ISO 15799:2003
(Main)Soil quality — Guidance on the ecotoxicological characterization of soils and soil materials
Soil quality — Guidance on the ecotoxicological characterization of soils and soil materials
ISO 15799:2003 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 and possible adverse effects on aquatic and soil-dwelling organisms, and habitat maintenance and the retention function of the soil.
Qualité du sol — Lignes directrices relatives à la caractérisation écotoxicologique des sols et des matériaux du sol
L'ISO 15799:2003 fournit les lignes directrices relatives à la sélection des méthodes expérimentales permettant l'évaluation du potentiel écotoxique des sols et des matériaux du sol (par exemple terres excavées ayant fait l'objet d'une remédiation, remblais, talus) par rapport à leur utilisation prévue et aux effets éventuellement défavorables pour les organismes vivant dans l'eau et le sol, et le maintien des fonctions d'habitat et de rétention du sol.
Kakovost tal – Navodilo za ekotoksikološko karakterizacijo tal in talnih materialov
General Information
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Buy Standard
Standards Content (Sample)
SLOVENSKI STANDARD
SIST ISO 15799:2006
01-december-2006
Kakovost tal – Navodilo za ekotoksikološko karakterizacijo tal in talnih materialov
Soil quality -- Guidance on the ecotoxicological characterization of soils and soil
materials
Qualité du sol -- Lignes directrices relatives à la caractérisation écotoxicologique des
sols et des matériaux du sol
Ta slovenski standard je istoveten z: ISO 15799:2003
ICS:
13.080.99 Drugi standardi v zvezi s Other standards related to
kakovostjo tal soil quality
SIST ISO 15799:2006 en
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
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SIST ISO 15799:2006
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SIST ISO 15799:2006
INTERNATIONAL ISO
STANDARD 15799
First edition
2003-11-15
Soil quality — Guidance on the
ecotoxicological characterization of soils
and soil materials
Qualité du sol — Lignes directrices relatives à la caractérisation
écotoxicologique des sols et des matériaux du sol
Reference number
ISO 15799:2003(E)
©
ISO 2003
---------------------- Page: 3 ----------------------
SIST ISO 15799:2006
ISO 15799:2003(E)
PDF disclaimer
This PDF file may contain embedded typefaces. In accordance with Adobe's licensing policy, this file may be printed or viewed but
shall not be edited unless the typefaces which are embedded are licensed to and installed on the computer performing the editing. In
downloading this file, parties accept therein the responsibility of not infringing Adobe's licensing policy. The ISO Central Secretariat
accepts no liability in this area.
Adobe is a trademark of Adobe Systems Incorporated.
Details of the software products used to create this PDF file can be found in the General Info relative to the file; the PDF-creation
parameters were optimized for printing. Every care has been taken to ensure that the file is suitable for use by ISO member bodies. In
the unlikely event that a problem relating to it is found, please inform the Central Secretariat at the address given below.
© ISO 2003
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 either ISO at the address below or
ISO's member body in the country of the requester.
ISO copyright office
Case postale 56 • CH-1211 Geneva 20
Tel. + 41 22 749 01 11
Fax + 41 22 749 09 47
E-mail copyright@iso.org
Web www.iso.org
Published in Switzerland
ii © ISO 2003 — All rights reserved
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SIST ISO 15799:2006
ISO 15799:2003(E)
Contents Page
Foreword. iv
Introduction . v
1 Scope. 1
2 Terms and definitions. 1
2.1 Types of soil and other soil materials. 1
2.2 Terms relating to soil characteristics . 2
2.3 Land and sites . 2
3 Field of application . 3
3.1 Soils and areas of soil use where ecotoxicological tests should be considered:. 3
3.2 Soils and areas of soil use where ecotoxicological tests are not necessary (provided
groundwater contamination can be excluded): . 3
4 Selection of tests according to use/re-use of soils and soil materials and soil functions . 3
4.1 Usefulness of ecotoxicity tests . 3
4.2 General criteria for selection of tests . 4
4.3 Considerations for the examination of soil functions. 4
5 Sampling, transport, storage and sample preparation . 7
6 Limitations of proposed biotests for soils/soil materials . 7
Annex A (informative) Standardized forms of recommended test systems. 8
Bibliography . 31
© ISO 2003 — All rights reserved iii
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SIST ISO 15799:2006
ISO 15799:2003(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 15799 was prepared by Technical Committee ISO/TC 190, Soil quality, Subcommittee SC 7, Soil and site
assessment.
iv © ISO 2003 — All rights reserved
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SIST ISO 15799:2006
ISO 15799:2003(E)
Introduction
The majority of existing ecotoxicological test methods (biotests) being internationally harmonized were
developed to describe the ecotoxic potential of a test substance when added to a soil or soil material. These
methods can be used, with some modification, for the ecotoxicological characterization of soils and soil
materials with respect to their function and depending on the intended use. However, in such cases, users of
the methods need to be aware that the validation of the methods is not complete.
For substances with properties resulting in toxic effects, biotests are a complement to conventional chemical
analysis. Results from chemical analysis can be used for ecotoxicological assessments based on information
on the substances identified, including properties of the chemicals, e.g. their bioaccumulation potential. This
information is often scarce (if it exists at all) and does not include possible interactions (synergy/antagonism)
between chemicals and the complex soil matrix. Furthermore, an exhaustive identification and quantification of
substances is impractical. Therefore, ecotoxicological testing of soils can be used for investigating the
potential toxicity of complex chemical mixtures. The extrapolation from laboratory tests to field conditions
requires adequate consideration of important environmental factors within the test conditions and the selection
of suitable ecotoxicological endpoints.
This International Standard is one of a series providing guidance on soils and soil materials in relation to
certain functions and uses, including wildlife conservation, and ought to be read in conjunction with those
other standards.
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SIST ISO 15799:2006
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SIST ISO 15799:2006
INTERNATIONAL STANDARD ISO 15799:2003(E)
Soil quality — Guidance on the ecotoxicological
characterization of soils and soil materials
1 Scope
This International Standard 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 and possible adverse effects on aquatic and soil-dwelling organisms, and
habitat maintenance and the retention function of the soil.
It does not cover tests for bioaccumulation. Genotoxicity tests using eukaryotic organisms in soils are not yet
available. It is not applicable to the ecological assessment of uncontaminated soils with a view to natural,
agricultural or horticultural use, such soils being of possible interest where they can serve as a reference for
the assessment of soils from contaminated sites. Nor is the interpretation of the results gained by application
of the proposed methods within its scope.
2 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
2.1 Types of soil and other soil materials
2.1.1
soil
upper layer of the Earth’s crust composed of mineral particles, organic matter, water, air and organisms
[ISO 11074-1:1996, definition 5.4]
2.1.2
soil material
excavated soil, dredged materials, manufactured soils, treated soils and fill materials
[ISO 15176:2002, definition 3.1.4]
2.1.3
excavated soil
any natural material excavated from the ground, including top soil, sub soil, altered parent rock and parent
rock itself
NOTE Excavated soil typically arises during construction works.
[ISO 15176:2002, definition 3.1.5]
2.1.4
standard soil
field-collected soil whose main properties (e.g. pH, texture, organic matter content) are within a known range
[34]
EXAMPLE Eurosoils .
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2.2 Terms relating to soil characteristics
2.2.1
habitat function
ability of soils/soil materials to serve as a habitat for micro-organisms, plants, soil-living animals and their
interactions (biocenoses)
2.2.2
retention function
ability of soils/soil materials to adsorb pollutants in such that they cannot be mobilized via the water pathway
and translocated into the food chain
NOTE The habitat and retention functions include the following soil functions according to ISO 11074-1:
control of substance and energy cycles as components of ecosystems;
basis for the life of plants, animals and humans;
carrier of genetic reservoir;
basis for the production of agricultural products;
buffer inhibiting movement of water, contaminants or other agents into the ground water.
2.2.3
pollutant
substance or agent present in the soil which due to its properties, amount or concentration causes adverse
impact on soil functions or soil use
cf. contaminant (2.2.4), potentially harmful substance (2.2.5)
[ISO 15176:2002, definition 3.2.7]
NOTE See Introduction to ISO 11074-1:1996.
2.2.4
contaminant
substance or agent present in soil as a result of human activity
cf. pollutant (2.2.3), potentially harmful substance (2.2.5)
NOTE There is no assumption in this definition that harm results from the presence of the contaminant.
[ISO 15176:2002, definition 3.2.6]
2.2.5
potentially harmful substance
substance which, when present in sufficient concentration or amount, may be harmful to humans or the
environment
NOTE It may be present as a result of human activity [contaminant (2.2.4)] or naturally.
[ISO 15176:2002, definition 3.2.8]
2.3 Land and sites
2.3.1
re-use
useful and harmless utilization of soil materials
NOTE In the context of this International Standard the re-use means the transfer of soil materials to another location
for use in agriculture, horticulture, forestry, gardens, recreational areas and construction sites.
[ISO 15176:2002, term 3.4.1]
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3 Field of application
3.1 Soils and areas of soil use where ecotoxicological tests should be considered:
assessment of the ability of a soil to sustain a natural biocenosis or agriculture;
assessment of the combined ecotoxicity of all bioavailable contaminants present in soils or soil materials;
assessment of the ecotoxicity of potentially harmful substances in cases where the soil or soil material
can affect the ground and surface water;
identification of soils or soil materials (refills, embankments) having a low degree of contamination —
usually within a depth of 1 m — and which can remain at the site without further treatment;
detection of potential ecotoxicity which could not be traced by chemical analysis;
monitoring and control of the success of soil treatment (off-site, on-site, in situ);
monitoring and control of soils/soil materials that have been decontaminated and are to be applied at the
surface.
3.2 Soils and areas of soil use where ecotoxicological tests are not necessary (provided
groundwater contamination can be excluded):
contaminated soils classified as hazardous waste or which can be characterized clearly by
chemical/analytical parameters, in which cases ecotoxicological testing could be useful for a final
investigation after remediation and for process control during biological remediation;
commercially/industrially used areas with no prospect of horticultural/agricultural use;
soil materials or backfilled materials in an area to be effectively sealed by covering with buildings or other
forms of low permeability cover such as concrete or tarmacadam or asphalt.
4 Selection of tests according to use/re-use of soils and soil materials and soil
functions
4.1 Use of ecotoxicity tests
Toxicants can affect different species (and in some cases genotypes) present within ecosystems at different
concentrations. The ideal approach for the precise ecotoxicological characterization of the soil toxicity is to
use a battery of tests with several species belonging to different taxonomic and trophic groups, in order to
avoid false negative results owing to an adaptation of a test system (genotypic shift) to a specific contaminant
as compared to uncontaminated soils. Studies using field or semi-field investigations are rarely carried out and
can be very expensive.
The ideal scheme can be rendered more practicable by the adoption of simpler testing strategies and the
application of safety factors to the results obtained. If, however, testing is performed on one species or
function only, the high diversity in the sensitivity of species to toxicants will result in a high level of uncertainty.
