1、November 2015 English price group 14No 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!%GNO“2364344www.din.d
2、eDIN EN ISO 11268-2S o i l q u a l i t y E f f e c t s o f p o l l u t a n t s o n e a r t h w o r m s P a r t 2 : D e t e r m i n a t i o n o f e f f e c t s o n r e p r o d u c t i o n o f E i s e n i a f e t i d a / E i s e n i a a n d r e i ( I S O 1 1 2 6 8 - 2 : 2 0 1 2 ) ;E n g l i s h v e r
3、s i o n E N I S O 1 1 2 6 8 - 2 : 2 0 1 5 ,English translation of DIN EN ISO 11268-2:2015-11B o d e n b e s c h a f f e n h e i t W i r k u n g e n v o n S c h a d s t o f f e n a u f R e g e n w r m e r T e i l 2 : B e s t i m m u n g d e r W i r k u n g a u f d i e R e p r o d u k t i o n s l e i
4、s t u n g v o n E i s e n i a f e t i d a / E i s e n i a a n d r e i ( I S O 1 1 2 6 8 - 2 : 2 0 1 2 ) ;E n g l i s c h e F a s s u n g E N I S O 1 1 2 6 8 - 2 : 2 0 1 5 ,Englische bersetzung von DIN EN ISO 11268-2:2015-11Q u a l i t d u s o l E f f e t s d e s p o l l u a n t s v i s - - v i s d e
5、 s v e r s d e t e r r e P a r t i e 2 : D t e r m i n a t i o n d e s e f f e t s s u r l a r e p r o d u c t i o n d e E i s e n i a f e t i d a / E i s e n i a a n d r e i ( I S O 1 1 2 6 8 - 2 : 2 0 1 2 ) ;V e r s i o n a n g l a i s e E N I S O 1 1 2 6 8 - 2 : 2 0 1 5 ,Traduction anglaise de DI
6、N EN ISO 11268-2:2015-11SupersedesD I N I S O 1 1 2 6 8 2 : 2 0 0 0 0 3www.beuth.deDocument comprises 29 pagesDTranslation by DIN-Sprachendienst.In case of doubt, the German-language original shall be considered authoritative.10.15 DIN EN ISO 11268-2:2015-11 2 A comma is used as the decimal marker.
7、National foreword The text of ISO 11268-2:2012 has been prepared by Technical Committee ISO/TC 190 “Soil quality” and has been taken over as EN ISO 11268-2:2015 by Technical Committee CEN/TC 345 “Characterization of soils” (Secretariat: NEN, Netherlands). Expert assistance and specialized laboratori
8、es 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), Working Committee NA 119-01-02-04 UA Biologische Verfahren of NA 119-01-02 AA Abfall- und
9、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 Part 2: Determination of effects on reproduction of Eisenia fetida/Eisenia andrei Part 3: G
10、uidance 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 10381-6 DIN ISO 10381-6 ISO 10390 DIN ISO 10390 ISO 10694 DIN EN 15936 ISO 11260 DIN EN ISO 11260 ISO 11268-1 DIN EN ISO 11268-1
11、 ISO 11277 DIN ISO 11277 ISO 11465 DIN EN 15934 ISO 15799 DIN ISO 15799 ISO 17616 DIN ISO 17616 Amendments This standard differs from DIN ISO 11268-2:2000-03 as follows: a) Eisenia andrei is now included as a test organism; b) boric acid is now the reference substance; c) Clause 10 “Statistical anal
12、ysis” has been included; d) Annex A “Determination of the acute toxicity of chemicals (in particular pesticides) on earthworms under tropical test conditions” has been included; e) former Annexes D and E have been deleted; f) E.4 (Annex F in the previous edition) “Results of tests using boric acid a
13、s reference substance” has been added; g) the standard has been editorially revised. DIN EN ISO 11268-2:2015-11 3 DIN ISO 11268-2: 2000-03 National Annex NA (informative) Bibliography DIN EN 15934, Sludge, treated biowaste, soil and waste Calculation of dry matter fraction after determination of dry
14、 residue or water content DIN EN 15936, Sludge, treated biowaste, soil and waste Determination of total organic carbon (TOC) by dry combustion DIN EN ISO 11268-1, Soil quality Effects of pollutants on earthworms Part 1: Determination of acute toxicity to Eisenia fetida/Eisenia andrei DIN EN ISO 1126
15、0, Soil quality Determination of effective cation exchange capacity and base saturation level using barium chloride solution DIN 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 processe
16、s, biomass and diversity in the laboratory 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 ISO 15799, Soil quality Guidance on the ecotoxicological characterizatio
17、n of soils and soil materials DIN ISO 17616, Soil quality Guidance on the choice and evaluation of bioassays for ecotoxicological characterization of soils and soil materials Previous editions DIN EN ISO 11268-2:2015-11 4 This page is intentionally blank EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE
18、NORM EN ISO 11268-2 August 2015 ICS 13.080.30 English Version Soil quality - Effects of pollutants on earthworms - Part 2: Determination of effects on reproduction of Eisenia fetida/Eisenia andrei (ISO 11268-2:2012) Qualit du sol - Effets des polluants vis-vis des vers de terre - Partie 2: Dterminat
19、ion des effets sur la reproduction de Eisenia fetida/Eisenia andrei (ISO 11268-2:2012) Bodenbeschaffenheit - Wirkungen von Schadstoffen auf Regenwrmer - Teil 2: Bestimmung der Wirkung auf die Reproduktionsleistung von Eisenia fetida/Eisenia andrei (ISO 11268-2:2012) This European Standard was approv
20、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
21、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
22、 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
23、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
24、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 11268-2:2015 EContents PageEuropean foreword . 3Introduction 41 Scope 52 Normative references . 53 Terms and definition
25、s . 64 Principle . 75 Reagents and material . 86 Apparatus 97 Procedure 107.1 Experimental design . 107.2 Preparation of test mixture . 117.3 Addition of the earthworms 127.4 Test conditions and measurements . 127.5 Reference substance 138 Calculation and expression of results . 138.1 Calculation 13
