SIST EN ISO 23611-6:2013

SLOVENSKI STANDARD
01-oktober-2013
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Soil quality - Sampling of soil invertebrates - Part 6: Guidance for the design of sampling
programmes with soil invertebrates (ISO 23611-6:2012)
Bodenbeschaffenheit - Probenahme von Wirbellosen im Boden - Teil 6: Anleitung für die
Aufstellung von Programmen für die Probenahme von Wirbellosen im Boden (ISO 23611
-6:2012)
Qualité du sol - Prélèvement des invertébrés du sol - Partie 6: Lignes directrices pour la
conception de programmes d'échantillonnage des invertébrés du sol (ISO 23611-6:2012)
Ta slovenski standard je istoveten z: EN ISO 23611-6:2013
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.

EUROPEAN STANDARD
EN ISO 23611-6
NORME EUROPÉENNE
EUROPÄISCHE NORM
July 2013
ICS 13.080.05; 13.080.30
English Version
Soil quality - Sampling of soil invertebrates - Part 6: Guidance
for the design of sampling programmes with soil invertebrates
(ISO 23611-6:2012)
Qualité du sol - Prélèvement des invertébrés du sol - Partie Bodenbeschaffenheit - Probenahme von Wirbellosen im
6: Lignes directrices pour la conception de programmes Boden - Teil 6: Anleitung für die Planung der Probenahme
d'échantillonnage des invertébrés du sol (ISO 23611- von Wirbellosen im Boden (ISO 23611-6:2012)
6:2012)
This European Standard was approved by CEN on 11 July 2013.

CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this European
Standard the status of a national standard without any alteration. Up-to-date lists and bibliographical references concerning such national
standards may be obtained on application to the CEN-CENELEC Management Centre or to any CEN member.

This European Standard exists in three official versions (English, French, German). A version in any other language made by translation
under the responsibility of a CEN member into its own language and notified to the CEN-CENELEC Management Centre has the same
status as the official versions.

CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia,
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Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and United
Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION

EUROPÄISCHES KOMITEE FÜR NORMUNG

Management Centre: Avenue Marnix 17, B-1000 Brussels
© 2013 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 23611-6:2013: E
worldwide for CEN national Members.

Contents Page
Foreword .3

Foreword
The text of ISO 23611-6:2012 has been prepared by Technical Committee ISO/TC 190 “Soil quality” of the
International Organization for Standardization (ISO) and has been taken over as EN ISO 23611-6:2013 by
Technical Committee CEN/TC 345 “Characterization of soils” the secretariat of which is held by NEN.
This European Standard shall be given the status of a national standard, either by publication of an identical
text or by endorsement, at the latest by January 2014, and conflicting national standards shall be withdrawn at
the latest by January 2014.
Attention is drawn to the possibility that some of the elements of this document may be the subject of patent
rights. CEN [and/or CENELEC] shall not be held responsible for identifying any or all such patent rights.
According to the CEN-CENELEC Internal Regulations, the national standards organizations of the following
countries are bound to implement this European Standard: Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech
Republic, Denmark, Estonia, Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece,
Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal,
Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom.
Endorsement notice
The text of ISO 23611-6:2012 has been approved by CEN as EN ISO 23611-6:2013 without any modification.