It is therefore recommended to test at least a microbial process, a species from the plant kingdom, and one
from the animal kingdom, usually a saprophagous/detritivorous species; if more than one animal species is
tested, a predatory species should be included in the test battery. The minimum number of species to be
tested depends on the regulations to which the test strategy must comply. This International Standard only
gives the basic principles for their use. Further considerations to the selection of tests using soil organisms are
given in 4.3.
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4.2 General criteria for selection of tests
Criteria for the selection of ecotoxicity tests have been established in the context of hazard assessment and
classification of chemicals. These criteria should also apply for the ecotoxicological characterization of
contaminated soils. Criteria reviewed were: scientific validity, ecological significance, practicability and
acceptability. See [30] and [31].
The basic requirements that test protocols must meet in order to be established in international standards
include reproducibility, statistical validity, general acceptance and performance.
The importance of a criterion is relative to the specific situation. Decisions have to be made as to which
criteria are the most important or which tests may have to be modified, and choices made between these and
more practical considerations — for example, easy culturing of test organisms in the laboratory or the
availability of life stages required for a test throughout the year.
The test methods recommended here were originally designed for hazard assessment of chemicals and were
in most cases internationally harmonized (e.g. by OECD, EU or ISO). In most of the methods, provisions have
been made to adapt the test design to come within the scope of this International Standard. Nevertheless, in
many cases experience still has to be gained using the test methods for characterizing soil quality. In addition,
the selection of ecotoxicological test methods for the assessment of soils or soil materials depends on their
intended use or re-use and on the soil functions to be protected, in particular the retention and habitat
functions.
Table 1 gives an example of a decision scheme based on the relevant function.
Table 1 — Relevance for ecotoxicological testing to the intended re-use of the soil
Soil function
Retention function Habitat function
Re-use of soils
Aquatic organisms Plant growth Soil biocenoses
Detection of biological effects
a
Below sealed areas low low
low
Commercially and industrially used unsealed areas high low low
Landfill covering high high low
Green areas, parks and recreation areas high high high
Areas used in horticulture or agriculture high high high
a
Applies only to the unsaturated soil zone.
4.3 Considerations for the examination of soil functions
4.3.1 Retention function
Transport via water of soluble, colloidal or particle fractions plays a dominant role in the risk assessment of
contaminated soils. This is true not only because water may mobilize contaminants, but also because
contaminants and metabolites in the water phase potentially have a severe effect on micro-organisms, plants
and soil fauna.
Aqueous eluates are useful for testing ecotoxic effects on organisms exposed via the water-mediated
transport. It should be taken into account that substances mobilized via water could be subjected to different
types of changes, (e.g. metabolism or hydrolization) when transported into the groundwater and from there
into surface waters, and that their concentrations are reduced by dilution. Moreover, substances may be
mobilized over time due to environmental changes (e.g. pH, chemical and biological transformation). Eluates
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may serve as early indicators for the contamination of pore and ground water prior to the exposure of surface
and drinking water.
With these aspects, the investigation of groundwater and eluates is of the utmost importance — regardless of
the proposed soil use.
4.3.2 Habitat function
4.3.2.1 Representativeness of organisms and processes
The suitability of the soil for living organisms can best be examined by means of test methods selected to
include organisms and processes representative of different taxonomic groups.
4.3.2.2 Soil material used as control for bioassays on solid matrices
As a general principle in ecotoxicological testing, any end points measured in a treatment are compared with
those measured in the control or controls.
When evaluating the suitability of the soil for soil-dwelling organisms, it is a prerequisite to compare the
contaminated soil or soil material with a control material, which may also be used for preparing dilution series
with the contaminated sample.
Several types of control material can be used:
an uncontaminated soil with comparable pedological properties to the sample being tested;
an inert material (e.g. quartz sand);
a certified natural soil (e.g. standard soil);
a standardized artificial soil (see ISO 11267, ISO 11268-1 and ISO 11268-2).
The choice between these control materials should be made depending entirely on the aims of the
ecotoxicological assessment, the type of biological test being carried out and the requirements of the test
organism. This recommendation cannot be generalized for all biological tests. Adding sand to a soil or a soil
material can create a compact mixture incompatible with the growth and development of many organisms (e.g.
plant growth tests). It is preferable to use a more complex control material (such as artificial soil) for dilution
where this would have the advantage of reproducing more closely the natural environment of the organisms
and even if it may interact with pollutants. Placing an organism in a medium that does not match the most
important characteristics of its natural habitat may cause stress.
If a dose-response curve is needed, one of the control materials mentioned above may be used to dilute
the contaminated substrate.
If the aim is to classify each sample of soil or soil materials in terms of ecotoxicity hazard, it is preferable
to use an inert material (e.g. quartz sand), which will not interact with the pollutants present in the sample
and whose composition and granulometry can be rigorously standardized.
The requirements of the control material must take into account the different soil uses and the type and origin
of the soil (e.g. undisturbed soil, refilling material, excavated soil, remediated soil). Nutrient deficiency, as well
as physical conditions, can cause differences in plant growth and animal behaviour that need not necessarily
be caused by the pollutant situation and the hazard potential.
If the aim is to evaluate the ecotoxicity of a soil or soil material sample from a contaminated site, the
preferred method would be to use an uncontaminated control material similar to the sample being tested.
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SIST ISO 15799:2006
ISO 15799:2003(E)
If the aim is to evaluate the ecotoxicity of soils or soil materials which may be re-employed for certain
specific uses, the preferred method would be to use as a control material any material which may in
future be mixed with the soil or soil material.
4.3.2.3 Soil as substrate (medium) for soil micro-organisms
The soil microflora comprises on average 80 % of the mass of organisms living in soil. In combination with the
microfauna, the main functions of the microflora are the decomposition and degradation of complex organic
substances to easily available nutrients, thereby maintaining the natural substance cycles of carbon, nitrogen,
phosphorus and sulfur.
Substrate-induced respiration provides an indicator of the microbial population density.
Nitrifying bacteria, which are responsible for the oxidation of ammonium to nitrite and from nitrite to nitrate, are
a very sensitive group of micro-organisms. Reduced nitrification need not necessarily lead to significant
changes in the ecosystem but can be used as a sensitive indicator for the inhibition of an essential soil
process.
The purpose of determining the microbial biomass or other microbial processes in soils is to allow assessment
of the continued maintenance of soil fertility, the potential ability to degrade organic compounds, and the
effects of added materials on the soil microbial community.
4.3.2.4 Soil as substrate for plant growth
After micro-organisms, plant roots constitute the largest biological surface in soil. Their contact area with soil
particles is increased by the presence of root hairs and mycorrhizal associations (VA-mycorrhiza with
cultivated plants and additional ectomycoorhizae with woody plants).
As with the other bioassays proposed, tests with higher plants are designed to assess the bioavailability and
effects of pollutants detected or not detected by chemical analysis, respectively. By applying a test period of at
least 14 days, short-term changes in the soil by the test plant itself are included.
The accumulation of pollutants in plants, their metabolism and their effects on consumers are not investigated
in these tests. They do not apply to the assessment of soil fertility and productivity.
4.3.2.5 Soil as substrate for soil living fauna
Soil animals generally fulfil the following four functions:
a) mechanical activities (drainage, aeration, mixing, mechanical comminution);
b) chemical changes (enhanced availability of nitrate and phosphate from excrements and accelerated
formation of clay-humus complexes, after the substrate has passed the gut);
c) biological changes (distribution of micro-organisms in the soil matrix, synergistic effects through
stimulation of microbial activity and organic matter decomposition);
d) significant links in the food web.
Short-term and long-term tests are available for examination of the effects of pollutants on soil fauna. For
testing the habitat function, characterization by sublethal test parameters is particularly recommended.
Since a single test method cannot adequately represent the vast number of very diverse invertebrates, a test
battery should be used. When selecting the individual test species, the following criteria should be considered:
a) trophic level — e.e. saprophagous and predatory species should be included;
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b) taxonomic/physiological groups — in order to cover the biodiversity of soil communities, at least
representatives of annelids and arthropoda have to be selected;
c) size class/exposure pathway: species of the micro-, meso- and macrofauna do not only represent various
size classes but also different life-styles and therefore exposure routes (e.g. pore water versus food
uptake);
d) ecological role — at least soil-dwelling and litter-inhabiting species are important to consider.
Only internationally standardized methods should be used.
5 Sampling, transport, storage and sample preparation
Before soil quality is assessed by any of the methods proposed, soil samples need to be collected from the
site under investigation. Soil sampling should be carried out by trained personnel with sufficient knowledge of
sampling, handling of samples and safety measures at contaminated sites and sampling locations. The
sampling strategy and handling should be determined by the site to be investigated, the kind of contamination
and the aim of the biological tests (e.g. quantities of soil samples could vary between 100 mg and 100 kg,
depending on the tests selected).
Record all data concerning sampling, transport and sample preparation. For instructions on the design of
sampling programmes, sampling techniques, safety, investigations of natural, cultivated, urban and industrial
sites and on the collection, handling and storage of soil for the assessment of aerobic/anaerobic microbial
processes in the laboratory, see ISO 10381-6.
6 Limitations of proposed biotests for soils/soil materials
Biological test systems are suitable for volatile pollutants only to a limited extent. Other methods should be
developed for this purpose. Similarly, the impact of organic contaminants, which are easily degradable under
aerobic conditions, may be detected incompletely by the methods described. In this case, alternative methods
for sampling and sample preparation should be applied.
NOTE The proposed terrestrial and aquatic test methods in A.1 and A.2 were developed to assess the ecotoxic
potential of chemicals. The characterization of soils or soil eluates was not their primary goal. Therefore, the methods
need to be adapted to the specific requirements of soil and site assessment.
© ISO 2003 — All rights reserved 7
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SIST ISO 15799:2006
ISO 15799:2003(E)
Annex A
(informative)
Standardized forms of recommended test systems
A.1 Terrestrial test methods
A.1.1 Soil fauna
A.1.1.1 Collembola — Effects on reproduction
See Table A.1.
Table A.1
1. Title of test Soil quality — Inhibition of reproduction of Collembola (Folsomia candida) by
soil pollutants
2. Harmonization International
3. Reference ISO 11267
4. Principle Determination of the effect on reproduction of springtails incubated over a
4 weeks test period
5. Test type Static subchronic
6. Test organism Springtails
Breeding stocks Folsomia candida Willem 1902
Age 10 d to 12 d
Feeding Dry yeast
7. Test substrate Artificial soil, contaminated soil
Volume 30 g (wet mass)/container
8. Test conditions
Test chamber Enclosures
Temperature 20 °C ± 2 °C
pH 6 ± 0,5
Light inte
...
