1、November 2015 English price group 12No part of this translation may be reproduced without prior permission ofDIN Deutsches Institut fr Normung e. V., Berlin. Beuth Verlag GmbH, 10772 Berlin, Germany,has the exclusive right of sale for German Standards (DIN-Normen).ICS 13.080.30!%GNQ“2364346www.din.d
2、eDIN EN ISO 11268-3Soil quality Effects of pollutants on earthworms Part 3: Guidance on the determination of effects in field situations (ISO 11268-3:2014);English version EN ISO 11268-3:2015,English translation of DIN EN ISO 11268-3:2015-11Bodenbeschaffenheit Wirkungen von Schadstoffen auf Regenwrm
3、er Teil 3: Anleitung fr die Bestimmung von Wirkungen unter Freilandbedingungen (ISO 11268-3:2014);Englische Fassung EN ISO 11268-3:2015,Englische bersetzung von DIN EN ISO 11268-3:2015-11Qualit du sol Effets des polluants vis-vis des vers de terre Partie 3: Lignes directrices relatives la dterminati
4、on des effets sur site (ISO 11268-3:2014);Version anglaise EN ISO 11268-3:2015,Traduction anglaise de DIN EN ISO 11268-3:2015-11SupersedesDIN ISO 11268-3:2000-03www.beuth.deDocument comprises 21 pagesDTranslation by DIN-Sprachendienst.In case of doubt, the German-language original shall be considere
5、d authoritative.10.15 DIN EN ISO 11268-3:2015-11 2 A comma is used as the decimal marker. National foreword The text of ISO 11268-3:2014 has been prepared by Technical Committee ISO/TC 190 “Soil quality” and has been taken over as EN ISO 11268-3:2015 by Technical Committee CEN/TC 345 “Characterizati
6、on of soils” (Secretariat: NEN, Netherlands). Expert assistance and specialized laboratories will be required to perform the analyses described in this standard. The responsible German body involved in its preparation was the DIN-Normenausschuss Wasserwesen (DIN Standards Committee Water Practice),
7、Working Committee NA 119-01-02-04 UA Biologische Verfahren of NA 119-01-02 AA Abfall- und Bodenuntersuchung. 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 P
8、art 2: Determination of effects on reproduction of Eisenia fetida/Eisenia andrei Part 3: Guidance on the determination of effects in field situations The DIN Standards corresponding to the International Standards referred to in this document are as follows: ISO 10390 DIN ISO 10390 ISO 10694 DIN EN 1
9、5936 ISO 11274 DIN EN ISO 11274 ISO 11277 DIN ISO 11277 ISO 23611-1 DIN EN ISO 23611-1 ISO 23611-2 DIN EN ISO 23611-2 ISO 23611-3 DIN EN ISO 23611-3 ISO 23611-4 DIN EN ISO 23611-4 ISO 23611-5 DIN EN ISO 23611-5 ISO 23611-6 DIN EN ISO 23611-6 Amendments This standard differs from DIN ISO 11268-3:2000
10、-03 as follows: a) ISO 11268-3 has been taken over as European Standard EN ISO 11268-3; b) the Introduction has been drawn up and added; c) the scope of the standard now also covers the application of chemicals by means of soil injection or drilled pelleted formulations; d) Clause 3 now also gives t
11、he unit for the concentration of test substances incorporated in the soil; e) reference is made to ISO 23611-1 on the sampling of earthworms; f) Clause 5 “Reagents and material” and Clause 6 “Apparatus” are now included; g) sampling is now done by hand-sorting and formalin extraction; the extraction
12、 method using mustard and electrical extraction are no longer included; h) Clause 7 “Procedure” and Clause 8 “Preparation for the test” have been restructured. DIN EN ISO 11268-3:2015-11 3 Previous editions DIN ISO 11268-3: 2000-03 National Annex NA (informative) Bibliography DIN EN 15936, Sludge, t
13、reated biowaste, soil and waste Determination of total organic carbon (TOC) by dry combustion DIN EN ISO 11274, Soil quality Determination of the water retention characteristics Laboratory methods DIN EN ISO 23611-1, Soil quality Sampling of soil invertebrates Part 1: Hand-sorting and formalin extra
14、ction of earthworms DIN EN ISO 23611-2, Soil quality Sampling of soil invertebrates Part 2: Sampling and extraction of micro-arthropods (Collembola and Acarina) DIN EN ISO 23611-3, Soil quality Sampling of soil invertebrates Part 3: Sampling and soil extraction of enchytraeids DIN EN ISO 23611-4, So
15、il quality Sampling of soil invertebrates Part 4: Sampling, extraction and identification of soil-inhabiting nematodes DIN EN ISO 23611-5, Soil quality Sampling of soil invertebrates Part 5: Sampling and extraction of soil macro-invertebrates DIN EN ISO 23611-6, Soil quality Sampling of soil inverte
