Soil quality - Guidance on the choice and evaluation of bioassays for ecotoxicological characterization of soils and soil materials (ISO 17616:2019)

This document is one of the family of standards (ISO 15799, ISO 19204) providing guidance on the characterization of soils and soil materials in relation to their retention and habitat functions and uses. It is appropriate to use it in conjunction with the two other standards in this family. It provides guidance on the choice and evaluation of tests applied for ecotoxicological characterization of soils and soil materials. Recommendations for test strategies with respect to the protection of ground and surface waters and the maintenance of the habitat function of soil are included. The tests recommended represent a minimum test battery that can be complemented by additional tests, or even be replaced by others, according to the intended uses or protection goals envisaged. The effect values indicated in this document do not refer to regulation but represent the lowest level at which an adverse effect is considered likely to occur.

Bodenbeschaffenheit - Anleitung für die Auswahl und Beurteilung von Biotestverfahren zur ökotoxikologischen Charakterisierung von Böden und Bodenmaterialien (ISO 17616:2019)

Dieses Dokument ist eine aus einer Normenfamilie (ISO 15799, ISO 19204) zur Verfügung stehende Anleitung zur Charakterisierung von Böden und Bodenmaterialien in Bezug auf ihre Rückhalte- und Lebensraumfunktion und Verwertung. Es ist für die Anwendung in Verbindung mit den anderen beiden Normen dieser Familie geeignet. Es gibt eine Anleitung für die Auswahl und Beurteilung von Prüfungen zur ökotoxikologischen Charakterisierung von Böden und Bodenmaterialien. Eingeschlossen sind Empfehlungen für Prüfstrategien hinsichtlich des Schutzes von Grundwasser und Oberflächenwasser und der Erhaltung der Lebensraumfunktion von Böden. Die empfohlenen Prüfungen stellen eine Mindesttestbatterie dar, die abhängig von der vorgesehenen Nutzung oder den angestrebten Schutzzielen durch zusätzliche Prüfverfahren ergänzt werden kann. Die in diesem Dokument angezeigten Wirkungsschwellen beziehen sich nicht auf gesetzliche Vorgaben, sondern stellen den niedrigsten Gehalt dar, bei dem wahrscheinlich ein schädlicher Effekt eintritt.

Qualité du sol - Lignes directrices relatives aux choix et à l'évaluation des essais appliqués pour la caractérisation écotoxicologique des sols et des matériaux de type sol (ISO 17616:2019)

Le présent document fait partie de la famille de normes (ISO 15799, ISO 19204) fournissant des recommandations relatives à la caractérisation des sols et des matériaux de type sol par rapport à leurs fonctions d'habitat et de rétention ainsi qu'à leurs utilisations. Il est approprié de l'utiliser conjointement avec les deux autres normes de cette famille. Il fournit des recommandations relatives au choix et à l'évaluation des essais appliqués pour la caractérisation écotoxicologique des sols et des matériaux de type sol. Des recommandations relatives à des stratégies d'essai pour la protection des eaux superficielles et souterraines et le maintien de la fonction d'habitat du sol sont incluses. Les essais recommandés représentent une batterie minimale d'essais pouvant être complétée par des essais supplémentaires ou même être remplacée par d'autres essais en fonction des utilisations prévues ou des objectifs de protection envisagés. Les valeurs d'effet indiquées dans le présent document ne se réfèrent pas à la réglementation, mais représentent le niveau le plus faible auquel une réponse est supposée entraîner un effet préjudiciable.

Kakovost tal - Navodilo za izbiro in vrednotenje bioloških preskusov za ekotoksikološko karakterizacijo tal in talnih materialov (ISO 17616:2019)

Ta dokument je iz družine standardov (ISO 15799, ISO 19204), ki zagotavljajo navodilo za karakterizacijo tal in talnih materialov v zvezi z njihovimi zadrževalnimi in habitatnimi funkcijami ter uporabo. Uporabljati ga je treba v povezavi z drugima dvema standardoma iz te družine. Zagotavlja navodilo za izbiro in vrednotenje preskusov, ki se uporabljajo za ekotoksikološko karakterizacijo tal in talnih materialov. Vključena so priporočila za preskusne strategije v zvezi z varstvom podzemnih in površinskih voda ter ohranjanjem habitatne funkcije tal. Priporočeni preskusi predstavljajo minimalni sklop preskusov, ki ga je mogoče dopolniti z dodatnimi preskusi ali celo nadomestiti z drugimi glede na predvideno uporabo ali predvidene cilje zaščite. Vrednosti učinka, navedene v tem dokumentu, se ne nanašajo na predpise, temveč predstavljajo najnižjo raven, pri kateri se šteje, da se bo škodljiv učinek verjetno pojavil.