INTERNATIONAL ISO
STANDARD 15799
First edition
2003-11-15
Soil quality — Guidance on the
ecotoxicological characterization of soils
and soil materials
Qualité du sol — Lignes directrices relatives à la caractérisation
écotoxicologique des sols et des matériaux du sol
Reference number
ISO 15799:2003(E)
©
ISO 2003
---------------------- Page: 1 ----------------------
ISO 15799:2003(E)
PDF disclaimer
This PDF file may contain embedded typefaces. In accordance with Adobe's licensing policy, this file may be printed or viewed but
shall not be edited unless the typefaces which are embedded are licensed to and installed on the computer performing the editing. In
downloading this file, parties accept therein the responsibility of not infringing Adobe's licensing policy. The ISO Central Secretariat
accepts no liability in this area.
Adobe is a trademark of Adobe Systems Incorporated.
Details of the software products used to create this PDF file can be found in the General Info relative to the file; the PDF-creation
parameters were optimized for printing. Every care has been taken to ensure that the file is suitable for use by ISO member bodies. In
the unlikely event that a problem relating to it is found, please inform the Central Secretariat at the address given below.
© ISO 2003
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 either ISO at the address below or
ISO's member body in the country of the requester.
ISO copyright office
Case postale 56 • CH-1211 Geneva 20
Tel. + 41 22 749 01 11
Fax + 41 22 749 09 47
E-mail copyright@iso.org
Web www.iso.org
Published in Switzerland
ii © ISO 2003 — All rights reserved
---------------------- Page: 2 ----------------------
ISO 15799:2003(E)
Contents Page
Foreword. iv
Introduction . v
1 Scope. 1
2 Terms and definitions. 1
2.1 Types of soil and other soil materials. 1
2.2 Terms relating to soil characteristics . 2
2.3 Land and sites . 2
3 Field of application . 3
3.1 Soils and areas of soil use where ecotoxicological tests should be considered:. 3
3.2 Soils and areas of soil use where ecotoxicological tests are not necessary (provided
groundwater contamination can be excluded): . 3
4 Selection of tests according to use/re-use of soils and soil materials and soil functions . 3
4.1 Usefulness of ecotoxicity tests . 3
4.2 General criteria for selection of tests . 4
4.3 Considerations for the examination of soil functions. 4
5 Sampling, transport, storage and sample preparation . 7
6 Limitations of proposed biotests for soils/soil materials . 7
Annex A (informative) Standardized forms of recommended test systems. 8
Bibliography . 31
© ISO 2003 — All rights reserved iii
---------------------- Page: 3 ----------------------
ISO 15799:2003(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 15799 was prepared by Technical Committee ISO/TC 190, Soil quality, Subcommittee SC 7, Soil and site
assessment.
iv © ISO 2003 — All rights reserved
---------------------- Page: 4 ----------------------
ISO 15799:2003(E)
Introduction
The majority of existing ecotoxicological test methods (biotests) being internationally harmonized were
developed to describe the ecotoxic potential of a test substance when added to a soil or soil material. These
methods can be used, with some modification, for the ecotoxicological characterization of soils and soil
materials with respect to their function and depending on the intended use. However, in such cases, users of
the methods need to be aware that the validation of the methods is not complete.
For substances with properties resulting in toxic effects, biotests are a complement to conventional chemical
analysis. Results from chemical analysis can be used for ecotoxicological assessments based on information
on the substances identified, including properties of the chemicals, e.g. their bioaccumulation potential. This
information is often scarce (if it exists at all) and does not include possible interactions (synergy/antagonism)
between chemicals and the complex soil matrix. Furthermore, an exhaustive identification and quantification of
substances is impractical. Therefore, ecotoxicological testing of soils can be used for investigating the
potential toxicity of complex chemical mixtures. The extrapolation from laboratory tests to field conditions
requires adequate consideration of important environmental factors within the test conditions and the selection
of suitable ecotoxicological endpoints.
This International Standard is one of a series providing guidance on soils and soil materials in relation to
certain functions and uses, including wildlife conservation, and ought to be read in conjunction with those
other standards.
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INTERNATIONAL STANDARD ISO 15799:2003(E)
Soil quality — Guidance on the ecotoxicological
characterization of soils and soil materials
1 Scope
This International Standard 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 and possible adverse effects on aquatic and soil-dwelling organisms, and
habitat maintenance and the retention function of the soil.
It does not cover tests for bioaccumulation. Genotoxicity tests using eukaryotic organisms in soils are not yet
available. It is not applicable to the ecological assessment of uncontaminated soils with a view to natural,
agricultural or horticultural use, such soils being of possible interest where they can serve as a reference for
the assessment of soils from contaminated sites. Nor is the interpretation of the results gained by application
of the proposed methods within its scope.
2 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
2.1 Types of soil and other soil materials
2.1.1
soil
upper layer of the Earth’s crust composed of mineral particles, organic matter, water, air and organisms
[ISO 11074-1:1996, definition 5.4]
2.1.2
soil material
excavated soil, dredged materials, manufactured soils, treated soils and fill materials
[ISO 15176:2002, definition 3.1.4]
2.1.3
excavated soil
any natural material excavated from the ground, including top soil, sub soil, altered parent rock and parent
rock itself
NOTE Excavated soil typically arises during construction works.
[ISO 15176:2002, definition 3.1.5]
2.1.4
standard soil
field-collected soil whose main properties (e.g. pH, texture, organic matter content) are within a known range
[34]
EXAMPLE Eurosoils .
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ISO 15799:2003(E)
2.2 Terms relating to soil characteristics
2.2.1
habitat function
ability of soils/soil materials to serve as a habitat for micro-organisms, plants, soil-living animals and their
interactions (biocenoses)
2.2.2
retention function
ability of soils/soil materials to adsorb pollutants in such that they cannot be mobilized via the water pathway
and translocated into the food chain
NOTE The habitat and retention functions include the following soil functions according to ISO 11074-1:
control of substance and energy cycles as components of ecosystems;
basis for the life of plants, animals and humans;
carrier of genetic reservoir;
basis for the production of agricultural products;
buffer inhibiting movement of water, contaminants or other agents into the ground water.
2.2.3
pollutant
substance or agent present in the soil which due to its properties, amount or concentration causes adverse
impact on soil functions or soil use
cf. contaminant (2.2.4), potentially harmful substance (2.2.5)
[ISO 15176:2002, definition 3.2.7]
NOTE See Introduction to ISO 11074-1:1996.
2.2.4
contaminant
substance or agent present in soil as a result of human activity
cf. pollutant (2.2.3), potentially harmful substance (2.2.5)
NOTE There is no assumption in this definition that harm results from the presence of the contaminant.
[ISO 15176:2002, definition 3.2.6]
2.2.5
potentially harmful substance
substance which, when present in sufficient concentration or amount, may be harmful to humans or the
environment
NOTE It may be present as a result of human activity [contaminant (2.2.4)] or naturally.
[ISO 15176:2002, definition 3.2.8]
2.3 Land and sites
2.3.1
re-use
useful and harmless utilization of soil materials
NOTE In the context of this International Standard the re-use means the transfer of soil materials to another location
for use in agriculture, horticulture, forestry, gardens, recreational areas and construction sites.
[ISO 15176:2002, term 3.4.1]
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ISO 15799:2003(E)
3 Field of application
3.1 Soils and areas of soil use where ecotoxicological tests should be considered:
assessment of the ability of a soil to sustain a natural biocenosis or agriculture;
assessment of the combined ecotoxicity of all bioavailable contaminants present in soils or soil materials;
assessment of the ecotoxicity of potentially harmful substances in cases where the soil or soil material
can affect the ground and surface water;
identification of soils or soil materials (refills, embankments) having a low degree of contamination —
usually within a depth of 1 m — and which can remain at the site without further treatment;
detection of potential ecotoxicity which could not be traced by chemical analysis;
monitoring and control of the success of soil treatment (off-site, on-site, in situ);
monitoring and control of soils/soil materials that have been decontaminated and are to be applied at the
surface.
3.2 Soils and areas of soil use where ecotoxicological tests are not necessary (provided
groundwater contamination can be excluded):
contaminated soils classified as hazardous waste or which can be characterized clearly by
chemical/analytical parameters, in which cases ecotoxicological testing could be useful for a final
investigation after remediation and for process control during biological remediation;
commercially/industrially used areas with no prospect of horticultural/agricultural use;
soil materials or backfilled materials in an area to be effectively sealed by covering with buildings or other
forms of low permeability cover such as concrete or tarmacadam or asphalt.
4 Selection of tests according to use/re-use of soils and soil materials and soil
functions
4.1 Use of ecotoxicity tests
Toxicants can affect different species (and in some cases genotypes) present within ecosystems at different
concentrations. The ideal approach for the precise ecotoxicological characterization of the soil toxicity is to
use a battery of tests with several species belonging to different taxonomic and trophic groups, in order to
avoid false negative results owing to an adaptation of a test system (genotypic shift) to a specific contaminant
as compared to uncontaminated soils. Studies using field or semi-field investigations are rarely carried out and
can be very expensive.
The ideal scheme can be rendered more practicable by the adoption of simpler testing strategies and the
application of safety factors to the results obtained. If, however, testing is performed on one species or
function only, the high diversity in the sensitivity of species to toxicants will result in a high level of uncertainty.
It is therefore recommended to test at least a microbial process, a species from the plant kingdom, and one
from the animal kingdom, usually a saprophagous/detritivorous species; if more than one animal species is
tested, a predatory species should be included in the test battery. The minimum number of species to be
tested depends on the regulations to which the test strategy must comply. This International Standard only
gives the basic principles for their use. Further considerations to the selection of tests using soil organisms are
given in 4.3.
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ISO 15799:2003(E)
4.2 General criteria for selection of tests
Criteria for the selection of ecotoxicity tests have been established in the context of hazard assessment and
classification of chemicals. These criteria should also apply for the ecotoxicological characterization of
contaminated soils. Criteria reviewed were: scientific validity, ecological significance, practicability and
acceptability. See [30] and [31].
The basic requirements that test protocols must meet in order to be established in international standards
include reproducibility, statistical validity, general acceptance and performance.
The importance of a criterion is relative to the specific situation. Decisions have to be made as to which
criteria are the most important or which tests may have to be modified, and choices made between these and
more practical considerations — for example, easy culturing of test organisms in the laboratory or the
availability of life stages required for a test throughout the year.
The test methods recommended here were originally designed for hazard assessment of chemicals and were
in most cases internationally harmonized (e.g. by OECD, EU or ISO). In most of the methods, provisions have
been made to adapt the test design to come within the scope of this International Standard. Nevertheless, in
many cases experience still has to be gained using the test methods for characterizing soil quality. In addition,
the selection of ecotoxicological test methods for the assessment of soils or soil materials depends on their
intended use or re-use and on the soil functions to be protected, in particular the retention and habitat
functions.
Table 1 gives an example of a decision scheme based on the relevant function.
