EN ISO 8199:2007
(Main)Water quality - General guidance on the enumeration of micro-organisms by culture (ISO 8199:2005)
Water quality - General guidance on the enumeration of micro-organisms by culture (ISO 8199:2005)
ISO 8199:2005 presents guidance for carrying out manipulations which are common to each technique for the microbiological examination of water, particularly the preparation of samples, culture media and apparatus. It also describes the various enumeration techniques available and the criteria for the choice of a particular technique. It is mainly intended for bacteria, yeasts and moulds. Some aspects are also applicable to viruses and parasites.
Wasserbeschaffenheit - Allgemeine Anleitung zur Zählung von Mikroorganismen durch Kulturverfahren (ISO 8199:2005)
Diese Internationale Norm gibt eine Anleitung zu Arbeitsweisen, die für jedes Verfahren für die mikrobiologische Untersuchung von Wasser, besonders für die Aufbereitung von Proben, Kulturmedien und Geräten üblich sind. Sie beschreibt die unterschiedlichen verfügbaren Zählverfahren und die Kriterien für die Auswahl eines bestimmten Verfahrens. Diese Internationale Norm ist hauptsächlich für Bakterien, Hefen und Pilze bestimmt. Einige Aspekte sind auch auf Viren und Parasiten anwendbar.
Qualité de l'eau - Lignes directrices générales pour le dénombrement des micro-organismes sur milieu de culture (ISO 8199:2005)
L'ISO 8199:2005 présente des lignes directrices concernant des manipulations communes à chaque technique d'examen microbiologique de l'eau, en particulier la préparation des échantillons, les milieux de culture et l'appareillage. Elle décrit également les diverses techniques de dénombrement possibles et les critères de choix d'une technique particulière. Elle concerne principalement les bactéries, les levures et les champignons. Certains aspects peuvent aussi s'appliquer aux virus et aux parasites.
Kakovost vode - Splošno navodilo za štetje mikroorganizmov v gojišču (ISO 8199:2005)
General Information
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Standards Content (Sample)
SLOVENSKI STANDARD
01-december-2007
1DGRPHãþD
SIST ISO 8199:1997
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Water quality - General guidance on the enumeration of micro-organisms by culture (ISO
8199:2005)
Wasserbeschaffenheit - Allgemeine Anleitung zur Zählung von Mikroorganismen durch
Kulturverfahren (ISO 8199:2005)
Qualité de l'eau - Lignes directrices générales pour le dénombrement des micro-
organismes sur milieu de culture (ISO 8199:2005)
Ta slovenski standard je istoveten z: EN ISO 8199:2007
ICS:
07.100.20 Mikrobiologija vode Microbiology of water
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
EUROPEAN STANDARD
EN ISO 8199
NORME EUROPÉENNE
EUROPÄISCHE NORM
October 2007
ICS 07.100.20
English Version
Water quality - General guidance on the enumeration of micro-
organisms by culture (ISO 8199:2005)
Qualité de l'eau - Lignes directrices générales pour le Wasserbeschaffenheit - Allgemeine Anleitung zur Zählung
dénombrement des micro-organismes sur milieu de culture von Mikroorganismen durch Kulturverfahren (ISO
(ISO 8199:2005) 8199:2005)
This European Standard was approved by CEN on 27 September 2007.
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. Up-to-date lists and bibliographical references concerning such national
standards may be obtained on application to the CEN Management Centre or to any CEN member.
This European Standard exists 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 Management Centre has the same status as the
official versions.
CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Cyprus, Czech Republic, Denmark, Estonia, Finland,
France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal,
Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and United Kingdom.
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EUROPÄISCHES KOMITEE FÜR NORMUNG
Management Centre: rue de Stassart, 36 B-1050 Brussels
© 2007 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 8199:2007: E
worldwide for CEN national Members.
Foreword
The text of ISO 8199:2005 has been prepared by Technical Committee ISO/TC 147 “Water quality” of the
International Organization for Standardization (ISO) and has been taken over as EN ISO 8199:2007 by
Technical Committee CEN/TC 230 “Water analysis” the secretariat of which is held by DIN.
This European Standard shall be given the status of a national standard, either by publication of an identical
text or by endorsement, at the latest by April 2008, and conflicting national standards shall be withdrawn at the
latest by April 2008.
According to the CEN/CENELEC Internal Regulations, the national standards organizations of the following
countries are bound to implement this European Standard: Austria, Belgium, Bulgaria, Cyprus, Czech
Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia,
Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain,
Sweden, Switzerland and the United Kingdom.
Endorsement notice
The text of ISO 8199:2005 has been approved by CEN as a EN ISO 8199:2007 without any modification.