General Information

Status
Published
Public Enquiry End Date
02-Aug-2022
Publication Date
12-Feb-2023
Technical Committee
Current Stage
6060 - National Implementation/Publication (Adopted Project)
Start Date
12-Jan-2023
Due Date
19-Mar-2023
Completion Date
13-Feb-2023

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Standards Content (Sample)

SLOVENSKI STANDARD
SIST EN ISO 17616:2023
01-marec-2023
Kakovost tal - Navodilo za izbiro in vrednotenje bioloških preskusov za
ekotoksikološko karakterizacijo tal in talnih materialov (ISO 17616:2019)
Soil quality - Guidance on the choice and evaluation of bioassays for ecotoxicological
characterization of soils and soil materials (ISO 17616:2019)
Bodenbeschaffenheit - Anleitung für die Auswahl und Beurteilung von Biotestverfahren
zur ökotoxikologischen Charakterisierung von Böden und Bodenmaterialien (ISO
17616:2019)
Qualité du sol - Lignes directrices relatives aux choix et à l'évaluation des essais
appliqués pour la caractérisation écotoxicologique des sols et des matériaux de type sol
(ISO 17616:2019)
Ta slovenski standard je istoveten z: EN ISO 17616:2022
ICS:
13.080.05 Preiskava tal na splošno Examination of soils in
general
SIST EN ISO 17616:2023 en,fr,de
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

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SIST EN ISO 17616:2023

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SIST EN ISO 17616:2023


EN ISO 17616
EUROPEAN STANDARD

NORME EUROPÉENNE

October 2022
EUROPÄISCHE NORM
ICS 13.080.05
English Version

Soil quality - Guidance on the choice and evaluation of
bioassays for ecotoxicological characterization of soils and
soil materials (ISO 17616:2019)
Qualité du sol - Lignes directrices relatives aux choix et Bodenbeschaffenheit - Anleitung für die Auswahl und
à l'évaluation des essais appliqués pour la Beurteilung von Biotestverfahren zur
caractérisation écotoxicologique des sols et des ökotoxikologischen Charakterisierung von Böden und
matériaux de type sol (ISO 17616:2019) Bodenmaterialien (ISO 17616:2019)
This European Standard was approved by CEN on 9 October 2022.

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-CENELEC 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-CENELEC Management
Centre has the same status as the official versions.

CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia,
Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway,
Poland, Portugal, Republic of North Macedonia, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Türkiye and
United Kingdom.





EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION

EUROPÄISCHES KOMITEE FÜR NORMUNG

CEN-CENELEC Management Centre: Rue de la Science 23, B-1040 Brussels
© 2022 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 17616:2022 E
worldwide for CEN national Members.

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SIST EN ISO 17616:2023
EN ISO 17616:2022 (E)
Contents Page
European foreword . 3

2

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SIST EN ISO 17616:2023
EN ISO 17616:2022 (E)
European foreword
The text of ISO 17616:2019 has been prepared by Technical Committee ISO/TC 190 "Soil quality” of the
International Organization for Standardization (ISO) and has been taken over as EN ISO 17616:2022 by
Technical Committee CEN/TC 444 “Environmental characterization of solid matrices” the secretariat of
which is held by NEN.
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 2023, and conflicting national standards shall be
withdrawn at the latest by April 2023.
Attention is drawn to the possibility that some of the elements of this document may be the subject of
patent rights. CEN shall not be held responsible for identifying any or all such patent rights.
Any feedback and questions on this document should be directed to the users’ national standards body.
A complete listing of these bodies can be found on the CEN website.
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,
Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Iceland,
Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Republic of
North Macedonia, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Türkiye and the
United Kingdom.
Endorsement notice
The text of ISO 17616:2019 has been approved by CEN as EN ISO 17616:2022 without any modification.

3

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SIST EN ISO 17616:2023

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SIST EN ISO 17616:2023
INTERNATIONAL ISO
STANDARD 17616
Second edition
2019-10
Soil quality — Guidance on the choice
and evaluation of bioassays for
ecotoxicological characterization of
soils and soil materials
Qualité du sol — Lignes directrices relatives aux choix et
à l'évaluation des essais appliqués pour la caractérisation
écotoxicologique des sols et des matériaux de type sol
Reference number
ISO 17616:2019(E)
©
ISO 2019

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SIST EN ISO 17616:2023
ISO 17616:2019(E)

COPYRIGHT PROTECTED DOCUMENT
© ISO 2019
All rights reserved. Unless otherwise specified, or required in the context of its implementation, no part of this publication may
be reproduced or utilized otherwise in any form or by any means, electronic or mechanical, including photocopying, or posting
on the internet or an intranet, without prior written permission. Permission can be requested from either ISO at the address
below or ISO’s member body in the country of the requester.
ISO copyright office
CP 401 • Ch. de Blandonnet 8
CH-1214 Vernier, Geneva
Phone: +41 22 749 01 11
Fax: +41 22 749 09 47
Email: copyright@iso.org
Website: www.iso.org
Published in Switzerland
ii © ISO 2019 – All rights reserved

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SIST EN ISO 17616:2023
ISO 17616:2019(E)

Contents Page
Foreword .iv
Introduction .v
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 1
3.1 Assessment . 1
3.2 Types of soil and other soil materials . 2
3.3 Terms relating to Soil characteristics . 2
3.4 Land and sites . 3
4 Principles and applications of test batteries . 3
5 Testing strategy and interpretation of test results according to the use and re-use of
soils / soil materials and soil functions . 4
5.1 Monitoring of soil treatment success . 4
5.2 Assessment of the ecotoxic potential of soils / soil materials . 4
5.2.1 General. 4
5.2.2 Choice of test battery and evaluation of test results for assessing retention
function . 5
5.2.3 Choice of test battery and evaluation of test results for assessing habitat
function . 8
Annex A (informative) Tests with soil eluate — Expression of results .10
Bibliography .11
© ISO 2019 – All rights reserved iii

