Soil quality - Inhibition of reproduction of Collembola (Folsomia candida) by soil contaminants (ISO 11267:2014)

ISO 11267:2014 specifies one of the methods for evaluating the habitat function of soils and determining effects of soil contaminants and substances on the reproduction of Folsomia candida Willem by dermal and alimentary uptake. This chronic test is applicable to soils and soil materials of unknown quality, e.g. from contaminated sites, amended soils, soils after remediation, industrial, agricultural or other sites of concern and waste materials.
Effects of substances are assessed using a standard soil, preferably a defined artificial soil substrate. For contaminated soils, the effects are determined in the soil to be tested and in a control soil. According to the objective of the study, the control and dilution substrate (dilution series of contaminated soil) are either an uncontaminated soil comparable to the soil to be tested (reference soil) or a standard soil (e.g. artificial soil).
ISO 11267:2014 provides information on how to use this method for testing substances under temperate conditions.
The method is not applicable to volatile substances, i.e. substances for which H (Henry's constant) or the air/water partition coefficient is greater than 1, or for which the vapour pressure exceeds 0,013 3 Pa at 25 °C.

Bodenbeschaffenheit - Hemmung der Reproduktion von Collembolen (Folsomia candida) durch Verunreinigungen (ISO 11267:2014)

Diese Internationale Norm legt eines der Verfahren zur Bewertung der Lebensraumfunktion von Böden und zur Bestimmung der Wirkungen von Bodenverunreinigungen und Substanzen auf die Reproduktion von Folsomia candida Willem bei Aufnahme über die Haut und durch die Nahrung fest. Diese chronische Prüfung ist anwendbar für Böden und Bodenmaterialien unbekannter Beschaffenheit, z. B. von verunreinigten Standorten, beaufschlagten Böden, Böden nach der Sanierung, industriellen, landwirtschaftlichen oder anderen betroffenen Standorten und Abfallmaterialien.
Die Wirkungen der Substanzen werden mit einem Standardboden, vorzugsweise mit einem festgelegten künstlichen Bodensubstrat, bewertet. Bei verunreinigten Böden werden die Wirkungen im zu prüfenden Boden und in einem Kontrollboden bestimmt. Entsprechend der Zielsetzung der Untersuchung sind die Kontrolle und das zur Verdünnung verwendete Substrat (Verdünnungsreihen des verunreinigten Bodens) entweder ein mit dem zu prüfenden Boden vergleichbarer, nicht verunreinigter Boden (Referenzboden) oder ein Standardboden (z. B. künstlicher Boden).
Diese Internationale Norm gibt Informationen, wie dieses Verfahren zur Prüfung von Substanzen unter gemäßigten Bedingungen anzuwenden ist.
Das Verfahren ist nicht auf flüchtige Substanzen anwendbar, d. h. Substanzen, bei denen H (Henry-Konstante) oder der Luft/Wasser-Verteilungskoeffizient größer als 1 ist oder bei denen der Dampfdruck 0,013 3 Pa bei 25 °C überschreitet.
ANMERKUNG Es kann nicht sichergestellt werden, dass die Prüfsubstanz während der gesamten Versuchsdauer stabil bleibt. Im Prüfverfahren wird die Persistenz der Prüfsubstanz nicht überwacht.

Qualité du sol - Inhibition de la reproduction de Collembola (Folsomia candida) par des contaminants du sol (ISO 11267:2014)

L'ISO 11267:2014 spécifie l'une des méthodes permettant d'évaluer la fonction d'habitat des sols et de déterminer les effets de contaminants du sol et de substances sur la reproduction de Folsomia candida Willem par absorption cutanée et ingestion. Cet essai chronique s'applique aux sols et matériaux du sol de qualité inconnue, par exemple provenant de sites contaminés, de sols amendés, de sols après remédiation, de sites industriels, agricoles ou d'autres sites d'intérêt et de déchets.
Les effets des substances sont évalués à l'aide d'un sol standard, de préférence un substrat de sol artificiel défini. Pour les sols contaminés, les effets sont déterminés dans le sol soumis à essai et dans un sol témoin. Selon l'objectif de l'étude, il convient que le substrat témoin et de dilution (gamme de dilutions d'un sol contaminé) soit un sol non contaminé comparable au sol à évaluer (sol de référence) ou un sol standard (par exemple un sol artificiel).
L'ISO 11267:2014 fournit des informations sur la manière d'utiliser cette méthode pour évaluer des substances dans des conditions tempérées.
La méthode ne s'applique pas aux substances volatiles, c'est-à-dire aux substances pour lesquelles H (constante de Henry) ou le coefficient de partage air/eau est supérieur à 1, ou pour lesquelles la pression de vapeur excède 0,013 3 Pa à 25 °C.

Kakovost tal - Zaviranje razmnoževanja vrste Folsomia candida iz rodu skakačev (Collembola) zaradi onesnaževal v tleh (ISO 11267:2014)

Standard EN-ISO 11267 določa eno od metod za ocenjevanje funkcije tal habitata in določevanje učinkov onesnaževal tal in snovi na razmnoževanje vrste Folsomia candida iz rodu skakačev (Collembola) z dermalnim ali prehranskim vnosom. Ta kronični preskus se uporablja za tla in talne materiale neznane kakovosti, na primer tla iz onesnaženih območij, obogatena tla, tla po sanaciji, industrijska, kmetijska in druga problematična mesta ter odpadne materiale. Vpliv snovi se določi z uporabo standardnih tal, najbolje z določenim umetnim talnim substratom. Za kontaminirana tla se učinki določijo v preskušenih in kontrolnih tleh. Glede na cilj raziskave sta kontrolni in zredčeni substrat (serija redčenja kontaminiranih tal) neokužena tla, primerljiva s tlemi, ki se jih preskuša (referenčna tla), ali standardna tla (npr. umetna tla). Ta mednarodni standard ponuja informacije o uporabi te metode za preskušanje snovi pod zmernimi pogoji. Ta metoda ni primerna za hlapljive snovi, npr. snovi, pri katerih je H (Henryjeva konstanta) ali koeficient deleža zraka/vode, večji od 1, ali pri katerih parni tlak pri 25 °C presega 0,013 3 Pa.

