prEN ISO 15799

SLOVENSKI STANDARD
01-februar-2026
Kakovost tal - Navodilo za ekotoksikološko karakterizacijo tal in talnih materialov
(ISO/DIS 15799:2025)
Soil quality - Guidance on the ecotoxicological characterization of soils and soil materials
(ISO/DIS 15799:2025)
Bodenbeschaffenheit - Anleitung zur ökotoxikologischen Charakterisierung von Böden
und Bodenmaterialien (ISO/DIS 15799:2025)
Qualité du sol - Lignes directrices relatives à la caractérisation écotoxicologique des sols
et des matériaux de type sol (ISO/DIS 15799:2025)
Ta slovenski standard je istoveten z: prEN ISO 15799
ICS:
13.080.99 Drugi standardi v zvezi s Other standards related to
kakovostjo tal soil quality
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

DRAFT
International
Standard
ISO/DIS 15799
ISO/TC 190/SC 4
Soil quality — Guidance on the
Secretariat: AFNOR
ecotoxicological characterization of
Voting begins on:
soils and soil materials
2025-12-23
Qualité du sol — Lignes directrices relatives à la caractérisation
Voting terminates on:
écotoxicologique des sols et des matériaux du sol
2026-03-17
ICS: 13.080.99
THIS DOCUMENT IS A DRAFT CIRCULATED
FOR COMMENTS AND APPROVAL. IT
IS THEREFORE SUBJECT TO CHANGE
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Reference number
ISO/DIS 15799:2025(en)
DRAFT
ISO/DIS 15799:2025(en)
International
Standard
ISO/DIS 15799
ISO/TC 190/SC 4
Soil quality — Guidance on the
Secretariat: AFNOR
ecotoxicological characterization of
Voting begins on:
soils and soil materials
Qualité du sol — Lignes directrices relatives à la caractérisation
Voting terminates on:
écotoxicologique des sols et des matériaux du sol
ICS: 13.080.99
THIS DOCUMENT IS A DRAFT CIRCULATED
FOR COMMENTS AND APPROVAL. IT
IS THEREFORE SUBJECT TO CHANGE
AND MAY NOT BE REFERRED TO AS AN
INTERNATIONAL STANDARD UNTIL
PUBLISHED AS SUCH.
This document is circulated as received from the committee secretariat.
IN ADDITION TO THEIR EVALUATION AS
BEING ACCEPTABLE FOR INDUSTRIAL,
© ISO 2025
TECHNOLOGICAL, COMMERCIAL AND
USER PURPOSES, DRAFT INTERNATIONAL
All rights reserved. Unless otherwise specified, or required in the context of its implementation, no part of this publication may
STANDARDS MAY ON OCCASION HAVE TO
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Published in Switzerland Reference number
ISO/DIS 15799:2025(en)
ii
ISO/DIS 15799:2025(en)
Contents Page
Foreword .iv
Introduction .v
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 1
3.1 Types of soil and other soil materials .1
3.2 Terms relating to soil characteristics .2
3.3 Land and sites .3
4 Field of application . 3
4.1 Soils and areas of soil use where ecotoxicity tests should be considered .3
4.2 Soils and areas of soil use where ecotoxicological tests are not necessary .3
5 Selection of tests according to the use/re-use of soils and soil materials and soil
functions . 3
5.1 Use of ecotoxicity tests .3
5.2 General criteria for selection of tests .4
5.3 Considerations for the examination of soil functions .5
5.3.1 Retention function .5
5.3.2 Habitat function .5
6 Sampling, transport, storage and sample preparation . 7
7 Limitations of proposed biotests for soils/soil materials . 7
Annex A (informative) Standardized forms of recommended test systems . 8
Bibliography .44