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SIST EN ISO 17616:2023
ISO 17616:2019(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.
The procedures used to develop this document and those intended for its further maintenance are
described in the ISO/IEC Directives, Part 1. In particular, the different approval criteria needed for the
different types of ISO documents should be noted. This document was drafted in accordance with the
editorial rules of the ISO/IEC Directives, Part 2 (see www .iso .org/ directives).
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. Details of
any patent rights identified during the development of the document will be in the Introduction and/or
on the ISO list of patent declarations received (see www .iso .org/ patents).
Any trade name used in this document is information given for the convenience of users and does not
constitute an endorsement.
For an explanation on the voluntary nature of standards, the meaning of ISO specific terms and
expressions related to conformity assessment, as well as information about ISO's adherence to the
World Trade Organization (WTO) principles in the Technical Barriers to Trade (TBT) see the following
URL: www .iso .org/ iso/ foreword .html.
This document was prepared by Technical Committee ISO/TC 190, Soil quality, Subcommittee SC 4,
Biological characterization.
This second edition cancels and replaces the first edition (ISO 17616:2008), which has been technically
revised. The main changes compared to the previous edition are as follows:
[10]
— definitions for “soil use” and “ecosystem service” have been added to Clause 3;
— end points of ecotoxicity tests (e.g. mortality, reproduction, growth, genotoxicity, and other
functional activities), as well as the overall principles and application of test batteries have been
clarified in Clause 4;
— sub-chronic toxicity tests have been added;
— Figure 1 was revised;
— Tables 1 and 2 (test batteries for retention and habitat function assessment, respectively) have been
revised (test categories, test organisms added/deleted, references updated).
Any feedback or questions on this document should be directed to the user’s national standards body. A
complete listing of these bodies can be found at www .iso .org/ members .html.
iv © ISO 2019 – All rights reserved

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SIST EN ISO 17616:2023
ISO 17616:2019(E)

Introduction
The characterization of contaminated soils can be based on strategies considering chemical analyses
and/or bioassays. ISO 15799 provides guidance on the selection of experimental methods for the
assessment of the ecotoxic potential of soils and soil materials (e.g. excavated and remediated soils,
refills, embankments) with respect to their intended use or re-use, and possible adverse effects on
aquatic and soil dwelling organisms.
An assessment strategy giving instructions for the choice and evaluation of test results is hence
proposed. The evaluation of the bioassays outcome is based on empirically-derived critical dilution
levels that take into account the sensitivity of the test system and the intended use/re-use of the site
under investigation. This approach intends to contribute to an effective and comparable assessment
[1]
within the ecotoxicological characterization of contaminated soil or soil materials . The test systems
included in this approach are not mandatory and may be replaced or accomplished by other test methods.
Nevertheless, the selected test systems have proved to appropriately characterize contaminated soils
[2],[3]
and soil materials with respect to their ecotoxic properties , both towards aquatic and terrestrial
organisms, the latter being responsible for maintaining essential soil functions.
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SIST EN ISO 17616:2023
INTERNATIONAL STANDARD ISO 17616:2019(E)
Soil quality — Guidance on the choice and evaluation of
bioassays for ecotoxicological characterization of soils and
soil materials
1 Scope
This document is one of the family of standards (ISO 15799, ISO 19204) providing guidance on the
characterization of soils and soil materials in relation to their retention and habitat functions and
uses. It is appropriate to use it in conjunction with the two other standards in this family. It provides
guidance on the choice and evaluation of tests applied for ecotoxicological characterization of soils
and soil materials. Recommendations for test strategies with respect to the protection of ground and
surface waters and the maintenance of the habitat function of soil are included. The tests recommended
represent a minimum test battery that can be complemented by additional tests, or even be replaced
by others, according to the intended uses or protection goals envisaged. The effect values indicated
in this document do not refer to regulation but represent the lowest level at which an adverse effect is
considered likely to occur.
2 Normative references
There are no normative references in this document.
3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
ISO and IEC maintain terminological databases for use in standardization at the following addresses:
— ISO Online browsing platform: available at https:// www .iso .org/ obp
— IEC Electropedia: available at http:// www .electropedia .org/
3.1 Assessment
3.1.1
soil-related assessment
assessment of the ecotoxic potential of soils (3.2.1), soil substrates and soil materials (3.2.2) based on
chemical analyses, biological tests and field inventories (monitoring) such as that mentioned in the
[4]
TRIAD approach
Note 1 to entry: TRIAD means an assessment approach based on a combination of chemical (i.e. residue analysis),
ecotoxicological (i.e. laboratory ecotoxic tests) and ecological (i.e. monitoring) data.
3.1.2
risk
expression of the probability that an adverse effect on soil (3.2.1) functions will occur under defined
conditions and the magnitude of the consequences of the effect occurring
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SIST EN ISO 17616:2023
ISO 17616:2019(E)