INTERNATIONAL ISO
STANDARD 8199
Second edition
2005-06-15
Water quality — General guidance on the
enumeration of micro-organisms by
culture
Qualité de l'eau — Lignes directrices générales pour le dénombrement
des micro-organismes sur milieu de culture
Reference number
ISO 8199:2005(E)
©
ISO 2005
ISO 8199:2005(E)
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ii © ISO 2005 – All rights reserved
ISO 8199:2005(E)
Contents Page
Foreword. iv
Introduction . v
1 Scope . 1
2 Normative references . 1
3 Principle. 1
4 General. 1
5 Diluents and culture media. 2
6 Sterilization of apparatus and glassware. 5
7 Samples . 5
8 Enumeration after inoculation of test portions of the sample in (or on) solid media . 6
9 Enumeration by inoculation in liquid media . 15
Annex A (informative) Criteria for the choice of enumeration technique. 22
Annex B (informative) MPN tables. 27
Bibliography . 37
ISO 8199:2005(E)
Foreword
ISO (the International Organization for Standardization) is a worldwide federation of national standards bodies
(ISO member bodies). The work of preparing International Standards is normally carried out through ISO
technical committees. Each member body interested in a subject for which a technical committee has been
established has the right to be represented on that committee. International organizations, governmental and
non-governmental, in liaison with ISO, also take part in the work. ISO collaborates closely with the
International Electrotechnical Commission (IEC) on all matters of electrotechnical standardization.
International Standards are drafted in accordance with the rules given in the ISO/IEC Directives, Part 2.
The main task of technical committees is to prepare International Standards. Draft International Standards
adopted by the technical committees are circulated to the member bodies for voting. Publication as an
International Standard requires approval by at least 75 % of the member bodies casting a vote.
Attention is drawn to the possibility that some of the elements of this document may be the subject of patent
rights. ISO shall not be held responsible for identifying any or all such patent rights.
ISO 8199 was prepared by Technical Committee ISO/TC 147, Water quality, Subcommittee SC 4,
Microbiological methods.
This second edition cancels and replaces the first edition (ISO 8199:1988), which has been technically revised.
iv © ISO 2005 – All rights reserved
ISO 8199:2005(E)
Introduction
Techniques for the isolation and enumeration of micro-organisms, based on their ability to grow on specified
culture media, are an important and widely used means of assessing the microbiological quality of water. The
purpose of this guide is to gather in a single document the information common to the various enumeration
techniques so as to avoid repetition of technical details in individual standards and to facilitate the choice of
the technique most suitable for a particular problem.
INTERNATIONAL STANDARD ISO 8199:2005(E)
Water quality — General guidance on the enumeration of micro-
organisms by culture
WARNING — Persons using this International Standard should be familiar with normal laboratory
practice. This standard 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 International
Standard be carried out by suitably trained staff.
1 Scope
This International Standard presents guidance for carrying out manipulations which are common to each
technique for the microbiological examination of water, particularly the preparation of samples, culture media
and apparatus. It also describes the various enumeration techniques available and the criteria for the choice
of a particular technique. This International Standard is mainly intended for bacteria, yeasts and moulds.
Some aspects are also applicable to viruses and parasites.
2 Normative references
The following referenced documents are indispensable for the application 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.
ISO 3696:1987, Water for analytical laboratory use — Specification and test methods
ISO 19458, Water quality — Sampling for microbiological analysis
3 Principle
The general principle of these techniques consists of inoculating a known volume of a water sample on or into
a culture medium (solid or liquid). It is assumed that after incubation each micro-organism present multiplies,
giving either a colony visible directly on the solid medium, or changes in observable properties of the liquid
medium. The choice of a particular method depends not only on the nature of the micro-organisms sought, but
also on the nature of the water and the reasons for the examination.
4 General
Uniformity of temperatures and (incubation) times: The following accepted ranges of temperatures and times
during incubation or storage are applied, when appropriate for the intended target organism, and unless
otherwise stated in the specific standard.
ISO 8199:2005(E)
Storage temperatures:
(− 70 ± 10) °C; (− 20 ± 5) °C; (5 ± 3) °C;
Incubation temperatures:
(22 ± 2) °C; (36 ± 2) °C;
Sterilization temperatures:
(115 ± 3) °C; (121 ± 3) °C; (170 ± 10) °C;
Incubation times:
(21 ± 3) h; (44 ± 4) h; (68 ± 4) h.
Other times and temperatures may be specified for specific methods when necessary.
The upper incubation temperature limits are very strict (they can have large influences on the growth). The
lower temperature limits may be exceeded for short periods, e.g. due to opening the door of an incubator, but
recovery shall be rapid.
Tolerances on volumes and masses: Unless otherwise stated, the accepted range of any measured value is:
stated value ± 5 %.
5 Diluents and culture media
5.1 General
5.1.1 Quality requirements
Use constituents of uniform quality and analytical-grade chemicals for the preparation of media. Other grades
of chemical may be used provided they can be shown to produce the same results. Alternatively, dehydrated
complete media or diluents may be used. Follow the manufacturer's instructions strictly.
Use glass-distilled or demineralized water which is free from substances that might affect growth of micro-
organisms under the test conditions. The water shall comply with the requirements of ISO 3696:1987, grade 3.
Unless otherwise stated, the ingredients are added to the volume of water instead of making up to a certain
volume, as is normal in the preparation of microbiological culture media.
Before use, check the quality of the media, diluents and filters, e.g. by following the procedures described in
ISO 7704, ISO/TS 11133-1 or ISO/TS 11133-2.
5.1.2 Sterilization
Dispense diluents and culture media in containers suitable for sterilization by autoclaving. For most purposes,
a temperature of (121 ± 3) °C for 15 min is adequate. However, a differ
...
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