iii
ISO/DIS 15799:2025(en)
Foreword
ISO (the International Organization for Standardization) is a worldwide federation of national standards
bodies (ISO member bodies). The work of preparing International Standards is normally carried out through
ISO technical committees. Each member body interested in a subject for which a technical committee
has been established has the right to be represented on that committee. International organizations,
governmental and non-governmental, in liaison with ISO, also take part in the work. ISO collaborates closely
with the International Electrotechnical Commission (IEC) on all matters of electrotechnical standardization.
The procedures used to develop this document and those intended for its further maintenance are described
in the ISO/IEC Directives, Part 1. In particular, the different approval criteria needed for the different types
of ISO documents should be noted. This document was drafted in accordance with the editorial rules of the
ISO/IEC Directives, Part 2 (see www.iso.org/directives).
Attention is drawn to the possibility that some of the elements of this document may be the subject of patent
rights. ISO shall not be held responsible for identifying any or all such patent rights. Details of any patent
rights identified during the development of the document will be in the Introduction and/or on the ISO list of
patent declarations received (see www.iso.org/patents).
Any trade name used in this document is information given for the convenience of users and does not
constitute an endorsement.
For an explanation of the voluntary nature of standards, the meaning of ISO specific terms and expressions
related to conformity assessment, as well as information about ISO's adherence to the World Trade
Organization (WTO) principles in the Technical Barriers to Trade (TBT) see www.iso.org/iso/foreword.html.
This document was prepared by Technical Committee ISO/TC 190, Soil quality, Subcommittee SC 4, Biological
characterization.
This third edition cancels and replaces the second edition (ISO 15799:2019), which has been technically
revised. The main changes compared to the previous edition are as follows:
— standardized forms of recommended test systems in Annex A have been amended and updated (e.g.
ISO 20130 was included and ISO 7346 was deleted).
Any feedback or questions on this document should be directed to the user’s national standards body. A
complete listing of these bodies can be found at www.iso.org/members.html.

iv
ISO/DIS 15799:2025(en)
Introduction
Most of the existing ecotoxicological test methods (biotests) that are being internationally harmonized
were developed to describe the ecotoxic potential of a test substance when added to a soil/soil material.
These methods can be used with some modifications for the ecotoxicological characterization of soils and
soil materials with respect to their function depending on the intended use. 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 it 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 parameters.
v
DRAFT International Standard ISO/DIS 15799:2025(en)
Soil quality — Guidance on the ecotoxicological
characterization of soils and soil materials
1 Scope
This document is one of a family of International Standards providing guidance on soils and soil materials in
relation to certain functions and uses including conservation of biodiversity. It applies in conjunction with
these other standards. It 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.
NOTE This is a reflection of the maintenance of the habitat and retention function of the soil. In fact, the methods
listed in this document are suitable for usage in a TRIAD approach, i.e. for an ecological assessment of potentially
contaminated soils (see Reference [18]).
This document does not cover tests for bioaccumulation.
The ecological assessment of uncontaminated soils with a view to natural, agricultural or horticultural use
is not within the scope of this document. Such soils can be of interest if they can serve as a reference for the
assessment of soils from contaminated sites (see References [29], [30] and [33]).
The interpretation of results gained by applying the proposed methods is not in the scope of this document.
2 Normative references
There are no normative references in this document.
3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
ISO and IEC maintain terminological databases for use in standardization at the following addresses:
— ISO Online browsing platform: available at https:// www .iso .org/ obp
— IEC Electropedia: available at https:// www .electropedia .org/