General Information

Status
Withdrawn
Publication Date
18-Feb-2014
Withdrawal Date
15-Aug-2023
Current Stage

Relations

Buy Standard

Standard
EN ISO 11267:2014
English language
27 pages
sale 10% off
Preview
sale 10% off
Preview
e-Library read for
1 day
Draft
prEN ISO 11267:2012
English language
28 pages
sale 10% off
Preview
sale 10% off
Preview
e-Library read for
1 day

Standards Content (Sample)

SLOVENSKI STANDARD
SIST EN ISO 11267:2014
01-maj-2014
.DNRYRVWWDO=DYLUDQMHUD]PQRåHYDQMDYUVWH)ROVRPLDFDQGLGDL]URGXVNDNDþHY
&ROOHPEROD ]DUDGLRQHVQDåHYDOYWOHK ,62
Soil quality - Inhibition of reproduction of Collembola (Folsomia candida) by soil
contaminants (ISO 11267:2014)
Bodenbeschaffenheit - Hemmung der Reproduktion von Collembolen (Folsomia candida)
durch Verunreinigungen (ISO 11267:2014)
Qualité du sol - Inhibition de la reproduction de Collembola (Folsomia candida) par des
contaminants du sol (ISO 11267:2014)
Ta slovenski standard je istoveten z: EN ISO 11267:2014
ICS:
13.080.30 Biološke lastnosti tal Biological properties of soils
SIST EN ISO 11267:2014 en,fr,de
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

---------------------- Page: 1 ----------------------

SIST EN ISO 11267:2014

---------------------- Page: 2 ----------------------

SIST EN ISO 11267:2014

EUROPEAN STANDARD
EN ISO 11267

NORME EUROPÉENNE

EUROPÄISCHE NORM
February 2014
ICS 13.080.30
English Version
Soil quality - Inhibition of reproduction of Collembola (Folsomia
candida) by soil contaminants (ISO 11267:2014)
Qualité du sol - Inhibition de la reproduction de Collembola Bodenbeschaffenheit - Hemmung der Reproduktion von
(Folsomia candida) par des contaminants du sol (ISO Collembolen (Folsomia candida) durch Verunreinigungen
11267:2014) (ISO 11267:2014)
This European Standard was approved by CEN on 4 January 2014.

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,
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 United
Kingdom.





EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION

EUROPÄISCHES KOMITEE FÜR NORMUNG

CEN-CENELEC Management Centre: Avenue Marnix 17, B-1000 Brussels
© 2014 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 11267:2014 E
worldwide for CEN national Members.

---------------------- Page: 3 ----------------------

SIST EN ISO 11267:2014
EN ISO 11267:2014 (E)
Contents Page
Foreword .3
2

---------------------- Page: 4 ----------------------

SIST EN ISO 11267:2014
EN ISO 11267:2014 (E)
Foreword
This document (EN ISO 11267:2014) has been prepared by Technical Committee ISO/TC 190 “Soil quality” in
collaboration with 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 August 2014, and conflicting national standards shall be withdrawn at
the latest by August 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 11267:2014 has been approved by CEN as EN ISO 11267:2014 without any modification.


3

---------------------- Page: 5 ----------------------

SIST EN ISO 11267:2014

---------------------- Page: 6 ----------------------

SIST EN ISO 11267:2014
INTERNATIONAL ISO
STANDARD 11267
Second edition
2014-02-15
Soil quality — Inhibition of
reproduction of Collembola (Folsomia
candida) by soil contaminants
Qualité du sol — Inhibition de la reproduction de Collembola
(Folsomia candida) par des contaminants du sol
Reference number
ISO 11267:2014(E)
©
ISO 2014

---------------------- Page: 7 ----------------------

SIST EN ISO 11267:2014
ISO 11267:2014(E)

COPYRIGHT PROTECTED DOCUMENT
© ISO 2014
All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized otherwise in any form
or by any means, electronic or mechanical, including photocopying, or posting on the internet or an intranet, without prior
written permission. Permission can be requested 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 2014 – All rights reserved

---------------------- Page: 8 ----------------------

SIST EN ISO 11267:2014
ISO 11267:2014(E)

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 . 4
6 Apparatus . 6
7 Procedure. 6
7.1 Experimental design . 6
7.2 Preparation of test mixture . 7
7.3 Addition of the biological material . 9
7.4 Test conditions and measurements . 9
7.5 Determination of surviving Collembola . 9
8 Calculation and expression of results . 9
8.1 Calculation . 9
8.2 Expression of results . 9
9 Validity of the test .10
10 Statistical analysis .10
10.1 General .10
10.2 Single-concentration tests .10
10.3 Multi-concentration tests.11
11 Test report .11
Annex A (informative) Techniques for rearing and breeding of Collembola .13
Annex B (informative) Determination of water-holding capacity .15
Annex C (informative) Guidance on adjustment of pH of artificial soil .16
Annex D (informative) Extraction and counting of Collembola .17
Bibliography .18
© ISO 2014 – All rights reserved iii

---------------------- Page: 9 ----------------------

SIST EN ISO 11267:2014
ISO 11267:2014(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.
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 on the meaning of ISO specific terms and expressions related to conformity
assessment, as well as information about ISO’s adherence to the WTO principles in the Technical Barriers
to Trade (TBT) see the following URL: Foreword - Supplementary information
The committee responsible for this document is ISO/TC 190, Soil quality, Subcommittee SC 4, Biological
methods.
This second edition cancels and replaces the first edition (ISO 11267:1999), which has been technically
revised.
iv © ISO 2014 – All rights reserved

---------------------- Page: 10 ----------------------

SIST EN ISO 11267:2014
ISO 11267:2014(E)

