EN ISO 5667-16:1998

Water quality - Sampling - Part 16: Guidance on biotesting of samples (ISO 5667-16:1998)

Biotesting requires special sampling and pretreatment provisions. It is intended to elaborate a guideline for these.

Wasserbeschaffenheit - Probenahme - Teil 16: Anleitung zur Probenahme und Durchführung biologischer Testverfahren (ISO 5667-16:1998)

Qualité de l'eau - Echantillonnage - Partie 16: Lignes directrices pour les essais biologiques des échantillons (ISO 5667-16:1998)

Kakovost vode – Vzorčenje – 16. del: Navodilo za biološke preskuse vzorcev (ISO 5667-16:1998)

General Information

• Status: Withdrawn
• Publication Date: 30-Sep-1998
• Withdrawal Date: 09-May-2017
• ICS: 13.060.10 - Water of natural resources
• Technical Committee: CEN/TC 230 - Water analysis
• Drafting Committee: CEN/TC 230 - Water analysis
• Current Stage: 9960 - Withdrawal effective - Withdrawal
• Completion Date: 10-May-2017
• Ref Project: SIST EN ISO 5667-16:2000 - Water quality - Sampling - Part 16: Guidance on biotesting of samples (ISO 5667-16:1998)

Relations

• Replaced By: EN ISO 5667-16:2017 - Water quality - Sampling - Part 16: Guidance on biotesting of samples (ISO 5667-16:2017)
• Effective Date: 17-May-2017

Standards Content (Sample)

SLOVENSKI STANDARD
01-januar-2000
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Water quality - Sampling - Part 16: Guidance on biotesting of samples (ISO 5667-
16:1998)
Wasserbeschaffenheit - Probenahme - Teil 16: Anleitung zur Probenahme und
Durchführung biologischer Testverfahren (ISO 5667-16:1998)
Qualité de l'eau - Echantillonnage - Partie 16: Lignes directrices pour les essais
biologiques des échantillons (ISO 5667-16:1998)
Ta slovenski standard je istoveten z: EN ISO 5667-16:1998
ICS:
13.060.70 Preiskava bioloških lastnosti Examination of biological
vode properties of water
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