3.1 Types of soil and other soil materials
3.1.1
soil
upper layer of the Earth’s crust composed of mineral particles, organic matter, water, air and organisms
[SOURCE: ISO 11074:2015, 2.1.11, modified — The definition has been slightly modified and the Note 1 to
entry has been deleted.]
3.1.2
soil material
material which includes excavated soil (3.1.3), dredged materials, manufactured soils, treated soils and fill
materials
ISO/DIS 15799:2025(en)
3.1.3
excavated soil
any natural material excavated from ground including top-soil, sub-soil, altered parent rock and parent
rock itself
Note 1 to entry: Excavated soil typically arises during construction works.
[SOURCE: ISO 15176:2002 (see Reference [15]), 3.1.5]
3.1.4
standard soil
field collected soil whose main properties (e.g. pH, texture, organic matter content) are within a known range
Note 1 to entry: An example for standard soils is “Eurosoils” (see Reference [28]).
3.2 Terms relating to soil characteristics
3.2.1
habitat function
ability of soils (3.1.1)/soil materials (3.1.2) to serve as a habitat for microorganisms, plants, soil living animals
and their interactions (biocenoses)
3.2.2
retention function
ability of soils (3.1.1)/soil materials (3.1.2) to adsorb pollutants (3.2.3) in such a way that they cannot be
mobilised via the water pathway and translocated into the food chain
Note 1 to entry: The habitat and retention functions include the following soil functions according to ISO 11074:2015
(see Reference [7]):
— control of substance and energy cycles as components of ecosystems;
— basis for the life of plants, animals and man;
— carrier of genetic reservoir;
— basis for the production of agricultural products;
— buffer inhibiting movement of water, contaminants or other agents into the ground water.
3.2.3
pollutant
substance which due to their properties, amount or concentration cause impacts on the soil function or soil use
Note 1 to entry: See also contaminant (3.2.4) and potentially harmful substance (3.2.5).
[SOURCE: ISO 11074:2015, 3.4.18, modified — Wording has been slightly modified and Note 1 to entry has
been added.]
3.2.4
contaminant
substance or agent present in the soil (3.1.1) as a result of human activity
Note 1 to entry: There is no assumption in this definition that harm results from the presence of the contaminant: see
also pollutant (3.2.3) and potentially harmful substance (3.2.5).
[SOURCE: ISO 15176:2002, definition 3.2.6, modified — The wording in the Note 1 to entry has been slightly
modified.]
ISO/DIS 15799:2025(en)
3.2.5
potentially harmful substance
substance which when present in sufficient concentration or amount may be harmful to humans or the
environment
Note 1 to entry: It may be the result of human activity [contaminant (3.2.4)] or naturally occurring.
[SOURCE: ISO 15176:2002, 3.2.8, modified — A Note 1 to entry has been added.]
3.3 Land and sites
3.3.1
re-use
useful and harmless utilisation of soil materials
Note 1 to entry: In the context of this International Standard, re-use means the transfer of soil materials to another
location for use in agriculture, horticulture, forestry, gardens, recreational areas and construction sites.
[SOURCE: ISO 15176:2002, 3.4.1]
4 Field of application
4.1 Soils and areas of soil use where ecotoxicity tests should be considered
Ecotoxicity tests should be considered in the following soils and areas of soil use:
— 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/soil material can
affect the ground and surface water.
— Identification of soils or soil materials (refills, embankments) with a low degree of contamination usually
within a depth of 1 m, 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 efficacy of soil treatment (excavated soil, in situ).
— Monitoring and control of soils/soil materials, which have been decontaminated and are expected to
remain functionally active on the surface (for a final investigation after remediation and for process
control during biological remediation).
4.2 Soils and areas of soil use where ecotoxicological tests are not necessary
Provided that groundwater contamination can be excluded, ecotoxicological testing is not necessary in the
following cases.
— Commercially/industrially used areas with no prospect of horticultural/agricultural use.
— Soil materials or backfilled materials in an area which is to be effectively sealed by covering with
buildings or other forms of low permeability cover such as concrete or tarmacadam or asphalt.
5 Selection of tests according to the use/re-use of soils and soil materials and soil
functions
5.1 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

