1、BSI Standards PublicationBS ISO 11268-3:2014Soil quality Effects ofpollutants on earthwormsPart 3: Guidance on the determination ofeffects in field situationsBS EN ISO 11268-3:2015BS EN ISO 11268-3:2015 BRITISH STANDARDNational forewordThis British Standard is the UK implementation of EN ISO 11268-3
2、:2015. It is identical to ISO 11268-3:2014. It supersedes BS ISO 11268-3:2014 which is withdrawn.The UK participation in its preparation was entrusted to Technical Committee EH/4, Soil quality.A list of organizations represented on this committee can be obtained on request to its secretary.This publ
3、ication does not purport to include all the necessary provisions of a contract. Users are responsible for its correct application. The British Standards Institution 2015. Published by BSI Standards Limited 2015ISBN 978 0 580 89039 0ICS 13.080.30Compliance with a British Standard cannot confer immuni
4、ty from legal obligations.This British Standard was published under the authority of the Standards Policy and Strategy Committee on 30 November 2014.Amendments/corrigenda issued since publicationDate Text affected31 August 2015 This corrigendum renumbers BS ISO 11268-3:2014 as BS EN ISO 11268-3:2015
5、EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN ISO 11268-3 August 2015 ICS 13.080.30 English Version Soil quality - Effects of pollutants on earthworms - Part 3: Guidance on the determination of effects in field situations (ISO 11268-3:2014) Qualit du sol - Effets des polluants vis-vis des ver
6、s de terre - Partie 3: Lignes directrices relatives la dtermination des effets sur site (ISO 11268-3:2014) Bodenbeschaffenheit - Wirkungen von Schadstoffen auf Regenwrmer - Teil 3: Anleitung fr die Bestimmung von Wirkungen unter Freilandbedingungen (ISO 11268-3:2014)This European Standard was approv
7、ed by CEN on 6 August 2015. 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 st
8、andards 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
9、 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, Hungar
10、y, 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 COMIT EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR NORMUNG CEN-CENELE
11、C Management Centre: Avenue Marnix 17, B-1000 Brussels 2015 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN ISO 11268-3:2015 EEN ISO 11268-3:2015 (E) 3 European foreword The text of ISO 11268-3:2014 has been prepared by Technical C
12、ommittee ISO/TC 190 “Soil quality” of the International Organization for Standardization (ISO) and has been taken over as EN ISO 11268-3:2015 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 n
13、ational standard, either by publication of an identical text or by endorsement, at the latest by February 2016, and conflicting national standards shall be withdrawn at the latest by February 2016. Attention is drawn to the possibility that some of the elements of this document may be the subject of
14、 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
15、, 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
16、 and the United Kingdom. Endorsement notice The text of ISO 11268-3:2014 has been approved by CEN as EN ISO 11268-3:2015 without any modification. BS EN ISO 11268-3:2015EN ISO 11268-3:2015 (E)Contents PageForeword ivIntroduction v1 Scope . 12 Normative references 13 Units . 14 Principle 15 Reagents
17、and material 26 Apparatus . 27 Procedure. 27.1 Sampling of earthworm populations . 27.2 Preservation 37.3 Determination of biomass . 38 Preparation for the test . 38.1 Test site 39 Procedure. 69.1 Application of test substance 69.2 Sampling dates 69.3 Reference substance . 710 Data assessment 710.1
18、End points . 710.2 Identification of earthworm species 810.3 Determination of biomass with gut content 811 Calculation and expression of results . 812 Validity of the test 813 Test report . 9Annex A (informative) Additional requirements of pesticide testing .10Annex B (informative) Information on sp
19、ecific earthworm species or communities in different climatic or geographic regions 11Bibliography .12 ISO 2014 All rights reserved iiiBS EN ISO 11268-3:2015 ISO 11268-3:2014(E)ForewordISO (the International Organization for Standardization) is a worldwide federation of national standards bodies (IS
20、O 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, gove
21、rnmental 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 mainten
22、ance 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).Attenti
23、on 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 a
24、nd/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
25、assessment, as well as information about ISOs adherence to the WTO principles in the Technical Barriers to Trade (TBT) see the following URL: Foreword - Supplementary informationThe committee responsible for this document is ISO/TC 190, Soil quality, Subcommittee SC 4, Biological methods.This second
26、 edition cancels and replaces the first edition (ISO 11268-3:1999), which has been technically revised.ISO 11268 consists of the following parts, under the general title Soil quality Effects of pollutants on earthworms: Part 1: Determination of acute toxicity to Eisenia fetida/Eisenia andrei Part 2:
27、 Determination of effects on reproduction to Eisenia fetida/Eisenia andrei Part 3: Guidance on the determination of effects in field situationsiv ISO 2014 All rights reservedBS EN ISO 11268-3:2015 ISO 11268-3:2014(E)IntroductionThe earthworm field test is based on a method being developed by the Ger
28、man Federal Biological Research Centre for Agriculture and Forestry for the testing of pesticides.6Later, it was internationally standardized by the International Organization for Standardization (ISO), taking into account results and recommendations of an international workshop in 1991 in Sheffield
29、, United Kingdom, 7“Ecotoxicology of Earthworms”, as a tool for characterizing soil quality. Growing experience has shown that the practical performance of the test can be improved. In two meetings organized by the Federal Biological Research Centre for Agriculture and Forestry (Braunschweig, 2002)