3.1.3
LID-value
lowest-ineffective-dilution value
lowest value of the dilution factor above which the test gives an ecotoxicological relevant reduction
(e.g. 20 % inhibition of luminescence)
EXAMPLE A LID 8 corresponds to a dilution of soil extract of 1 : 8 (i.e 1 part of soil extract: 7 parts of
dilution water).
Note 1 to entry: The exact definitions are given in the standard of the respective bioassay. According to
[5] [6]
ISO 13829 for the umu-test, it is the “DLi value” (explanation in Annex D). According to ISO 16240 3.4, it is the
“decisive D value”.
min
Note 2 to entry: For further information on results expression and interpretation based on LID values see
Annex A of this document.
3.2 Types of soil and other soil materials
3.2.1
soil
upper layer of the Earth’s crust composed of mineral particles, organic matter, water, air and organisms
[SOURCE: ISO 15799:2019, 3.1.1]
3.2.2
soil material
material coming from soil (3.2.1) and displaced and/or modified by human activity, including excavated
soil, dredged materials, manufactured soils, remediated treated soils or fill materials
[SOURCE: ISO 17402:2008, 3.16]
3.3 Terms relating to Soil characteristics
3.3.1
habitat function
ability of soils (3.2.1)/ soil materials (3.2.2) to serve as a habitat for microorganisms, plants, soil-living
animals and their interactions (i.e. biocenose)
[SOURCE: ISO 15799:2019, 3.2.1]
3.3.2
retention function
ability of soils (3.2.1)/ soil materials (3.2.2) to adsorb pollutants in such a way that they cannot be
mobilised via the water pathway and translocated into the food chain
Note 1 to entry: The habitat and retention functions include the following soil functions according to
[7]
ISO 11074:2015 , 3.3.31:
— control of substance and energy cycles as components of ecosystems;
— basis for the life of plants, animals and man;
— carrier of genetic reservoir;
— basis for the production of agricultural products;
— buffer inhibiting movement of water, contaminants or other agents into the ground water.
[SOURCE: ISO 15799:2019, 3.2.2]
2 © ISO 2019 – All rights reserved

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ISO 17616:2019(E)

3.3.3
contaminant
substance or agent present in the soil (3.2.1) as a result of human activity
Note 1 to entry: There is no assumption in this definition that harm results from the presence of the contaminant.
Note 2 to entry: See also pollutant (3.2.3) and potentially harmful substance (3.2.5) definitions in ISO 15799:2019
[9]
[SOURCE: ISO 15176:2002 , 3.2.6, modified — Note 2 to entry added.]
3.3.4
pollutant
substances, which due to their properties, amount or concentration cause negative impacts on the soil
(3.2.1) function or soil use
[9]
Note 1 to entry: Adapted from ISO 15176:2002 3.2.7.
3.4 Land and sites
3.4.1
re-use
useful and harmless utilisation of soil materials (3.2.2)
Note 1 to entry: In the context of this document re-use means the transfer of soil materials to another location for
use in agriculture, horticulture, forestry, gardens, recreational areas and construction sites.
[9]
[SOURCE: ISO 15176:2002 , 3.4.1; ISO 15799:2019, 3.3.1]
3.4.2
soil use
use of the ecosystem services (3.4.3) that soil (3.2.1) provides
3.4.3
ecosystem service
service that is (directly or indirectly) provided by an ecosystem to benefit people
[SOURCE: Based on Millennium Ecosystem Assessment]
4 Principles and applications of test batteries
The sensitivity of organisms (e.g. bacteria, plants, animals) to toxicants may vary significantly from one
species to another. Thereby, it is admitted that only the results of several ecotoxicity tests can give a
clear indication of the toxic effects of soil or soil materials. As such, the combination of ecotoxicity tests,
defined as a battery, shall include organisms belonging to various trophic levels, several biological
responses or end points (e.g. mortality, reproduction, growth, genotoxicity, as well as other functional
activities), in order to take into account the variability of species sensitivity within the studied
compartment.
The ecotoxicity tests included in batteries should at least have the following characteristics:
— sensitivity;
— practicability;
— compliance with standardized methods;
— high cost efficiency;
— representativeness of the soil ecosystem and/or of the selected application scenario (i.e. habitat or
retention functions).
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ISO 17616:2019(E)