INTERNATIONAL ISO
STANDARD 5667-16
First edition
1998-10-01
Water quality — Sampling —
Part 16:
Guidance on biotesting of samples
Qualité de l’eau — Échantillonnage
Partie 16: Lignes directrices pour les essais biologiques des échantillons
A
Reference number
ISO 5667-16:1998(E)
ISO 5667-16:1998(E)
Contents Page
1 Scope . 1
2 Normative references. 1
3 Sampling. 1
4 Transport. 2
5 Preservation and storage. 3
6 Apparatus and equipment . 3
7 Pretreatment and preparation of samples . 4
8 Treatment of samples during the test . 10
9 General guidance regarding test design. 11
10 Special guidance regarding test performance . 13
11 Special biological assays . 16
12 Evaluation. 20
13 Presentation of results. 23
14 Test report. 24
15 Basic principles of quality assurance for biotesting . 25
Annex A Lowest Ineffective Dilution (LID) . 28
Annex B Bibliography . 29
©  ISO 1998
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 • CH-1211 Genève 20 • Switzerland
Internet [e-mail]
Printed in Switzerland
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ISO ISO 5667-16:1998(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.
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 ISO 5667-16 was prepared by Technical Committee
ISO/TC 147, Water quality, Subcommittee SC 6, Sampling.
ISO 5667 consists of the following parts, under the general title Water
quality — Sampling:
 Part 1: Guidance on the design of sampling programmes
 Part 2: Guidance on sampling techniques
 Part 3: Guidance on the preservation and handling of samples
 Part 4: Guidance on sampling from lakes, natural and man-made
 Part 5: Guidance on sampling of drinking water and water used for
food and beverage processing
 Part 6: Guidance on sampling of rivers and streams
 Part 7: Guidance on sampling of water and steam in boiler plants
 Part 8: Guidance on the sampling of wet deposition
 Part 9: Guidance on sampling from marine waters
 Part 10: Guidance on sampling of waste waters
 Part 11: Guidance on sampling of groundwaters
 Part 12: Guidance on sampling of bottom sediments
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ISO 5667-16:1998(E) ISO
 Part 13: Guidance on sampling of water, wastewater and related
sludges
 Part 14: Guidance on quality assurance of environmental water
sampling and handling
 Part 15: Guidance on preservation and handling of sludge and
sediment samples
 Part 16: Guidance on biotesting of samples.
Annexes A and B of this part of ISO 5667 are for information only.
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ISO ISO 5667-16:1998(E)
Introduction
Biological tests are suitable for determining the effect of chemical and
physical parameters on test organisms under specific experimental
conditions. In principle, the methods of chemical analysis are not suitable
for determining the biological effects. These effects can be enhancing or
inhibiting, and can be determined by the reaction of the organisms, e.g.
death, growth, proliferation, morphological, physiological and histological
changes. Inhibiting effects are triggered by toxic water constituents or by
other noxious influences.
Effects can refer to various levels, e.g. proceeding from (sub)cellular
structures or enzyme systems, concerning the whole organism, and
eventually the supra-organism or community level.
In the context of this part of ISO 5667, toxicity is the ability of a substance
to exert a deleterious effect on organisms or biocenoses due to its
chemical properties and its concentration.
The deleterious potential of a toxic substance can be counteracted by the
protective potential of the biological system, for instance by metabolic
detoxification and excretion. The apparent toxicity measurable in the
biological test is the result of the interaction between the substance and the
biological system.
Apart from the direct toxic effect of one or more water constituents,
damaging biological effects can be exerted by the combined action of all
noxious substances, e.g. by substances which are not toxic per se but
affect the chemical or physical properties of the medium and,
consequently, the living conditions for the organisms. This applies for
instance to oxygen-depleting substances, coloured substances or turbid
matter which reduce light exposure. It also includes non-substance-related
effects such as impairment or damage due to extreme temperature.
Biological tests also include those tests which examine the effect of
organisms on substances, e.g. microbial degradation studies.
The results of the biological tests refer primarily to the organisms used in
the test and the conditions stipulated in the test procedure. A harmful effect
stated by means of standardized tests can justify concern that aquatic
organisms and biocenoses might be endangered. The results, however, do
not permit direct or extrapolative conclusions as to the occurrence of
similar effects in the aquatic environment. This applies in particular to sub-
organism systems, as important properties and physiological functions of
intact organisms (e.g. protective integuments, repair mechanisms) are
removed or deactivated.
In principle there is no organism and no biocenosis which can be used to
test all the effects on the ecosystem possible under the various
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ISO 5667-16:1998(E) ISO
constellations of abiotic and biotic conditions. Only a few ("model") species
representing relevant ecological functions can be tested in practice.
Besides these fundamental and practical limitations in the selection of test
organisms, the sample to be tested can also pose experimental problems
on biotesting. Waters, in particular waste waters, are complex mixtures and