30、and by the German Federal Agency for Consumer Protection and Food Safety (Lille, 2005), an ad-hoc working group of experts from various countries and institutions proposed recommendations that should be taken into account if revision has been approved by voting in the periodical review. A report of
31、the discussions, comments, and recommendations has been published.8In cases where earthworms and other organisms are used as bioindicators to assess the soil quality of a site as a habitat for soil organisms, guidance for extraction procedures and advice for planning a survey is given in ISO 23611-1
32、 to ISO 23611-6. ISO 2014 All rights reserved vBS EN ISO 11268-3:2015 ISO 11268-3:2014(E)BS ISO 11268-3:2014BS EN ISO 11268-3:2015BS ISO 11268-3:2014 BS ISO 11268-3:2014Soil quality Effects of pollutants on earthworms Part 3: Guidance on the determination of effects in field situations1 ScopeThis pa
33、rt of ISO 11268 specifies techniques for determining the effects of substances on earthworms in the field and provides a basis for determining the effects of chemicals applied to or incorporated into soil, including soil injections or drilled pelleted formulations.2 Normative referencesThe following
34、 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 10390, Soil quali
35、ty Determination of pHISO 10694, Soil quality Determination of organic and total carbon after dry combustion (elementary analysis)ISO 11274, Soil quality Determination of the water-retention characteristic Laboratory methodsISO 11277, Soil quality Determination of particle size distribution in miner
36、al soil material Method by sieving and sedimentationISO 23611-1, Soil quality Sampling of soil invertebrates Part 1: Hand-sorting and formalin extraction of earthworms3 UnitsRates of application of test substances are expressed in kilograms per hectare (kg/ha) or litres per hectare (l/ha) of the sub
37、stance applied. When this is a formulated material, the application rate is expressed in terms of the amount of active ingredient applied.The concentrations of test substances incorporated in the soil are given in mg active ingredient (a.i.)/kg soil dry mass, dm. The same units are used when compari
38、ng the results of this field test with those gained in laboratory studies.4 PrincipleSpecies, numbers, and biomass of earthworms collected by sampling plots treated with a test substance are compared with those collected from treated control and reference plots. Sampling is performed as specified in
39、 ISO 23611-1. The duration of the study depends on the characteristics of the test substance but is usually of one years duration. Sampling dates are chosen to lie within the periods of activity of the earthworms.The test is of a randomized complete block design with four replicates per treatment. S
40、tatistical analysis of numbers of each species collected at each sampling occasion is used to determine the effects of treatments by comparing abundance, biomass, and diversity between control and treated plots.NOTE The test also generates samples of earthworms from treated plots for residue analysi
41、s where such information is appropriate.INTERNATIONAL STANDARD ISO 11268-3:2014(E) ISO 2014 All rights reserved 1BS EN ISO 11268-3:20155 Reagents and material5.1 Formalin formaldehyde solution, 4 % (volume fraction).5.2 Formalin formaldehyde solution, 37 % (volume fraction).5.3 Ethanol, 70 % (volume
42、 fraction).5.4 Carbendazim, e.g. applied as Derosal1)formulation (360 g a.i./l) as reference substance.6 ApparatusUse standard laboratory equipment and the following.6.1 Plastic vessel, (250 ml and 500 ml) for storing the worms.6.2 Plastic hand gloves.6.3 Forceps.6.4 Piece of thick plastic (1 m2to 2
43、 m2).6.5 Spade or shovel.6.6 Dissecting microscope, with low magnification (10 to 40).6.7 Balance (0,01 g to 200 g).6.8 Water-can, (preferably 20 l) with water (20 l per sampling plot).6.9 Pencil, note book, water resistant marker, labels.6.10 Thermometer, e.g. for measuring air temperature.6.11 Dry
44、ing cabinet, for soil water content determination.7 Procedure7.1 Sampling of earthworm populationsSampling of earthworms is done by a combination of two different methods: hand-sorting and formalin extraction. Based on several comparative studies, this combination is clearly recommended in the vario
45、us reviews on earthworm ecology (e.g. 9, 10, 13). For details of extraction procedures, see ISO 23611-1.Sampling should be done at times of the year where the animals are not forced by the environmental conditions (i.e. low soil water content and/or extremely high or low temperatures) into diapause
46、(i.e. are not reacting to formalin).Due to the individual size of the worms, a large plot shall be identified: a square of 50 cm 50 cm is often sufficient in the Holarctic where most adult earthworms have a length approximately between 1) Derosal is an example of a suitable product available commerc
47、ially. This information is given for the convenienceof users of this document and does not constitute an endorsement by ISO of this product.2 ISO 2014 All rights reservedBS EN ISO 11268-3:2015 ISO 11268-3:2014(E)1 cm and 20 cm. However, at places with a low density of earthworms e.g. soils with low
48、pH (10 % dominance and/or at least 10 individuals/m2to 15 individuals/m2). If at a sampling date no species is dominant in one of the endogeic or anecic groups, earthworms belonging to the same ecological group (e.g. endogeics) or belonging to the same morphological groups (e.g. Tanylobous and Epilo
49、bous) can be combined. Evaluation shall be performed at the species level or for each of the respective ecological or morphological groups; abundance and biomass of juvenile and adult earthworms belonging to non-dominant species as described above. Since juvenile earthworms of the same genus can often not be identified to the species level, the evaluation can be restricted to morphological groups (Tanylobous and Epilobous).NOTE 1 Despite the fact that the selection of the test site was based on the occurrenc
