CEN/TR 16193:2013

SLOVENSKI STANDARD
01-julij-2013
Blato, obdelani biološki odpadki in tla - Ugotavljanje prisotnosti in števila
Escherichia coli
Sludge, treated biowaste and soil - Detection and enumeration of Escherichia coli
Schlamm, behandelter Bioabfall und Boden - Nachweis und Zählung von Escherichia coli
Boue, biodéchet traité et sol - Recherche et dénombrement des Escherichia coli
Ta slovenski standard je istoveten z: CEN/TR 16193:2013
ICS:
13.030.20 7HNRþLRGSDGNL%ODWR Liquid wastes. Sludge
13.080.30 Biološke lastnosti tal Biological properties of soils
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

TECHNICAL REPORT
CEN/TR 16193
RAPPORT TECHNIQUE
TECHNISCHER BERICHT
May 2013
ICS 13.030.01
English Version
Sludge, treated biowaste and soil - Detection and enumeration
of Escherichia coli
Boue, biodéchet traité et sol - Recherche et dénombrement Schlamm, behandelter Bioabfall und Boden - Nachweis und
des Escherichia coli Zählung von Escherichia coli

This Technical Report was approved by CEN on 1 March 2011. It has been drawn up by the Technical Committee CEN/TC 400.

CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia,
Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania,
Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and United
Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION

EUROPÄISCHES KOMITEE FÜR NORMUNG

Management Centre: Avenue Marnix 17, B-1000 Brussels
© 2013 CEN All rights of exploitation in any form and by any means reserved Ref. No. CEN/TR 16193:2013: E
worldwide for CEN national Members.

Contents Page
Foreword . 4
Introduction . 5
1 Scope . 6
2 Normative references . 6
3 Terms and definitions . 6
4 Abbreviations . 7
5 Quality assurance . 7
6 Method A — Membrane filtration method for quantification . 7
6.1 Scope . 7
6.2 Principle. 8
6.3 Reagents, diluents and culture media . 8
6.4 Apparatus . 9
6.5 Sampling . 10
6.5.1 General . 10
6.5.2 Storage . 11
6.5.3 Handling . 11
6.6 Procedure . 11
6.6.1 Sample preparation . 11
6.6.2 Sample dilution . 12
6.6.3 Membrane filtration . 12
6.6.4 Resuscitation and enumeration of colonies on chromogenic agar . 13
6.6.5 Confirmation of colony identity . 13
6.6.6 Determination of the dry residue content . 13
6.7 Calculation and expression of results . 13
6.8 Performance data of the interlaboratory comparison — Method A . 14
6.8.1 Material used in the interlaboratory comparison study . 14
6.8.2 First assessment of the precision of the method . 15
6.8.3 Interlaboratory comparison results . 16
6.9 Pre-filtration and centrifugation — Comparison tests . 17
7 Method B — Miniaturised method (Most Probable Number) by inoculation in liquid
medium . 19
7.1 Scope . 19
7.2 Principle. 19
7.3 Reagents, diluents and culture media . 19
7.4 Apparatus . 21
7.5 Sampling . 22
7.5.1 General . 22
7.5.2 Storage . 22
7.5.3 Handling . 22
7.6 Procedure . 22
7.6.1 Sample preparation . 22
7.6.2 Analysis . 23
7.6.3 Determination of the dry residue content . 24
7.7 Expression of results . 24
7.7.1 Determination of the characteristic number . 24
7.7.2 Calculation of the MPN and its confidence interval . 25
7.8 Performance data . 27
7.8.1 MPN Statistical table . 27
7.8.2 Performance data of the interlaboratory comparison .3 5
7.8.3 First assessment of the precision of the method .3 6
7.8.4 Interlaboratory comparison results .3 7
7.9 Preparation of synthetic sea salt .3 9
7.9.1 Major ion composition of a convenient ocean synthetic sea salt . 39
7.9.2 Example for preparation from defined substances .3 9
7.10 Quality criteria for the manufacturing of the medium in microtitre plates (E. coli) . 40
8 Method C — Macromethod (Most Probable Number) in liquid medium . 41
8.1 Scope .4 1
8.2 Principle .4 1
8.3 Reagents, diluents and culture media .4 1
8.4 Apparatus .4 2
8.5 Sampling .4 3
8.5.1 General .4 3
8.5.2 Sample storage.4 3
8.5.3 Sample handling .4 3
8.6 Procedure .4 3
8.6.1 Sample preparation .4 3
8.6.2 Analysis .4 4
8.6.3 Determination of the dry residue content .4 4
8.7 Expression of the results .4 4
8.8 Performance data .4 6
8.8.1 MPN Statistical table for 3-tubes MPN procedure .4 6
8.8.2 Repeatability and reproducibility .4 7
8.8.3 First assessment of the precision of the method .4 7
8.8.4 Interlaboratory comparison results .4 8
9 Test report .5 0
Bibliography .5 1

Foreword
This document (CEN/TR 16193:2013) has been prepared by Technical Committee CEN/TC 400 “Project
Committee - Horizontal standards in in the fields of sludge, biowaste and soil”, 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 [and/or CENELEC] shall not be held responsible for identifying any or all such patent rights.
This document has been prepared under a mandate given to CEN by the European Commission and the
European Free Trade Association.
This document is part of a modular horizontal approach in which this document belongs to the analytical step.
The preparation of this document by CEN is based on a mandate by the European Commission (Mandate
M/330). The mandate considers standards on sampling and analytical methods for hygienic and biological
parameters as well as inorganic and organic determinants. It was the aim of the mandate to develop
standards that are applicable to sludge, treated biowaste and soil and lead to equivalent results as far as this
is technically feasible.
Until now, test methods determining properties of materials within the environmental area were prepared in
Technical Committees (TCs) working on specific products/matrices (soil, waste, sludge etc). However, it is
recognised that many steps in test procedures can be used in test procedures for other products/matrices. By
careful determination of these steps and selection of specific questions wi
...