Table 1 — Relevance for ecotoxicological testing to the intended re-use of the soil
Soil function
Retention function Habitat function
Re-use of soils
Aquatic organisms Plant growth Soil biocenoses
Detection of biological effects
a
Below sealed areas low low
low
Commercially and industrially used unsealed areas high low low
Landfill covering high high low
Green areas, parks and recreation areas high high high
Areas used in horticulture or agriculture high high high
a
Applies only to the unsaturated soil zone.
4.3 Considerations for the examination of soil functions
4.3.1 Retention function
Transport via water of soluble, colloidal or particle fractions plays a dominant role in the risk assessment of
contaminated soils. This is true not only because water may mobilize contaminants, but also because
contaminants and metabolites in the water phase potentially have a severe effect on micro-organisms, plants
and soil fauna.
Aqueous eluates are useful for testing ecotoxic effects on organisms exposed via the water-mediated
transport. It should be taken into account that substances mobilized via water could be subjected to different
types of changes, (e.g. metabolism or hydrolization) when transported into the groundwater and from there
into surface waters, and that their concentrations are reduced by dilution. Moreover, substances may be
mobilized over time due to environmental changes (e.g. pH, chemical and biological transformation). Eluates
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ISO 15799:2003(E)
may serve as early indicators for the contamination of pore and ground water prior to the exposure of surface
and drinking water.
With these aspects, the investigation of groundwater and eluates is of the utmost importance — regardless of
the proposed soil use.
4.3.2 Habitat function
4.3.2.1 Representativeness of organisms and processes
The suitability of the soil for living organisms can best be examined by means of test methods selected to
include organisms and processes representative of different taxonomic groups.
4.3.2.2 Soil material used as control for bioassays on solid matrices
As a general principle in ecotoxicological testing, any end points measured in a treatment are compared with
those measured in the control or controls.
When evaluating the suitability of the soil for soil-dwelling organisms, it is a prerequisite to compare the
contaminated soil or soil material with a control material, which may also be used for preparing dilution series
with the contaminated sample.
Several types of control material can be used:
an uncontaminated soil with comparable pedological properties to the sample being tested;
an inert material (e.g. quartz sand);
a certified natural soil (e.g. standard soil);
a standardized artificial soil (see ISO 11267, ISO 11268-1 and ISO 11268-2).
The choice between these control materials should be made depending entirely on the aims of the
ecotoxicological assessment, the type of biological test being carried out and the requirements of the test
organism. This recommendation cannot be generalized for all biological tests. Adding sand to a soil or a soil
material can create a compact mixture incompatible with the growth and development of many organisms (e.g.
plant growth tests). It is preferable to use a more complex control material (such as artificial soil) for dilution
where this would have the advantage of reproducing more closely the natural environment of the organisms
and even if it may interact with pollutants. Placing an organism in a medium that does not match the most
important characteristics of its natural habitat may cause stress.
If a dose-response curve is needed, one of the control materials mentioned above may be used to dilute
the contaminated substrate.
If the aim is to classify each sample of soil or soil materials in terms of ecotoxicity hazard, it is preferable
to use an inert material (e.g. quartz sand), which will not interact with the pollutants present in the sample
and whose composition and granulometry can be rigorously standardized.
The requirements of the control material must take into account the different soil uses and the type and origin
of the soil (e.g. undisturbed soil, refilling material, excavated soil, remediated soil). Nutrient deficiency, as well
as physical conditions, can cause differences in plant growth and animal behaviour that need not necessarily
be caused by the pollutant situation and the hazard potential.
If the aim is to evaluate the ecotoxicity of a soil or soil material sample from a contaminated site, the
preferred method would be to use an uncontaminated control material similar to the sample being tested.
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ISO 15799:2003(E)
If the aim is to evaluate the ecotoxicity of soils or soil materials which may be re-employed for certain
specific uses, the preferred method would be to use as a control material any material which may in
future be mixed with the soil or soil material.
4.3.2.3 Soil as substrate (medium) for soil micro-organisms
The soil microflora comprises on average 80 % of the mass of organisms living in soil. In combination with the
microfauna, the main functions of the microflora are the decomposition and degradation of complex organic
substances to easily available nutrients, thereby maintaining the natural substance cycles of carbon, nitrogen,
phosphorus and sulfur.
Substrate-induced respiration provides an indicator of the microbial population density.
Nitrifying bacteria, which are responsible for the oxidation of ammonium to nitrite and from nitrite to nitrate, are
a very sensitive group of micro-organisms. Reduced nitrification need not necessarily lead to significant
changes in the ecosystem but can be used as a sensitive indicator for the inhibition of an essential soil
process.
The purpose of determining the microbial biomass or other microbial processes in soils is to allow assessment
of the continued maintenance of soil fertility, the potential ability to degrade organic compounds, and the
effects of added materials on the soil microbial community.
4.3.2.4 Soil as substrate for plant growth
After micro-organisms, plant roots constitute the largest biological surface in soil. Their contact area with soil
particles is increased by the presence of root hairs and mycorrhizal associations (VA-mycorrhiza with
cultivated plants and additional ectomycoorhizae with woody plants).
As with the other bioassays proposed, tests with higher plants are designed to assess the bioavailability and
effects of pollutants detected or not detected by chemical analysis, respectively. By applying a test period of at
least 14 days, short-term changes in the soil by the test plant itself are included.
The accumulation of pollutants in plants, their metabolism and their effects on consumers are not investigated
in these tests. They do not apply to the assessment of soil fertility and productivity.
4.3.2.5 Soil as substrate for soil living fauna
Soil animals generally fulfil the following four functions:
a) mechanical activities (drainage, aeration, mixing, mechanical comminution);
b) chemical changes (enhanced availability of nitrate and phosphate from excrements and accelerated
formation of clay-humus complexes, after the substrate has passed the gut);
c) biological changes (distribution of micro-organisms in the soil matrix, synergistic effects through
stimulation of microbial activity and organic matter decomposition);
d) significant links in the food web.
Short-term and long-term tests are available for examination of the effects of pollutants on soil fauna. For
testing the habitat function, characterization by sublethal test parameters is particularly recommended.
Since a single test method cannot adequately represent the vast number of very diverse invertebrates, a test
battery should be used. When selecting the individual test species, the following criteria should be considered:
a) trophic level — e.e. saprophagous and predatory species should be included;
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ISO 15799:2003(E)
b) taxonomic/physiological groups — in order to cover the biodiversity of soil communities, at least
representatives of annelids and arthropoda have to be selected;
c) size class/exposure pathway: species of the micro-, meso- and macrofauna do not only represent various
size classes but also different life-styles and therefore exposure routes (e.g. pore water versus food
uptake);
d) ecological role — at least soil-dwelling and litter-inhabiting species are important to consider.
Only internationally standardized methods should be used.
5 Sampling, transport, storage and sample preparation
Before soil quality is assessed by any of the methods proposed, soil samples need to be collected from the
site under investigation. Soil sampling should be carried out by trained personnel with sufficient knowledge of
sampling, handling of samples and safety measures at contaminated sites and sampling locations. The
sampling strategy and handling should be determined by the site to be investigated, the kind of contamination
and the aim of the biological tests (e.g. quantities of soil samples could vary between 100 mg and 100 kg,
depending on the tests selected).
Record all data concerning sampling, transport and sample preparation. For instructions on the design of
sampling programmes, sampling techniques, safety, investigations of natural, cultivated, urban and industrial
sites and on the collection, handling and storage of soil for the assessment of aerobic/anaerobic microbial
processes in the laboratory, see ISO 10381-6.
6 Limitations of proposed biotests for soils/soil materials
Biological test systems are suitable for volatile pollutants only to a limited extent. Other methods should be
developed for this purpose. Similarly, the impact of organic contaminants, which are easily degradable under
aerobic conditions, may be detected incompletely by the methods described. In this case, alternative methods
for sampling and sample preparation should be applied.
NOTE The proposed terrestrial and aquatic test methods in A.1 and A.2 were developed to assess the ecotoxic
potential of chemicals. The characterization of soils or soil eluates was not their primary goal. Therefore, the methods
need to be adapted to the specific requirements of soil and site assessment.
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ISO 15799:2003(E)
Annex A
(informative)
Standardized forms of recommended test systems
A.1 Terrestrial test methods
A.1.1 Soil fauna
A.1.1.1 Collembola — Effects on reproduction
See Table A.1.
Table A.1
1. Title of test Soil quality — Inhibition of reproduction of Collembola (Folsomia candida) by
soil pollutants
2. Harmonization International
3. Reference ISO 11267
4. Principle Determination of the effect on reproduction of springtails incubated over a
4 weeks test period
5. Test type Static subchronic
6. Test organism Springtails
Breeding stocks Folsomia candida Willem 1902
Age 10 d to 12 d
Feeding Dry yeast
7. Test substrate Artificial soil, contaminated soil
Volume 30 g (wet mass)/container
8. Test conditions
Test chamber Enclosures
Temperature 20 °C ± 2 °C
pH 6 ± 0,5
Light intensity/quality Between 400 lx and 800 lx
Photoperiod 12 h:12 h or 16 h:8 h
Soil moisture 40 % to 60 % of total water holding capacity
9. No. replicates 4
10. Test duration/incubation 28 d
11. Neg. control/ dilution soil Artificial soil
12. Validity criteria Control: mortality < 20 %, min. reproduction 100 juveniles, CV u 30 %
13. Pos. control/reference toxicant E 605 forte (a.i. 507,5 g/l) Betanal plus (a.i. 160 g/l)
Mean EC50, CV LOEC: 0,18 mg/kg to 0,32 mg/kg; 100 mg/kg to 200 mg/kg
14. Statistics Multiple t-test, u-test, regression analysis
15. Test parameter(s) Mortality of adults, inhibition of reproduction
16. End points EC (x = % effect level, e.g. 10, 50), NOEC
x
17. Limitations/comments
The test was originally designed for testing substances added to an artificial soil. To compare or to monitor soil quality,
the method has to be adapted. Care should be taken that any control soil used meets the biological requirements of the
test species. The number of replicates might have to be increased because of the heterogeneity of field samples.
8 © ISO 2003 — All rights reserved
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...