INTERNATIONAL ISO
STANDARD 23611-6
First edition
2012-09-15
Soil quality — Sampling of soil
invertebrates —
Part 6:
Guidance for the design of sampling
programmes with soil invertebrates
Qualité du sol — Prélèvement des invertébrés du sol —
Partie 6: Lignes directrices pour la conception de programmes
d’échantillonnage des invertébrés du sol
Reference number
ISO 23611-6:2012(E)
©
ISO 2012
ISO 23611-6:2012(E)
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
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Published in Switzerland
ii © ISO 2012 – All rights reserved

ISO 23611-6:2012(E)
Contents Page
Foreword .iv
Introduction . v
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 2
3.1 Soil biology . 2
3.2 Soil protection . 3
3.3 Methods . 4
4 Principle . 4
4.1 General . 4
4.2 Question to be answered when planning a field study . 5
5 Objectives of sampling . 6
5.1 General . 6
5.2 General remarks . 6
5.3 Pre-conditions . 7
5.4 The performance of the site-specific assessment of contaminated land . 7
5.5 The study of potential side effects of anthropogenic impacts . 7
5.6 The biological classification and assessment of soils in order to determine the biological
quality of soils . 7
5.7 Biogeographical monitoring in nature protection or restauration . 8
6 Samples and sampling points . 8
6.1 General . 8
6.2 Sampling patterns . 8
6.3 Selecting and identifying the sampling location . 9
6.4 Preparation of the sampling site . 9
6.5 Further general advice on sampling performance .10
7 Practical considerations for the biological sampling of soils .10
7.1 General .10
7.2 Formal preparations .10
7.3 Requirements on sampling personnel and safety precautions .10
7.4 Preliminary survey . 11
7.5 Main study .12
8 Design options for sampling soil invertebrates .12
8.1 Introduction .12
8.2 Description of possible sampling strategies .13
8.3 Recommendations from the European programme ENVASSO (Environmental Assessment of
Soil for Monitoring) .17
9 Sampling report .19
10 Quality control and quality assurance .20
Annex A (informative) Case studies .21
Bibliography .37
ISO 23611-6:2012(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 23611-6 was prepared by Technical Committee ISO/TC 190, Soil quality, Subcommittee SC 4,
Biological methods.
ISO 23611 consists of the following parts, under the general title Soil quality — Sampling of soil invertebrates:
— Part 1: Hand-sorting and formalin extraction of earthworms
— Part 2: Sampling and extraction of micro-arthopods (Collembola and Acarina)
— Part 3: Sampling and soil extraction of enchytraeids
— Part 4: Sampling extraction and identification of soil-inhabiting nematodes
— Part 5: Sampling and extraction of soil macro-invertebrates
— Part 6: Guidance for the design of sampling programmes with soil invertebrates
iv © ISO 2012 – All rights reserved

ISO 23611-6:2012(E)
Introduction
The biodiversity of soil fauna is tremendous. Soil harbours species-rich communities, which regulate ecosystem
processes such as organic matter decomposition, nutrient flows or soil fertility in general, References [40], [45].
All terrestrial animal phyla can be found in soils, Reference [16]. In addition to thousands of bacterial and fungal
“species“, more than 1 000 species of invertebrates in abundances of up to 1,5 million individuals can be found
within a square metre of soil, References [3], [5]. This diversity can only be reliably estimated by investigation
of the soil community itself, since other parameters like climate are not or only weakly correlated with species
richness, Reference [24].
The composition of this community, as well as the abundance and biomass of the individual species and
groups is a valuable source of information, since they integrate various abiotic and biotic effects such as soil
properties and conditions, climate, competition or biogeographical influences, Reference [68]. For this reason,
the evaluation of the biodiversity of soil invertebrate communities becomes more and more important for the
classification and assessment of biological soil quality, Reference [51]. However, this work is only possible
if data collection (i.e. sampling of the soil fauna) is carried out according to standardized methods. For this
reason, a number of ISO guidelines have been prepared covering the sampling of the most important soil
organism groups.
In the individual parts of ISO 23611, the practical work concerning the respective animal group is described
in detail. However, (nearly) nothing is said about how to plan the use of such methods or how to evaluate the
results. Despite the fact that sampling for any field study can be different depending on the individual purpose,
guidance is needed for monitoring studies in a legal context. Such studies can include the following:
— site-specific risk assessment of contaminated land;
— study of potential side effects of anthropogenic impacts (e.g. the application of chemicals or the
building of roads);
— the biological classification and assessment of soils in order to determine the biological quality of soils;
— long-term biogeographical monitoring in the context of nature protection or restoration, including global
change [e.g. as in the long-term ecological research project (LTER)].
Spatial studies focusing on environmental and ecological questions require a carefully designed strategy for
collecting data (References [31], [65]). Before identifying the optimal design, two issues have to be clarified:
what is the objective of the study and what is already known about the survey area? Afterwards, one may select
one of the well-known design patterns (e.g. grid sampling, random sampling, clustered sampling or random
transects) or prepare a study-specific design. In any case, the field sampling design has to be practical, e.g. the
volume of soil to be sampled, depending on the size and distribution of the organisms, has to be manageable
(i.e. the smaller the individual animal, the smaller the size), and cost effective.