Depending on the use of the soil or soil material (e.g. the agricultural use of waste) or the protection
goal (i.e. habitat or retention functions), the applied test battery can differ (see Clause 5 in ISO 15799
[1]
for guidance on test selection criteria) . Nevertheless, the selected tests should allow the identification
of the most sensitive trophic level(s) and give information on the toxic effects induced by solid samples.
The evaluation of results from the ecotoxicological tests should take into consideration the application
purposes (further details on the field of application in ISO 15799, Clause 4), which can broadly be for:
— monitoring and control of the success of soil treatment (off-site, on-site, in situ) (see 5.1),
— assessment of soil / soil material quality or contamination effects according to its use or re-use
(see 5.2).
Irrespective of the application purpose, the ecotoxicological characterization of soils and soil materials
[1]
depends on the soil use/re-use and soil functions requiring protection , as aforementioned. Overall, it
can essentially rely on the:
— assessment of mobile and bioavailable potentially harmful substances, in cases where the soil/
soil material (see 3.2.1 and 3.2.2) can affect the ground and/or surface water, as well as in cases
where pollutants are added to soils (e.g. agricultural use of wastes like sludge, composts, etc.) (i.e.
retention function), and/or
— assessment of the ecotoxic potential of soils and soil materials (see 3.2.1 and 3.2.2) and possible
adverse effects on soil-dwelling organisms (i.e. habitat function).
5 Testing strategy and interpretation of test results according to the use and re-
use of soils / soil materials and soil functions
5.1 Monitoring of soil treatment success
For evaluating the efficiency of soil treatments, the procedure below may be followed at different stages:
a) before the treatment – perform the ecotoxicological assessment of the soil sample(s) using a test
battery (5.2);
b) monitoring during the soil treatment process – perform a simple test selected from the battery
above [e.g. the most sensitive and practical test used among the battery applied in 5.1 a)];
c) at the end of the treatment – perform a new assessment with the same test battery used in 5.1 a), in
order to judge the success of the treatment.
5.2 Assessment of the ecotoxic potential of soils / soil materials
5.2.1 General
If soils or soil materials are assessed with respect to their intended use or re-use, the tests (see
ISO 15799) appropriate to evaluate their quality regarding the retention (see Table 1) and/or habitat
functions (see Table 2) should be applied. A strategy for the assessment of the ecotoxicological
characterization of soils and soil materials is proposed in Figure 1. The chemical characterization of
soils should always be conducted to increase the reliability of interpretations of the ecotoxicological
results obtained upon the selected test battery.
The test battery usually includes a set of acute and chronic/sub-chronic toxicity tests. If acute toxicity
is detected, it is not necessary to perform other tests. On the other hand, if no acute effect is detected,
chronic/sub-chronic toxicity and genotoxicity test(s) shall be conducted.
The assessment of soil and soil materials may be influenced by the collection, handling and storage
methods followed. Thereby, standardized procedures should be conducted in accordance with
[11]
ISO 18400-206 .
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ISO 17616:2019(E)

5.2.2 Choice of test battery and evaluation of test results for assessing retention function
5.2.2.1 Acute and chronic/sub-chronic assays for assessing the ecotoxic potential of soil/soil
materials
In a first approach, acute and sub-chronic tests using luminescent bacteria, algae and daphnids (see
Table 1) are recommended to assess the retention function of soils through the testing of soil/soil
material eluates. Depending on the legal requirements, other standardized bioassays may be selected
(see Table 1 or Annex A of ISO 15799 for choosing additional assays).
For the assessment of effects, toxicity criteria are also given in Table 1. The toxicity criteria are
provided as LID-values, percentages of inhibition or mortality. If these values are exceeded, then an
ecotoxicological potential is highlighted, thereby indicating that soil pollutants are soluble in water,
bio-available, and can be transported via the water path. If at least one positive test result is obtained,
the use of the soil or soil material is limited, or the requirements of remediation are not fulfilled (see
Figure 1).
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ISO 17616:2019(E)

Figure 1 — Flowchart describing a strategy for assessing the ecotoxicological potential of soils
and soil materials
6 © ISO 2019 – All rights reserved

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SIST EN ISO 17616:2023
ISO 17616:2019(E)

Table 1 — Biological tests for the assessment of ecotoxic and genotoxic potential of liquid-
extractable compounds possibly present in soils or soil materials (retention function)
Significant
Test Trophic Test
a,b
Reference End point LID value biological
category level species
b
effect
Inhibition of light
c [12]
Decomposer Vibrio fischeri ISO 11348 LID > 8 20 % Inhibition
emission
Acute ecotoxic
tests
Primary
[13]
Daphnia magna ISO 6341 Immobilization LID > 4 20 % Inhibition
consumer
[14]
Lemna minor ISO 20079 Growth inhibition — 25 % Inhibition
Primary
Raphidocelis
producer [15]
ISO 8692 Growth inhibition LID > 4 25 % Inhibition
subcapitata
Chronic/
20 % Mortality
sub-chronic Ceriodaphnia Mortality and
[16]
ISO 20665 —
dubia reproduction
ecotoxic tests
30 % Inhibition
Primary
consumer
20 % Mortality
Brachionus calicy- Mortality and
[17]
ISO 20666 —
florus reproduction
30 % Inhibition
Salmonella chol-
Induction rate of
[5]
eraesuis subsp. ISO 13829 D W 3 —
Li
umuC gene
choleraesuis
Induction rate
Salmonella chol- of umuC gene in
[5]
Decomposer eraesuis subsp. ISO 13829 concentrated water D W 3 —
Li
choleraesuis extract 1 : 15 (water
: soil)
Genotoxicity
Salmonella typh- Deci-
Induction rate of
tests [6]
imurium TA 98 ISO 16240 sive D —
min
mutant colonies
and TA 100 value W 2
Statistically sig-
nificant increase
Micronucleus on micronuleus
Primary
[18]
Vicia faba ISO 29200 frequency — frequency in the
producer
increase soil / soil material
relatively to the
negative control.
NOTE  Tests given in bold letters are part of a minimum test battery.
a
LID-values are relevant for a liquid/soil (L/S) ratio of 2 : 1. Further guidance on the preparation of soil eluates can be
[19] [20] [21]
obtained in EN ISO/TS 21268-1 , EN ISO TS 21268-2 or EN 14735 .
b
Toxicity criteria established on the basis of testing a wide variety of contaminated and uncontaminated soils. It should
[22] [23]
be guaranteed that no false positive assessment is obtained , .
c [24]
Reclassified as Aliivibrio fischeri (Beijerinck 1889) Urbanczyk et al. .
Nutrients extracted from soils and certain soil materials can interfere with the effects of microalgae
growth inhibitors, since a surplus of essential macronutrients, like nitrates and phosphates may
stimulate microalgae metabolism and physiological performance. Therefore, it shall be taken into
account that the toxicity of pollutants can be masked by substances favouring algal growth, and that soil
composition shall be considered when analysing results. The i
...

