CEN/TS 17804:2022

SLOVENSKI STANDARD
01-junij-2024
Organska, organsko-mineralna in anorganska gnojila - Ugotavljanje prisotnosti
enterokokov (Enterocococaceae)
Organic, organo-mineral and inorganic fertilizers - Detection of Enterocococaceae
Organische, organisch-mineralische und anorganische Düngemittel - Nachweis von
Enterococcaceae
Engrais organiques, organo-minéraux et inorganiques - Recherche des Enterococaceae
Ta slovenski standard je istoveten z: CEN/TS 17804:2022
ICS:
65.080 Gnojila Fertilizers
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

CEN/TS 17804
TECHNICAL SPECIFICATION
SPÉCIFICATION TECHNIQUE
May 2022
TECHNISCHE SPEZIFIKATION
ICS 65.080
English Version
Organic, organo-mineral and inorganic fertilizers -
Detection of Enterocococaceae
Engrais organiques, organo-minéraux et inorganiques Organische, organisch-mineralische und
- Recherche des Enterococaceae anorganische Düngemittel - Nachweis von
Enterococcaceae
This Technical Specification (CEN/TS) was approved by CEN on 13 April 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 17804:2022 E
worldwide for CEN national Members.

Contents Page
European foreword . 3
Introduction . 4
1 Scope . 5
2 Normative references . 5
3 Terms and definitions . 5
4 Principle . 6
5 Reagents . 7
5.1 General. 7
5.2 Diluents . 7
5.2.1 General. 7
5.2.2 Basic phosphate buffer . 7
5.2.3 Double-buffered phosphate buffer . 7
5.3 Selective media . 7
5.3.1 Slanetz Bartley agar . 7
5.3.2 Bile Esculin Azide agar . 8
6 Equipment and consumables . 8
7 Sampling . 9
8 Preparation of test sample . 9
9 Procedure (see Figure A.1 in Annex A) . 9
9.1 Preparation of the initial suspension and decimal dilutions . 9
9.2 Inoculation and incubation . 9
9.2.1 General. 9
9.2.2 Procedure . 10
9.3 Enumeration of colonies . 10
9.4 Confirmation (optional) . 10
9.4.1 General. 10
9.4.2 Confirmation on the second selective medium . 10
9.4.3 Catalase Test . 11
10 Expression of results . 11
11 Test report . 12
Annex A (normative) Diagram of the procedure . 13
Annex B (normative) Composition and preparation of culture media and reagents . 14
B.1 General. 14
B.2 Basic phosphate buffer . 14
B.3 Double-buffered phosphate buffer . 15
B.4 Slanetz Bartley agar . 15
B.5 Bile Esculin Azide agar . 16
Bibliography . 17
European foreword
This document (CEN/TS 17804:2022) has been prepared by Technical Committee CEN/TC 260
“Fertilizers and liming materials”, the secretariat of which is held by DIN.
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 methodology has been developed to detect and enumerate Enterococcaceae in organic, organo-
mineral and inorganic fertilizers in order to be able to control certain hygienic requirements in
Regulation (EU) 2019/1009 [1].
Enterococcaceae in the sense of this document include several species of the genus Enterococcus (3.7, 3.8)
with a faecal origin. Consequently, it can be used as an indicator of faecal contamination. It can also be
used to monitor the effectiveness of pasteurization or disinfection treatments. Compared to E.coli, they
have a higher tenacity and can therefore better reflect the behaviour of all pathogens in fertilizers.
Because of the large variety of fertilizers, this method is not appropriate in every detail for certain
products. In this case, different methods which are specific to these products can be used if absolutely
necessary for justified technical reasons. Nevertheless, every attempt should be made to apply this
method as far as possible.
Mineral components in fertilizers can have a negative impact on the survivability of microorganisms
when they go into solution. In addition to an unfavourable shift in the pH value, the products can have a
strong osmotic effect or be toxic to cells themselves (e.g. copper). Therefore, it can be necessary to test
the inhibitory effect of the fertilizers to be investigated in a pre-test.
The method is validated in an interlaboratory study for the following products (Enterococcaceae were
investigated in both native and spiked test material):
Table 1 — Product groups and matrices for which the methods described in this method are
applicable and tested in a validation trial
Product group matrix
Organic fertilizers to be determined at an international ring trial
Organo-mineral fertilizers to be determined at an international ring trial
Inorganic fertilizers to be determined at an international ring trial
International ring trials will be conducted on the basis of this document.
1 Scope
This document specifies a method for the detection and enumeration of Enterococcaceae in fertilizers of
the following Product Function Categories (PFCs) of EU fertilizing products, as described in Regulation
(EU) 2019/1009 [1]:
• PFC 1(A): Organic fertilizer;
• PFC 1(B): Organo-mineral fertilizer;
• PFC 1(C): Inorganic fertilizer, which contains more than 1 % by mass of organic carbon, other than
organic carbon from chelating or complexing agents, nitrification inhibitors, denitrification
inhibitors or urease inhibitors, coating agents, urea or calcium cyanamide. The present method was
validated on products known as present on the market in April 2021 and conform to Regulation (EU)
2019/1009 [1] that are inorganic fertilizers with more than 1 % of organic carbon such as poultry
manure and struvite with low level of organic matter. In case that other products would be developed
having other physical and chemical characteristics, it might become necessary to develop different
methods to correctly account for pathogens they might contain.
This document specifies a colony-count technique on selective media, Slanetz Bartley agar or Bile Esculin
Azide agar, respectively. The method is based on EN ISO 7899-2:2000.
2 Normative references
There are no normative references in this document.
3 Terms and definitions
For the purposes of this document, the following terms and definitions 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
laboratory sample
sample intended for laboratory inspection or testing
3.2
test sample
sample prepared from the laboratory sample (3.1) and from which test portions (3.3) will be taken
3.3
test portion
quantity of material taken from the test sample (or if both are the same, from the laboratory sample) and
on which the test is carried out
3.4
initial suspension
primary dilution obtained after a weighed or measured quantity of the product under examination (or of
a test sample prepared from the product) has been mixed with, normally, a nine-fold quantity of diluent
Note 1 to entry: A closer ratio between the diluent and the quantity of product 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,
suitable for the inoculation of culture media, is obtained
Note 1 to entry: Ten-fold dilutions are normally used to produce
...