ISO/DIS 15799:2025(en)
to use a battery of tests with several species belonging to different taxonomic and trophic groups to avoid
false-negative results due to an adaption 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
may 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/detritivores species. If more than
one animal species are 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 shall comply. This
document only gives the basic principles for their use. Further considerations to the selection of tests using
soil organisms are given in 5.3.
5.2 General criteria for selection of tests
Criteria for the selection of ecotoxicity tests were 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 References [27] and [28]).
Basic requirements which test protocols shall fulfil in order to be laid down 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 between which
criteria are most important or tests which may have to be modified by more practical considerations, such
as 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 (see Annex A) in this document 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 them provisions have been made to adapt the test design for the purposes within the scope of
this document. In addition, the selection of ecotoxicological test methods for the assessment of soils/soil
materials depends on their intended use/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
Re-use of soils Soil function
Retention function Habitat function
Aquatic organisms Plant growth Soil biocenoses
Detection of biological effects
a
Below sealed areas low low low
Commercially and industrially used unsealed high low low
areas
Landfill covering high high low
Green areas, parks and recreation areas high high high
a
Applies only to the unsatured soil zone.

ISO/DIS 15799:2025(en)
5.3 Considerations for the examination of soil functions
5.3.1 Retention function
Transport via water of soluble, colloidal or particle fractions play a dominant role in the risk assessment
of contaminated soils. This is true not only because water can mobilize contaminants, but also because
contaminants and metabolites in the water phase potentially have a severe effect on microorganisms, plants
and soil fauna.
Aqueous eluates (for preparation see Clause 6, ISO 18772, ISO 21268 series, EN 14735) 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 can be subjected to different types of changes, such as metabolism
or hydrolysis when transported into the groundwater and from there into surface waters, and that
their concentrations are reduced by dilution. Moreover, substances can be mobilised over time due to
environmental changes (e.g. pH, chemical and biological transformation). Eluates can 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 utmost importance regardless of the
proposed soil use.
For ecotoxicology tests working with aqueous soil extracts and aquatic test organisms it shall be considered
that nutrient ions and compounds are easily dissolved in water (at least easier than hydrophobic pollutants)
and can substantially interfere during the test.
5.3.2 Habitat function
5.3.2.1 General
The suitability of the soil for living organisms can best be examined by means of test methods which are
selected to include organisms and processes representative of different taxonomic and ecological groups.
5.3.2.2 Soil material used as control for bioassays on solid matrices
As a general principle in ecotoxicological testing, any parameter measured in a treatment is compared with
the one measured in the control(s).
In order to evaluate 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 11268-1 and ISO 11268-2, ISO 11267).
The choice between these control materials depends entirely on the aims of the ecotoxicological assessment,
the type of biological test being carried out and the requirements of the test organism (see Reference [31]).
This recommendation cannot be generalized for all biological tests. Adding sand to a soil or a soil material
can create a compact mixture which is 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 which 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.
ISO/DIS 15799:2025(en)
— 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 shall 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 (see Reference [49]).
— 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 that is similar to the sample
being tested.
— 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 any material which may in future be
mixed with soil or soil material.
5.3.2.3 Soil as substrate (medium) for soil microorganisms
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 microorganisms. Decreased 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.
5.3.2.4 Soil as substrate for plant growth
After microorganisms, 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 ectomycorrhizal 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.
5.3.2.5 Soil as substrate for soil-inhabiting fauna
Soil animals generally fulfil the following four functions:
— mechanical activities (drainage, aeration, mixing, mechanical comminution);
— chemical changes (enhanced availability of nitrate and phosphate from excrements and accelerated
formation of clay-humus complexes, after the substrate has passed the gut);
— biological changes (distribution of microorganisms in the soil matrix, synergistic effects through
stimulation of microbial activity and organic matter decomposition);
— significant links in the food web.

ISO/DIS 15799:2025(en)
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 sub-lethal test parameters is particularly recommended as
usually more relevant.
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:
— trophic level: e.g. saprophagous and predatory species should be included;
— taxonomic/physiological groups: in order to cover the biodiversity of soil communities, at least
representatives of Annelida and Arthropoda have to be selected;
— 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);
— ecological role: at least soil-dwelling and litter-inhabiting species are important to be considered.
Only internationally standardized methods should be used.
6 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 (see ISO 23611-6, Reference [43]). 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 can
vary between 100 g and 100 kg depending on the tests selected).
Record all data concerning sampling, transport and sample preparation. 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 biological functional
and structural parameters in the laboratory are given in ISO 18400-206 (see Reference [44]). For the
preparation of eluates for testing the retention function with aquatic test methods ISO 18772, ISO 21268
series and EN 14735 are recommended (see References [22] and [23]).
7 Limitations of proposed biotests for soils/soil materials
Biological test systems are only suitable to a limited extent for volatile pollutants. 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.
The proposed terrestrial and aquatic test methods in Annex A (A.1 and A.2) were developed initially to
assess the ecotoxic potential of chemicals. More recently, most of the methods listed in Annex A have been
adapted to the specific requirements of soil, water and site assessment.

ISO/DIS 15799:2025(en)
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
1. Title of the test: Soil quality — Inhibition of reproduction of Collembola (Folsomia
candida) by soil contaminants
2. Harmonization International
3. References ISO 11267 (see Reference [8])
4. Principle Determination of the effect on reproduction of springtails incubated
over a four 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 Enclosure capable of being temperature-controlled
Temperature 20 °C ± 2 °C
pH 6 ± 0,5 for testing contaminated soils do not adjust pH
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 At least 4
10. Test duration/incubation 28 d
11. Neg. control/ dilution soil Reference or standard soil (e.g. artificial soil)
12. Validity criteria Control: mortality < 20 %, min. reproduction 100 juveniles,
CV ≤ 30 %
13. Pos. Control/reference Boric acid or Betosip (a.i. 157 g/l Phenmedipham)
toxicant Mean EC50, CV
14. Statistics ANOVA, multiple t-test, u-test, regression analysis

