ISO 16387:2023

INTERNATIONAL ISO
STANDARD 16387
Third edition
2023-03
Soil quality — Effects of contaminants
on Enchytraeidae (Enchytraeus sp.) —
Determination of effects on
reproduction
Qualité du sol — Effets des contaminants sur les Enchytraeidae
(Enchytraeus sp.) — Détermination des effets sur la reproduction
Reference number
© ISO 2023
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ii
Contents Page
Foreword .iv
Introduction .v
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 1
4 Principle . 3
5 Reagents and material . 3
6 Apparatus . 6
7 Test environment . 6
8 Procedure .7
8.1 Experimental design . 7
8.1.1 General . 7
8.1.2 Range-finding test . 7
8.1.3 Definitive test . 7
8.2 Preparation of test mixtures . 8
8.2.1 Testing contaminated soil . 8
8.2.2 Testing substances added to the test substrate . 8
8.2.3 Preparation of control container . 9
8.3 Addition of food . 9
8.4 Addition of the biological material . 9
8.5 Test conditions and measurements . 9
8.6 Reference substance . 9
9 Calculation and expression of results .10
9.1 Calculation . 10
9.2 Expression of results . . 10
10 Validity of the test .10
11 Statistical analysis .10
11.1 General . 10
11.2 Range-finding test . 10
11.3 Definitive test . 11
12 Test report .11
Annex A (informative) Conditions for culture of Enchytraeus sp .13
Annex B (informative) Test procedure using other Enchytraeus species .15
Annex C (normative) Determination of total water holding capacity .17
Annex D (informative) Detailed description of extraction techniques .18
Annex E (informative) Overview of the statistical assessment of data (NOEC determination) .20
Bibliography .21
iii
Foreword
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described in the ISO/IEC Directives, Part 1. In particular, the different approval criteria needed for the
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www.iso.org/iso/foreword.html.
This document was prepared by Technical Committee ISO/TC 190, Soil quality, Subcommittee SC 4,
Biological characterization, in collaboration with the European Committee for Standardization (CEN)
Technical Committee CEN/TC 444, Environmental characterization of solid matrices, in accordance with
the Agreement on technical cooperation between ISO and CEN (Vienna Agreement).
This third edition cancels and replaces the second edition (ISO 16387:2014), which has been technically
revised.
The main changes are as follows:
— correction of the vapor pressure value to update Clause 1 according to the recommendations of
ECHA/OECD;
— addition (in Annex D) of an extraction method of enchytraeids with colloidal silica, suitable for
Enchytraeus crypticus.
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
Introduction
Ecotoxicological test systems are applied to obtain information about the effects of contaminants in
[38]
soil and are proposed to complement conventional chemical analysis. ISO 15799 includes a list and
short characterization of recommended and standardized test systems. Aquatic test systems with soil
eluate are applied to obtain information about the fraction of contaminants potentially reaching the
groundwater by the water path (retention function of soils), whereas terrestrial test systems are used
to assess the habitat function of soils. For the latter, a standardized test system using Enchytraeidae (a
chronic test with end-point reproduction) is proposed.
This document describes a method that is based on the determination of acute and sublethal effects of
contaminated soils to adult Enchytraeidae of the genus Enchytraeus. Optionally, the method can be used
for testing substances added to standard soils (e.g. artificial soil) for their sublethal hazard potential to
Enchytraeidae.
Soil-dwelling annelids of the genus Enchytraeus are ecologically relevant, i.e. they are abundant in
many soils where earthworms are scarce but can also reach high population densities in soils well
inhabited by earthworms. Enchytraeidae can be used in laboratory tests as well as in semi-field and
field studies. From a practical point of view, many Enchytraeus species are easy to handle and breed,
and their generation time is significantly shorter than that of earthworms [the test duration for a
reproduction test with Enchytraeidae is four weeks to six weeks, compared to eight weeks (12 weeks
including synchronization) with earthworms]. In addition, a much smaller volume of soil is needed in
the enchytraeid test compared to the amount needed in earthworm tests.
This document has been drawn up taking into consideration test procedures recommended by the
[24],[25]
Organization for Economic Co-operation and Development (OECD ).
v
INTERNATIONAL STANDARD ISO 16387:2023(E)
Soil quality — Effects of contaminants on Enchytraeidae
(Enchytraeus sp.) — Determination of effects on
reproduction
1 Scope
This document specifies one of the methods for evaluating the habitat function of soils and determining
effects of soil contaminants and substances on the reproduction of Enchytraeus sp. by dermal and
alimentary uptake in a chronic test. It is applicable to soils and soil materials of unknown quality, for
example, from contaminated sites, amended soils, soils after remediation, agricultural or other sites
under concern and waste materials.
This document provides information on how to use this method for testing substances under temperate
conditions.
The method is not applicable to substances, for which the air/soil partition coefficient is greater than 1,
or to substances for which the vapour pressure exceeds 300 Pa at 25 °C.
NOTE No provision is made in the test method for monitoring the persistence of the substance under test.
2 Normative references
The following documents are referred to in the text in such a way that some or all of their content
constitutes requirements of this document. For dated references, only the edition cited applies. For
undated references, the latest edition of the referenced document (including any amendments) applies.
ISO 10390, Soil, treated biowaste and sludge – Determination of pH
ISO 10694, Soil quality — Determination of organic and total carbon after dry
...