SLOVENSKI STANDARD
oSIST prEN ISO 17616:2022
01-julij-2022
Kakovost tal - Navodilo za izbiro in vrednotenje bioloških preskusov za
ekotoksikološko karakterizacijo tal in talnih materialov (ISO 17616:2019)
Soil quality - Guidance on the choice and evaluation of bioassays for ecotoxicological
characterization of soils and soil materials (ISO 17616:2019)
Qualité du sol - Lignes directrices relatives aux choix et à l'évaluation des essais
appliqués pour la caractérisation écotoxicologique des sols et des matériaux de type sol
(ISO 17616:2019)
Ta slovenski standard je istoveten z: prEN ISO 17616
ICS:
13.080.05 Preiskava tal na splošno Examination of soils in
general
oSIST prEN ISO 17616:2022 en,fr,de
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

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oSIST prEN ISO 17616:2022

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oSIST prEN ISO 17616:2022
INTERNATIONAL ISO
STANDARD 17616
Second edition
2019-10
Soil quality — Guidance on the choice
and evaluation of bioassays for
ecotoxicological characterization of
soils and soil materials
Qualité du sol — Lignes directrices relatives aux choix et
à l'évaluation des essais appliqués pour la caractérisation
écotoxicologique des sols et des matériaux de type sol
Reference number
ISO 17616:2019(E)
©
ISO 2019

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oSIST prEN ISO 17616:2022
ISO 17616:2019(E)

COPYRIGHT PROTECTED DOCUMENT
© ISO 2019
All rights reserved. Unless otherwise specified, or required in the context of its implementation, no part of this publication may
be reproduced or utilized otherwise in any form or by any means, electronic or mechanical, including photocopying, or posting
on the internet or an intranet, without prior written permission. Permission can be requested from either ISO at the address
below or ISO’s member body in the country of the requester.
ISO copyright office
CP 401 • Ch. de Blandonnet 8
CH-1214 Vernier, Geneva
Phone: +41 22 749 01 11
Fax: +41 22 749 09 47
Email: copyright@iso.org
Website: www.iso.org
Published in Switzerland
ii © ISO 2019 – All rights reserved

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oSIST prEN ISO 17616:2022
ISO 17616:2019(E)

Contents Page
Foreword .iv
Introduction .v
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 1
3.1 Assessment . 1
3.2 Types of soil and other soil materials . 2
3.3 Terms relating to Soil characteristics . 2
3.4 Land and sites . 3
4 Principles and applications of test batteries . 3
5 Testing strategy and interpretation of test results according to the use and re-use of
soils / soil materials and soil functions . 4
5.1 Monitoring of soil treatment success . 4
5.2 Assessment of the ecotoxic potential of soils / soil materials . 4
5.2.1 General. 4
5.2.2 Choice of test battery and evaluation of test results for assessing retention
function . 5
5.2.3 Choice of test battery and evaluation of test results for assessing habitat
function . 8
Annex A (informative) Tests with soil eluate — Expression of results .10
Bibliography .11
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oSIST prEN ISO 17616:2022
ISO 17616:2019(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.
The procedures used to develop this document and those intended for its further maintenance are
described in the ISO/IEC Directives, Part 1. In particular, the different approval criteria needed for the
different types of ISO documents should be noted. This document was drafted in accordance with the
editorial rules of the ISO/IEC Directives, Part 2 (see www .iso .org/ directives).
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. Details of
any patent rights identified during the development of the document will be in the Introduction and/or
on the ISO list of patent declarations received (see www .iso .org/ patents).
Any trade name used in this document is information given for the convenience of users and does not
constitute an endorsement.
For an explanation on the voluntary nature of standards, the meaning of ISO specific terms and
expressions related to conformity assessment, as well as information about ISO's adherence to the
World Trade Organization (WTO) principles in the Technical Barriers to Trade (TBT) see the following
URL: www .iso .org/ iso/ foreword .html.
This document was prepared by Technical Committee ISO/TC 190, Soil quality, Subcommittee SC 4,
Biological characterization.
This second edition cancels and replaces the first edition (ISO 17616:2008), which has been technically
revised. The main changes compared to the previous edition are as follows:
[10]
— definitions for “soil use” and “ecosystem service” have been added to Clause 3;
— end points of ecotoxicity tests (e.g. mortality, reproduction, growth, genotoxicity, and other
functional activities), as well as the overall principles and application of test batteries have been
clarified in Clause 4;
— sub-chronic toxicity tests have been added;
— Figure 1 was revised;
— Tables 1 and 2 (test batteries for retention and habitat function assessment, respectively) have been
revised (test categories, test organisms added/deleted, references updated).
Any feedback or questions on this document should be directed to the user’s national standards body. A
complete listing of these bodies can be found at www .iso .org/ members .html.
iv © ISO 2019 – All rights reserved

