FprEN 13040-2

• Abstract:

  This document specifies the general requirements for the preparation of samples, initial suspensions and further dilutions prior to microbiological examinations of soil improvers and growing media. This method is applicable especially for sample preparation prior to microbiological examinations of e.g. Escherichia coli (E. coli), Salmonella spp. and enterococci.; If a laboratory receives a material/product that is not listed in Annex B but has external characteristics (i.e. hardness, fibrous nature, particle size) like one of the materials/products listed in Annex B, then the procedural steps for sample preparation (pre-treatment, treatment and dilution) are carried out in the same way as listed for the material/product which has comparable external characteristics.; Any special diluents or practices required for particular materials or microorganisms in specific standard methods take priority over the general requirements listed in this document.

• Status: Not Published
• Publication Date: 06-Jan-2027
• ICS: 65.080 - Fertilizers
• Technical Committee: CEN/TC 223 - Soil improvers and growing media
• Current Stage: 5020 - Submission to Vote - Formal Approval
• Start Date: 17-Sep-2026
• Due Date: 29-Jan-2026
• Completion Date: 17-Sep-2026
• Directive: Harmonized Standard2019/1009 - 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 and amending Regulations (EC) No 1069/2009 and (EC) No 1107/2009 and repealing Regulation (EC) No 2003/2003 (Text with EEA relevance)
• Mandate: M/564 - Fertilising products in support of Regulation (EU) 2019/1009
• Mandate: M/564 Amd 1 - amending Implementing Decision C(2020) 612 as regards certain harmonised standards and standardisation deliverables, deadlines for their adoption and the duration of the validity of the standardisation request
• Mandate: M/564 Amd 2 - Commission Implementing decision amending Implementing Decision C(2020)612 as regards harmonised standards, the deadlines for the adoption of those standards and the duration of the validity of the standardisation request to the European Committee for Standardisation as regards the EU fertilising products in support of Regulation (EU) 2019/1009 of the European Parliament and of the Council
• Ref Project: oSIST prEN 13040-2:2026 - Soil improvers and growing media - Sample preparation - Part 2: Sample preparation for microbiological examination

Overview

FprEN 13040-2: Soil Improvers and Growing Media - Sample Preparation for Microbiological Examination is a European standard developed by CEN (European Committee for Standardization). This document specifies the essential requirements for preparing samples, including the creation of initial suspensions and subsequent dilutions, prior to conducting microbiological examinations of soil improvers and growing media. It is particularly relevant for laboratory testing for microorganisms such as Escherichia coli (E. coli), Salmonella spp., and enterococci. Proper and standardized sample preparation is crucial to obtaining reliable, repeatable, and comparable results across laboratories and jurisdictions.

Key Topics

• Sample Handling: Details best practices for handling, homogenization, and subdivision of samples to ensure representativity and avoid contamination.
• Pre-treatment and Treatment: Outlines procedures depending on material type, considering characteristics like hardness, fibrousness, or particle size.
• Initial Suspension Preparation: Describes methods for preparing and homogenizing suspensions with recommended diluents (e.g., phosphate buffer, buffered peptone water), addressing pH adjustments where necessary.
• Further Dilutions: Provides instructions on creating additional dilutions to achieve suitable concentrations for microbiological analysis.
• Equipment and Reagents: Specifies the apparatus, such as homogenizers, pipettes, and sterilization devices, as well as types of culture media and diluents for optimal sample processing.
• Quality and Safety: Emphasizes aseptic techniques, storage at appropriate temperatures, and guidelines for safe laboratory practice.

Applications

Implementation of FprEN 13040-2 yields significant benefits in several contexts:

• Laboratory Testing: Ensures uniformity and comparability when analyzing soil improvers and growing media for microbial contamination or compliance with health standards.
• Quality Control in Production: Supports producers in certifying the microbiological safety of their products, such as compost, peat, bark, digestates, coco coir, and other substrates.
• Regulatory Compliance: Assists laboratories and manufacturers in meeting European legal and safety requirements related to microbiological content in soil and growing media products.
• Research and Development: Facilitates reliable and repeatable microbiological investigations within research settings, allowing precise comparison of new soil improvers or growing media.
• Import/Export Procedures: Offers an internationally recognized framework for sample preparation, crucial in global trade where documented testing processes are required.