ISO/DIS 15799:2025(en)
15. Test parameter(s) Mortality of adults, inhibition of reproduction
16. End points EC (x = % effect level, e.g. 10, 50), NOEC, NOER
x
17. Limitations/Comments Originally the test was designed for testing substances added to an
artificial soil. The test is also applicable to soils and soil materials of
unknown quality, e.g. from contaminated sites, amended soils, soils
after remediation, industrial, agricultural or other sites under con-
cern as well as waste materials.
A.1.1.2 Earthworms — Acute toxicity
1. Title of the test: Soil quality — Effects of pollutants on earthworms — Part 1:
Determination of acute toxicity to Eisenia fetida/Eisenia andrei
2. Harmonization International
3. References ISO 11268-1 (see Reference [9])
4. Principle The percent mortality of adult earthworms (species: Eisenia fetida or
Eisenia andrei) exposed to the test soil are compared to those observed
for test organisms exposed to a control soil.
5. Test type Acute, static
6. Test organism Earthworms
Breeding stocks Eisenia fetida Savigny, E. andrei Bouché
Age > 2 months
Feeding No
7. Test substrate Test soil, standard soil (e.g. Artificial Soil)
Volume 500 g (dry mass)
8. Test conditions
Test chamber Enclosure capable of being controlled
Temperature 20 °C ± 2 °C
pH 6 ± 0,5, Field-collected soils, soil or waste materials: Soil pH should not
be corrected as it can influence bioavailability of soil contaminants.
Light intensity/quality 400 lx to 800 lx
Photoperiod Between 12 h: 12 h or 16 h: 8 h
Soil moisture 40 % to 60 % water holding capacity
9. No. Replicates 4
10. Test duration/ incubation 14 days
11. Neg. control/dilution soil Standard soil or artificial soil
12. Validity criteria Control: mortality < 10 %, biomass loss ≤ 20 %
13. Positive control/ reference Boric acid. Significant effects on mortality should be observed between
toxicant 3 000 mg and 4 500 mg of boric acid per kilogram of artificial soil (dry
mass).
14. Statistics Fisher’s exact test ANOVA
15. Test parameter(s) Mortality, biomass
16. End points LC50 – 14 d
17. Limitations/Comments Originally the test was designed for testing substances added to an
artificial soil. The test is also applicable to soils and soil materials of
unknown quality, e.g. from contaminated sites, amended soils, soils
after remediation, agricultural or other sites concerned, and waste ma-
terials. In a new Annex the test performance under tropical conditions
is described. The test has been used successfully for the assessment of
wastes. (see References [25] and [27])

ISO/DIS 15799:2025(en)
A.1.1.3 Earthworms — Effects on reproduction
1. Title of the test: Soil quality — Effects of pollutants on earthworms — Part 2: Determi-
nation of effects on reproduction of Eisenia fetida/Eisenia andrei
2. Harmonization International
3. References ISO 11268-2 (see Reference [10])
4. Principle The effects on reproduction of adult earthworms (species: Eisenia
fetida or Eisenia andrei) exposed to the test soil are compared to those
observed for
test organisms exposed to a control soil.
5. Test type Subchronic, static
6. Test organism Earthworms
Breeding stocks Eisenia fetida Savigny, E. andrei Bouché
Age between 2 months and 1 year
Feeding Cow dung
7. Test substrate Test soil, standard soil (e.g. Artificial Soil)
Volume 500 g to 600 g dry mass
8. Test conditions
Test chamber Enclosure capable of being controlled
Temperature 20 °C ± 2 °C
pH 6 ± 0,5 Field-collected soils, soil or waste materials: Soil pH should not
be corrected as it can influence bioavailability of soil contaminants.
Light intensity/quality 400 lx to 800 lx
Photoperiod Between 12 h: 12 h or 16 h: 8 h
Soil moisture 40 % to 60 % water holding capacity
9. No. Replicates 4
10. Test duration/ incubation 8 weeks
11. Neg. control/ dilution soil Standard soil or artificial soil
12. Validity criteria Control: 30 juveniles/container, CV ≤ 30 %, adult mortality ≤ 10 %
13. Pos. Control/reference Boric acid. Effects on reproduction should be observed at concentra-
toxicant tions of between 200 mg and 600 mg of boric acid per kilogram of artifi-
cial soil (dry mass).
14. Statistics Multiple t-test, u – test, regression analysis
15. Test parameter(s) Mortality, growth, reproduction
16. End points EC50, NOEC
17. Limitations/Comments Same as for ISO 11268-1. In a new Annex the test performance under
tropical conditions is described. The test has been used successfully for
the assessment of wastes. (see Reference [27])