NORME ISO
INTERNATIONALE 15799
Première édition
2003-11-15
Qualité du sol — Lignes directrices
relatives à la caractérisation
écotoxicologique des sols et des
matériaux du sol
Soil quality — Guidance on the ecotoxicological characterization of soils
and soil materials
Numéro de référence
ISO 15799:2003(F)
©
ISO 2003
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ISO 15799:2003(F)
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ISO 15799:2003(F)
Sommaire Page
Avant-propos. iv
Introduction . v
1 Domaine d'application. 1
2 Termes et définitions . 1
2.1 Types des sols et autres matériaux du sol. 1
2.2 Termes relatifs aux caractéristiques du sol. 2
2.3 Terrain et sites. 3
3 Applications des essais écotoxicologiques.3
3.1 Utilisation des sols et zones de sols où il convient d’envisager des essais
écotoxicologiques: . 3
3.2 Utilisation de sols et zones de sols où les essais écotoxicologiques ne sont pas
nécessaires (sous réserve de pouvoir exclure toute contamination des eaux
souterraines): . 3
4 Choix des essais suivant l’utilisation/la réutilisation des sols et des matériaux du sol et
selon les fonctions du sol. 4
4.1 Utilisation d’essais d’écotoxicité . 4
4.2 Critères généraux pour le choix des essais. 4
4.3 Considérations relatives à l’examen des fonctions du sol. 5
5 Échantillonnage, transport, stockage et préparation des échantillons . 8
6 Limites des essais biologiques proposés pour les sols/matériaux du sol. 8
Annexe A (informative) Formes normalisées des systèmes d'essai recommandés. 9
Bibliographie . 32
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ISO 15799:2003(F)
Avant-propos
L'ISO (Organisation internationale de normalisation) est une fédération mondiale d'organismes nationaux de
normalisation (comités membres de l'ISO). L'élaboration des Normes internationales est en général confiée
aux comités techniques de l'ISO. Chaque comité membre intéressé par une étude a le droit de faire partie du
comité technique créé à cet effet. Les organisations internationales, gouvernementales et non
gouvernementales, en liaison avec l'ISO participent également aux travaux. L'ISO collabore étroitement avec
la Commission électrotechnique internationale (CEI) en ce qui concerne la normalisation électrotechnique.
Les Normes internationales sont rédigées conformément aux règles données dans les Directives ISO/CEI,
Partie 2.
La tâche principale des comités techniques est d'élaborer les Normes internationales. Les projets de Normes
internationales adoptés par les comités techniques sont soumis aux comités membres pour vote. Leur
publication comme Normes internationales requiert l'approbation de 75 % au moins des comités membres
votants.
L'attention est appelée sur le fait que certains des éléments du présent document peuvent faire l'objet de
droits de propriété intellectuelle ou de droits analogues. L'ISO ne saurait être tenue pour responsable de ne
pas avoir identifié de tels droits de propriété et averti de leur existence.
L'ISO 15799 a été élaborée par le comité technique ISO/TC 190, Qualité du sol, sous-comité SC 7, Évaluation
des sols et des sites.
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ISO 15799:2003(F)
Introduction
La plupart des méthodes d’essai écotoxicologique existantes (essais biologiques) en cours d’harmonisation
au plan international ont été mises au point pour décrire le potentiel écotoxique d’une substance lorsqu’on
l’ajoute à un sol/matériau du sol. Ces méthodes peuvent être utilisées sous réserve de quelques modifications
pour la caractérisation écotoxicologique des sols/matériaux du sol pour ce qui concerne leur fonction, suivant
l’utilisation prévue. Néanmoins, dans ce contexte, il convient que les utilisateurs des présentes méthodes
gardent à l’esprit que la validation de celles-ci n’est pas terminée.
Les essais biologiques complètent les analyses chimiques classiques pour les substances ayant des
propriétés toxiques. Les résultats obtenus par une analyse chimique peuvent être utilisés pour effectuer des
évaluations écotoxicologiques à partir de données relatives aux substances identifiées, y compris
les propriétés des produits chimiques telles que leur potentiel de bioaccumulation. Ces données sont
souvent rares (pour autant qu’elles existent) et ne comprennent pas les éventuelles interactions
(synergie/antagonisme) entre les produits chimiques et la matrice complexe du sol. En outre, une
identification et une quantification exhaustives des substances sont irréalisables. Par conséquent, pour
étudier l’éventuelle toxicité de mélanges chimiques complexes dans les sols, il est possible d’utiliser les
essais écotoxicologiques des sols. L’extrapolation des essais de laboratoire aux conditions de terrain
nécessite une prise en compte adéquate des facteurs environnementaux essentiels dans les conditions
d’essai ainsi que le choix de critères d’effet écotoxicologiques appropriés.
La présente Norme internationale fait partie d’une série de normes qui donnent des recommandations
relatives aux sols et aux matériaux du sol eu égard à certaines fonctions et utilisations, y compris la
préservation des organismes vivants. Il convient de la lire conjointement avec ces autres normes.
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NORME INTERNATIONALE ISO 15799:2003(F)
Qualité du sol — Lignes directrices relatives à la caractérisation
écotoxicologique des sols et des matériaux du sol
1 Domaine d'application
La présente Norme internationale fournit les lignes directrices relatives à la sélection des méthodes
expérimentales permettant l’évaluation du potentiel écotoxique des sols et des matériaux du sol (par exemple
terres excavées ayant fait l’objet d’une remédiation, remblais, talus) par rapport à leur utilisation prévue et aux
effets éventuellement défavorables pour les organismes vivant dans l’eau et le sol, et le maintien des
fonctions d’habitat et de rétention du sol.
La présente Norme internationale ne traite pas des essais relatifs à la bioaccumulation. Les essais de
génotoxicité utilisant des organismes eucaryotes du sol ne sont pas encore disponibles. La présente Norme
internationale ne s’applique pas à l’évaluation écologique des sols non contaminés en vue d’une utilisation
naturelle, agricole ou horticole, de tels sols pouvant être intéressants s’ils peuvent servir de référence pour
l’évaluation de sols provenant de sites contaminés. De même, elle ne concerne pas l’interprétation des
résultats obtenus d’après les méthodes proposées dans son domaine d’application.
2 Termes et définitions
Pour les besoins du présent document, les termes et définitions suivants s'appliquent.
2.1 Types des sols et autres matériaux du sol
2.1.1
sol
couche supérieure de la croûte terrestre composée de particules minérales, de matière organique, d’eau, d’air
et d’organismes
[ISO 11074-1:1996, définition 5.4]
2.1.2
matériau du sol
ensemble des terres excavées, des matériaux de dragage, des sols artificiels, des sols traités et des
matériaux de remblai
[ISO 15176:2002, définition 3.1.4]
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ISO 15799:2003(F)
2.1.3
terre excavée
tout type de matériau naturel extrait du sol, y compris le sol superficiel, le sous-sol, la roche mère altérée et la
roche mère elle-même
NOTE La terre excavée est généralement mise à jour lors de travaux de construction.
[ISO 15176:2002, définition 3.1.5]
2.1.4
sol étalon
sol prélevé sur le terrain dont les propriétés principales (par exemple pH, texture, teneur en matières
organiques) se situent dans une plage connue
[34]
EXEMPLE Euro-sols .
2.2 Termes relatifs aux caractéristiques du sol
2.2.1
fonction d’habitat
aptitude des sols/matériaux du sol à assurer l’habitat des micro-organismes, des plantes, des animaux vivant
dans le sol, et leurs interactions (biocénoses)
2.2.2
fonction de rétention
aptitude des sols/matériaux du sol à adsorber les polluants de manière à ce qu’ils ne soient pas mobilisés via
l’eau et transférés vers la chaîne alimentaire
NOTE Les fonctions d’habitat et de rétention comprennent les fonctions du sol suivantes conformément à
l’ISO 11074-1:
contrôle des cycles des substances et de l’énergie en tant que compartiments d’écosystèmes;
base de la vie pour les plantes, les animaux et l’homme;
vecteur de réserve génétique;
base de production de produits agricoles;
tampon empêchant le transfert via l’eau, de contaminants ou d’autres agents vers les nappes phréatiques.
2.2.3
polluant
substance ou agent présent dans le sol et qui, de par ses propriétés, quantités ou concentration, a des effets
négatifs (nocifs) sur les fonctions du sol ou l’utilisation du sol
cf. contaminant (2.2.4), substance potentiellement dangereuse (2.2.5)
[ISO 15176:2002, définition 3.2.7]
NOTE Voir l’Introduction de l’ISO 11074-1:1996.
2.2.4
contaminant
substance ou agent présent dans le sol résultant de l’activité humaine
cf. polluant (2.2.3), substance potentiellement dangereuse (2.2.5)
NOTE Cette définition n’implique nullement que la présence de contaminant a un effet négatif.
[ISO 15176:2002, définition 3.2.6]
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ISO 15799:2003(F)
2.2.5
substance potentiellement dangereuse
substance pouvant être dangereuse pour les êtres humains ou l’environnement lorsqu’elle est présente en
quantité ou concentration suffisante
NOTE Sa présence peut être due à l’activité humaine [contaminant (2.2.4)] ou à une cause naturelle.
[ISO 15176:2002, définition 3.2.8]
2.3 Terrain et sites
2.3.1
réutilisation
utilisation sans risque et avec profit des matériaux du sol
NOTE Dans le cadre de la présente Norme internationale, le terme «réutilisation» signifie le transfert des matériaux
du sol vers d’autres lieux pour une utilisation dans l’agriculture, l’horticulture, l’exploitation forestière, les jardins, les zones
de loisirs et les chantiers.
[ISO 15176:2002, définition 3.4.1]
3 Applications des essais écotoxicologiques
3.1 Utilisation des sols et zones de sols où il convient d’envisager des essais
écotoxicologiques:
évaluation de l’aptitude d’un sol à supporter la biocénose naturelle ou l’agriculture;
évaluation de l’écotoxicité cumulée de tous les contaminants biodisponibles présents dans les sols ou les
matériaux du sol;
évaluation de l’écotoxicité des substances potentiellement dangereuses dans les cas où le sol/les
matériaux du sol peuvent affecter les eaux souterraines et superficielles;
identification des sols ou des matériaux du sol (remblais, talus) faiblement contaminés (généralement
jusqu’à une profondeur de 1 m) qui peuvent demeurer sur le site sans traitement supplémentaire;
détection de l’écotoxicité potentielle n’ayant pu être mise en évidence par une analyse chimique;
surveillance et contrôle du succès du traitement d’un sol (hors site, sur site, in situ);
surveillance et contrôle des sols/matériaux du sol décontaminés et devant être mis en place sous forme
de remblais.
3.2 Utilisation de sols et zones de sols où les essais écotoxicologiques ne sont pas
nécessaires (sous réserve de pouvoir exclure toute contamination des eaux souterraines):
sols contaminés classés comme déchets dangereux ou pouvant être clairement caractérisés par des
paramètres analytiques/chimiques; dans de tels cas, les essais écotoxicologiques peuvent être utiles en
vue d’une étude finale après remédiation et pour le contrôle du procédé pendant la remédiation
biologique;
zones à usage commercial/industriel sans perspective d’utilisation horticole/agricole;
matériaux du sol ou matériaux de remblayage sur un terrain qui devra être rendu imperméable car il sera
recouvert de bâtiments ou de toute autre forme de revêtement à faible perméabilité comme le béton, le
macadam ou l’asphalte.