In studies focusing on soil invertebrates, it is not possible to observe the entire population. Therefore, sampling
is done only at a limited number of locations. The main reason for using statistical sound sampling schemes
is that such sampling guarantees scientific objectivity and avoids forms of bias such as those caused by
judgement sampling. This is especially valuable if the objective is to obtain data that are representative for the
whole area. At the same time, statistics-based sampling schemes ensure standardized sampling methods over
time, i.e., if the same area is to be re-sampled in the future, the results will be comparable.
The rational for this guidance s on the design of field sampling methods for soil invertebrates takes into
consideration the guidance provided in ISO 10381-1 describing soil sampling in general.
The design of microbiological studies is already covered by ISO 10381-6, ISO 14240-1 and ISO 14240-2.
INTERNATIONAL STANDARD ISO 23611-6:2012(E)
Soil quality — Sampling of soil invertebrates —
Part 6:
Guidance for the design of sampling programmes with soil
invertebrates
1 Scope
This part of ISO 23611 provides guidance for the design of field studies with soil invertebrates (e.g. for the
monitoring of the quality of a soil as a habitat for organisms). Detailed information on the sampling of the
most important soil organisms is provided in the other parts of this International Standard (ISO 23611-1 to
ISO 23611-5).
This part of ISO 23611 is used for all terrestrial biotopes in which soil invertebrates occur. Basic information on
the design of field studies in general is already laid down in ISO 10381-1. This information can vary according
to the national requirements or the climatic/regional conditions of the site to be sampled.
NOTE While this part of ISO 23611 aims to be applicable globally for all terrestrial sites that are inhabited by soil
invertebrates, the existing information refers mostly to temperate regions. However, the (few) studies from other (tropical
and boreal) regions, as well as theoretical considerations, allow the conclusion that the principles laid down in this part of
ISO 23611 are generally valid, References [4], [6], [40], [21].
This part of ISO 23611 gives information on site-specific risk assessment of contaminated land, study of
potential side effects of anthropogenic impacts (e.g. the application of chemicals or the building of roads), the
biological classification and assessment of soils in order to determine the biological quality of soils, and long-
term biogeographical monitoring in the context of nature protection or restoration, including global change (e.g.
as in long-term ecological research projects).
2 Normative references
The following referenced documents are indispensable for the application 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 10381-1:2002, Soil quality — Sampling — Part 1: Guidance on the design of sampling programmes
ISO 10381-2, Soil quality — Sampling — Part 2: Guidance on sampling techniques
ISO 10381-3, Soil quality — Sampling — Part 3: Guidance on safety
ISO 10381-4, Soil quality — Sampling — Part 4: Guidance on the procedure for investigation of natural, near-
natural and cultivated sites
ISO 10381-5, Soil quality — Sampling — Part 5: Guidance on the procedure for the investigation of urban and
industrial sites with regard to soil contamination
ISO 10381-6, Soil quality — Sampling — Part 6: Guidance on the collection, handling and storage of soil under
aerobic conditions for the assessment of microbiological processes, biomass and diversity in the laboratory
ISO 10390, Soil quality — Determination of pH
ISO 10694, Soil quality — Determination of organic and total carbon after dry combustion (elementary analysis)
ISO 11074, Soil quality — Vocabulary
ISO 23611-6:2012(E)
ISO 11260, Soil quality — Determination of effective cation exchange capacity and base saturation level using
barium chloride solution
ISO 11272, Soil quality — Determination of dry bulk density
ISO 11274, Soil quality — Determination of the water-retention characteristic — Laboratory methods
ISO 11277, Soil quality — Determination of particle size distribution in mineral soil material — Method by
sieving and sedimentation
ISO 11461, Soil quality — Determination of soil water content as a volume fraction using coring sleeves —
Gravimetric method
ISO 11465, Soil quality — Determination of dry matter and water content on a mass basis — Gravimetric method
ISO 11466, Soil quality — Extraction of trace elements soluble in aqua regia
ISO 13878, Soil quality — Determination of total nitrogen content by dry combustion (“elemental analysis“)