Related Standards

• EN 12579: Soil improvers and growing media - Sampling
• EN ISO 7218: Microbiology of the food chain - General requirements and guidance for microbiological examinations
• EN ISO 11133: Microbiology of food, animal feed and water - Preparation, production, storage, and performance testing of culture media
• prEN 17732: Soil improvers and growing media - Terminology

These standards collectively establish a comprehensive system for the sampling, preparation, and examination of soil improvers and growing media. Laboratories and stakeholders are encouraged to consult all relevant documents to ensure full compliance and quality assurance.

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Adhering to FprEN 13040-2 enables consistency and reliability in microbiological testing of soil-related products, supporting public health, agricultural productivity, and international trade. For organizations working within the soil improver and growing media sectors, following this standard is crucial for maintaining trust, safety, and compliance in their operations.

SLOVENSKI STANDARD
01-april-2026
Izboljševalci tal in rastni substrati - Priprava vzorcev - 2. del: Priprava vzorcev za
mikrobiološke preskuse
Soil improvers and growing media - Sample preparation - Part 2: Sample preparation for
microbiological examination
Bodenverbesserungsmittel und Kultursubstrate - Probenvorbereitung - Teil 2:
Probenvorbereitung für mikrobiologische Untersuchungen
Amendements du sol et supports de culture - Préparation des échantillons - Partie 2 :
Préparation des échantillons pour l’examen microbiologique
Ta slovenski standard je istoveten z: prEN 13040-2
ICS:
65.080 Gnojila Fertilizers
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

DRAFT
EUROPEAN STANDARD
prEN 13040-2
NORME EUROPÉENNE
EUROPÄISCHE NORM
December 2025
ICS 65.080
English Version
Soil improvers and growing media - Sample preparation -
Part 2: Sample preparation for microbiological
examination
Amendements du sol et supports de culture - Bodenverbesserungsmittel und Kultursubstrate -
Préparation des échantillons - Partie 2 : Préparation Probenvorbereitung - Teil 2: Probenvorbereitung für
des échantillons pour l'examen microbiologique mikrobiologische Untersuchungen
This draft European Standard is submitted to CEN members for second enquiry. It has been drawn up by the Technical
Committee CEN/TC 223.
If this draft becomes a European Standard, 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.

This draft European Standard was established by CEN 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.
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 European Standard. It is distributed for review and comments. It is subject to change without
notice and shall not be referred to as a European Standard.

EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION

EUROPÄISCHES KOMITEE FÜR NORMUNG

CEN-CENELEC Management Centre: Rue de la Science 23, B-1040 Brussels
© 2025 CEN All rights of exploitation in any form and by any means reserved Ref. No. prEN 13040-2:2025 E
worldwide for CEN national Members.