Introduction
Ecotoxicological test systems are applied to obtain information about the effects of contaminants in soil
and are proposed to complement conventional chemical analysis (see [2] and [4]). Reference [2] includes
a list and short characterization of recommended and standardized test systems and [4] gives guidance
on the choice and evaluation of the bioassays. 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.
Soil-dwelling Collembola are ecologically relevant species for ecotoxicological testing. Springtails are
prey animals for a variety of endogeic and epigeic invertebrates and they contribute to decomposition
processes in soils. In acidic soils they may be the most important soil invertebrates besides enchytraeids
[19]
with respect to that function, since earthworms are typically absent. Additionally, Collembola
represent arthropod species with a different route and a different rate of exposure compared to
[1] [3]
earthworms and enchytraeids. Various species were used in bioassays of which four species were
used most commonly, Folsomia candida, Folsomia fimetaria, Onychiurus armatus, and Orchesella cincta.
[20]
Numerous soil toxicity tests supported by Environment Canada (EC) resulted in the development
and standardization of a biological test method for determining the lethal and sublethal toxicity of
[10]
samples of contaminated soil to Collembola. The method prepared by EC includes three species,
Orthonychiurus folsomi, Folsomia candida, and Folsomia fimetaria. As standardized test systems using
Collembola as indicator organisms for the habitat function of soil, another two methods exist. One is
designed for assessing the effects of substances on the reproductive output of the Collembola, Folsomia
[19] [21]
fimetaria L. and Folsomia candida Willem in soil, , , and the other method described here, focuses on
testing contaminated soil. Optionally the method can be used for testing substances added to standard
soils (e.g. artificial soil) for their sublethal hazard potential to Collembola.
This International Standard describes a method that is based on the determination of sublethal effects
of contaminated soils to adult Collembola of the species Folsomia candida Willem. The species is
[10] [19]
distributed worldwide. It plays a similar ecological role to Folsomia fimetaria. , Folsomia candida
reproduces parthenogenetically and is an easily accessible species as it is commercially available and
easy to culture. Folsomia candida is considered to be a representative of soil arthropods and Collembola
in particular. Background information on the ecology of springtails and their use in ecotoxicological
[22]
testing is available.
© ISO 2014 – All rights reserved v

---------------------- Page: 11 ----------------------

SIST EN ISO 11267:2014

---------------------- Page: 12 ----------------------

SIST EN ISO 11267:2014
INTERNATIONAL STANDARD ISO 11267:2014(E)
Soil quality — Inhibition of reproduction of Collembola
(Folsomia candida) by soil contaminants
1 Scope
This International Standard specifies one of the methods for evaluating the habitat function of soils and
determining effects of soil contaminants and substances on the reproduction of Folsomia candida Willem
by dermal and alimentary uptake. This chronic test is applicable to soils and soil materials of unknown
quality, e.g. from contaminated sites, amended soils, soils after remediation, industrial, agricultural or
other sites of concern and waste materials.
Effects of substances are assessed using a standard soil, preferably a defined artificial soil substrate. For
contaminated soils, the effects are determined in the soil to be tested and in a control soil. According to
the objective of the study, the control and dilution substrate (dilution series of contaminated soil) are
either an uncontaminated soil comparable to the soil to be tested (reference soil) or a standard soil (e.g.
artificial soil).
This International Standard provides information on how to use this method for testing substances
under temperate conditions.
The method is not applicable to volatile substances, i.e. substances for which H (Henry’s constant) or the
air/water partition coefficient is greater than 1, or for which the vapour pressure exceeds 0,013 3 Pa at
25 °C.
NOTE The stability of the test substance cannot be ensured over the test period. No provision is made in the
test method for monitoring the persistence of the substance under test.
2 Normative references
The following documents, in whole or in part, are normatively referenced in this document and are
indispensable for its application. For dated references, only the edition cited applies. For undated
references, the latest edition of the referenced document (including any amendments) applies.
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 10694, Soil quality — Determination of organic and total carbon after dry combustion (elementary
analysis)
ISO 10390, Soil quality — Determination of pH
ISO 11260, Soil quality — Determination of effective cation exchange capacity and base saturation level
using barium chloride solution
ISO 11277, Soil quality — Determination of particle size distribution in mineral soil material — Method by
sieving and sedimentation
ISO 11465, Soil quality — Determination of dry matter and water content on a mass basis — Gravimetric
method
3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
© ISO 2014 – All rights reserved 1

---------------------- Page: 13 ----------------------

SIST EN ISO 11267:2014
ISO 11267:2014(E)

3.1
contaminant
substance or agent present in the soil as a result of human activity
3.2
ECx
effect concentration for x % effect
concentration (mass fraction) of a test substance that causes x % of an effect on a given end-point within
a given exposure period when compared with a control
EXAMPLE An EC50 is a concentration estimated to cause an effect on a test end-point in 50 % of an exposed
population over a defined exposure period.
Note 1 to entry: The ECx is expressed as a percentage of soil to be tested (dry mass) per soil mixture (dry mass).
When substances are tested, the ECx is expressed as mass of the test substance per dry mass of soil in milligrams
per kilogram.
3.3
ERx
effect rate
rate of a soil to be tested that causes an x % of an effect on a given end-point within a given exposure
period when compared with a control
3.4
limit test
single concentration test consisting of at least four replicates each, the soil to be tested without any
dilution or the highest concentration of test substance mixed into the control soil and the control
3.5
LOEC
lowest observed effect concentration
lowest test substance concentration that has a statistically significant effect (probability p < 0,05)
Note 1 to entry: In this test, the LOEC is expressed as a mass of test substance per dry mass of the soil to be
tested. All test concentrations above the LOEC should usually show an effect that is statistically different from
the control.
3.6
LOER
lowest observed effect rate
lowest rate of a soil to be tested in a control soil at which a statistically significant effect is observed
3.7
NOEC
no observed effect concentration
highest test substance concentration immediately below the LOEC at which no effect is observed
Note 1 to entry: In this test, the concentration corresponding to the NOEC has no statistically significant effect
(probability p < 0,05) within a given exposure period when compared with the control.
3.8
NOER
no observed effect rate
lowest rate of a soil to be tested immediately below the LOER which, when compared to the control, has
no statistically significant effect (probability p < 0,05) within a given exposure period
3.9
reference soil
uncontaminated soil with comparable pedological properties (nutrient concentrations, pH, organic
carbon content and texture) to the soil being studied
2 © ISO 2014 – All rights reserved

