EN ISO 10705-1:2001
(Main)Water quality - Detection and enumeration of bacteriophages - Part 1: Enumeration of F-specific RNA bacteriophages (ISO 10705-1:1995)
Water quality - Detection and enumeration of bacteriophages - Part 1: Enumeration of F-specific RNA bacteriophages (ISO 10705-1:1995)
Specifies a method for the detection and enumeration of F-specific ribonucleic acid bacteriophages. The sample is incubated with an appropriate host strain. Applicable to all kinds of water, sediments and sludges, even for shellfish extracts. The presence of F-specific RNA bacteriophages in a water sample indicates pollution by wastewater contaminated by human or animal faeces.
Wasserbeschaffenheit - Nachweis und Zählung von Bakteriophagen - Teil 1: Zählung von F-spezifischen RNA-Bakteriophagen (ISO 10705-1:1995)
Dieser Teil von ISO 10705 legt ein Verfahren zum Nachweis und zur Zählung
von F-spezifischen Ribonucleinsäure-(RNA-)Bakteriophagen durch Bebrütung
der Probe mit einem geeigneten Wirtsstamm fest. Das Verfahren kann für alle
Arten von Wasser, Sedimenten und Schlämmen angewandt werden, wenn
notwendig nach Verdünnung. Im Falle von geringen Anzahlen kann eine Auf-konzentration
erforderlich sein, für die ein gesonderter Teil von ISO 10705
entwickelt werden wird. Das Verfahren kann auch für Schalentierextrakte ange-wandt
werden. In Abhängigkeit von der relativen Häufigkeit der F-spezifischen
RNA-Bakteriophagen gegenüber Hintergrundorganismen können zusätzliche
Bestätigungstests notwendig sein, die ebenfalls in diesem Teil von ISO 10705
festgelegt werden.
Das Vorhandensein von F-spezifischen RNA-Bakteriophagen in einer Wasser-probe
zeigt im Allgemeinen eine Verunreinigung durch Abwasser an, das mit
menschlichen oder tierischen Fäkalien kontaminiert ist. Ihr Überleben in der
Umwelt, die Entfernung durch gängige Wasseraufbereitungsprozesse und die
Anreicherung oder Zurückhaltung in Schalentieren ähneln denen von nah-rungs-
und wasserbürtigen enteralen Viren des Menschen, z. B. Enteroviren,
Hepatitis-A-Virus und Rotaviren.
Qualité de l'eau - Détection et dénombrement des bactériophages - Partie 1: Dénombrement des bactériophages ARN F spécifiques (ISO 10705-1:1995)
La présente partie de l'ISO 10705 prescrit une méthode de détection et de dénombrement des bactériophages ARN F spécifiques par incubation de l'échantillon avec une souche-hôte appropriée. La méthode peut être appliquée à tous les types d'eaux, de sédiments et de boues, si nécessaire après dilution. Dans le cas de faible population, une étape de préconcentration peut être nécessaire pour laquelle une partie séparée sera élaborée. La méthode peut également être appliquée aux extraits de coquillage. Des essais supplémentaires de confirmation des résultats, également spécifiés dans la présente partie de l'ISO 10705, peuvent être nécessaires, suivant le nombre plus ou moins élevé des bactériophages ARN F spécifiques par rapport aux autres organismes présents. La présence de bactériophages ARN F spécifiques dans un échantillon d'eau indique en général une pollution par des eaux usées contaminées par des matières fécales d'origine animale ou humaine. Les caractéristiques de leur résistance dans l'environnement, de leur élimination par des procédés courants de traitement de l'eau, ainsi que de leur concentration ou rétention par les coquillages, ressemble à celle des virus entériques humains présents dans l'eau et dans les aliments, par exemple les entérovirus, le virus de l'hépatite A et les rotavirus.