ISO 14869-1, Soil quality — Dissolution for the determination of total element content — Part 1: Dissolution
with hydrofluoric and perchloric acids
ISO 15709, Soil quality — Soil water and the unsaturated zone — Definitions, symbols and theory
ISO 15799, Soil quality — Guidance on the ecotoxicological characterization of soils and soil materials
ISO 17616, Soil quality — Guidance on the choice and evaluation of bioassays for ecotoxicological
characterization of soils and soil materials
ISO 23611-1:2006, Soil quality — Sampling of soil invertebrates — Part 1: Hand-sorting and formalin extraction
of earthworms
ISO 23611-2, Soil quality — Sampling of soil invertebrates — Part 2: Sampling and extraction of micro-
arthropods (Collembola and Acarina)
ISO 23611-3, Soil quality — Sampling of soil invertebrates — Part 3: Sampling and soil extraction of enchytraeids
ISO 23611-4, Soil quality — Sampling of soil invertebrates — Part 4: Sampling, extraction and identification of
free-living stages of nematodes
ISO 23611-5, Soil quality — Sampling of soil invertebrates — Part 5: Sampling and extraction of soil macro-
invertebrates
3 Terms and definitions
For the purposes of this document, the terms and definitions given in ISO 11074 and the following apply.
3.1 Soil biology
3.1.1
biodiversity
variability among living organisms on the earth, including the variability within and between species, and within
and between ecosystems
NOTE Also often used as the number and variety of organisms found within a specified geographic region.
3.1.2
community
association of organisms, belonging to different species, families, etc. living at the same time at the same
place, i.e. the living portion of an ecosystem
See Reference [42].
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ISO 23611-6:2012(E)
3.1.3
invertebrate
term embracing all organisms except the chordates and microflora
NOTE This is not a taxonomic term.
3.1.4
microfauna, mesofauna and macrofauna
way of classifying the soil fauna according to the size (length, diameter) of the individual animals
See Reference [66].
EXAMPLE Important examples of the microfauna are protozoans and nematodes, for the mesofauna collembolans,
mites and enchytraeids, and for the macrofauna earthworms and snails.
3.1.5
taxocoenosis
total number of species belonging to the same higher taxonomic unit (e.g. family, order) within a community
3.2 Soil protection
3.2.1
soil quality
capacity of a specific kind of soil to function, within natural or managed ecosystem boundaries, to sustain plant
and animal productivity, maintain or enhance water and air quality, and support human health and habitation
See References [16], [30].
NOTE In more recent definitions, the natural functions of soil are specifically listed: soil as a habitat for organisms, as
part of natural systems (in particular nutrient cycles) and for decomposition, retention and filtration, Reference [6].
3.2.2
habitat
sum of the environment of a particular species or community (e.g. in terms of soil properties, land use, climate)
3.2.3
habitat function
ability of soils/soil materials to serve as a habitat for microorganisms, plants, and soil-living animals, and
support their interactions (community or biocenosis)
3.2.4
contamination
substance(s) or agent(s) present in the soil as a result of human activity
NOTE There is no assumption in this definition that harm results from the presence of the contaminant.
3.2.5
pollutant
substances which, due to their properties, amount or concentration, cause impacts on soil functions or soil use
3.2.6
reference soil
uncontaminated soil with comparable pedological properties to the soil being studied except that it is free of
contamination
ISO 23611-6:2012(E)
3.3 Methods
3.3.1
Geographical Information Systems
GIS
in the strictest sense, a computer system capable of assembling, storing, manipulating, and displaying
geographically referenced information, i.e. data identified according to their locations
NOTE Practitioners also regard the total GIS as including operating personnel and the data that go into the system
(US. Geological Survey, 2006).
3.3.2
site-specific assessment
evaluation of the quality of a specific-site by using chemical, biological or other methods
3.3.3
environmental risk assessment
process of identifying and quantifying risk (probability that an effect occurs) to non-human organisms and
determining the acceptability of these risks
3.3.4
soil function
property of (specific) soils, often used in legal documents
NOTE Usually natural soil functions (e.g. the soil as a habitat for organisms) and anthropogenic soil functions (e.g.
soil as a substrate for crop production) are distinguished.