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Introduction
The characterization of contaminated soils can be based on strategies considering chemical analyses
and/or bioassays. ISO 15799 provides guidance on the selection of experimental methods for the
assessment of the ecotoxic potential of soils and soil materials (e.g. excavated and remediated soils,
refills, embankments) with respect to their intended use or re-use, and possible adverse effects on
aquatic and soil dwelling organisms.
An assessment strategy giving instructions for the choice and evaluation of test results is hence
proposed. The evaluation of the bioassays outcome is based on empirically-derived critical dilution
levels that take into account the sensitivity of the test system and the intended use/re-use of the site
under investigation. This approach intends to contribute to an effective and comparable assessment
[1]
within the ecotoxicological characterization of contaminated soil or soil materials . The test systems
included in this approach are not mandatory and may be replaced or accomplished by other test methods.
Nevertheless, the selected test systems have proved to appropriately characterize contaminated soils
[2],[3]
and soil materials with respect to their ecotoxic properties , both towards aquatic and terrestrial
organisms, the latter being responsible for maintaining essential soil functions.
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oSIST prEN ISO 17616:2022
INTERNATIONAL STANDARD ISO 17616:2019(E)
Soil quality — Guidance on the choice and evaluation of
bioassays for ecotoxicological characterization of soils and
soil materials
1 Scope
This document is one of the family of standards (ISO 15799, ISO 19204) providing guidance on the
characterization of soils and soil materials in relation to their retention and habitat functions and
uses. It is appropriate to use it in conjunction with the two other standards in this family. It provides
guidance on the choice and evaluation of tests applied for ecotoxicological characterization of soils
and soil materials. Recommendations for test strategies with respect to the protection of ground and
surface waters and the maintenance of the habitat function of soil are included. The tests recommended
represent a minimum test battery that can be complemented by additional tests, or even be replaced
by others, according to the intended uses or protection goals envisaged. The effect values indicated
in this document do not refer to regulation but represent the lowest level at which an adverse effect is
considered likely to occur.
2 Normative references
There are no normative references in this document.
3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
ISO and IEC maintain terminological databases for use in standardization at the following addresses:
— ISO Online browsing platform: available at https:// www .iso .org/ obp
— IEC Electropedia: available at http:// www .electropedia .org/
3.1 Assessment
3.1.1
soil-related assessment
assessment of the ecotoxic potential of soils (3.2.1), soil substrates and soil materials (3.2.2) based on
chemical analyses, biological tests and field inventories (monitoring) such as that mentioned in the
[4]
TRIAD approach
Note 1 to entry: TRIAD means an assessment approach based on a combination of chemical (i.e. residue analysis),
ecotoxicological (i.e. laboratory ecotoxic tests) and ecological (i.e. monitoring) data.
3.1.2
risk
expression of the probability that an adverse effect on soil (3.2.1) functions will occur under defined
conditions and the magnitude of the consequences of the effect occurring
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ISO 17616:2019(E)