Kakovost vode - Ugotavljanje prisotnosti in števila bakteriofagov - 1. del - Ugotavljanje števila F-specifičnih RNA bakteriofagov (ISO 10705-1:1995)
General Information
Standards Content (Sample)
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.Wasserbeschaffenheit - Nachweis und Zählung von Bakteriophagen - Teil 1: Zählung von F-spezifischen RNA-Bakteriophagen (ISO 10705-1:1995)Qualité de l'eau - Détection et dénombrement des bactériophages - Partie 1: Dénombrement des bactériophages ARN F spécifiques (ISO 10705-1:1995)Water quality - Detection and enumeration of bacteriophages - Part 1: Enumeration of F-specific RNA bacteriophages (ISO 10705-1:1995)07.100.20Mikrobiologija vodeMicrobiology of waterICS:Ta slovenski standard je istoveten z:EN ISO 10705-1:2001SIST EN ISO 10705-1:2001en01-december-2001SIST EN ISO 10705-1:2001SLOVENSKI
STANDARDSIST ISO 10705-1:19981DGRPHãþD
EUROPEAN STANDARDNORME EUROPÉENNEEUROPÄISCHE NORMEN ISO 10705-1August 2001ICS 07.100.20English versionWater quality - Detection and enumeration of bacteriophages -Part 1: Enumeration of F-specific RNA bacteriophages (ISO10705-1:1995)Qualité de l'eau - Détection et dénombrement desbactériophages - Partie 1: Dénombrement desbactériophages ARN F spécifiques (ISO 10705-1:1995)Wasserbeschaffenheit - Nachweis und Zählung vonBakteriophagen - Teil 1: Zählung von F-spezifischen RNA-Bakteriophagen (ISO 10705-1:1995)This European Standard was approved by CEN on 29 June 2001.CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this EuropeanStandard the status of a national standard without any alteration. Up-to-date lists and bibliographical references concerning such nationalstandards may be obtained on application to the 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 translationunder the responsibility of a CEN member into its own language and notified to the Management Centre has the same status as the officialversions.CEN members are the national standards bodies of Austria, Belgium, Czech Republic, Denmark, Finland, France, Germany, Greece,Iceland, Ireland, Italy, Luxembourg, Netherlands, Norway, Portugal, Spain, Sweden, Switzerland and United Kingdom.EUROPEAN COMMITTEE FOR STANDARDIZATIONCOMITÉ EUROPÉEN DE NORMALISATIONEUROPÄISCHES KOMITEE FÜR NORMUNGManagement Centre: rue de Stassart, 36
B-1050 Brussels© 2001 CENAll rights of exploitation in any form and by any means reservedworldwide for CEN national Members.Ref. No. EN ISO 10705-1:2001 ESIST EN ISO 10705-1:2001
INTERNATIONAL STANDARD IS0 10705-I First edition 1995-08-01 Water quality - Detection and enumeration of bacteriophages - Part 1: Enumeration of F-specific RNA bacteriophages Qualit6 de I’eau - D6tection et dhnombrement des bactkiophages - Partie 1: Dhnombrement des bactkiophages ARN F spkifiques Reference number IS0 10705-I :I 995(E) SIST EN ISO 10705-1:2001
IS0 10705=1:1995(E) Foreword IS0 (the International Organization for Standardization) is a worldwide federation of national standards bodies (IS0 member bodies). The work of preparing International Standards is normally carried out through IS0 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. IS0 collaborates closely with the International Electrotechnical Commission (IEC) on all matters of electrotechnical standardization. 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. International Standard IS0 10705-l was prepared by Technical Committee lSO/TC 147, Water quality, Subcommittee SC 4, Microbiological methods. IS0 10705 consists of the following parts, under the general title Water quality - Detection and enumeration of bacteriophages: - Part 7: Enumeration of F-specific RNA bacteriophages - Part 2: Enumeration of somatic coliphages Annex A forms an integral part of this part of IS0 10705. Annexes B and C are for information only. 0 IS0 1995 All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying and microfilm, without permission in writing from the publisher. International Organization for Standardization Case Postale 56 l CH-1211 Geneve 20 l Switzerland Printed in Switzerland ii SIST EN ISO 10705-1:2001