---------------------- Page: 14 ----------------------

SIST EN ISO 11267:2014
ISO 11267:2014(E)

3.10
standard soil
field-collected soil or artificial soil whose main properties (pH, texture, organic matter content) are
within a known range
EXAMPLE Euro soils, artificial soil, LUFA Standard soil.
Note 1 to entry: The properties of standard soils can differ from the soil to be tested.
3.11
control soil
reference or standard soil used as a control and as a medium for preparing dilution series with soils to
be tested or a reference substance, which fulfils the validity criteria
Note 1 to entry: In the case of natural soil, it is advisable to demonstrate its suitability for a test and for achieving
the test validity criteria before using the soil in a definitive test.
3.12
test mixture
mixture of contaminated soil or the test substance (e.g. chemical, biosolid, waste) with control soil
3.13
test mixture ratio
ratio between the soil to be tested and the control soil in a test mixture
4 Principle
The effects on reproduction of 10 d to 12 d old Collembola (Folsomia candida) exposed to the soil to be
tested are compared to those observed in a control soil. If appropriate, effects based on exposure to a
test mixture of contaminated soil and control soil or a range of concentrations of a test substance mixed
into control soil are determined. Test mixtures are prepared at the start of the test and are not renewed
within the test period.
The Collembola are incubated until offspring (F ) emerge from eggs laid by mature adults, and the
1
number of offspring is determined. Usually offspring emerge within 28 d in control experiments. The
results obtained from the tests are compared with a control or, if appropriate, are used to determine
the concentrations which cause no effects on mortality and reproduction (NOER/NOEC) and the
concentration resulting in x% reduction of juveniles hatched from eggs compared to the control
(ERx/ECx, 28 d) respectively.
If testing a concentration series, all test dilutions/concentrations above the LOER/LOEC have a harmful
effect equal to or greater than that observed at the LOER/LOEC. Where there is no prior knowledge of
the concentration of the soil to be tested or the test substance likely to have an effect, then it is useful to
conduct the test in two steps:
— An acute toxicity test (range-finding test) is carried out, to give an indication of the effect
dilution/concentration, and the dilution/concentration giving no mortality (NOER/NOEC).
Dilutions/concentrations to be used in the definitive test can then be selected;
— the definitive test on reproduction to determine sublethal effects of (dilutions of) contaminated
soil or the concentration of a substance which, when evenly mixed into the standard soil, causes
no significant effects on numbers of offspring hatched from eggs compared with the control
(NOER/NOEC), and the lowest concentration causing effects (LOER/LOEC).
NOTE The use of a reference soil is an essential requirement to demonstrate the present status of the test
population, and to avoid misinterpretation of results.
© ISO 2014 – All rights reserved 3

---------------------- Page: 15 ----------------------

SIST EN ISO 11267:2014
ISO 11267:2014(E)

5 Reagents and material
5.1 Biological material, in this test, 10 d to 12 d old juvenile springtails of the species Folsomia candida
Willem are used (see A.1 for details on synchronization of breeding).
5.2 Test mixture, which may consist of field-collected soil or control soil amended by the test substance.
5.2.1 Field-collected soil or waste
The sample(s) can be field-collected soil from an industrial, agricultural or other site of concern, or
waste materials (e.g. dredged material, municipal sludge from a wastewater treatment plant, composed
material, or manure) under consideration for possible land disposal.
The field-collected soils used in the test shall be passed through a sieve of 4 mm square mesh to remove
coarse fragments and thoroughly mixed. If necessary, soil may be air-dried without heating before
sieving. Storage of soil to be tested should be as short as possible. The soil shall be stored in accordance
with ISO 10381-6 using containers that minimize losses of soil contaminants by volatilization and
sorption to the container walls. If soils or test mixtures have been stored, they should be mixed a second
time immediately before use. Soil pH should not be corrected as it can influence bioavailability of soil
contaminants.
For interpretation of test results, the following characteristics shall be determined for each soil sampled
from a field site:
a) pH in accordance with ISO 10390,
b) texture (sand, loam, silt) in accordance with ISO 11277
c) water content in accordance with ISO 11465,
d) water-holding capacity according to Annex B,
e) cationic exchange capacity in accordance with ISO 11260,
f) organic carbon in accordance with ISO 10694,
g) percentage of material removed by the 4 mm sieve
NOTE It is important to measure the water holding capacity of all mixtures used in the test.
5.2.2 Control soil, either a) reference soil (3.9) or b) standard soil (3.10) that allows the presence of
Collembola. Control soil and soil used for dilution shall not differ in one test (either a) or b)).
a) If reference soils from uncontaminated areas near a contaminated site are available, they should
be treated and characterized like the soils to be tested. If a toxic contamination or unusual soil
properties cannot be ruled out, standard control soils should be preferred.
b) For testing the effects of substances mixed into soil, standard soils (e.g. artificial soil, LUFA) shall be
used as test substrate. The properties of the field-collected standard soil shall be reported.
4 © ISO 2014 – All rights reserved

---------------------- Page: 16 ----------------------

SIST EN ISO 11267:2014
ISO 11267:2014(E)

