1、BSI Standards PublicationBS EN ISO 11268-1:2015Soil quality Effects ofpollutants on earthwormsPart 1: Determination of acute toxicityto Eisenia fetida/Eisenia andrei (ISO11268-1:2012)BS EN ISO 11268-1:2015 BRITISH STANDARDNational forewordThis British Standard is the UK implementation of EN ISO11268
2、-1:2015. It is identical to ISO 11268-1:2012. It supersedes BS7755-4.2.1:1994 which is withdrawn.The UK participation in its preparation was entrusted to TechnicalCommittee EH/4, Soil quality.A list of organizations represented on this committee can beobtained on request to its secretary.This public
3、ation does not purport to include all the necessaryprovisions of a contract. Users are responsible for its correctapplication. The British Standards Institution 2015. Published by BSI StandardsLimited 2015ISBN 978 0 580 89038 3ICS 13.080.30Compliance with a British Standard cannot confer immunity fr
4、omlegal obligations.This British Standard was published under the authority of theStandards Policy and Strategy Committee on 31 August 2015.Amendments issued since publicationDate Text affectedEUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN ISO 11268-1 August 2015 ICS 13.080.30 English Version
5、Soil quality - Effects of pollutants on earthworms - Part 1: Determination of acute toxicity to Eisenia fetida/Eisenia andrei (ISO 11268-1:2012) Qualit du sol - Effets des polluants vis-vis des vers de terre - Partie 1: Dtermination de la toxicit aigu vis-vis de Eisenia fetida/Eisenia andrei (ISO 11
6、268-1:2012) Bodenbeschaffenheit - Wirkungen von Schadstoffen auf Regenwrmer - Teil 1: Bestimmung der akuten Toxizitt auf Eisenia fetida/Eisenia andrei (ISO 11268-1:2012) This European Standard was approved by CEN on 6 August 2015. CEN members are bound to comply with the CEN/CENELEC Internal Regulat
7、ions 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.
8、 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
9、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, Po
10、rtugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and United Kingdom. EUROPEAN COMMITTEE FOR STANDARDIZATION COMIT EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR NORMUNG CEN-CENELEC Management Centre: Avenue Marnix 17, B-1000 Brussels 2015 CEN All rights of exploitation in any
11、 form and by any means reserved worldwide for CEN national Members. Ref. No. EN ISO 11268-1:2015 EBS EN ISO 11268-1:2015EN ISO 11268-1:2015 (E) 3 European foreword The text of ISO 11268-1:2012 has been prepared by Technical Committee ISO/TC 190 “Soil quality” of the International Organization for St
12、andardization (ISO) and has been taken over as EN ISO 11268-1:2015 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 national standard, either by publication of an identical text or by endorsem
13、ent, 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 patent rights. CEN and/or CENELEC shall not be held responsible for identi
14、fying 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 Yugosl
15、av 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 11268-1:2012 ha
16、s been approved by CEN as EN ISO 11268-1:2015 without any modification. BS EN ISO 11268-1:2015ISO 11268-1:2012(E) ISO 2012 All rights reserved iiiContents PageForeword ivIntroduction v1 Scope 12 Normative references . 13 Terms and definitions . 24 Principle . 35 Reagents and material . 36 Apparatus
17、57 Procedure 57.1 Experimental design . 57.2 Preparation of test mixture . 67.3 Addition of the earthworms 87.4 Test conditions and measurements . 87.5 Reference substance 88 Calculation and expression of results . 88.1 Calculation 88.2 Expression of results . 89 Validity of the test 910 Statistical
18、 analysis 910.1 General . 910.2 Single-concentration tests 910.3 Multi-concentration tests 1011 Test report . 11Annex A (informative) Determination of the acute toxicity of chemicals (in particular pesticides) on earthworms under tropical test conditions 12Annex B (informative) Culturing of Eisenia
19、fetida and Eisenia andrei .14Annex C (informative) Determination of water holding capacity of artificial soil 15Annex D (informative) Background information on the acute effect of boric acid on earthworms . 16Bibliography .17BS EN ISO 11268-1:2015ISO 11268-1:2012(E)ForewordISO (the International Org
20、anization 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 establi
21、shed 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 standardizatio
22、n.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. Pu
23、blication 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
24、 rights.ISO 11268-1 was prepared by Technical Committee ISO/TC 190, Soil quality, Subcommittee SC 4, Biological methods.This second edition cancels and replaces the first edition (ISO 11268-1:1993), which has been technically revised.ISO 11268 consists of the following parts, under the general title
25、 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: Guidance on the determination of effects in field situationsiv ISO 2012 All rights res