3.3.5
soil organism function
activity provided by individual species or, more often, by interaction of several species or the whole soil
community, e.g. nitrogen fixation or organic-matter breakdown
4 Principle
4.1 General
The design of field studies for the investigation of soil invertebrates differs significantly depending on the
respective aim. However, in all cases, it is necessary to take samples since the site and biological populations
to be studied are usually too large to be studied in total. In addition, most soil invertebrates live hidden within the
soil and/or are too small to be studied directly. The samples collected should be as representative as possible of
the site to be characterized but destruction should be kept at a minimum. In addition, the occurrence of material
not naturally belonging to the study site (e.g. waste or chemicals) can cause problems when taking samples in
multiphase systems such as soils, which contains water, gases, mineral solids and biological material.
The study design (e.g. the position and density of sampling points, time of sampling, and the sampling method)
depends mainly on the objectives of the study and on the amount and quality of information already available
from the study site (e.g. historical data, personal experience). The design also depends on whether information
is needed as an average value (sampling for the spatial mean, e.g. the average number of nematodes) or as
a spatial distribution (e.g. sampling for a map showing nematode abundances in relation to soil properties).
In addition, the sheer size and the heterogeneity of soil properties, as well as those of the organisms to be
sampled shall be taken into consideration. In any case, a list of measurement end points should be compiled
for the respective organism group(s) and the main limitations of the sampling method(s) shall also be known.
The latter refers mainly to the high natural variability of invertebrate data. The normal statistical tests used by
those who take composite samples (microflora, soil properties) or many samples (soil properties) which can be
processed more or less automatically, cannot be applied here.
Some consideration should also be given to the degree of detail and precision that is required and also the
manner in which the results are to be expressed (e.g. maximum and minimum values in a table, graphical
presentations or maps). Appropriate statistical methods for the evaluation of area-related data (including the
use of GIS methods) shall be identified as well. It can often be necessary to carry out an exploratory sampling
4 © ISO 2012 – All rights reserved

ISO 23611-6:2012(E)
programme before the final study design can be defined in detail. The main points on which decisions shall be
made are listed in 4.2, reflecting the logical order of how to proceed a study.
NOTE This clause was written in close consideration with ISO 10381-1.
4.2 Question to be answered when planning a field study
The objective of a study can be established by the following questions:
— Why is such a study going to be performed?
— What information is necessary to answer the questions asked and how can this information be clearly presented?
— Which approach is used for the interpretation of the results?
— How can the study outcome be tailored to the needs of the study sponsor (or stakeholder)?
The preliminary information can be defined by the following questions:
— What is already known about present and historical (especially land-use, management) site and soil
characteristics?
— What information is missing? Can it be made available?
— Who is to be contacted for certain (e.g. historical) sources?
— Are there any legal problems such as entering the sites?
— Shall other than biological parameters be measured at the same site and time, i.e. are (negative) interactions
of the various sampling programmes to be expected?
— Has the site been visited already?
The strategy of a study can be developed by the following questions:
— How are the delineations in time and space of the area(s) to be investigated determined?
— Which organism groups and measurement end points are appropriate to reach the study objective?
— Which sampling patterns, sampling points, sampling times, depths of sampling should be used?
— Can methods specified in International Standards be employed for all activities?
The decision on sampling and analysis can be made by answering the following questions:
— Can the sampling be done according to the respective International Standard or is there any deviation?
— How is the communication with the personnel responsible for sample presentation and analysis coordinated?
— Which statistical evaluation methods are being employed?
— Does sampling correspond to later data analyses?
— Is it possible to address the right taxonomic level when studying the biological material?
— How is the documentation organized?
The following questions on safety should be answered:
— Are all necessary safety precautions at that site considered?
— Is information concerning landowners, local authorities etc. secured?
— Are the requirements of ISO 10381-3, covering guidance on safety in sampling programmes, as well as
those safety issues listed in other parts of this International Standard (ISO 23611-1 to ISO 23611-5 fulfilled?
ISO 23611-6:2012(E)
The following questions on the sampling report should be answered:
— Is there any deviation from the basic content of a study report as specified in this part of ISO 23611?