The substrate called artificial soil can be used as a standard soil and has the following composition:
Percentage expressed on dry mass basis
   − Sphagnum peat finely ground [a particle size of          10 %
(2 ± 1) mm is acceptable] and with no visible plant remains
   − Kaolinite clay containing not less than 30 % kaolinite          20 %
   − Industrial quartz sand (dominant fine sand with more          69 %
than 50 % of particle size 0,05 mm to 0,2 mm)
Approximately 0,3 % to 1,0 % calcium carbonate (CaCO , pulverized, analytical grade) are necessary to
3
get a pH of 6,0 ± 0,5.
NOTE 1 Taking the properties of highly non-polar (log K > 2) or ionizing substances into account, 5 % of peat
ow
have proven to be sufficient for maintaining the desired structure of the artificial soil.
NOTE 2 It has been demonstrated that Folsomia candida can comply with the validity criteria even on
reproduction when tested in field soils with lower organic carbon content (e.g. 2,7 %), and there is experience
that this can be achieved in artificial soil with 5 % peat. Therefore, it is not necessary before using such a soil in a
definitive test to demonstrate the suitability of the artificial soil for allowing the test to comply with the validity
criteria unless the peat contents is lower than specified above.
Prepare the artificial soil at least three days prior to the start of the test, by mixing the dry constituents
listed above thoroughly in a large-scale laboratory mixer. A portion of the deionized water required
is added during mixing. Allowance should be made for any water that is used
...

SLOVENSKI STANDARD
oSIST prEN ISO 11267:2012
01-november-2012
Kakovost tal - Zaviranje razmnoževanja Collembole (Folsomia candida) zaradi
onesnaževal v tleh (ISO/DIS 11267:2012)
Soil quality - Inhibition of reproduction of Collembola (Folsomia candida) by soil
pollutants (ISO/DIS 11267:2012)
Bodenbeschaffenheit - Hemmung der Reproduktion von Collembolen (Folsomia candida)
durch Bodenschadstoffe (ISO/DIS 11267:2012)
Qualité du sol - Inhibition de la reproduction de Collembola (Folsomia candida) par des
polluants du sol (ISO/DIS 11267:2012)
Ta slovenski standard je istoveten z: prEN ISO 11267
ICS:
13.080.30 Biološke lastnosti tal Biological properties of soils
oSIST prEN ISO 11267:2012 en,fr,de
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

---------------------- Page: 1 ----------------------
oSIST prEN ISO 11267:2012

---------------------- Page: 2 ----------------------
oSIST prEN ISO 11267:2012


EUROPEAN STANDARD
DRAFT
prEN ISO 11267
NORME EUROPÉENNE

EUROPÄISCHE NORM

September 2012
ICS 13.080.30
English Version
Soil quality - Inhibition of reproduction of Collembola (Folsomia
candida) by soil pollutants (ISO/DIS 11267:2012)
Qualité du sol - Inhibition de la reproduction de Collembola Bodenbeschaffenheit - Hemmung der Reproduktion von
(Folsomia candida) par des polluants du sol (ISO/DIS Collembolen (Folsomia candida) durch Bodenschadstoffe
11267:2012) (ISO/DIS 11267:2012)
This draft European Standard is submitted to CEN members for parallel enquiry. It has been drawn up by the Technical Committee
CEN/TC 345.

If this draft becomes a European Standard, 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.

This draft European Standard was established by CEN 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,
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 United
Kingdom.

Recipients of this draft are invited to submit, with their comments, notification of any relevant patent rights of which they are aware and to
provide supporting documentation.

Warning : This document is not a European Standard. It is distributed for review and comments. It is subject to change without notice and
shall not be referred to as a European Standard.


EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION

EUROPÄISCHES KOMITEE FÜR NORMUNG

Management Centre: Avenue Marnix 17, B-1000 Brussels
© 2012 CEN All rights of exploitation in any form and by any means reserved Ref. No. prEN ISO 11267:2012: E
worldwide for CEN national Members.

---------------------- Page: 3 ----------------------
oSIST prEN ISO 11267:2012
prEN ISO 11267:2012 (E)
Contents Page
Foreword .3

2

---------------------- Page: 4 ----------------------
oSIST prEN ISO 11267:2012
prEN ISO 11267:2012 (E)
Foreword
This document (prEN ISO 11267:2012) has been prepared by Technical Committee ISO/TC 190 “Soil quality”
in collaboration with Technical Committee CEN/TC 345 “Characterization of soils” the secretariat of which is
held by NEN.
This document is currently submitted to the parallel Enquiry.
Endorsement notice
The text of ISO/DIS 11267:2012 has been approved by CEN as a prEN ISO 11267:2012 without any
modification.
3

---------------------- Page: 5 ----------------------
oSIST prEN ISO 11267:2012

---------------------- Page: 6 ----------------------
oSIST prEN ISO 11267:2012

DRAFT INTERNATIONAL STANDARD ISO/DIS 11267
ISO/TC 190/SC 4 Secretariat: AFNOR
Voting begins on Voting terminates on

2012-09-06 2013-02-06
INTERNATIONAL ORGANIZATION FOR STANDARDIZATION    МЕЖДУНАРОДНАЯ ОРГАНИЗАЦИЯ ПО СТАНДАРТИЗАЦИИ    ORGANISATION INTERNATIONALE DE NORMALISATION


Soil quality — Inhibition of reproduction of Collembola
(Folsomia candida) by soil pollutants
Qualité du sol — Inhibition de la reproduction de Collembola (Folsomia candida) par des polluants du sol
[Revision of first edition (ISO 11267:1999)]
ICS 13.080.30


ISO/CEN PARALLEL PROCESSING
This draft has been developed within the International Organization for Standardization (ISO), and
processed under the ISO-lead mode of collaboration as defined in the Vienna Agreement.
This draft is hereby submitted to the ISO member bodies and to the CEN member bodies for a parallel
five-month enquiry.
Should this draft be accepted, a final draft, established on the basis of comments received, will be
submitted to a parallel two-month approval vote in ISO and formal vote in CEN.


