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OSI nternational Standard 6461 / 1 INTERNATIONAL ORGANIZATION FOR STANDARDIZATION ME XAYHAPOAHAA OPTAHW3AUWR nO CTAHAAPTM3AUWW-ORGANISATION INTE RNATIONALE DE NORMALISATION Water quality - Detection and enumeration of the spores of sulfite-reducing anaerobes (clostridia) Part 1 : Method by enrichment in a liguid medium Qualite de I'eau - Recherche et denombrement des spores de micro-organismes anaerobies sulfito-reducteurs (clostridia) - Partie 1: Méthode par enrichissement dans un milieu liquide First edition - 1986-02-01 ISO 6461/1-1986 (E) UDC 543.39 :579.852.13 Ref. No. ISO 6461/1-1986 (E) Price based on 3 pages Foreword national standards bodies (iSO member bodies). The work of preparing International Standards is normally carried out through ISO technical committees. Each member the right to be represented on that committee. International organizations, govern- mental and non-governmental, in liaison with IsO, also take part in the work. Draft International Standards adopted by the technical committees are circulated to the member bodies for approval before their acceptance as International Standards by the ISo Council. They are approved in accordance with ISO procedures requiring at least 75 % approval by the member bodies voting. International Standard ISO 6461/1was prepared by Technical Committee ISO/TC 147, Water quality. Users should note that all International Standards undergo revision from time to time latest edition, unless otherwise stated. C International OrganizationforStandardization,1986 Printed in Switzerland ISO6461/1-1986 (E) INTERNATIONAL STANDARD Water quality -- Detection and enumeration of the spores of sulfite-reducing anaerobes (clostridia) Part 1: Method by enrichment in a liquid medium 0 Introduction 4 Definition The spores of sulfite-reducing anaerobes (clostridia) are wide- For the purpose of this part of isO 6461, the following defi- spread in the environment. They are present in human and nition applies. animal faecal matter,inwaste water and in soil.Unlike Escherichia coli and other coliform organisms, the spores sur- ciostridia: Suifite-reducing, spore-forming, anaerobic micro- vive in water for long periods as they are more resistant than organisms which belong to the Bacillaceae family and the vegetative forms to the action of chemical and physical factors. genus Clostridium. They may thus give an indication of remote or intermittent pollution. They may even be resistant to chlorination at levels which are normally used for the treatment of water, and they are thus useful for control purposes. Principle ISo 6461 consists of the following parts : The detection of spores of sulfite-reducing anaerobes (clostridia) in a specified volume of a water sample requires the Part 1: Method by enrichment in a liquid medium. following steps. Part 2: Method by membrane filtration. 5.1 Selection of spores 1 Scope Selection of spores in the sample by applying heat for a period of time sufficient to destroy vegetative bacteria. This part of ISo 6461 specifies a method for the detection and enumeration of the spores of sulfite-reducing anaerobes (clostridia)by enrichment ina liquid medium. 5.2 Enrichment culture Detection and enumeration of spores of sulfite-reducing 2 Field of application anaerobes by inoculating volumes of the sample into liquid enrichment media, followed by incubation at 37 ± 1 C for The method is applicable to all types of water, including turbid 44 ± 4 h in anaerobic conditions. water. Culture media and reagents 3 References Iso 3696, Water for laboratory use -- Specifications. 6.1 Basic materials ISo 5667, Water quality - Sampling In order to improve the reproducibility of the results, it is recommended that,forthe preparation of the diluents and Part2:Guidanceonsamplingtechniques. culture media, dehydrated basic components or complete Part 3: Guidance on the preservation and handling of dehydrated media be used. Similarly, commercially prepared samples. reagents may also be used. T

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