oSIST prEN ISO 10705-3:2024

Water quality - Detection and enumeration of bacteriophages - Part 3: Validation of methods for concentration of bacteriophages from water (ISO 10705-3:2003)

This part of ISO 10705 specifies the general principles for assessing the performance of methods for the concentration of bacteriophages from water. Concentration is recommended for those water samples expected to contain < 3 pfp (plaque-forming particles) per millilitre. Concentration methods can be applied to all kinds of water provided that the amount and nature of suspended solids and/or dissolved matter do not interfere with the concentration procedure.
This part of ISO 10705 does not give specific details of concentration methods, but outlines the fundamental
principles for evaluating the suitability of a particular method for a given type and volume of water. Annex A
gives examples of methods that have been found satisfactory and their fields of application.

Wasserbeschaffenheit - Nachweis und Zählung von Bakteriophagen - Teil 3: Validierung von Verfahren für die Konzentration von Bakteriophagen in Wasser (ISO 10705-3:2003)

Dieser Teil von ISO 10705 legt die allgemeinen Grundsätze für die Bewertung der Leistungsfähigkeit von Verfahren für die Aufkonzentrierung von Bakteriophagen aus Wasser fest. Eine Aufkonzentrierung wird für Wasserproben empfohlen, die voraussichtlich < 3 pfp (Plaque formende Partikel) je Milliliter enthalten. Aufkonzentrierungsverfahren können bei allen Arten von Wasser angewendet werden, sofern die Menge und Beschaffenheit der suspendierten und/oder gelösten Stoffe das Aufkonzentrierungsverfahren nicht beeinträchtigen.
Dieser Teil von ISO 10705 enthält keine detaillierten Angaben zu Aufkonzentrierungsverfahren, sondern umreißt die grundlegenden Prinzipien zur Bewertung der Eignung eines bestimmten Verfahrens für eine bestimmte Art und Menge von Wasser. Anhang A enthält Beispiele für Verfahren, die sich als zufriedenstellend erwiesen haben, und deren Anwendungsbereiche.
WARNHINWEIS — Anwender dieses Teils von ISO 10705 sollten mit der üblichen Laborpraxis vertraut sein. In diesem Teil von ISO 10705 wird nicht der Anspruch erhoben, alle bei seiner Anwendung möglicherweise auftretenden Sicherheitsprobleme zu behandeln. Es liegt in der Verantwortung des Arbeitsgebers, geeignete Sicherheits- und Gesundheitspraktiken vorzusehen und die Einhaltung etwaiger nationaler Bestimmungen sicherzustellen.
WICHTIG — Das mit der Validierung von Verfahren für die Aufkonzentrierung von Bakteriophagen aus Wasser befasste Laborpersonal muss unbedingt über einschlägige Erfahrungen mit den Verfahren zur Zählung von Bakteriophagen verfügen (siehe ISO/TR 13843 [1]).

Qualité de l'eau - Détection et dénombrement des bactériophages - Partie 3: Validation des méthodes de concentration des bactériophages dans l'eau (ISO 10705-3:2003)

L'ISO 10705-3:2003 spécifie les principes généraux qui permettent d'évaluer les performances des méthodes de concentration des bactériophages dans l'eau. La concentration est recommandée pour les échantillons d'eau supposés contenir moins de 3 pfp (particules formant plage) par millilitre. Les méthodes de concentration peuvent être appliquées à tous les types d'eau, sous réserve que la quantité et la nature des matières en suspension et/ou des matières dissoutes n'interfèrent pas avec le mode opératoire de concentration.
L'ISO 10705-3:2003 ne donne pas de détails spécifiques quant aux méthodes de concentration, mais esquisse les principes fondamentaux pour l'évaluation de l'adéquation d'une méthode particulière pour un type et un volume d'eau donnés. L'Annexe A donne des exemples de méthodes qui ont été jugées satisfaisantes ainsi que leur domaine d'application.