To expedite distribution, this document is circulated as received from the committee
secretariat. ISO Central Secretariat work of editing and text composition will be undertaken at
publication stage.
Pour accélérer la distribution, le présent document est distribué tel qu'il est parvenu du
secrétariat du comité. Le travail de rédaction et de composition de texte sera effectué au
Secrétariat central de l'ISO au stade de publication.



THIS DOCUMENT IS A DRAFT CIRCULATED FOR COMMENT AND APPROVAL. IT IS THEREFORE SUBJECT TO CHANGE AND MAY NOT BE
REFERRED TO AS AN INTERNATIONAL STANDARD UNTIL PUBLISHED AS SUCH.
IN ADDITION TO THEIR EVALUATION AS BEING ACCEPTABLE FOR INDUSTRIAL, TECHNOLOGICAL, COMMERCIAL AND USER PURPOSES,
DRAFT INTERNATIONAL STANDARDS MAY ON OCCASION HAVE TO BE CONSIDERED IN THE LIGHT OF THEIR POTENTIAL TO BECOME
STANDARDS TO WHICH REFERENCE MAY BE MADE IN NATIONAL REGULATIONS.
RECIPIENTS OF THIS DRAFT ARE INVITED TO SUBMIT, WITH THEIR COMMENTS, NOTIFICATION OF ANY RELEVANT PATENT RIGHTS OF WHICH
THEY ARE AWARE AND TO PROVIDE SUPPORTING DOCUMENTATION.
©  International Organization for Standardization, 2012

---------------------- Page: 7 ----------------------
oSIST prEN ISO 11267:2012
ISO/DIS 11267

Copyright notice
This ISO document is a Draft International Standard and is copyright-protected by ISO. Except as permitted
under the applicable laws of the user’s country, neither this ISO draft nor any extract from it may be
reproduced, stored in a retrieval system or transmitted in any form or by any means, electronic,
photocopying, recording or otherwise, without prior written permission being secured.
Requests for permission to reproduce should be addressed to 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
Reproduction may be subject to royalty payments or a licensing agreement.
Violators may be prosecuted.

ii © ISO 2012 – All rights reserved

---------------------- Page: 8 ----------------------
oSIST prEN ISO 11267:2012
ISO/DIS 11267
Contents Page
Foreword . iv
Introduction . v
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 2
4 Principle . 3
5 Reagents . 4
6 Apparatus . 6
7 Procedure . 6
8 Calculation and expression of results . 9
9 Validity of the test . 10
10 Statistical analysis . 10
11 Test report . 11
Annex A (informative) Techniques for rearing and breeding of Collembola . 13
Annex B (informative) Extraction and counting of Collembola . 15
Annex C (informative) Determination of water-holding capacity . 16
Annex D (informative) Guidance on adjustment of pH of artificial soil . 17
Bibliography . 18

© ISO 2012 – All rights reserved iii

---------------------- Page: 9 ----------------------
oSIST prEN ISO 11267:2012
ISO/DIS 11267
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 11267 was prepared by Technical Committee ISO/TC 190, Soil quality, Subcommittee SC 4, Biological
methods.
This second/third/. edition cancels and replaces the first/second/. edition (), [clause(s) / subclause(s) /
table(s) / figure(s) / annex(es)] of which [has / have] been technically revised.

iv © ISO 2012 – All rights reserved

---------------------- Page: 10 ----------------------
oSIST prEN ISO 11267:2012
ISO/DIS 11267
Introduction
Ecotoxicological test systems are applied to obtain information about the effects of contaminants in soil and
are proposed to complement conventional chemical analysis (see ISO 15799 and ISO 17616). ISO 15799
includes a list and short characterisation of recommended and standardized test systems and ISO 17616
gives guidance on the choice and evaluation of the bioassays. 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.
Soil dwelling Collembola are ecologically relevant species for ecotoxicological testing. Springtails are prey
animals for a variety of endogeic and epigeic invertebrates and they contribute to decomposition processes in
soils. In acidic soils they may be the most important soil invertebrates besides enchytraeids with respect to
that function, since earthworms are typically absent [23]. Additionally, Collembola represent arthropod species
with a different route and a different rate of exposure compared to earthworms [3] and enchytraeids [8].
Various species were used in bioassays of which four species were used most commonly, Folsomia candida,
Folsomia fimetaria, Onychiurus armatus, and Orchesella cincta [24]. Numerous soil toxicity tests supported by
Environment Canada (EC) resulted in the development and standardization of a a biological test method for
determining the lethal and sublethal toxicity of samples of contaminated soil to Collembola [14]. The method
prepared by EC includes 3 species, Orthonychiurus folsomi, Folsomia candida, and Folsomia fimetaria. As
standardized test systems using Collembola as indicator organisms for the habitat function of soil, another two
methods exist. One is designed for assessing the effects of chemicals on the reproductive output of the
Collembola, Folsomia fimetaria L. and Folsomia candida Willem in soil [23], [25], and the other method being
described here, focuses on testing contaminated soil. Optionally the method can be used for testing chemicals
added to standard soils (e.g. artificial soil) for their sublethal hazard potential to Collembola.
This International Standard describes a method that is based on the determination of sublethal effects of
contaminated soils to adult Collembola of the species Folsomia candida (Willem). The species is distributed
world wide. It plays a similar ecological role as Folsomia fimetaria [14], [23]. Folsomia candida reproduces
parthenogenetically and is an easy accessible species as it is commercially available and easy to culture.
Folsomia candida is considered to be a representative of soil arthropods and Collembola in particular.
Background information on the ecology of springtails and their use in ecotoxicological testing is available [26].
© ISO 2012 – All rights reserved v

---------------------- Page: 11 ----------------------
oSIST prEN ISO 11267:2012

---------------------- Page: 12 ----------------------
oSIST prEN ISO 11267:2012
DRAFT INTERNATIONAL STANDARD ISO/DIS 11267