often contain sparingly soluble, volatile, unstable, coloured substances
and/or suspended, sometimes colloidal, particles. The complexity and
heterogeneity of materials give rise to a variety of experimental problems
when performing biotests.
Special problems are related to the instability of the test material due to
reactions and processes such as:
 physical (e.g. phase separation, sedimentation, volatilization);
 chemical (e.g. hydrolysis, photodegradation, precipitation); and/or
 biological (e.g. biodegradation, biotransformation, biological uptake in
organisms).
Other problems, especially if spectrometric measurements are applied,
relate to turbidity and colour.
This part of ISO 5667 is one of a group of International Standards dealing
with the sampling of waters. It should be read in conjunction with the other
parts and in particular with ISO 5667-1, ISO 5667-2 and ISO 5667-3.
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INTERNATIONAL STANDARD  © ISO ISO 5667-16:1998(E)
Water quality — Sampling —
Part 16:
Guidance on biotesting of samples
1 Scope
This part of ISO 5667 gives practical guidance on sampling, pretreatment, performance and evaluation of waters in
the context of biotesting. Information is given on how to cope with the problems for biotesting arising from the nature
of the water sample and the suitability of the test design.
It is intended to convey practical experience concerning precautions to be taken by describing methods successfully
proven to solve or to circumvent some of the experimental problems of biotesting of waters.
Reference has been made as far as possible to existing International Standards and guidelines. Information taken
from published papers or oral communication is utilized as well.
Primarily dealt with are substance-related problems concerning sampling, pretreatment and preparation of water
samples for biotesting and treatment of samples during the test, especially when performing tests with waters and
waste waters containing unstable or removable ingredients. Basic principles of quality assurance, evaluation of data
and presentation of results are outlined.
Special emphasis is laid on ecotoxicological testing with organisms ('single-species biotests'). Some features
addressed in this general guidance apply as well to biodegradation and/or bioaccumulation studies as far as
sampling and sample preparations is concerned. Preparation of poorly soluble substances and testing beyond the
water-solubility limit is also addressed.
This part of ISO 5667 is not applicable to bacteriological examination of water. Appropriate methods are described
in other International Standards.
2 Normative references
The following standards contain provisions which, through reference in this text, constitute provisions of this part of
ISO 5667. 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 ISO 5667 are encouraged to investigate the possibility applying the
most recent editions of the standards indicated below. Members of IEC and ISO maintain registers of currently valid
International Standards.
ISO 5667-3 :1994, Water quality — Sampling — Part 3: Guidance on the preservation and handling of samples.
ISO 5667-10 :1992, Water quality — Sampling — Part 10: Guidance on sampling of waste waters.
3 Sampling
3.1 General
The choice of representative sampling points, frequency of sampling, type of samples taken, etc. is dependent on
the objective of the study. In general, the sampling approach for chemical analysis is compatible with the purpose of
biotesting.
© ISO
ISO 5667-16:1998(E)
Some tests, however, require the water and waste water to be handled and kept in a particular way.
Depending on the type of investigation (e.g. toxicity or biodegradation tests) and the way the samples are to be
processed, it is necessary to divide a sample into different portions which are preserved and/or stored under
different conditions and processed in different ways.
If several samples have been taken (e.g. from different locations or at several times) they may be combined to
achieve greater representativity. These samples should be thoroughly mixed and, if necessary, divided into
subsamples. To obtain subsamples of equal quality, it should be ensured that the bulk sample maintains
homogeneity during the subsampling process, e.g. by continuous shaking or stirring. This holds particularly in the
case of two-phase mixtures, e.g. waters containing suspended particles, algal suspensions. It is recommended to
use cooling sampling apparatus when several samples taken at several times are combined.
3.2 Samplers/vessels/containers
The volume, shape and material of the vessels are dependent on the nature of the sample (e.g.
degradability/stability), the number of replicates, the volume required for these tests and the necessity of preserving
and storing the samples prior to further processing.
The time required for freezing and thawing should be minimized by reducing the sample volume, i.e. the size of the
vessel. In general it is appropriate to use one-litre vessels for freezing. For tests requiring larger volumes, the
sample should be divided into vessels holding not more than 10 l.
The total sample volume taken should be sufficient to cover any supplementary or repeated testing. Remaining
subsamples stored frozen separately should be saved until the final evaluation has been made.
The material of vessels should be chemically inert, easily cleaned and resistant to heating and freezing. Glassware,
polyethene or polytetrafluoroethene (PTFE) vessels are recommended.
3.3 Filling status of containers
It should be decided whether the containers should be fi
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