Plant biostimulants - Determination of microorganisms' concentration

This document specifies general rules to determine the concentration of microorganisms present in plant biostimulant products.
The method is applicable to microbial plant biostimulants for verifying that the concentration of microorganisms does not exceed the respective limits outlined in the EU Regulation on Fertilising Products [1].
This horizontal method might not be appropriate in every detail for certain products. In this case, it is necessary to refer to the methodology of specific determination and quantification of the microorganisms.

Pflanzen-Biostimulanzien - Bestimmung der Konzentration von Mikroorganismen

Dieses Dokument legt allgemeine Regeln für die Bestimmung der Konzentration von Mikroorganismen in Biostimulansprodukten fest.
Das Verfahren ist auf mikrobielle Pflanzen-Biostimulanzien anwendbar, und zwar, um zu verifizieren, dass die Konzentration von Mikroorganismen die entsprechenden Grenzwerte der EU Düngeprodukte-Verordnung [1] nicht überschreitet.
Dieses horizontale Verfahren ist für bestimmte Produkte nicht unbedingt bis ins Detail geeignet. In diesem Fall ist es notwendig, auf die Methodik der spezifischen Bestimmung und Quantifizierung der Mikro-organismen zurückzugreifen.

Biostimulants des végétaux - Détermination de la concentration en microorganismes

Le présent document spécifie les règles générales pour déterminer la concentration en microorganismes présents dans les produits biostimulants des végétaux.
La méthode est applicable aux biostimulants des végétaux microbiens pour vérifier que la concentration en microorganismes ne dépasse pas les limites correspondantes indiquées dans le Règlement UE sur les fertilisants [1].
Cette méthode horizontale pourrait ne pas être appropriée dans chaque détail pour certains produits. Dans ce cas, il est nécessaire de se référer à la méthodologie de détermination et de quantification spécifique des microorganismes.

Rastlinski biostimulanti - Določevanje koncentracije mikroorganizmov

Ta dokument je bil namenjen opredelitvi splošnih pravil za določanje koncentracije mikroorganizmov, prisotnih v rastlinskih biostimulantih.
Metoda se uporablja za mikrobne rastlinske biostimulante za preverjanje, da koncentracija mikroorganizmov ne presega ustreznih mejnih vrednosti, navedenih v Uredbi EU o sredstvih za gnojenje [1].
Ta vodoravna metoda v nekaterih podrobnostih morda ne bo primerna za nekatere izdelke. V tem primeru se je treba sklicevati na metodologijo specifičnega določanja in kvantifikacije mikroorganizmov.

General Information

Status
Published
Publication Date
22-Mar-2022
Current Stage
6060 - Definitive text made available (DAV) - Publishing
Start Date
23-Mar-2022
Due Date
29-Aug-2022
Completion Date
23-Mar-2022

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SLOVENSKI STANDARD
01-februar-2023
Rastlinski biostimulanti - Določevanje koncentracije mikroorganizmov
Plant biostimulants - Determination of microorganisms' concentration
Pflanzen-Biostimulanzien - Bestimmung der Konzentration von Mikroorganismen
Biostimulants des végétaux - Détermination de la concentration en microorganismes
Ta slovenski standard je istoveten z: CEN/TS 17714:2022
ICS:
65.080 Gnojila Fertilizers
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

CEN/TS 17714
TECHNICAL SPECIFICATION
SPÉCIFICATION TECHNIQUE
March 2022
TECHNISCHE SPEZIFIKATION
ICS 65.080
English Version
Plant biostimulants - Determination of microorganisms'
concentration
Biostimulants des végétaux - Détermination de la Pflanzen-Biostimulanzien - Bestimmung der
concentration en microorganismes Konzentration von Mikroorganismen
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 17714:2022 E
worldwide for CEN national Members.

