prEN ISO 23611-1

SLOVENSKI STANDARD
01-september-2025
Kakovost tal - Vzorčenje nevretenčarjev v tleh - 1. del: Ročno razvrščanje
deževnikov in njihova ekstrakcija (ISO/DIS 23611-1:2025)
Soil quality - Sampling of soil invertebrates - Part 1: Hand-sorting and extraction of
earthworms (ISO/DIS 23611-1:2025)
Bodenbeschaffenheit - Probenahme von Wirbellosen im Boden - Teil 1: Handauslese
und Extraktion von Regenwürmern (ISO/DIS 23611-1:2025)
Qualité du sol - Prélèvement des invertébrés du sol - Partie 1: Tri manuel et extraction
des vers de terre (ISO/DIS 23611-1:2025)
Ta slovenski standard je istoveten z: prEN ISO 23611-1
ICS:
13.080.30 Biološke lastnosti tal Biological properties of soils
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

DRAFT
International
Standard
ISO/DIS 23611-1
ISO/TC 190/SC 4
Soil quality — Sampling of soil
Secretariat: AFNOR
invertebrates —
Voting begins on:
Part 1: 2025-06-20
Hand-sorting and extraction of
Voting terminates on:
2025-09-12
earthworms
Qualité du sol — Prélèvement des invertébrés du sol —
Partie 1: Tri manuel et extraction des vers de terre
ICS: 13.080.05; 13.080.30
THIS DOCUMENT IS A DRAFT CIRCULATED
FOR COMMENTS AND APPROVAL. IT
IS THEREFORE SUBJECT TO CHANGE
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This document is circulated as received from the committee secretariat.
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Reference number
ISO/DIS 23611-1:2025(en)
DRAFT
ISO/DIS 23611-1:2025(en)
International
Standard
ISO/DIS 23611-1
ISO/TC 190/SC 4
Soil quality — Sampling of soil
Secretariat: AFNOR
invertebrates —
Voting begins on:
Part 1:
Hand-sorting and extraction of
Voting terminates on:
earthworms
Qualité du sol — Prélèvement des invertébrés du sol —
Partie 1: Tri manuel et extraction des vers de terre
ICS: 13.080.05; 13.080.30
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
ISO/CEN PARALLEL PROCESSING
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Published in Switzerland Reference number
ISO/DIS 23611-1:2025(en)
ii
ISO/DIS 23611-1:2025(en)
Contents Page
Foreword .iv
Introduction .v
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 1
4 Principle . 2
5 Reagents . 3
6 Apparatus . 3
7 Procedure . 4
7.1 Sampling of the earthworms.4
7.1.1 General .4
7.1.2 Hand-sorting .4
7.1.3 AITC extraction .5
7.2 Preservation .6
7.3 Determination of biomass .6
7.4 Preparation of a composite sample for community DNA metabarcoding .7
8 Data assessment . 7
9 Test report . 7
Annex A (informative) Other methods for sampling . 9
Annex B (informative) Species identification in earthworms .10
Annex C (normative) Determination of maximum water-holding capacity .11
Bibliography .12

iii
ISO/DIS 23611-1:2025(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
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constitute an endorsement.
For an explanation on the meaning of ISO specific terms and expressions related to conformity assessment,
as well as information about ISO’s adherence to the World Trade Organization (WTO) principles in the
Technical Barriers to Trade (TBT) see the following URL: www.iso.org/iso/foreword.html.
This document was prepared by Technical Committee ISO/TC 190, Soil quality, Subcommittee SC 4, Biological
characterization.
This third edition cancels and replaces the second edition (ISO 23611-1:2018), which has been technically
revised. The main changes are:
— addition of a definition for ecological categories of earthworms;
— addition of a section on the preparation of composite samples for community DNA metabarcoding;
— removal of the informative Annex C on the modified TSBF method;
— removal of the outdated informative Annex E with examples of earthworm monitoring programmes
(including presentation of their results).
A list of all parts in the ISO 23611 series can be found on the ISO website.

iv
ISO/DIS 23611-1:2025(en)
Introduction
This Standard has been originally drawn up since there was a growing need for the standardization of
terrestrial zoological field methods. Such methods, mainly covering the sampling, extraction and handling
of soil invertebrates, are necessary for the following purposes:
[9][19][30][33][45]
— biological classification of soils including soil quality assessment ;
[2][15][17][39][44]
— terrestrial bio-indication and long-term monitoring ;
[29]
— evaluation of the effects of chemicals on soil animals in the field .
Data for these purposes are gained by standardized methods since they can form the basis for far-reaching
decisions (e.g. whether a given site should be remediated or not). In fact, the lack of such standardized
methods is one of the most important reasons why bio-classification and bio-assessment in terrestrial (i.e.
soil) habitats has so far relatively rarely been used in comparison to aquatic sites.
Since it is neither possible nor useful to standardize methods for all soil organisms, the most important ones
have been selected. In this document sampling of earthworms is described.
Originally, the methods described in this document were developed for taxonomical and ecological studies,
investigating the role of earthworms (macrofauna) in various soil ecosystems. These animals are without
[21][23][35]
doubt the most important soil invertebrates in temperate regions. This does not necessarily imply
that earthworms are less important in tropical soils. In fact, there is wide evidence that earthworms may
be as important as or even more important than the abundant termites and ants in many locations of the
[22]
tropical region. Since Darwin (1881) (see Reference [11]), their influence on soil structure (e.g. aeration,
[14]
water holding capacity) and soil functions like litter decomposition and nutrient cycling is well-known.
Due to their often very high biomass they are also important in many terrestrial food-webs.
In earlier versions of this document the chemical formalin was recommended as extraction fluid. Since then,
evidence increased that formalin has properties of concern, mainly in terms of human toxicity. According
to the European Chemicals Agency (ECHA) chemical database, this chemical is labelled as carcinogenic
[13]
and suspected to be mutagenic (Categories Carc. 1B and Muta. 2). In addition, negative effects on non-
target organisms (including soil microorganisms, mesofauna and plants) have been reported (e.g. see
Reference [10]). Therefore, this substance has been replaced.
Due to the growing reservations against the use of formalin, several alternatives have been studied. In
Reference [46] allyl isothiocyanate (AITC) was tested for its effectiveness as a chemical expellant for
sampling earthworms. AITC is a natural breakdown product of glucosinolates in many Cruciferae, i.e. it is
the component imparting the sharp taste of mustard. In the ECHA chemical database, properties of concern
[12]
‘skin sensitising’ and ‘respiratory sensitising’ are listed. It is also toxic and hazardou
...