1107/2009, Article 3, point 15]
1
As impacted by EN ISO 7218:2007/A1:2013.
2
As impacted by EN ISO 11133:2014/A1:2018 and EN ISO 11133:2014/A2:2020.
6
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SIST-TS CEN/TS 17719
...
SLOVENSKI STANDARD
kSIST-TS FprCEN/TS 17719:2021
01-november-2021
[Not translated]
Plant biostimulants - Determination of the anaerobic plate count
Biostimulanzien für die pflanzliche Anwendung - Bestimmung der anaeroben Keimzahl
Ta slovenski standard je istoveten z: FprCEN/TS 17719
ICS:
65.080 Gnojila Fertilizers
kSIST-TS FprCEN/TS 17719:2021 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 17719:2021
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kSIST-TS FprCEN/TS 17719:2021
FINAL DRAFT
TECHNICAL SPECIFICATION
FprCEN/TS 17719
SPÉCIFICATION TECHNIQUE
TECHNISCHE SPEZIFIKATION
September 2021
ICS 65.080
English Version
Plant biostimulants - Determination of the anaerobic plate
count
Biostimulanzien für die pflanzliche Anwendung -
Bestimmung der anaeroben Keimzahl
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 17719:2021 E
worldwide for CEN national Members.
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kSIST-TS FprCEN/TS 17719:2021
FprEN 17719:2021 (E)
Contents Page
European foreword . 3
1 Scope . 5
2 Normative references . 5
3 Terms and definitions . 5
4 Principle . 6
4.1 Brief description . 6
5 Culture media and reagents . 6
6 Equipment and consumables . 6
7 Preparation of test sample . 7
7.1 General. 7
7.2 Liquid - water based- formulations . 7
7.3 Liquid - oil based (emulsifiable concentrate - EC) formulations . 7
7.4 Solid - Wettable Powder (WP) formulations . 7
7.5 Solid - Water dispersible granules (WDG) formulations . 7
7.6 Solid - Pellets, granules, microgranules (slow release) formulations . 7
7.7 Solid substrates . 7
8 Procedure . 8
8.1 Test portion, initial suspension and dilutions . 8
8.2 Inoculation and incubation . 8
8.3 Critical Control Point . 8
9 Calculation . 8
10 Expression of results . 8
11 Performance characteristics of the method . 9
11.1 Interlaboratory studies . 9
11.2 Sensitivity . 9
11.3 Specificity . 9
12 Test report . 9
13 Quality assurance . 9
Annex A (informative) Guidelines for calculating and reporting the Anaerobic Plate Counts
(ANPC) . 10
Annex B (normative) Composition and preparation of culture media and reagents . 11
Bibliography . 13
2
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kSIST-TS FprCEN/TS 17719:2021
FprEN 17719:2021 (E)
European foreword
This document (FprCEN/TS 17719:2021) 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 Standardization Request given to CEN by the European
Commission and the European Free Trade Association.
3
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kSIST-TS FprCEN/TS 17719:2021
FprEN 17719:2021 (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: 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 Fertilizing Products Regulation
2019/1009 [1].
The method is applicable to microbial plant biostimulants except those composed of aerobic bacterium
to verify that the concentration of anaerobes does not exceed the respective limits described in the EU
Fertilizers Regulation.
Table 1 summarizes many of the agro-ecological principles and the role played by biostimulants.
Table 1 — Agro-ecological principles and the role played by biostimulants
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
— for ex growth, metabolism, 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.
IMPORTANT — It is absolutely essential that tests conducted in accordance with this document be
carried out by suitably trained staff.
4
---------------------- Page: 6 ----------------------
kSIST-TS FprCEN/TS 17719:2021
FprEN 17719:2021 (E)
1 Scope
This document provides a horizontal method for enumeration of microorganisms that are able to grow
and form colonies in a solid medium after anaerobic incubation at 30 °C.
The method is applicable to microbial plant biostimulants for verifying that the concentration of
anaerobes does not exceed the respective limits outlined in the EU Regulation on Fertilising Products [1].
This method does not apply to the microbiological monitoring of the environment in which microbial
plant biostimulants are manufactured.
No information about potential human pathogens can be inferred from anaerobic plate counts.
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.
FprCEN/TS 17724, Plant Biostimulants – Terminology
1
EN ISO 7218:2007, Microbiology of food and animal feeding stuffs - General requirements and guidance
for microbiological examinations
FprCEN/TS 17708, Plant biostimulants – Preparation of sample for microbial analysis
EN ISO 11133:2014, Microbiology of food, animal feed and water – Preparation, production, storage and
2
performance testing of culture media
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
microorganism
any microbiological entity, including lower fungi, bacteria and viruses, cellular or non-cellular, capable of
replication or of transferring genetic material
[SOURCE: Regulation (EC) No 1107/2009, Article 3, point 15]
3.2
facultative organism
microorganism capable of both aerobic and anaerobic metabolism
1
As impacted by EN ISO 7218:2007/A1:2013.
2
As impacted by EN ISO 11133:2014/A1:2018 and EN ISO 11133:2014/A2:2020.
5
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kSIST-TS FprCEN/TS 17719:2021
FprEN 17719:2021 (E)
[SOURCE: EN ISO 11139:2018, 3.114]
3.3
obligate anaerobe
organism that only lives and grows in the absence of molecular oxygen
[SOURCE: EN ISO 11139:2018, 3.186]
3.4
viable count
value established from enumeration of recoverable colony-forming units
[SOURCE: EN ISO 11139:2018, 3.186]
4 Principle
4.1 General
Viable anaerobic bacteria are enumerated by the plate count technique under an anaerobic atmosphere
[2]. Caution shall be exercised when applying the method since isolates may be pathogenic.
4.2 Brief description
A fixed amount of a dilution of the test sample is placed in an empty Petri dish and mixed in a specified
melted agar culture medium to create a pour plate.
The plates are incubated at 30 °C ± 1 °C under anaerobic conditions for 48-72 h.
The number of microorganisms per gram or millilitre of test sample is calculated as specified in Clause 8.
5 Culture media and reagents
2
For current laboratory practices,
...
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