26、8.2 Expression of results . 139 Validity of the test 1310 Statistical analysis 1310.1 General . 1310.2 Single-concentration tests 1410.3 Multi-concentration tests 1411 Test report . 15Annex A (informative) Determination of the chronic toxicity of chemicals on earthworms under tropical conditions 17A
27、nnex B (informative) Culturing of Eisenia fetida and Eisenia andrei .19Annex C (informative) Determination of water holding capacity of artificial soil 20Annex D (informative) Techniques for counting juvenile worms hatched from cocoons.21Annex E (informative) Performance of the method .22Bibliograph
28、y .24DIN EN ISO 11268-2:2015-11 EN ISO 11268-2:2015 (E) 2 3 European foreword The text of ISO 11268-2:2012 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 11268-2:2015 by Technical Commit
29、tee 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 February 2016, and conflicting national standards shall be withd
30、rawn 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 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
31、, 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, L
32、atvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom. Endorsement notice The text of ISO 11268-2:2012 has been approved by CEN as EN ISO 11268-2:2015 without any modification. DIN EN ISO 112
33、68-2:2015-11 EN ISO 11268-2:2015 (E) 3 IntroductionEcotoxicological 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 34and ISO 17616 35). ISO 15799 includes a list and short character
34、ization 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 (retentio
35、n function of soils), whereas terrestrial test systems are used to assess the habitat function of soils. As standardized test systems using earthworms as indicator organisms for the habitat function of soil, an acute test for survival and a chronic test for reproduction are available.This part of IS
36、O 11268 describes a method that is based on the determination of sublethal effects of contaminated soils on adult earthworms of the species Eisenia fetida (Savigny 1826) and Eisenia andrei (Andr 1963). Optionally, the method can be used for testing chemicals added to standard soils (e.g. artificial
37、soil) for their sublethal hazard potential to earthworms. Finally, information is provided on how to use this method for testing chemicals under tropical conditions (see Annex A).Eisenia fetida and Eisenia andrei are considered to be representatives of soil fauna and earthworms in particular. Backgr
38、ound information on the ecology of earthworms and their use in ecotoxicological testing is available. Other species, e.g. Aporrectodea caliginosa, Lumbricus rubellus and Lumbricus terrestris, have also been used as test organisms. These or other species have not been proven to be more sensitive in g
39、eneral, and the database and experience in testing soils is small 1617.This part of ISO 11268 has been drawn up taking into consideration test procedures adopted by the Organization for Economic Cooperation and Development 2728and by the European Union 11.DIN EN ISO 11268-2:2015-11 EN ISO 11268-2:20
40、15 (E) 4 WARNING Contaminated soils may contain unknown mixtures of toxic, mutagenic, or otherwise harmful chemicals or infectious microorganisms. Occupational health risks may arise from dust or evaporated chemicals during handling and incubation. Precautions should be taken to avoid skin contact.1
41、 ScopeThis part of ISO 11268 specifies one of the methods for evaluating the habitat function of soils and determining the effects of soil contaminants and chemicals on the reproduction of Eisenia fetida/Eisenia andrei by dermal and alimentary uptake. This chronic test is applicable to soils and soi
42、l materials of unknown quality, e.g. from contaminated sites, amended soils, soils after remediation, agricultural or other sites concerned, and waste materials.Effects of substances are assessed using a standard soil, preferably a defined artificial soil substrate. For contaminated soils, the effec
43、ts 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. ar
44、tificial soil).Information is provided on how to use this method for testing chemicals under temperate as well as under tropical conditions.The method is not applicable to volatile substances, i.e. substances for which H (Henrys constant) or the air/water partition coefficient is greater than 1, or
45、for which the vapour pressure exceeds 0,013 3 Pa at 25 C.This method does not take into account the persistence of the substance during the test.2 Normative referencesThe following referenced documents are indispensable for the application of this document. For dated references, only the edition cit
46、ed 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 a
47、nd diversity in the laboratoryISO 10390, Soil quality Determination of pHISO 10694, Soil quality Determination of organic and total carbon after dry combustion (elementary analysis)ISO 11260, Soil quality Determination of effective cation exchange capacity and base saturation level using barium chlo
48、ride solutionISO 11268-1, Soil quality Effects of pollutants on earthworms Part 1: Determination of acute toxicity to Eisenia fetida/Eisenia andreiISO 11277, Soil quality Determination of particle size distribution in mineral soil material Method by sieving and sedimentationISO 11465, Soil quality Determination of dry matter and water content on a mass basis Gravimetric methodDIN EN ISO 11268-2:2015-11