— Is additional information required?
— How is it ensured that any later deviation from this part of ISO 23611 or the study plan is documented
and distributed?
Answers to these questions are given in Clauses 5 to 8.
5 Objectives of sampling
5.1 General
Biological soil investigations address a number of different questions related to the status of invertebrates
living in or on the soil (including many different species belonging to different trophic, taxonomic, physiological
or functional groups and size classes), often after or under some kind of anthropogenic impact. In the case of
ecotoxicological questions, usually laboratory tests are used to study the effects of the impact (e.g. chemicals
added to the soil) on invertebrates and thus on the soil quality in general. Such methods are presented in
ISO 15799, while the assessment of the test results is given in ISO 17616. Further guidance on sampling,
collection, handling and preparation of contaminated soil for biological (i.e. ecotoxicological) testing has currently
been prepared by Reference [21]. This is particularly important for the identification and characterization of
field reference soils which are necessary for the determination of biological reference values. Examples are
provided in Annex A (case studies).
5.2 General remarks
As stated in the Introduction, the principal objectives of sampling soil invertebrates can be distinguished as follows:
— the performance of the site-specific characterization and assessment of contaminated land;
— the study of potential side effects of anthropogenic impacts (e.g. the application of chemicals or the
building of roads);
— the biological classification and assessment of soils in order to determine the biological quality of soils;
— long-term biogeographical monitoring in the context of nature protection or restoration, including global
change (e.g. as in the long-term ecological research project (LTER)).
To a different degree, all four objectives include the determination of biological reference (or base-line) values,
meaning that it shall be clarified which community of soil organisms occurs in a specific soil assuming that
there is no anthropogenic impact. Since this precondition is, in many if not all soils, not fulfilled any more, such
a “normal” state shall be defined, e.g. by sampling of reference soils. These soils have been selected based on
criteria like being representative for certain regions or land-use forms or lack of contamination, Reference [14].
The use of the soil and site are of varying importance depending on the primary objective of an investigation.
The results obtained from sampling can indicate a need for further investigation, e.g. detected contamination
can indicate a need for identification and assessment of potential hazards and risks. However, assessment of
such hazards or risks is not covered by this part of ISO 23611. In addition, capture-recapture methods – while
often used in ecology for terrestrial above-ground invertebrates (e.g. spiders, Reference [26]) are rarely used
in general monitoring schemes and thus will not be covered in this part of ISO 23611.
Often soil invertebrates are a part of an entire monitoring effort that includes other biological (mainly microbial),
as well as pedological, climatic and possibly also agricultural parameters. If such monitoring programmes are
performed at regular intervals, permanent sampling sites shall be set up. In such a case, additional efforts are
mandatory in order to secure an effective exchange of information. Sampling is usually carried out within the
main rooting zone (rarely at greater depths since most soil invertebrates live within the uppermost 30 cm of the
soil). Soil horizons or layers may or may not be separately sampled (samples shall be labelled accordingly).
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ISO 23611-6:2012(E)
To adequately support legal or regulatory action, particular attention should be paid to all aspects of quality
assurance. The guidance given in ISO 10381-5 is particularly relevant. After clarifying the most important pre-
conditions, the four groups of main objectives as given above are briefly presented in the following subclauses.
However, it should be kept in mind that, in reality, one specific study can fit into more than one of these groups.
5.3 Pre-conditions
Before designing a field study with soil invertebrates, it is highly recommended to characterize the respective
area pedologically, Reference [43]. Depending on the principal objectives, it is usually necessary to determine
for the body of soil or part thereof
— the nature, concentrations and distribution of naturally occurring substances,
— the nature, concentrations and distribution of contaminants,
— the physical and chemical properties and variations,
— the anthropogenic impact at that site, in particular the land use (including vegetation cover).
It is often necessary to take into account changes in the above-mentioned variables with time and space
(vertically, horizontally), caused by either natural (e.g. climatic) or anthropogenic activities.