16、brates Part 6: Guidance for the design of sampling programmes with soil invertebrate DIN ISO 10390, Soil quality Determination of pH DIN ISO 11277, Soil quality Determination of particle size distribution in mineral soil material Method by sieving and sedimentation DIN EN ISO 11268-3:2015-11 4 This
17、page is intentionally blank 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 d
18、es polluants vis-vis des vers 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
19、 European Standard was approved 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 reference
20、s 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 mem
21、ber 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, Fr
22、ance, 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 C O M I T EUR O P EN D E NOR M A L I S ATI
23、O N EUR O P I S C HES KOM I T E E F R NOR M UNG CEN-CENELEC 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 E Contents PageEuropean foreword .3Introduction
24、41 Scope . 52 Normative references 53 Units . 54 Principle 55 Reagents and material 66 Apparatus . 67 Procedure. 67.1 Sampling of earthworm populations . 67.2 Preservation 77.3 Determination of biomass . 78 Preparation for the test . 78.1 Test site 79 Procedure 109.1 Application of test substance .
25、109.2 Sampling dates . 109.3 Reference substance 1110 Data assessment. 1110.1 End points 1110.2 Identification of earthworm species . 1210.3 Determination of biomass with gut content . 1211 Calculation and expression of results 1212 Validity of the test . 1213 Test report 13Annex A (informative) Add
26、itional requirements of pesticide testing .14Annex B (informative) Information on specific earthworm species or communities in different climatic or geographic regions 15Bibliography .16DIN EN ISO 11268-3:2015-11 EN ISO 11268-3:2015 (E) 2 European foreword The text of ISO 11268-3:2014 has been prepa
27、red by Technical Committee 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
28、 the status of a national 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
29、 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,
30、 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, S
31、witzerland, Turkey 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. DIN EN ISO 11268-3:2015-11 EN ISO 11268-3:2015 (E) 3 IntroductionThe earthworm field test is based on a method being developed by the G
32、erman 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 Sheffie
33、ld, 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
34、) 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 o
35、f 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
36、-1 to ISO 23611-6.DIN EN ISO 11268-3:2015-11 EN ISO 11268-3:2015 (E) 4 1 ScopeThis part 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
37、 soil injections or drilled pelleted formulations.2 Normative referencesThe 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 editio
38、n of the referenced document (including any amendments) applies.ISO 10390, Soil quality 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 Laborator
39、y methodsISO 11277, Soil quality Determination of particle size distribution in mineral 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 a
40、re expressed in kilograms per hectare (kg/ha) or litres per hectare (l/ha) of the substance 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 m
41、g active ingredient (a.i.)/kg soil dry mass, dm. The same units are used when comparing 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 coll
42、ected from treated control and reference plots. Sampling is performed as specified in 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.Th
43、e test is of a randomized complete block design with four replicates per treatment. Statistical 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
44、 The test also generates samples of earthworms from treated plots for residue analysis where such information is appropriate.DIN EN ISO 11268-3:2015-11 EN ISO 11268-3:2015 (E) 5 5 Reagents and material5.1 Formalin formaldehyde solution, 4 % (volume fraction).5.2 Formalin formaldehyde solution, 37 %
45、(volume fraction).5.3 Ethanol, 70 % (volume 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 F
46、orceps.6.4 Piece of thick plastic (1 m2to 2 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,
47、e.g. for measuring air temperature.6.11 Drying 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 comb
48、ination is clearly recommended in the various 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 (i.e. are not reacting to formalin).Due