prEN 13040-2:2025 (E)
Contents Page
European foreword . 3
1 Scope . 4
2 Normative references . 4
3 Terms and definitions . 4
4 Principle . 5
5 Reagents . 5
6 Apparatus and equipment . 5
7 Transportation and storage of samples . 6
8 Procedure . 7
8.1 Preparation of the test portion . 7
8.2 Preparation of the initial suspension . 7
8.2.1 General. 7
8.2.2 For detection (Salmonella) . 8
8.2.3 For enumeration (E. coli and enterococci) . 8
8.2.4 Combined procedure for quantitative and qualitative methods . 8
8.3 Further dilutions . 9
Annex A (normative) Culture media and reagents . 11
Annex B (informative) Examples of product types with their proposed corresponding
procedural steps for sample preparation . 13
Bibliography . 18
prEN 13040-2:2025 (E)
European foreword
This document (prEN 13040-2:2025) has been prepared by Technical Committee CEN/TC 223 “Soil
improvers and growing media”, the secretariat of which is held by NEN.
nd
This document is currently submitted to the CEN 2 Enquiry.
This document has been prepared under a standardization request addressed to CEN by the European
Commission. The Standing Committee of the EFTA States subsequently approves these requests for its
Member States.
A list of all parts in this series can be found on the CEN website: www.cencenelec.eu.
EN 13040, Soil improvers and growing media — Sample preparation, is currently composed with the
following parts:
— Part 1: Sample preparation for chemical and physical tests, determination of dry matter content,
moisture content and laboratory bulk density;
— Part 2: Sample preparation for microbiological examination.
prEN 13040-2:2025 (E)
1 Scope
This document specifies the general requirements for the preparation of samples, initial suspensions and
further dilutions prior to microbiological examinations of soil improvers and growing media. This
method is intended especially for sample preparation prior to microbiological examinations of e.g.
Escherichia coli (E. coli), Salmonella spp. and enterococci.
If a laboratory receives a material/product that is not listed in Annex B but has external characteristics
(i.e. hardness, fibrous nature, particle size) like one of the materials/products listed in Annex B, then the
procedural steps for sample preparation (pre-treatment, treatment and dilution) are carried out in the
same way as listed for the material/product which has comparable external characteristics.
Any special diluents or practices required for particular materials or microorganisms in specific standard
methods take priority over the general requirements listed in this document.
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 12579, Soil improvers and growing media - Sampling
EN ISO 7218, Microbiology of the food chain - General requirements and guidance for microbiological
examinations (ISO 7218)
prEN 17732, Soil improvers and growing media - Terminology
3 Terms and definitions
For the purposes of this document, the terms and definitions given in EN 12579, prEN 17732 and the
following apply.
ISO and IEC maintain terminology databases for use in standardization at the following addresses:
— ISO Online browsing platform: available at https://www.iso.org/obp/
— IEC Electropedia: available at https://www.electropedia.org/
3.1
laboratory sample
in relation to chemical and physical testing, a final sample intended for laboratory testing and in relation
to microbiological testing, each separate segment sample intended for laboratory testing
3.2
test sample
sample prepared from the laboratory sample (3.1) and from which test portions (3.3) are taken
Note 1 to entry: A test sample is only necessary when the laboratory sample is used for multiple analysis (e.g.
microbiological and chemical analysis). When multiple analyses are performed (e.g. microbiological and chemical
analysis), appropriately sized test sample needs to be taken first for microbiological analysis in order to avoid
contamination. The remaining test sample material shall be stored in a cool place for possible subsequent analyses.
The remaining laboratory sample is thereafter passed on for chemical analyses, if necessary.