3.1.3
LID-value
lowest-ineffective-dilution value
lowest value of the dilution factor above which the test gives an ecotoxicological relevant reduction
(e.g. 20 % inhibition of luminescence)
EXAMPLE A LID 8 corresponds to a dilution of soil extract of 1 : 8 (i.e 1 part of soil extract: 7 parts of
dilution water).
Note 1 to entry: The exact definitions are given in the standard of the respective bioassay. According to
[5] [6]
ISO 13829 for the umu-test, it is the “DLi value” (explanation in Annex D). According to ISO 16240 3.4, it is the
“decisive D value”.
min
Note 2 to entry: For further information on results expression and interpretation based on LID values see
Annex A of this document.
3.2 Types of soil and other soil materials
3.2.1
soil
upper layer of the Earth’s crust composed of mineral particles, organic matter, water, air and organisms
[SOURCE: ISO 15799:2019, 3.1.1]
3.2.2
soil material
material coming from soil (3.2.1) and displaced and/or modified by human activity, including excavated
soil, dredged materials, manufactured soils, remediated treated soils or fill materials
[SOURCE: ISO 17402:2008, 3.16]
3.3 Terms relating to Soil characteristics
3.3.1
habitat function
ability of soils (3.2.1)/ soil materials (3.2.2) to serve as a habitat for microorganisms, plants, soil-living
animals and their interactions (i.e. biocenose)
[SOURCE: ISO 15799:2019, 3.2.1]
3.3.2
retention function
ability of soils (3.2.1)/ soil materials (3.2.2) to adsorb pollutants in such a way that they cannot be
mobilised via the water pathway and translocated into the food chain
Note 1 to entry: The habitat and retention functions include the following soil functions according to
[7]
ISO 11074:2015 , 3.3.31:
— control of substance and energy cycles as components of ecosystems;
— basis for the life of plants, animals and man;
— carrier of genetic reservoir;
— basis for the production of agricultural products;
— buffer inhibiting movement of water, contaminants or other agents into the ground water.
[SOURCE: ISO 15799:2019, 3.2.2]
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3.3.3
contaminant
substance or agent present in the soil (3.2.1) as a result of human activity
Note 1 to entry: There is no assumption in this definition that harm results from the presence of the contaminant.
Note 2 to entry: See also pollutant (3.2.3) and potentially harmful substance (3.2.5) definitions in ISO 15799:2019
[9]
[SOURCE: ISO 15176:2002 , 3.2.6, modified — Note 2 to entry added.]
3.3.4
pollutant
substances, which due to their properties, amount or concentration cause negative impacts on the soil
(3.2.1) function or soil use
[9]
Note 1 to entry: Adapted from ISO 15176:2002 3.2.7.
3.4 Land and sites
3.4.1
re-use
useful and harmless utilisation of soil materials (3.2.2)
Note 1 to entry: In the context of this document re-use means the transfer of soil materials to another location for
use in agriculture, horticulture, forestry, gardens, recreational areas and construction sites.
[9]
[SOURCE: ISO 15176:2002 , 3.4.1; ISO 15799:2019, 3.3.1]
3.4.2
soil use
use of the ecosystem services (3.4.3) that soil (3.2.1) provides
3.4.3
ecosystem service
service that is (directly or indirectly) provided by an ecosystem to benefit people
[SOURCE: Based on Millennium Ecosystem Assessment]
4 Principles and applications of test batteries
The sensitivity of organisms (e.g. bacteria, plants, animals) to toxicants may vary significantly from one
species to another. Thereby, it is admitted that only the results of several ecotoxicity tests can give a
clear indication of the toxic effects of soil or soil materials. As such, the combination of ecotoxicity tests,
defined as a battery, shall include organisms belonging to various trophic levels, several biological
responses or end points (e.g. mortality, reproduction, growth, genotoxicity, as well as other functional
activities), in order to take into account the variability of species sensitivity within the studied
compartment.
The ecotoxicity tests included in batteries should at least have the following characteristics:
— sensitivity;
— practicability;
— compliance with standardized methods;
— high cost efficiency;
— representativeness of the soil ecosystem and/or of the selected application scenario (i.e. habitat or
retention functions).
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Depending on the use of the soil or soil material (e.g. the agricultural use of waste) or the protection
goal (i.e. habitat or retention functions), the applied test battery can differ (see Clause 5 in ISO 15799
[1]
for guidance on test selection criteria) . Nevertheless, the selected tests should allow the identification
of the most sensitive trophic level(s) and give information on the toxic effects induced by solid samples.
The evaluation of results from the ecotoxicological tests should take into consideration the application
purposes (further details on the field of application in ISO 15799, Clause 4), which can broadly be for:
— monitoring and control of the success of soil treatment (off-site, on-site, in situ) (see 5.1),
— assessment of soil / soil material quality or contamination effects according to its use or re-use
(see 5.2).
Irrespective of the application purpose, the ecotoxicological characterization of soils and soil materials
[1]
depends on the soil use/re-use and soil functions requiring protection , as aforementioned. Overall, it
can essentially rely on the:
— assessment of mobile and bioavailable potentially harmful substances, in cases where the soil/
soil material (see 3.2.1 and 3.2.2) can affect the ground and/or surface water, as well as in cases
where pollutants are added to soils (e.g. agricultural use of wastes like sludge, composts, etc.) (i.e.
retention function), and/or
— assessment of the ecotoxic potential of soils and soil materials (see 3.2.1 and 3.2.2) and possible
adverse effects on soil-dwelling organisms (i.e. habitat function).
5 Testing strategy and interpretation of test results according to the use and re-
use of soils / soil materials and soil functions
5.1 Monitoring of soil treatment success
For evaluating the efficiency of soil treatments, the procedure below may be followed at different stages:
a) before the treatment – perform the ecotoxicological assessment of the soil sample(s) using a test
battery (5.2);
b) monitoring during the soil treatment process – perform a simple test selected from the battery
above [e.g. the most sensitive and practical test used among the battery applied in 5.1 a)];
c) at the end of the treatment – perform a new assessment with the same test battery used in 5.1 a), in
order to judge the success of the treatment.
5.2 Assessment of the ecotoxic potential of soils / soil materials
5.2.1 General
If soils or soil materials are assessed with respect to their intended use or re-use, the tests (see
ISO 15799) appropriate to evaluate their quality regarding the retention (see Table 1) and/or habitat
functions (see Table 2) should be applied. A strategy for the assessment of the ecotoxicological
characterization of soils and soil materials is proposed in Figure 1. The chemical characterization of
soils should always be conducted to increase the reliability of interpretations of the ecotoxicological
results obtained upon the selected test battery.
The test battery usually includes a set of acute and chronic/sub-chronic toxicity tests. If acute toxicity
is detected, it is not necessary to perform other tests. On the other hand, if no acute effect is detected,
chronic/sub-chronic toxicity and genotoxicity test(s) shall be conducted.
The assessment of soil and soil materials may be influenced by the collection, handling and storage
methods followed. Thereby, standardized procedures should be conducted in accordance with
[11]
ISO 18400-206 .
4 © ISO
...

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