Kakovost vode - Ugotavljanje prisotnosti in števila bakteriofagov - 3. del: Validacija metod za koncentracijo bakteriofagov v vodi (ISO 10705-3:2003)

General Information

• Status: Not Published
• Public Enquiry End Date: 02-Jun-2024
• ICS: 07.100.20 - Microbiology of water
• Technical Committee: KAV - Water quality
• Current Stage: 4020 - Public enquire (PE) (Adopted Project)
• Start Date: 19-Mar-2024
• Due Date: 06-Aug-2024
• Completion Date: 21-Jun-2024
• Ref Project: EN ISO 10705-3:2024 - Water quality - Detection and enumeration of bacteriophages - Part 3: Validation of methods for concentration of bacteriophages from water (ISO 10705-3:2003)

Standards Content (Sample)

SLOVENSKI STANDARD
01-maj-2024
Kakovost vode - Ugotavljanje prisotnosti in števila bakteriofagov - 3. del:
Validacija metod za koncentracijo bakteriofagov v vodi (ISO 10705-3:2003)
Water quality - Detection and enumeration of bacteriophages - Part 3: Validation of
methods for concentration of bacteriophages from water (ISO 10705-3:2003)
Wasserbeschaffenheit - Nachweis und Zählung von Bakteriophagen - Teil 3: Validierung
von Verfahren für die Konzentration von Bakteriophagen in Wasser (ISO 10705-3:2003)
Qualité de l'eau - Détection et dénombrement des bactériophages - Partie 3: Validation
des méthodes de concentration des bactériophages dans l'eau (ISO 10705-3:2003)
Ta slovenski standard je istoveten z: prEN ISO 10705-3
ICS:
07.100.20 Mikrobiologija vode Microbiology of water
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

INTERNATIONAL ISO
STANDARD 10705-3
First edition
2003-10-01
Water quality — Detection and
enumeration of bacteriophages —
Part 3:
Validation of methods for concentration
of bacteriophages from water
Qualité de l'eau — Détection et dénombrement des bactériophages —
Partie 3: Validation des méthodes de concentration des bactériophages
dans l'eau
Reference number
ISO 10705-3:2003(E)
©
ISO 2003
ISO 10705-3:2003(E)
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ii © ISO 2003 — All rights reserved

ISO 10705-3:2003(E)
Contents Page
Foreword. iv
1 Scope. 1
2 Normative references. 1
3 Terms and definitions. 2
4 Principle. 2
5 Reagents. 2
6 Apparatus and glassware. 3
7 Sampling. 3
8 Preparation of sewage samples for spiking. 3
9 Procedure. 4
10 Calculation. 5
11 Analytical quality control . 6
12 Test report. 7
Annex A (informative) Recommended methods for concentration of bacteriophages from water
depending on the volume, turbidity and particle content. 8
Annex B (informative) Example of a validation process of a concentration method. 10
Bibliography . 13

ISO 10705-3:2003(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 10705-3 was prepared by Technical Committee ISO/TC 147, Water quality, Subcommittee SC 4,
Microbiological methods.
ISO 10705 consists of the following parts, under the general title Water quality — Detection and enumeration
of bacteriophages:
 Part 1: Enumeration of F-specific RNA bacteriophages
 Part 2: Enumeration of somatic coliphages
 Part 3: Validation of methods for concentration of bacteriophages from water
 Part 4: Enumeration of bacteriophages infecting Bacteroides fragilis

iv © ISO 2003 — All rights reserved

INTERNATIONAL STANDARD ISO 10705-3:2003(E)