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ISO 15799:2003(F)
4 Choix des essais suivant l’utilisation/la réutilisation des sols et des matériaux du
sol et selon les fonctions du sol
4.1 Utilisation d’essais d’écotoxicité
Les toxiques peuvent altérer différentes espèces (et dans certains cas, les génotypes) présentes dans les
écosystèmes à diverses concentrations. L’approche idéale pour effectuer une caractérisation
écotoxicologique précise de la toxicité du sol consiste à utiliser une batterie d’essais faisant intervenir
plusieurs espèces appartenant à différents groupes taxonomiques et trophiques et ce, afin d’éviter des
résultats négatifs erronés dus à une adaptation d’un système d’essai (décalage génotypique) à un
contaminant spécifique par comparaison avec des sols non contaminés. Des études de terrain ou mixtes sont
rarement effectuées et peuvent s’avérer très onéreuses.
La réalisation d’un programme idéal peut être facilitée par l’adoption de stratégies d’essai simples et par
l’application de coefficients de sécurité aux résultats obtenus. Cependant, la diversité de la sensibilité des
espèces aux toxiques est telle qu’elle engendre un degré élevé d’incertitude si les essais ne portent que sur
une seule espèce ou fonction. Il est donc recommandé de soumettre à l’essai au moins un processus
microbien, une espèce du règne végétal et une du règne animal, généralement une espèce
saprophage/détritivore ; si plusieurs espèces animales font l’objet d’essais, il convient d’inclure une espèce
prédatrice dans la batterie d’essais. Le nombre minimal d’espèces à étudier dépend des réglementations
auxquelles la stratégie d’essai doit satisfaire. La présente Norme internationale n’indique que les principes de
base en vue de leur utilisation. On trouvera en 4.3 d’autres critères de choix d’essais utilisant des organismes
vivant dans le sol.
4.2 Critères généraux pour le choix des essais
Les critères de choix des essais d’écotoxicologie ont été établis dans le contexte de l’évaluation des dangers
et de la classification des produits chimiques. Il convient que ces critères s’appliquent également à la
caractérisation écotoxicologique des sols contaminés. Les critères étudiés sont les suivants: validité
scientifique, importance écologique, faisabilité et acceptabilité (voir [30] et [31]).
Les exigences fondamentales que les protocoles d’essai doivent satisfaire afin d’être stipulés dans des
Normes internationales comprennent la reproductibilité, la validité statistique, l’acceptation générale et les
performances.
L'importance d'un critère est spécifique à chaque situation. Selon le cas, des règles de sélection des critères
les plus importants doivent être déterminées, de même que doivent être définis les essais qui nécessitent des
ajustements en fonction de considérations pratiques telles que la facilité de mise en culture des organismes
au laboratoire, ou la disponibilité continue des stades de croissance appropriés à la réalisation des essais tout
au long de l'année.
Les méthodes d’essai recommandées dans le présent document ont été conçues initialement pour
l’évaluation des dangers liés aux produits chimiques et ont été, dans la plupart des cas, harmonisées au
niveau international (par exemple par l’ISO, l’OCDE ou l’UE). Dans la plupart d’entre elles, des dispositions
ont été prises pour adapter la conception de l’essai aux besoins énoncés dans le domaine d’application de la
présente Norme internationale. Cependant, dans de nombreux cas, il faut encore acquérir de l’expérience en
ce qui concerne l’application de ces méthodes d’essai à la caractérisation de la qualité du sol. En outre, le
choix des méthodes d’essai écotoxicologique pour l’évaluation des sols/matériaux du sol dépend de
l’utilisation/de la réutilisation prévue et des fonctions du sol à protéger, en particulier ses fonctions de rétention
et d’habitat.
Le Tableau 1 donne un exemple de programme décisionnel fondé sur la fonction pertinente.
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ISO 15799:2003(F)
Tableau 1 — Pertinence des essais écotoxicologiques vis-à-vis de la réutilisation du sol prévue
Fonction du sol
Fonction de rétention Fonction d’habitat
Réutilisation des sols
organismes aquatiques croissance des végétaux biocénoses du sol
Détection des effets biologiques
a
Sous les zones imperméables faible faible faible
Dans les zones non imperméables, à élevé faible faible
usage commercial et industriel
Couverture de décharge élevé élevé faible
Zones vertes, parcs et zones de élevé élevé élevé
loisirs
Zones utilisées en horticulture ou élevé élevé élevé
agriculture
a
S’applique uniquement à une zone non saturée du sol.
4.3 Considérations relatives à l’examen des fonctions du sol
4.3.1 Fonction de rétention
Le transport de fractions solubles, colloïdales ou particulaires par l’eau joue un rôle prédominant dans
l’évaluation des risques induits par les sols contaminés. En effet, non seulement l’eau peut mobiliser les
contaminants mais ces derniers et leurs métabolites peuvent avoir de graves effets sur les micro-organismes,
les plantes et la faune du sol.
Les éluats aqueux sont utiles pour déterminer les effets écotoxiques sur les organismes exposés du fait d’un
transport par l’eau. Il convient de tenir compte, d’une part, du fait que les substances mobilisées par l’eau
peuvent être soumises à différents types de modifications, par exemple le métabolisme ou l’hydrolyse,
lorsqu’elles sont transportées vers les eaux souterraines ou les eaux superficielles et d’autre part, du fait que
leur concentration peut également diminuer au cours du temps par dilution. De plus, des substances peuvent
être mobilisées dans le temps en raison de variations environnementales (par exemple pH, transformation
chimique et biologique). Les éluats peuvent servir d’indicateurs précoces de la contamination des eaux
interstitielles et souterraines avant que cette contamination atteigne les eaux de surface et les eaux potables.
Au vu de ces considérations, l’étude des eaux souterraines et des éluats est de la plus haute importance,
quelle que soit l’utilisation prévue pour le sol.
4.3.2 Fonction d’habitat
4.3.2.1 Représentativité des processus et organismes
La meilleure façon d’étudier l’aptitude d’un sol à héberger des organismes vivants consiste à mettre en œuvre
des méthodes d’essai comprenant des processus et organismes représentatifs des différents groupes
taxonomiques.
4.3.2.2 Matériaux du sol utilisés comme témoins pour les essais biologiques effectués sur des
matrices solides
Comme principe général de tous les essais écotoxicologiques, tous les critères d’effet mesurés lors d’un
traitement sont comparés à ceux mesurés sur le ou les témoins.
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ISO 15799:2003(F)
Pour évaluer l’aptitude du sol à héberger des organismes vivants, il est d’abord nécessaire de comparer le sol
ou les matériaux du sol contaminés avec un matériau témoin qui peut également servir à préparer les séries
de dilution avec l’échantillon contaminé.
Il est possible d’utiliser différents types de matériaux témoins:
un sol non contaminé ayant des propriétés pédologiques comparables à celles de l’échantillon étudié;
un matériau inerte (par exemple, du sable quartzeux);
un sol naturel certifié (par exemple un sol étalon);
un sol artificiel normalisé (voir l’ISO 11267, l’ISO 11268-1 et l’ISO 11268-2).
Le choix entre ces matériaux témoins dépend entièrement des objectifs de l’évaluation écotoxicologique, du
type d’essai biologique effectué et des exigences de l’organisme d’essai. Cette recommandation ne peut pas
être généralisée à tous les essais biologiques. L’ajout de sable à un sol ou à des matériaux du sol peut créer
un mélange compact incompatible avec le développement et la croissance de nombreux organismes (par
exemple des essais de croissance de plante). Il est préférable d’utiliser un matériau témoin plus complexe (tel
que du sol artificiel) pour la dilution, si cela peut offrir l’avantage de reproduire de manière plus fidèle
l’environnement naturel des organismes, même s’il peut interagir avec les polluants. L’introduction d’un
organisme dans un milieu qui ne correspond pas aux principales caractéristiques de son habitat naturel peut
engendrer un stress.
Si l’on a besoin d’une courbe dose-réponse, il est permis d’utiliser l’un des matériaux témoins
susmentionnés pour diluer le substrat contaminé.
Si l’objectif est de classer chaque échantillon de sol ou de matériaux du sol en termes de risque
d’écotoxicité, il est préférable d’utiliser un matériau inerte (par exemple du sable quartzeux) qui
n’interagira pas avec les polluants présents dans l’échantillon et dont la composition et la granulométrie
peuvent être rigoureusement normalisées.
Les exigences du matériau témoin doivent tenir compte des différentes utilisations du sol ainsi que de son
type et de son origine (par exemple sol non perturbé, matériau de remblayage, terre excavée, sol ayant fait
l’objet d’une remédiation). Une carence en nutriments, de même que des conditions physiques inadaptées,
peut engendrer des différences de croissance entre les végétaux et des différences de comportement entre
les animaux qui ne sont pas nécessairement dues à la situation liée aux polluants et au potentiel de danger.
Si l’objectif est d’évaluer l’écotoxicité d’un échantillon de sol ou de matériaux du sol, prélevé dans un site
contaminé, il serait préférable d’utiliser un matériau témoin non contaminé similaire à l’échantillon étudié.
Si l’objectif est d’évaluer l’écotoxicité de sols ou de matériaux du sol pouvant être réemployés pour
certaines utilisations spécifiques, il serait préférable d’utiliser comme matériau témoin tout matériau apte
à être ultérieurement mélangé avec le sol ou les matériaux du sol en question.
4.3.2.3 Sol utilisé comme substrat (milieu) pour les micro-organismes présents dans le sol
La microflore du sol représente en moyenne 80 % de la masse des organismes qui vivent dans le sol. En
combinaison avec la microfaune, les principales fonctions de la microflore sont la décomposition et la
dégradation des substances organiques complexes en nutriments facilement disponibles, maintenant ainsi les
cycles naturels des substances carbonée, azotée, phosphorée et soufrée.
La respiration induite par le substrat fournit un indicateur de la densité de la population microbienne.
Les bactéries nitrifiantes, qui sont responsables de l’oxydation de l’ammonium en nitrites et des nitrites en
nitrates, constituent un groupe très sensible de micro-organismes. Une réduction de la nitrification ne conduit
pas nécessairement à des changements significatifs dans l’écosystème. Toutefois, elle peut être utilisée
comme un indicateur sensible de l’inhibition d’un processus essentiel du sol.
6 © ISO 2003 — Tous droits réservés
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ISO 15799:2003(F)
La détermination de la biomasse microbienne ou de tout autre processus microbien dans les sols a pour objet
de permettre l’évaluation du maintien en continu de la fertilité du sol, de l’aptitude potentielle à dégrader les
composés organiques et des effets induits par les matériaux ajoutés sur la communauté microbienne
tellurique.
4.3.2.4 Sol utilisé comme substrat pour la croissance de végétaux
Après les micro-organismes, les racines des plantes représentent la plus grande surface biologique dans le
sol. Leur surface de contact avec les particules du sol est accrue par la présence de filaments racinaires et
d’associations mycorhiziennes (mycorhizes à vésicules et arbuscules avec les plantes cultivées et
ectomycorhizes supplémentaires avec les plantes ligneuses).