INTERNATIONAL STANDARD 0 IS0 IS0 10705-1:1995(E) Water quality - Detection and enumeration of bacteriophages - Part 1: Enumeration of F-specific RNA bacteriophages 1 Scope This part of IS0 10705 specifies a method for the detection and enumeration of F-specific ribonucleic acid (RNA) bacteriophages by incubating the sample with an appropriate host strain. The method can be applied to all kinds of water, sediments and sludges, where necessary after dilution. In the case of low numbers, a preconcentration step may be necessary for which a separate part of IS0 10705 will be devel- oped. The method can also be applied to shellfish extracts. Depending on the relative abundance of F- specific RNA bacteriophages to background organisms, additional confirmatory tests may be necessary and are also specified in this part of IS0 10705. The presence of F-specific RNA bacteriophages in a water sample generally indicates pollution by wastewater contaminated by human or animal faeces. Their survival in the environment, removal by widely used water treatment processes and concentration or retention by shellfish resembles that of foodborne and waterborne human enteric viruses, for example the enteroviruses, hepatitis A virus and rotaviruses. 2 Normative references The following standards contain provisions which, through reference in this text, constitute provisions of this part of IS0 10705. At the time of publication, the editions indicated were valid. All standards are subject to revision, and parties to agreements based on this part of IS0 10705 are encouraged to investi- gate the possibility of applying the most recent edi- tions of the standards indicated below. Members of IEC and IS0 maintain registers of currently valid International Standards. IS0 3696: 1987, Water for analytical laboratory use - Specification and test methods. IS0 5667-l : 1980, Water quality - Sampling - Part I: Guidance on the design of sampling pro- grammes. IS0 5667-2: 1991, Water quality - Sampling - Part 2: Guidance on sampling techniques. IS0 5667-3: 1994, Water quality - Sampling - Part 3: Guidance on the preservation and handling of samples. IS0 6887: 1983, Microbiology - General guidance for the preparation of dilutions for microbiological exam- ina tion. I SO 8199: 1988, Water quality - General guide to the enumeration of micro-organisms by culture. 3 Definition For the purposes of this part of IS0 10705, the fol- lowing definition applies. 3.1 F-specific RNA bacteriophages: Bacterial vi- ruses which are capable of infecting a specified host strain with F-pili or sex-pili to produce visible plaques (clearance zones) on a confluent lawn grown under appropriate culture conditions, whereas the infectious process is inhibited in the presence of a concentration 1 SIST EN ISO 10705-1:2001
IS0 10705-1:1995(E) . 0 IS0 of 40 (occasionally 400) Clglml of RNase in the plating medium. 6.3 Reagents 6.3.1 RNase from bovine pancreas, specific ac- tivity approximately 50 units/mg (Kunitz). 4 Principle The sample is mixed with a small volume of semi- solid nutrient medium. A culture of host strain is added and plated on a solid nutrient medium. After this, incubation and reading of plates for visible plaques takes place. Where necessary, simultaneous examination of parallel plates with added RNase for confirmation by differential counts is carried out. The results are expressed as the number concentration of plaque-forming particles (Cpfp) per unit of volume. 5 Safety precautions The host strain used is a Salntonella fyphimurium mutant of low pathogenicity and should be hand- led in accordance with the appropriate national or international safety procedures for this bac- terial species. F-specific RNA bacteriophages are non-pathogenic for man and animals, but are very resistant to drying. Appropriate precautions should therefore be taken to prevent cross- contamination of test materials, particularly when examining or handling cultures of high titre or when inoculating cultures of the host strains. Such procedures must be carried out in a biohazard cabinet or a separate area of the lab- oratory. 