Water quality — Detection and enumeration of
bacteriophages —
Part 3:
Validation of methods for concentration of bacteriophages from
water
WARNING — Persons using this part of ISO 10705 should be familiar with normal laboratory practice.
This part of ISO 10705 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 imperative that personnel involved in validation of methods for concentration of
bacteriophages from water have relevant experience with the methods of enumeration of
[1]
bacteriophages (see ISO/TR 13843 ).
1 Scope
This part of ISO 10705 specifies the general principles for assessing the performance of methods for the
concentration of bacteriophages from water. Concentration is recommended for those water samples
expected to contain < 3 pfp (plaque-forming particles) per millilitre. Concentration methods can be applied to
all kinds of water provided that the amount and nature of suspended solids and/or dissolved matter do not
interfere with the concentration procedure.
This part of ISO 10705 does not give specific details of concentration methods, but outlines the fundamental
principles for evaluating the suitability of a particular method for a given type and volume of water. Annex A
gives examples of methods that have been found satisfactory and their fields of application.
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 6887-1, Microbiology of food and animal feeding stuffs — Preparation of test samples, initial suspension
and decimal dilutions for microbiological examination — Part 1: General rules for the preparation of the initial
suspension and decimal dilutions
ISO 8199, Water quality — General guide to the enumeration of micro-organisms by culture
ISO 10705-1, Water quality — Detection and enumeration of bacteriophages — Part 1: Enumeration of
F-specific RNA bacteriophages
ISO 10705-2, Water quality — Detection and enumeration of bacteriophages — Part 2: Enumeration of
somatic coliphages
ISO 10705-3:2003(E)
ISO 10705-4, Water quality — Detection and enumeration of bacteriophages — Part 4: Enumeration of
bacteriophages infecting Bacteroides fragilis
ISO/IEC Guide 2, Standardization and related activities — General vocabulary
3 Terms and definitions
For the purposes of this document, the terms and definitions given in ISO/IEC Guide 2 and the following
apply:
3.1
bacteriophages
bacterial viruses which are capable of infecting selected host strains
NOTE Bacteriophages produce visible plaques (clearance zones) in a confluent lawn of the host strain grown under
appropriate culture conditions.
4 Principle
The sample is treated according to a method of choice, by which the bacteriophages are concentrated from a
relatively large volume of sample (100 ml up to several litres) to a smaller volume (typically from a few to
20 ml). The concentrated sample is then analysed for bacteriophages according to an International Standard
method or other suitable protocol.
The concentration method to be evaluated should be carefully described in a protocol, following ISO standard
layout as much as possible. The description should include the target group(s) of bacteriophages and their
detection method(s), the types of water and ranges of volumes to be analysed, as well as exceptions to the
field of application, e.g. turbidity.
The method is validated according to principles laid down in this part of ISO 10705. The validation procedure
consists of determining the recovery of bacteriophages from a series of samples, seeded with naturally
polluted water (raw or treated sewage). The recovery is studied in a range of volumes, and particular attention
is paid to its reproducibility.
5 Reagents
Use ingredients of uniform quality and chemicals of analytical grade for the preparation of culture media. For
information on storage see ISO 8199, except where indicated in this part of ISO 10705. Alternatively, use
dehydrated complete media and follow strictly the manufacturer's instructions.
Other grades of chemicals may be used provided they can be shown to lead to the same results.
5.1 Water, for the preparation of media, glass-distilled water or de-ionized water free from substances that
might inhibit bacterial growth under the conditions of the test, and at least Grade 3 as specified in ISO 3696.
5.2 Diluent, for making dilutions, peptone-saline solution or another suitable diluent in accordance with
ISO 6887-1 or ISO 8199.
5.3 Culture media and reference cultures, as specified in the corresponding standard method of
ISO 10705-1, ISO 10705-2 and ISO 10705-4 for the phage assay.
5.4 Glycerol (ρ = 870 g/l), autoclaved at (121 ± 3) ºC for 15 min and stored in the dark at room temperature
for a period no longer than 1 year.
2 © ISO 2003 — All rights reserved

ISO 10705-3:2003(E)
6 Apparatus and glassware
SAFETY PRECAUTIONS — Field apparatus should be disinfected before use. Apply safety
precautions appropriate to the disinfectant solution used. Some stages of the concentration process
may involve the application of hydrostatic or pneumatic pressure. Observe relevant safety
precautions.
Use usual microbiological laboratory equipment as specified in the method for the phage assay (Clause 8),
and the protocol for the concentration method.
7 Sampling
Samples up to 10 l can conveniently be transported to the laboratory. Take the samples and deliver them to
[2]
the laboratory as specified in ISO 8199 (see also ISO 19458 ). For larger samples, it is advisable to perform
the first step of the concentration procedure in the field. This process may take up to several hours. If parallel
examination for indicator bacteria or other micro-organisms is carried out, take a time-proportional sample for
these analyses, preferably by filling a sample bottle with a side flow from the concentration apparatus. Filters,
precipitates or other products from the first concentration step may be further treated in the field, or may be
transported to the laboratory. Include the transport and storage conditions of intermediate stages of the
process in the validation procedure.
8 Preparation of sewage samples for spiking
Obtain a sample of primary or secondary (biologically treated) sewage and centrifuge at 1 000 g for 20 min or
filter through an 8 µm to 12 µm membrane filter. Store supernatant or filtrate on melting ice. Enumerate the
target bacteriophages in 1 ml volu
...