Contents Page
European foreword . 3
Introduction . 4
1 Scope . 6
2 Normative references . 6
3 Terms and definitions . 6
4 Principe of the method . 7
5 Sampling . 7
6 Preparation of sample for microbial analysis . 7
7 Method for enumeration of microorganism . 7
7.1 General. 7
7.2 Enumeration using a solid media. 8
7.3 Enumeration and quantification using a liquid medium . 16
8 Expression of results . 18
9 Test report . 18
10 Quality assurance . 18
Bibliography . 19

European foreword
This document (CEN/TS 17714:2022) 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.
Biostimulants used in agriculture can be applied in multiple ways: on soil, on plant, 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 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
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
- 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
This document defines the general rules for the determination of microorganism concentration in a
sample of biostimulant product.
The specific concentrations of microorganisms required in specific standard methods take precedence
over the general rules listed in this document.

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 general rules to determine the concentration of microorganisms present in plant
biostimulant products.
The method is applicable to microbial plant biostimulants for verifying that the concentration of
microorganisms does not exceed the respective limits outlined in the EU Regulation on Fertilising
Products [1].
This horizontal method might not be appropriate in every detail for certain products. In this case, it is
necessary to refer to the methodology of specific determination and quantification of the
microorganisms.
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
3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
3.1
plant biostimulant
product stimulating plant nutrition processes independently of the product's nutrient content with the
sole aim of improving one or more of the following characteristics of the plant or the plant rhizosphere:
a) nutrient use efficiency;
b) tolerance to abiotic stress;
c) crop quality traits;
d) availability of confined nutrients in soil or rhizosphere
3.2
microorganism
entity of microscopic size, encompassing bacteria, fungi, protozoa and viruses
[SOURCE: EN ISO 11139:2018, 3.176]
3.3
colony
localized visible accumulation of microbial mass (such as prokaryotes, bacteria, micromycetes, yeast and
fungi) or organisms (such as Dreissena species) developed on or in a solid nutrient medium from a viable
particle or organism
Note 1 to entry: Frequently, micro colonies from nearby viable particles, before becoming visible, fuse into one
macro colony. The number of visible colonies is, therefore, usually and underestimate of the number of viable
particles.
[SOURCE: ISO 6107-6:2021, [5], modified]
3.4
product
portion of an identified plant biostimulant product received in the laboratory for testing
3.5
sample
portion of the product (3.4) (at least 1 g or 1 ml) that is used in the test to prepare the initial suspension
3.6
initial suspension
suspension (or solution) of the sample (3.5) in a defined volume of an appropriate diluent
4 Principe of the method
The method of determining the concentration of microorganisms has been developed to provide a
general method for the enumeration of the microorganisms present in the plant biostimulant product.
The results are expressed as the number of active units by volume or weight, or in any other way relevant
to the microorganism, for example colony forming units per gram or the UPM for Mycorrhizae.
5 Sampling
Sampling is not part of the method specified in this document (see CEN/TS 17702-1). If there is no specific
International or European Standard, it is recommended that the parties concerned come to an agreement
on this subject.
It is important that the laboratory receives a sample which is representative and has not been damaged
or changed during transport or storage.
6 Preparation of sample for microbial analysis
Prepare the test sample from the laboratory sample in accordance with CEN/TS 17708. If there is no
specific International or European Standard, it is recommended that the parties concerned come to an
agreement on this subject.
7 Method for enumeration of microorganisms
7.1 General
When assessing the microbiological quality of biostimulants products, it is often not enough to know only
which microorganisms are present. In most cases, the quantitative aspect is equally important, which
brings about the need to enumerate microorganisms. This may be achieved in various ways: through
direct examination (microscopy), by inoculating solid or liquid media. However, this document only
covers enumeration using solid and liquid media.
Enumeration on solid media is based on the capacity of many microorganisms to produce colonies in or
on agar media that can be recognized as such with the naked eye or with the aid of a simple magnifying
glass.
If the level of bacteria is expected to be very low (less than 10 colonies in or on a plate at the lowest
dilution), enumeration using liquid media is recommended (e.g. MPN) to improve the statistical reliability
of the results.
7.2 Enumeration using a solid media
7.2.1 General
The different steps for enumeration using a solid media (number of petri dishes/dilution, Pour plate
techniques, surface inoculation, Incubat
...

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