6 Diluent, culture media and reagents 6.1 Basic materials Use ingredients of uniform quality and chemicals of analytical grade for the preparation of culture media and reagents, and follow the instructions given in an- nex A. For information on storage see IS0 8199, ex- cept where indicated in this part of IS0 10705. Alternatively, use dehydrated complete media and follow strictly the manufacturer’s instructions. For the preparation of media, use glass-distilled water or deionized water free from substances which might inhibit bacterial growth under the conditions of the test, and in accordance with IS0 3696. 6.2 Diluent For making sample dilutions, use peptone saline sol- ution as indicated in A.8. 6.3.2 Antibiotic discs, for susceptibility testing with nalidixic acid (130 pg; 9 mm) and kanamycin (100 pg; 9 mm). 6.3.3 Glycerol, 870 g/litre. 6.4 Microbiological reference cultures Salmonella typhimurium strain WG49, phage type 3 Nal’ (F’ 42 /ac::Tn5), NCTC 12484. Bacteriophage MS2, NCTC 12487 or ATCC 15597-Bl . Escherichia co/; K-12 Hfr from appropriate culture col- lection, e.g. NCTC 12486 or ATCC 23631. NOTE 1 The NCTC strains are available from the National Collection of Type Cultures, 61 Colindale Avenue, London NW9 6HT, England. The ATCC strains are available from the American Type Culture Collection, 12301 Parklawn Drive, Rockville, Maryland, U.S.A. 7 Apparatus and glassware Usual microbiological laboratory equipment, including 7.1 Hot-air oven for dry-heat sterilization and an autoclave. Apart from apparatus supplied sterile, glassware and other equipment shall be sterilized ac- cording to the instructions given in IS0 8199. 7.2 Incubator or water bath, thermostatically controlled at 37 “C + 1 “C. - 7.3 Incubator or water bath, thermostatically controlled at 37 “C +_ 1 “C and equipped with a rotary platform at 100 min-’ + 10 min-‘. - 7.4 Water bath, thermostatically controlled at 45 “C + 1 “C. - 7.5 Water bath or equivalent device, for melting agar media. 7.6 pH-meter. 7.7 Counting apparatus, with indirect, oblique light. 7.8 Deep freezer, thermostatically controlled at - 20 “C + 5 “C. - 2 SIST EN ISO 10705-1:2001
0 IS0 7.9 Deep freezer, thermostatically controlled at - 7o”c+100c. - 7.10 Spectrometer, capable of holding 1 cm cuvettes or side-arm of nephelometric flasks (7.17) and equipped with a filter in the range 500 nm to 650 nm with a maximum bandwidth of + 10 nm. Usual sterile, microbiological laboratory glassware or disposable plastics ware according to IS0 8199 and including the following. 7.11 Petri dishes, of diameter 9 cm or 15 cm. 7.12 Graduated pipettes, of capacities 1 ml, 5 ml and 10 ml. 7.13 Glass bottles, of suitable volumes. I 7.14 Culture tubes, with cap: 10705=1:1995(E) 7.15 Measuring cylinders, of suitable capacity. 7.16 Conical flasks, of capacity 250 ml to 300 ml, with cotton wool plugs or suitable alternatives. 7.17 Cuvettes, of optical path length 1 cm or nephelometric conical flasks, of capacity 250 ml to 300 ml, with cylindrical side-arms which can be fitted to the spectrometer (7.10) and with cotton wool plugs or suitable alternatives. (See figure 1.) 7.18 Membrane filter units, for sterilization, pore size 0,2 pm. 7.19 Plastics vials, lidded, of capacity I,5 ml to 2 ml. Figure 1 - Nephelometric conical flasks for culturing the host strain SIST EN ISO 10705-1:2001
IS0 10705-1:1995(E) 8 Sampling Take samples and deliver them to the laboratory in accordance with IS0 8199, IS0 5667-1, IS0 5667-2, and IS0 5667-3. 9 Preparation of test materials 9.1 Culturing and maintenance of host strains WG49 and E. co/i K12 Hfr The culturing and maintenance of host strains in- volves several stages which are summarized in figure2. The figure also indica
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