Comme dans le cas des autres essais biologiques proposés, les essais effectués avec des plantes
supérieures sont conçus pour permettre l’évaluation de la biodisponibilité et des effets des polluants,
respectivement décelés ou non par une analyse chimique. En retenant une période d’essai d’au moins
14 jours, les modifications à court terme produites dans le sol par la plante en essai proprement dite sont
prises en compte.
L’accumulation des polluants dans les plantes, leur métabolisme et leurs effets sur les consommateurs ne
sont pas étudiés dans le cadre de ces essais. Ils ne s’appliquent pas à l’évaluation de la fertilité et de la
productivité du sol.
4.3.2.5 Sol utilisé comme substrat pour la faune vivant dans le sol
La faune du sol remplit généralement les quatre fonctions suivantes:
a) activités mécaniques (drainage, aération, mélange, fragmentation mécanique);
b) modifications chimiques (augmentation de la disponibilité des nitrates et phosphates provenant des
excréments et formation accélérée de complexes argilo-humiques une fois que le substrat est issu de la
digestion);
c) modifications biologiques (répartition des micro-organismes dans la matrice du sol, effets synergiques par
stimulation de l’activité microbienne et décomposition de la matière organique);
d) maillons significatifs de la chaîne alimentaire.
On dispose d’essais à court et long terme pour examiner les effets des polluants sur la faune du sol. Pour
contrôler la fonction d’habitat, il est particulièrement recommandé de procéder à une caractérisation au moyen
de paramètres d’essais sublétaux.
Il convient d’utiliser une batterie d’essais car une seule méthode d’essai ne peut représenter correctement la
multitude des invertébrés extrêmement différents. Lors du choix d’une espèce particulière pour essai, il
convient de prendre en compte les critères suivants:
a) niveau trophique — il convient d’inclure, par exemple, les espèces saprophages et prédatrices;
b) groupes taxonomiques/physiologiques — il faut au moins choisir des représentants des annélides et des
arthropodes afin de couvrir la biodiversité des communautés du sol;
c) classe de taille/voie d’exposition: les espèces de la micro-, méso- et macro-faune ne représentent pas
seulement diverses classes de tailles mais également des styles
...
INTERNATIONAL ISO
STANDARD 15799
First edition
2003-11-15
Soil quality — Guidance on the
ecotoxicological characterization of soils
and soil materials
Qualité du sol — Lignes directrices relatives à la caractérisation
écotoxicologique des sols et des matériaux du sol
Reference number
ISO 15799:2003(E)
©
ISO 2003
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ISO 15799:2003(E)
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ii © ISO 2003 — All rights reserved
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ISO 15799:2003(E)
Contents Page
Foreword. iv
Introduction . v
1 Scope. 1
2 Terms and definitions. 1
2.1 Types of soil and other soil materials. 1
2.2 Terms relating to soil characteristics . 2
2.3 Land and sites . 2
3 Field of application . 3
3.1 Soils and areas of soil use where ecotoxicological tests should be considered:. 3
3.2 Soils and areas of soil use where ecotoxicological tests are not necessary (provided
groundwater contamination can be excluded): . 3
4 Selection of tests according to use/re-use of soils and soil materials and soil functions . 3
4.1 Usefulness of ecotoxicity tests . 3
4.2 General criteria for selection of tests . 4
4.3 Considerations for the examination of soil functions. 4
5 Sampling, transport, storage and sample preparation . 7
6 Limitations of proposed biotests for soils/soil materials . 7
Annex A (informative) Standardized forms of recommended test systems. 8
Bibliography . 31
© ISO 2003 — All rights reserved iii
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ISO 15799:2003(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 15799 was prepared by Technical Committee ISO/TC 190, Soil quality, Subcommittee SC 7, Soil and site
assessment.
iv © ISO 2003 — All rights reserved
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ISO 15799:2003(E)
Introduction
The majority of existing ecotoxicological test methods (biotests) being internationally harmonized were
developed to describe the ecotoxic potential of a test substance when added to a soil or soil material. These
methods can be used, with some modification, for the ecotoxicological characterization of soils and soil
materials with respect to their function and depending on the intended use. However, in such cases, users of
the methods need to be aware that the validation of the methods is not complete.
For substances with properties resulting in toxic effects, biotests are a complement to conventional chemical
analysis. Results from chemical analysis can be used for ecotoxicological assessments based on information
on the substances identified, including properties of the chemicals, e.g. their bioaccumulation potential. This
information is often scarce (if it exists at all) and does not include possible interactions (synergy/antagonism)
between chemicals and the complex soil matrix. Furthermore, an exhaustive identification and quantification of
substances is impractical. Therefore, ecotoxicological testing of soils can be used for investigating the
potential toxicity of complex chemical mixtures. The extrapolation from laboratory tests to field conditions
requires adequate consideration of important environmental factors within the test conditions and the selection
of suitable ecotoxicological endpoints.
This International Standard is one of a series providing guidance on soils and soil materials in relation to
certain functions and uses, including wildlife conservation, and ought to be read in conjunction with those
other standards.
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INTERNATIONAL STANDARD ISO 15799:2003(E)
Soil quality — Guidance on the ecotoxicological
characterization of soils and soil materials
1 Scope
This International Standard 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 and possible adverse effects on aquatic and soil-dwelling organisms, and
habitat maintenance and the retention function of the soil.
It does not cover tests for bioaccumulation. Genotoxicity tests using eukaryotic organisms in soils are not yet
available. It is not applicable to the ecological assessment of uncontaminated soils with a view to natural,
agricultural or horticultural use, such soils being of possible interest where they can serve as a reference for
the assessment of soils from contaminated sites. Nor is the interpretation of the results gained by application
of the proposed methods within its scope.
2 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
2.1 Types of soil and other soil materials
2.1.1
soil
upper layer of the Earth’s crust composed of mineral particles, organic matter, water, air and organisms
[ISO 11074-1:1996, definition 5.4]
2.1.2
soil material
excavated soil, dredged materials, manufactured soils, treated soils and fill materials
[ISO 15176:2002, definition 3.1.4]
2.1.3
excavated soil
any natural material excavated from the ground, including top soil, sub soil, altered parent rock and parent
rock itself
NOTE Excavated soil typically arises during construction works.
[ISO 15176:2002, definition 3.1.5]
2.1.4
standard soil
field-collected soil whose main properties (e.g. pH, texture, organic matter content) are within a known range
[34]
EXAMPLE Eurosoils .
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ISO 15799:2003(E)
2.2 Terms relating to soil characteristics
2.2.1
habitat function
ability of soils/soil materials to serve as a habitat for micro-organisms, plants, soil-living animals and their
interactions (biocenoses)
2.2.2
retention function
ability of soils/soil materials to adsorb pollutants in such that they cannot be mobilized via the water pathway
and translocated into the food chain
NOTE The habitat and retention functions include the following soil functions according to ISO 11074-1:
control of substance and energy cycles as components of ecosystems;
basis for the life of plants, animals and humans;
carrier of genetic reservoir;
basis for the production of agricultural products;
buffer inhibiting movement of water, contaminants or other agents into the ground water.
2.2.3
pollutant
substance or agent present in the soil which due to its properties, amount or concentration causes adverse
impact on soil functions or soil use
cf. contaminant (2.2.4), potentially harmful substance (2.2.5)
[ISO 15176:2002, definition 3.2.7]
NOTE See Introduction to ISO 11074-1:1996.
2.2.4
contaminant
substance or agent present in soil as a result of human activity
cf. pollutant (2.2.3), potentially harmful substance (2.2.5)
NOTE There is no assumption in this definition that harm results from the presence of the contaminant.
[ISO 15176:2002, definition 3.2.6]
2.2.5
potentially harmful substance
substance which, when present in sufficient concentration or amount, may be harmful to humans or the
environment
NOTE It may be present as a result of human activity [contaminant (2.2.4)] or naturally.
[ISO 15176:2002, definition 3.2.8]
2.3 Land and sites
2.3.1
re-use
useful and harmless utilization of soil materials
NOTE In the context of this International Standard the re-use means the transfer of soil materials to another location
for use in agriculture, horticulture, forestry, gardens, recreational areas and construction sites.
[ISO 15176:2002, term 3.4.1]
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3 Field of application
3.1 Soils and areas of soil use where ecotoxicological tests should be considered:
assessment of the ability of a soil to sustain a natural biocenosis or agriculture;
assessment of the combined ecotoxicity of all bioavailable contaminants present in soils or soil materials;
assessment of the ecotoxicity of potentially harmful substances in cases where the soil or soil material
can affect the ground and surface water;
identification of soils or soil materials (refills, embankments) having a low degree of contamination —
usually within a depth of 1 m — and which can remain at the site without further treatment;
detection of potential ecotoxicity which could not be traced by chemical analysis;
monitoring and control of the success of soil treatment (off-site, on-site, in situ);
monitoring and control of soils/soil materials that have been decontaminated and are to be applied at the
surface.
3.2 Soils and areas of soil use where ecotoxicological tests are not necessary (provided
groundwater contamination can be excluded):
contaminated soils classified as hazardous waste or which can be characterized clearly by
chemical/analytical parameters, in which cases ecotoxicological testing could be useful for a final
investigation after remediation and for process control during biological remediation;
commercially/industrially used areas with no prospect of horticultural/agricultural use;
soil materials or backfilled materials in an area to be effectively sealed by covering with buildings or other
forms of low permeability cover such as concrete or tarmacadam or asphalt.
4 Selection of tests according to use/re-use of soils and soil materials and soil
functions
4.1 Use of ecotoxicity tests
Toxicants can affect different species (and in some cases genotypes) present within ecosystems at different
concentrations. The ideal approach for the precise ecotoxicological characterization of the soil toxicity is to
use a battery of tests with several species belonging to different taxonomic and trophic groups, in order to
avoid false negative results owing to an adaptation of a test system (genotypic shift) to a specific contaminant
as compared to uncontaminated soils. Studies using field or semi-field investigations are rarely carried out and
can be very expensive.
The ideal scheme can be rendered more practicable by the adoption of simpler testing strategies and the
application of safety factors to the results obtained. If, however, testing is performed on one species or
function only, the high diversity in the sensitivity of species to toxicants will result in a high level of uncertainty.
It is therefore recommended to test at least a microbial process, a species from the plant kingdom, and one
from the animal kingdom, usually a saprophagous/detritivorous species; if more than one animal species is
tested, a predatory species should be included in the test battery. The minimum number of species to be
tested depends on the regulations to which the test strategy must comply. This International Standard only
gives the basic principles for their use. Further considerations to the selection of tests using soil organisms are
given in 4.3.
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4.2 General criteria for selection of tests
Criteria for the selection of ecotoxicity tests have been established in the context of hazard assessment and
classification of chemicals. These criteria should also apply for the ecotoxicological characterization of
contaminated soils. Criteria reviewed were: scientific validity, ecological significance, practicability and
acceptability. See [30] and [31].