In addition, pH, particle size distribution, C/N ratio, organic matter and organic carbon content, total nitrogen,
cation exchange capacity and water holding capacity of the soil should be measured in accordance with
ISO 10390, ISO 10694, ISO 11260, ISO 11272, ISO 11274, ISO 11277, ISO 13878, ISO 11461, ISO 11465,
ISO 15709, ISO 17616.
5.4 The performance of the site-specific assessment of contaminated land
When land is contaminated with chemicals and other substances that are potentially acting as pollutants to the
environment, it can be necessary to carry out an investigation as a part of a hazard and/or risk assessment.
This includes to determine the nature and extent of contamination, to identify hazards associated with the
contamination, to identify potential targets and routes of exposure, and to evaluate the environmental risks
relating to the current and future use of the site and neighbouring land. A sampling programme for risk
assessment can also comply with legal or regulatory requirements and careful attention to sample integrity is
recommended. An extensive overview of the benefits and limitations of biological parameters as a component
of contaminated land assessment is given in Reference [21].
5.5 The study of potential side effects of anthropogenic impacts
Sampling can be required following an anthropogenic effect such as the input of undesirable material (mainly
chemicals) which can be from a point source or from a diffuse source. Another example can be the building of
roads. The study design needs again to be developed on a site-specific basis. Sampling can also be required
to establish base-line conditions prior to an activity, which might affect the composition or quality of soil.
NOTE Such base-line sampling can also be performed as part of a biological soil classification and assessment (see 5.4).
5.6 The biological classification and assessment of soils in order to determine the biologi-
cal quality of soils
This is typically carried out at (irregular) time intervals to determine the biological quality of a soil for a particular
purpose (e.g. as part of a large-scale screening programme or in the context of a local planning activity). While
it has rarely been done so far in terrestrial habitats (except with plants), the information gained here can be
used for the preparation of biological soil maps, Reference [8].
NOTE The study of the biological soil quality can also be used for the determination of “base-line conditions” in the
context of the assessment of anthropogenic impacts (see 5.3) or of long-term changes such as global warming (5.7).
ISO 23611-6:2012(E)
5.7 Biogeographical monitoring in nature protection or restauration
Finally, the information gained in sampling programmes extends the knowledge on the biogeography of soil
organisms, which is necessary in the context of nature protection and conservation, in particular concerning
long-term changes like global warming. So far, only few soil invertebrates (mainly beetles or other insects
which in their larval stage live in the soil) have been put on the Red List of endangered species. Also there is
little proof that such species have been eradicated in modern times. However, in both cases, this fact is mainly
caused by the poor level of knowledge on these species; many species can have died out without notice.
Sampling programmes can also determine whether soil-biological assemblages (site-specifically) expected
in a region become established during nature restoration or after remediation measures (control of success).
6 Samples and sampling points
6.1 General
The selection, location and preparation of the sampling points depend on the objectives of the investigation,
the preliminary information available and the on-site conditions. Soil properties, the occurrence of organisms
and contamination vary continuously in space; the values at locations close together are more similar than
those farther apart and this spatial dependence can be described by the use of geostatistics, Reference
[43]. Geostatistics are used in the development of sampling strategies and are also used to analyse the data
generated from the soil sampled, Reference [41]. In this clause which closely follows the terminology used in
ISO 10381-1:2002, Annex C, several (standard) options and issues to be considered are given.
6.2 Sampling patterns
Sampling patterns are based on the estimation of the distribution of the soil invertebrates in the area to be
sampled. Several distribution patterns can be distinguished (of course with intermediate types):
— no specific distribution (i.e. random),
— homogenous distribution (probably very rare),
— clumped distribution,
— distribution varying according to an underlying gradient (linear or concentric).
Sampling design should be adjusted to the (theoretically expected) distribution pattern or observable local conditions
which make some patterns more probable. If the area to be sampled shows differences in important properties,
such as land use, soil conditions, geomorphology, vegetation patterns, the site should be subdivided according
to these differences and separate samples should be taken from “homogenous” sub-areas (stratified sampling).
In agricultural or forestry sampling, a small number of convenient sampling patterns are established in order
to obtain information from larger areas. Examples of such patterns are briefly described in the following (for
details see ISO 10381-4):
— Systematic patterns (irregular sampling):
— Assuming a relatively homogenous distribution, such sampling ca
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