ISO/DIS 15799:2025(en)
A.1.1.4 Enchytraeid — Effects on reproduction
1. Title of the test Soil quality — Effects of contaminants on Enchytraeidae (Enchytraeus
sp.) — Determination of effects on reproduction
2. Harmonization International
3. References ISO 16387 (see Reference [45])
4. Principle The effects on reproduction of adult enchytraeids (species: Enchytraeus sp.)
exposed to the test soil are compared to those observed for test organisms
exposed to a control soil.
5. Test type Subchronic, static
6. Test organism Enchytraeids
Breeding stocks Enchytraeus albidus Henle and other species of the Enchytraeus sp., in par-
ticular E. crypticus see Reference [37]
Age Adult worms with eggs in the clitellum region
Feeding Rolled oats
7. Test substrate Test soil, field soils or Artificial soil
Volume 20 g dry mass/container
8. Test conditions
Test chamber Enclosure capable of being controlled
Temperature 20 °C ± 2 °C
pH 6 ± 0,5
Light intensity/ quality 400 lx to 800 lx
Photoperiod Preferably 16 h: 8 h
Soil moisture 40 % to 60 % water holding capacity
9. No. Replicates 2 to 4 depending on the test design (NOEC/EC )
x
10. Test duration/ 6 weeks (final test)
incubation
11. Neg. control/ dilution Artificial soil, field soil (e.g. LUFA 2.2)
soil
12. Validity criteria Control: Mort. ≤ 20 %, min. no. of juveniles 25/vessel CV ≤ 50 %
13. Pos. Control/ Carbendazim EC (1,2 ± 0,8) mg a.i./kg
reference toxicant
Mean EC50, CV
14. Statistics Multiple t-test, regression analysis, probit analysis
15. Test parameter(s) Mortality, reproduction
16. End points LC50, NOEC, EC
x
17. Limitations/ The test has been used successfully for the assessment of wastes.
Comments
(see Reference [36])
ISO/DIS 15799:2025(en)
A.1.1.5 Nematoda — Effects on growth, fertility and reproduction
1. Title of the test Water quality and soil quality — Determination of the toxic effect of sedi-
ment and soil samples on growth, fertility and reproduction of
Caenorhabditis elegans (Nematoda)
2. Harmonization International
3. References ISO 10872 (see Reference [46])
4. Principle The effects on growth, fertility and reproduction of juvenile nematods (spe-
cies: C. elegans) exposed to the test soil are compared to those observed for
test organisms exposed to a control soil.
5. Test type Chronic, static
6. Test organism Nematods
Breeding stocks Caenorhabditis elegans (Maupas, 1899)
Age First stage juveniles (J1; exposed test organisms)
Feeding Defined food medium, containing the bacterium Escherichia coli
7. Test substrate Field soils
Volume 0,500 ± 0,010 g (dry mass) per test well
8. Test conditions
Test chamber Multidishes, with 12 wells, 3,5 cm /well
Temperature 20 °C ± 2 °C (room temperature)
pH Not specified (5,4 to 6,5 values show the range of control soils)
Light intensity/ quality Not specified
Photoperiod In the dark
Soil moisture 80 % water holding capacity
9. No. Replicates At least 4
10. Test duration/ 96 h
incubation
11. Neg. control/ dilution LUFA 2.2 or Artificial soil
soil
12. Validity criteria Mean recovery of exposed test organisms from the control is ≥ 80 %
and ≤ 120 %;
Mean percentage of males in the control is ≤ 10 %; the percentage of males
in a single control replicate is ≤ 20 %;
Mean fertility in the control is ≥ 80 %;
Mean reproduction in the control is ≥ 50 offspring per exposed test organ-
ism.
13. Pos. Control/ Benzylcetyldimethylammonium chloride monohydrate (BAC C16), tested in
reference toxicant water. At 15 mg/l growth should be inhibited in a range of 20 % to 80 %.
Mean EC50, CV
Additionally, the EC of BAC shall be determined at least every 12 months.
The EC (growth) in water shall be in the range of 8 mg to 22 mg BAC/l
14. Sta
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