Under preparation. Stage at time of publication: prEN 17732:2025.
prEN 13040-2:2025 (E)
3.3
test portion
quantity of material drawn from the test sample or directly from the laboratory sample and on which the
tests and observations are actually carried out
3.4
initial suspension
primary dilution obtained after the test portion (3.2) has been treated with, normally, a nine-fold quantity
of diluent
Note 1 to entry: A closer ratio between the diluent and the quantity of material is often not recommended because
of possible inhibiting influences of the matrix.
3.5
further dilution
suspension or solution obtained by mixing a measured volume of the initial suspension (3.4) with an x-
fold volume of diluent and by repeating this operation with further dilutions until a dilution series is
obtained that is suitable for the inoculation of culture media
Note 1 to entry: Normally ten-fold dilutions are used to yield a decimal dilution series, but other ratios may be
required for specific purposes.
4 Principle
Sample preparation prior to microbiologal examination consists of two consecutive steps, for which the
second step depends on the intent of the subsequent microbiological examination, meaning enumeration
or detection.
1) Removal of a representative test portion (3.3) out of the test sample (3.2) or laboratory sample (3.1)
after thorough homogenisation
2) Addition of the homogenic test portion to the dilution or enrichment medium and homogenisation
2a) For detection: Preparation of a suspension for the first enrichment in a way to obtain an as
uniform as possible distribution of microorganisms contained in the test portion
2b) For enumeration: Preparation of an initial suspension in a way to obtain an as uniform as
possible distribution of microorganisms contained in the test portion. Preparation, if necessary,
of further dilutions in order to reduce the number of microorganisms per unit volume to allow
counting of colonies on enumeration plates after incubation
5 Reagents
Follow current laboratory practices in accordance with standards comparable to EN ISO 7218. The
composition of culture media and reagents and their preparation are specified in Annex A. For
performance testing of culture media, it is advised to follow the procedures of EN ISO 11133.
6 Apparatus and equipment
Disposable equipment is an acceptable alternative to reusable glassware if it has suitable specifications.
The usual microbiological laboratory equipment (see EN ISO 7218) and, in particular, the following shall
be used.
6.1 Apparatus for dry sterilization (oven) or wet sterilization (autoclave).
prEN 13040-2:2025 (E)
6.2 Equipment for size reduction; apparatuses (aseptic) may include:
— scissors;
— tweezers;
— spoons;
— scalpels;
— knives;
— hammers (used with a sterile bag around the sample);
— saws;
— spatulas;
— mortar and pestle.
6.3 Homogenization devices; apparatuses (aseptic) may include:
— a magnetic stirrer with vessels and stirring bars ;
— orbital shaker, with bottles (glass beads may be used in the case of viscous or thick materials);
— peristaltic blender (paddle blender), with sterile plastic bags, preferable with a filter;
— vortex mixer (for dilutions).
6.4 Other laboratory equipment; may include:
— pipettes or pipettor and sterile tips, to dispense 1 ml to 10 ml with tips (with large opening if
necessary);
— scales and gravimetric dilution controllers, capable of weighing to 1 % of the mass;
— aseptic containers (bowls, wide-mouth containers, developing tray) for homogenization of the
laboratory sample.
7 Transportation and storage of samples
Sampling is not part of the method specified in this document. EN 12579 shall be followed for sampling
soil improvers and growing media for microbiological examination. It is important that the laboratory
receives a sample that is representative of the product under consideration. The sample shall not have
been damaged or changed during transport or storage.
The samples as submitted to the laboratory, shall be stored and treated so that they shall not undergo
any further decomposition, physical damage, hydration or dehydration. Samples shall be stored at
5 °C ± 3 °C, but not frozen.
It is recommended to examine the samples within 72 h after reception as the microbiome can change
over time.
prEN 13040-2:2025 (E)
8 Procedure
8.1 Preparation of the test portion
WARNING — In order to safeguard the health of laboratory personnel, it is essential that the procedures
described in this document are only undertaken in properly equipped laboratories, under the supervision
of a skilled microbiologist, and that great care is taken in the disposal of all incubated materials. Persons
using this document shall be familiar with normal laboratory practice. This document does not purport
to address all of the safety aspects, if any, associated with its use. It is the responsibility of the user to
establish appropriate safety and health practices.
All operations before and after opening the packaged material shall be carried out under aseptic
conditions, to avoid any external contamination.
The types of packaged materials sent to the laboratory can be in flexible packaging. This shall be removed
or opened aseptically.
NOTE There is no need to disinfect the packaging if the contents can be removed aseptically after opening
without any risk of external contamination.
Before taking the test portion, the sample material of the laboratory sample or test sample shall be
thoroughly homogenized so that representative sampling is possible. Liquid samples should be
homogenized by shaking the container or stirring with an aseptic object. If possible, solid samples should
be mixed in the packaging with an aseptic device. If this is not possible, the laboratory sample shall be
transferred to an aseptic container (bowl, developing tray) and mixed.
Remove any incidental pieces (e.g. stones, glass, plastic pieces) aseptically.
The following special features shall be used:
— For pre-shaped materials (e.g. mineral wool slabs, plugs): The test portion (3.3) is taken directly
from the material, without “reconstitution” by addition of water.
— For solid rigid materials (e.g. compost, bark): Solid materials shall be reduced in size to obtain a
representative test portion (3.3) under aseptic conditions (6.1). The dimension of the pieces shall not
exceed approximately 2 cm in all dimensions.
— For block form (compressed non-pre shaped) materials (e.g. blocks of compressed coir,
blocks of compressed peat): The material shall be crushed and homogenized using suitable tools
and weighed out in its original dry state. It shall not be reconstituted (EN 12579).
8.2 Preparation of the initial suspension
8.2.1 General
Table B.1 provides an overview of exemplary materials and recommended equipment as well as a
description of the operating procedure for sample preparation.
To avoid damage to microorganisms by sudden temperature changes, the temperature of all diluents shall
be approximately equal to the ambient temperature of the laboratory, unless otherwise specified for
particular materials.
Some materials can significantly change the pH value of the initial suspension, which can negatively affect
the viability of the microorganisms under investigation. In general, a phosphate buffer or buffered
peptone water is sufficient to prepare the initial suspension.
When testing bog peat, Sphagnum, alk
...