Soil quality — Inhibition of reproduction of Collembola
(Folsomia candida) by soil pollutants
1 Scope
This International Standard specifies one of the methods for evaluating the habitat function of soils and
determining effects of soil contaminants and chemicals to the reproduction of Folsomia candida (Willem) by
dermal and alimentary uptake. This chronic test is applicable to soils and soil materials of unknown quality
e. g. from contaminated sites, amended soils, soils after remediation, agricultural or other sites under concern
and waste materials.
Effects of substances are assessed using a standard soil, preferably a defined artificial soil substrate. For
contaminated soils, the effects are determined in the test soil and in a control soil. According to the objective
of the study, the control and dilution substrate (dilution series of contaminated soil) should be either an
uncontaminated soil comparable to the soil sample to be tested (reference soil) or a standard soil (e. g.
artificial soil).
Information is provided how to use this method for testing chemicals. The method is not applicable to volatile
substances, i. e. substances for which H (Henry's constant) or the air/water partition coefficient is greater than
1, or for which the vapour pressure exceeds 0,013 3 Pa at 25 °C. The stability of the test substance cannot be
assured over the test period. No allowance is made in the test method described for possible degradation of
the test substance over the course of the experiment.
WARNING — Contaminated soils may contain unknown mixtures of toxic, mutagenic, or otherwise
harmful chemicals or infectious micro-organisms. Occupational health risks may arise from dust or
evaporated chemicals during handling and incubation. Precautions should be taken to avoid skin
contact.
2 Normative references

The following documents, in whole or in part, are normatively referenced in this document and are
indispensable for its application. For dated references, only the edition cited applies. For undated references,
the latest edition of the referenced document (including any amendments) applies.
ISO 10381-6, Soil quality — Sampling — Part 6: Guidance on the collection, handling and storage of soil
under aerobic conditions for the assessment of microbial processes, biomass and diversity in the laboratory
ISO 10694, Soil quality — Determination of organic and total carbon after dry combustion (elementary
analysis)
ISO 10390:2005, Soil quality — Determination of pH
ISO 11260, Soil quality — Determination of effective cation exchange capacity and base saturation level using
barium chloride solution
ISO/DIS 11268-2:2011, Soil quality — Effects of pollutants on earthworms — Part 2: Determination of effects
on reproduction to Eisenia fetida/Eisenia andrei
© ISO 2012 – All rights reserved 1

---------------------- Page: 13 ----------------------
oSIST prEN ISO 11267:2012
ISO/DIS 11267
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 15176:2002, Soil quality — Characterization of excavated soil and other soil materials intended for re-use
3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
3.1
contaminant
substance or agent present in the soil as a result of human activity
[SOURCE: ISO 15176:2002].
3.2
ERx (effective rate) or ECx (effective concentration)
x % effect rate or concentration of the test soil or test substance at which reproduction is reduced by x %
compared to the control
3.3
limit test
single concentration test consisting of at least four replicates each, the test soil without any dilution or the
highest concentration of test substance mixed into the control soil and the control.
3.4
lowest observed effect rate (LOER) or effect concentration (LOEC)
lowest tested percentage of a test soil in a control soil or concentration of a substance at which a statistically
significant effect is observed
Note 1 to entry: The LOEC is expressed as percentage of test soil dry mass per test mixture dry mass. All test
mixtures above the LOEC have a harmful effect equal or greater than observed at the LOEC. If this condition cannot be
satisfied, an explanation should be given for how the LOEC and NOEC (3.4) have been selected.
3.5
NOER (No Observed Effective Rate) or NOEC (No Observed Effect Concentration)
test soil percentage immediately below the LOER/LOEC or, highest tested concentration of a test substance
which when compared to the control has no statistically significant lethal or other effect such as reduced
reproduction or mass alteration (p < 0,05)
Note 1 to entry: The NOEC is expressed as percentage of test soil dry mass per test mixture dry mass.
3.6
reference soil
uncontaminated site-specific soil (e. g. collected in the vicinity of a contaminated site) with similar properties
(nutrient concentrations, pH, organic carbon content and texture) as the test soil
2 © ISO 2012 – All rights reserved

---------------------- Page: 14 ----------------------
oSIST prEN ISO 11267:2012
ISO/DIS 11267
3.7
standard soil
field collected soil or artificial soil whose main properties (e. g. pH, texture, organic matter content) are within a
known range
EXAMPLE Euro-Soils [20 ], artificial soil [22], LUFA Standard Soil [21].
Note 1 to entry: The properties of standard soils can differ from the test soil.
3.8
control soil
reference or standard soil used as a control and as medium for preparing dilution series with test soils or a
reference substance, that allows the presence of Collembola. For natural soil it is advisable to demonstrate its
suitability for a test and for achieving the test validity criteria before using the soil in a definitive test.

3.9
test mixture
mixture of contaminated soil or the test substance with control soil (3.8)
Note 1 to entry: Test mixtures are given in per cent contaminated soil based on soil dry mass.
3.10
test mixture ratio
ratio between the test soil and the control soil in a test mixture
Note 1 to entry: Different ratios may be applied in a dilution series to establish a dose-response relationship.
4 Principle
The effects on reproduction of 9- to 12-day old Collembola (Folsomia candida) exposed to the test soil are
compared to those observed to samples exposed to a control soil. If appropriate, effects based on exposure to
a test mixture of contaminated soil and control soil or a range of concentrations of a test substance mixed into
control soil are determined. Test mixtures are prepared at the start of the test and are not renewed within the
test period.
The Collembola are incubated until offspring (F ) emerge from eggs laid by mature adults, and the number of
1
offspring is determined. Normally offspring emerge within 28 days in control experiments. The results obtained
from the tests are compared with a control soil or, if appropriate, are used to determine the concentrations
which cause no effects on mortality and reproduction (NOER/NOEC) and the concentration resulting in x %
reduction of juveniles hatched from eggs compared to the control (ERx/ECx, 28 d) respectively.
In case of testing a concentration series all test concentrations above the LOER/LOEC shall have a harmful
effect equal to, or greater than that observed at the LOER/LOEC. Where there is no prior knowledge of the
concentration of the test soil or the test substance likely to have an effect, then it is useful to conduct the test
in two steps:
a preliminary test is carried out to give an indication of the effect dilution/concentration, and the
concentration giving no mortality (NOER/NOEC). Concentrations to be used in the definitive test can then
be selected;
the definitive test to determine sublethal effects of (dilutions of) contaminated soil or the concentration of
a chemical which, when evenly mixed into the standard soil, causes no significant effects on numbers of
offspring hatched from eggs compared with the control (NOER/NOEC), and the lowest concentration
causing effects (LOER/LOEC).
© ISO 2012 – All rights reserved 3