The basic requirements that test protocols must meet in order to be established in international standards
include reproducibility, statistical validity, general acceptance and performance.
The importance of a criterion is relative to the specific situation. Decisions have to be made as to which
criteria are the most important or which tests may have to be modified, and choices made between these and
more practical considerations — for example, easy culturing of test organisms in the laboratory or the
availability of life stages required for a test throughout the year.
The test methods recommended here were originally designed for hazard assessment of chemicals and were
in most cases internationally harmonized (e.g. by OECD, EU or ISO). In most of the methods, provisions have
been made to adapt the test design to come within the scope of this International Standard. Nevertheless, in
many cases experience still has to be gained using the test methods for characterizing soil quality. In addition,
the selection of ecotoxicological test methods for the assessment of soils or soil materials depends on their
intended use or re-use and on the soil functions to be protected, in particular the retention and habitat
functions.
Table 1 gives an example of a decision scheme based on the relevant function.
Table 1 — Relevance for ecotoxicological testing to the intended re-use of the soil
Soil function
Retention function Habitat function
Re-use of soils
Aquatic organisms Plant growth Soil biocenoses
Detection of biological effects
a
Below sealed areas low low
low
Commercially and industrially used unsealed areas high low low
Landfill covering high high low
Green areas, parks and recreation areas high high high
Areas used in horticulture or agriculture high high high
a
Applies only to the unsaturated soil zone.
4.3 Considerations for the examination of soil functions
4.3.1 Retention function
Transport via water of soluble, colloidal or particle fractions plays a dominant role in the risk assessment of
contaminated soils. This is true not only because water may mobilize contaminants, but also because
contaminants and metabolites in the water phase potentially have a severe effect on micro-organisms, plants
and soil fauna.
Aqueous eluates are useful for testing ecotoxic effects on organisms exposed via the water-mediated
transport. It should be taken into account that substances mobilized via water could be subjected to different
types of changes, (e.g. metabolism or hydrolization) when transported into the groundwater and from there
into surface waters, and that their concentrations are reduced by dilution. Moreover, substances may be
mobilized over time due to environmental changes (e.g. pH, chemical and biological transformation). Eluates
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may serve as early indicators for the contamination of pore and ground water prior to the exposure of surface
and drinking water.
With these aspects, the investigation of groundwater and eluates is of the utmost importance — regardless of
the proposed soil use.
4.3.2 Habitat function
4.3.2.1 Representativeness of organisms and processes
The suitability of the soil for living organisms can best be examined by means of test methods selected to
include organisms and processes representative of different taxonomic groups.
4.3.2.2 Soil material used as control for bioassays on solid matrices
As a general principle in ecotoxicological testing, any end points measured in a treatment are compared with
those measured in the control or controls.
When evaluating the suitability of the soil for soil-dwelling organisms, it is a prerequisite to compare the
contaminated soil or soil material with a control material, which may also be used for preparing dilution series
with the contaminated sample.
Several types of control material can be used:
an uncontaminated soil with comparable pedological properties to the sample being tested;
an inert material (e.g. quartz sand);
a certified natural soil (e.g. standard soil);
a standardized artificial soil (see ISO 11267, ISO 11268-1 and ISO 11268-2).
The choice between these control materials should be made depending entirely on the aims of the
ecotoxicological assessment, the type of biological test being carried out and the requirements of the test
organism. This recommendation cannot be generalized for all biological tests. Adding sand to a soil or a soil
material can create a compact mixture incompatible with the growth and development of many organisms (e.g.
plant growth tests). It is preferable to use a more complex control material (such as artificial soil) for dilution
where this would have the advantage of reproducing more closely the natural environment of the organisms
and even if it may interact with pollutants. Placing an organism in a medium that does not match the most
important characteristics of its natural habitat may cause stress.
If a dose-response curve is needed, one of the control materials mentioned above may be used to dilute
the contaminated substrate.
If the aim is to classify each sample of soil or soil materials in terms of ecotoxicity hazard, it is preferable
to use an inert material (e.g. quartz sand), which will not interact with the pollutants present in the sample
and whose composition and granulometry can be rigorously standardized.
The requirements of the control material must take into account the different soil uses and the type and origin
of the soil (e.g. undisturbed soil, refilling material, excavated soil, remediated soil). Nutrient deficiency, as well
as physical conditions, can cause differences in plant growth and animal behaviour that need not necessarily
be caused by the pollutant situation and the hazard potential.
If the aim is to evaluate the ecotoxicity of a soil or soil material sample from a contaminated site, the
preferred method would be to use an uncontaminated control material similar to the sample being tested.
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If the aim is to evaluate the ecotoxicity of soils or soil materials which may be re-employed for certain
specific uses, the preferred method would be to use as a control material any material which may in
future be mixed with the soil or soil material.
4.3.2.3 Soil as substrate (medium) for soil micro-organisms
The soil microflora comprises on average 80 % of the mass of organisms living in soil. In combination with the
microfauna, the main functions of the microflora are the decomposition and degradation of complex organic
substances to easily available nutrients, thereby maintaining the natural substance cycles of carbon, nitrogen,
phosphorus and sulfur.
Substrate-induced respiration provides an indicator of the microbial population density.
Nitrifying bacteria, which are responsible for the oxidation of ammonium to nitrite and from nitrite to nitrate, are
a very sensitive group of micro-organisms. Reduced nitrification need not necessarily lead to significant
changes in the ecosystem but can be used as a sensitive indicator for the inhibition of an essential soil
process.
The purpose of determining the microbial biomass or other microbial processes in soils is to allow assessment
of the continued maintenance of soil fertility, the potential ability to degrade organic compounds, and the
effects of added materials on the soil microbial community.
4.3.2.4 Soil as substrate for plant growth
After micro-organisms, plant roots constitute the largest biological surface in soil. Their contact area with soil
particles is increased by the presence of root hairs and mycorrhizal associations (VA-mycorrhiza with
cultivated plants and additional ectomycoorhizae with woody plants).
As with the other bioassays proposed, tests with higher plants are designed to assess the bioavailability and
effects of pollutants detected or not detected by chemical analysis, respectively. By applying a test period of at
least 14 days, short-term changes in the soil by the test plant itself are included.
The accumulation of pollutants in plants, their metabolism and their effects on consumers are not investigated
in these tests. They do not apply to the assessment of soil fertility and productivity.
4.3.2.5 Soil as substrate for soil living fauna
Soil animals generally fulfil the following four functions:
a) mechanical activities (drainage, aeration, mixing, mechanical comminution);
b) chemical changes (enhanced availability of nitrate and phosphate from excrements and accelerated
formation of clay-humus complexes, after the substrate has passed the gut);
c) biological changes (distribution of micro-organisms in the soil matrix, synergistic effects through
stimulation of microbial activity and organic matter decomposition);
d) significant links in the food web.
Short-term and long-term tests are available for examination of the effects of pollutants on soil fauna. For
testing the habitat function, characterization by sublethal test parameters is particularly recommended.
Since a single test method cannot adequately represent the vast number of very diverse invertebrates, a test
battery should be used. When selecting the individual test species, the following criteria should be considered:
a) trophic level — e.e. saprophagous and predatory species should be included;
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b) taxonomic/physiological groups — in order to cover the biodiversity of soil communities, at least
representatives of annelids and arthropoda have to be selected;
c) size class/exposure pathway: species of the micro-, meso- and macrofauna do not only represent various
size classes but also different life-styles and therefore exposure routes (e.g. pore water versus food
uptake);
d) ecological role — at least soil-dwelling and litter-inhabiting species are important to consider.
Only internationally standardized methods should be used.
5 Sampling, transport, storage and sample preparation
Before soil quality is assessed by any of the methods proposed, soil samples need to be collected from the
site under investigation. Soil sampling should be carried out by trained personnel with sufficient knowledge of
sampling, handling of samples and safety measures at contaminated sites and sampling locations. The
sampling strategy and handling should be determined by the site to be investigated, the kind of contamination
and the aim of the biological tests (e.g. quantities of soil samples could vary between 100 mg and 100 kg,
depending on the tests selected).
Record all data concerning sampling, transport and sample preparation. For instructions on the design of
sampling programmes, sampling techniques, safety, investigations of natural, cultivated, urban and industrial
sites and on the collection, handling and storage of soil for the assessment of aerobic/anaerobic microbial
processes in the laboratory, see ISO 10381-6.
6 Limitations of proposed biotests for soils/soil materials
Biological test systems are suitable for volatile pollutants only to a limited extent. Other methods should be
developed for this purpose. Similarly, the impact of organic contaminants, which are easily degradable under
aerobic conditions, may be detected incompletely by the methods described. In this case, alternative methods
for sampling and sample preparation should be applied.
NOTE The proposed terrestrial and aquatic test methods in A.1 and A.2 were developed to assess the ecotoxic
potential of chemicals. The characterization of soils or soil eluates was not their primary goal. Therefore, the methods
need to be adapted to the specific requirements of soil and site assessment.
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Annex A
(informative)
Standardized forms of recommended test systems
A.1 Terrestrial test methods
A.1.1 Soil fauna
A.1.1.1 Collembola — Effects on reproduction
See Table A.1.
Table A.1
1. Title of test Soil quality — Inhibition of reproduction of Collembola (Folsomia candida) by
soil pollutants
2. Harmonization International
3. Reference ISO 11267
4. Principle Determination of the effect on reproduction of springtails incubated over a
4 weeks test period
5. Test type Static subchronic
6. Test organism Springtails
Breeding stocks Folsomia candida Willem 1902
Age 10 d to 12 d
Feeding Dry yeast
7. Test substrate Artificial soil, contaminated soil
Volume 30 g (wet mass)/container
8. Test conditions
Test chamber Enclosures
Temperature 20 °C ± 2 °C
pH 6 ± 0,5
Light intensity/quality Between 400 lx and 800 lx
Photoperiod 12 h:12 h or 16 h:8 h
Soil moisture 40 % to 60 % of total water holding capacity
9. No. replicates 4
10. Test duration/incubation 28 d
11. Neg. control/ dilution soil Artificial soil
12. Validity criteria Control: mortality < 20 %, min. reproduction 100 juveniles, CV u 30 %
13. Pos. control/reference toxicant E 605 forte (a.i. 507,5 g/l) Betanal plus (a.i. 160 g/l)
Mean EC50, CV LOEC: 0,18 mg/kg to 0,32 mg/kg; 100 mg/kg to 200 mg/kg
14. Statistics Multiple t-test, u-test, regression analysis
15. Test parameter(s) Mortality of adults, inhibition of reproduction
16. End points EC (x = % effect level, e.g. 10, 50), NOEC
x
17. Limitations/comments
The test was originally designed for testing substances added to an artificial soil. To compare or to monitor soil quality,
the method has to be adapted. Care should be taken that any control soil used meets the biological requirements of the
test species. The number of replicates might have to be increased because of the heterogeneity of field samples.
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