---------------------- Page: 15 ----------------------
oSIST prEN ISO 11267:2012
ISO/DIS 11267
NOTE The use of a reference soil is an essential requirement to demonstrate the present status of the test
population, and to avoid misinterpretation of results.
5 Reagents
5.1 Biological material
In this test, 9- to 12-day old juvenile springtails of the species Folsomia candida (Willem) are used (see A.1 for
details on synchronization of breeding).
5.2 Test sample may consist of field-collected soil or control soil amended by the test substance.
5.2.1 Field-collected soils, soil or waste materials
The sample(s) can be field-collected soil from an industrial, agricultural or other site of concern, or waste
materials (e. g. dredged material, municipal sludge from a wastewater treatment plant, composed material, or
manure) under consideration for possible land disposal.
Test soils shall be sieved by 4 mm mesh and thoroughly mixed. If necessary, soil may be air dried without
heating before sieving. Storage of test soils should be as short as possible. Store the soil in accordance with
ISO 10381-6 using containers that minimise losses of soil contaminants by volatilisation and sorption to the
container walls. If soils or soil mixtures have been stored, they should be mixed a second time immediately
before use. Soil pH should not be corrected as it can influence bioavailability of soil contaminants.
For interpretation of test results, the following characteristics shall be determined for each soil sampled from a
field site:
a) pH in accordance with ISO 10390,
b) texture (sand, loam, silt) in accordance with ISO 11277
c) water content in accordance with ISO 11465,
d) water holding capacity according to Annex D,
e) cationic exchange capacity in accordance with ISO 11260,
f) organic carbon in accordance with ISO 10694,
g) percentage of material removed by the 4 mm sieve
NOTE It is important to measure the water holding capacity of all mixtures used in the test.
5.2.2 Control soil, either a) reference soil (3.6) or b) standard soil (3.7) that allows the presence of
Collembola.
a) If reference soils from uncontaminated areas near a contaminated site are available, they should be
treated and characterised like the test soils. If a toxic contamination or unusual soil properties cannot be
ruled out, standard control soils should be preferred.
b) For testing the effects of substances mixed into soil or making dilutions of the test soil, standard soils
shall be used to prepare the test sample. The properties of the field collected standard soil shall be
reported.
The substrate called artificial soil can be used as a standard soil and has the following composition:
Percentage expressed on
4 © ISO 2012 – All rights reserved

---------------------- Page: 16 ----------------------
oSIST prEN ISO 11267:2012
ISO/DIS 11267
dry mass basis
Sphagnum peat finely ground and with no visible plant remains 10 %
Kaolinite clay containing not less than 30 % Kaolinite 20 %
Industrial quartz sand (dominant fine sand with more than 69 %
50 % of particle size 0,05 mm to 0,2 mm)
Approximately 0,3 % to 1,0 % calcium carbonate (CaCO , pulverised, analytical grade) are necessary to get a
3
pH of 6,0 0,5.
NOTE 1 Taking the properties of highly non-polar (log K > 2) or ionizing substances into account, 5 % of peat have
ow
proven to be sufficient for maintaining the desired structure of the artificial soil.
NOTE 2 It has been demonstrated that Folsomia candida can comply with the validity criteria even on reproduction
when tested in field soils with lower organic carbon content (e. g. 2,7 %), and there is experience that this can be achieved
in artificial soil with 5 % peat. Therefore, it is not necessary before using such a soil in a definitive test to demonstrate the
suitability of the artificial soil for allowing the test to comply with the validity criteria unless the peat contents lowered more
than specified above.
Prepare the artificial soil at least three days prior to start the test, by mixing the dry constituents listed above
thoroughly in a large-scale laboratory mixer. A portion of the deionized water required is added while mixing is
continued. Allowance should be made for any water that is used for introducing the test substance into the
soil. The amount of calcium carbonate required can vary, depending on properties of the individual batch of
sphagnum peat and should be determined by measuring sub-samples immediately before the test. Store the
mixed artificial soil at room temperature for at least two days to equilibrate acidity. To determine pH and the
maximum water holding capacity, the dry artificial soil is pre-moistened one or two days before starting the
test by adding deionised water to obtain approximately half of the required final water content of 40 % to 60 %
of the maximum water holding capacity.
The total water holding capacity is determined according to Annex D, the pH is determined according to
ISO 10390.
5.3 Reference substance
To ensure the quality of the test system, tests should be performed regularly (once or twice a year) with a
reference substance.
Boric acid and the agricultural chemical Betosip (a.i. 157 g/l Phenmedipham) have been tested in a ring test,
and are recommended as reference substances.
WARNING — When handling these chemicals, appropriate precautions should be taken to avoid
ingestion or skin contact.
NOTE 1 Boric acid: Effects on reproduction were observed at concentrations (IC ) of 147 mg boric acid/kg dry weight
50
of artificial soil and 169 mg boric acid/kg clay-loam soil dry weight) [9], [25].
NOTE 2 Betosip: Effects on reproduction ( 0,05) were observed at concentrations of between 100 mg and 200 mg
of the product per kilogram dry mass of the substrate.
5.4 Feeding
As a suitable food source, a sufficient amount, e.g. 2-10 mg, of granulated dried baker’s yeast, commercially
available for household use, is added to
...

Questions, Comments and Discussion

Ask us and Technical Secretary will try to provide an answer. You can facilitate discussion about the standard in here.