26、ervedBS EN ISO 11268-1:2015ISO 11268-1:2012(E)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 1579933 and ISO 1761634). ISO 15799 includes a list and short cha
27、racterization 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 (r
28、etention 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 par
29、t of ISO 11268 describes a method that is based on the determination of the acute toxicity of contaminated soils to 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 a standard soil (e.g. ar
30、tificial soil) for their acute toxic 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. Ba
31、ckground 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
32、in general, and the database and experience in testing soils is small 151623.This part of ISO 11268 has been drawn up taking into consideration test procedures adopted by the Organization for Economic Cooperation and Development 2627and by the European Union 9. ISO 2012 All rights reserved vBS EN IS
33、O 11268-1:2015BS EN ISO 11268-1:2015Soil quality Effects of pollutants on earthworms Part 1: Determination of acute toxicity to Eisenia fetida/Eisenia andreiWARNING Contaminated soils may contain unknown mixtures of toxic, mutagenic, or otherwise harmful chemicals or infectious microorganisms. Occup
34、ational health risks may arise from dust or evaporated chemicals during handling and incubation. Precautions should be taken to avoid skin contact.1 ScopeThis part of ISO 11268 specifies one of the methods for evaluating the habitat function of soils and determining the acute toxicity of soil contam
35、inants and chemicals to Eisenia fetida/Eisenia andrei by dermal and alimentary uptake. It is applicable to soils and soil materials of unknown quality, e.g. from contaminated sites, amended soils, soils after remediation, agricultural or other sites concerned, and waste materials.Effects of substanc
36、es are assessed using a standard soil, preferably a defined artificial soil substrate. For contaminated soils, the effects on survival 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 s
37、oil) 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 on how to use this method for testing chemicals under temperate as well as under tropical conditions.The method is not applicable
38、 to volatile substances, i.e. substances for which H (Henrys 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.This method does not take into account the possible degradation of the substances or contaminants during the te
39、st.2 Normative referencesThe following referenced documents are indispensable for the application of this document. For dated references, only the edition cited applies. For undated references, the latest edition of the referenced document (including any amendments) applies.ISO 10381-6, Soil quality
40、 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 laboratoryISO 10390, Soil quality Determination of pHISO 10694, Soil quality Determination of organic and total carbon aft
41、er dry combustion (elementary analysis)ISO 11260, Soil quality Determination of effective cation exchange capacity and base saturation level using barium chloride solutionISO 11277, Soil quality Determination of particle size distribution in mineral soil material Method by sieving and sedimentationI
42、SO 11465, Soil quality Determination of dry matter and water content on a mass basis Gravimetric methodINTERNATIONAL STANDARD ISO 11268-1:2012(E) ISO 2012 All rights reserved 1BS EN ISO 11268-1:2015ISO 11268-1:2012(E)3 Terms and definitionsFor the purposes of this document, the following terms and d
43、efinitions apply.3.1contaminantsubstance or agent present in the soil as a result of human activityISO 15176:20023.2survivalpercentage of living worms at the end of the test period3.3mortalitypercentage of dead or missing worms at the end of the test period3.4LC50lethal concentrationmedian lethal pe
44、rcentage of a test sample in a reference or a standard control soil, or concentration of a substance in the test sample, which kills 50 % of the test animals within the test periodNOTE The LC50is expressed as a percentage of test-soil dry mass per test-mixture dry mass.3.5limit testsingle concentrat
45、ion test consisting of at least four replicates each, the test sample without any dilution or the highest concentration of test substance mixed into the control soil and the control3.6lowest observed effect rate (LOER) or effect concentration (LOEC)lowest tested percentage of a test sample in a cont
46、rol soil or concentration of a substance at which a statistically significant effect is observedNOTE The LOEC is expressed as a percentage of test-soil dry mass per test-mixture dry mass. All test mixtures above the LOEC have a harmful effect equal to or greater than that observed at the LOEC. If th
47、is condition cannot be satisfied, an explanation should be given for how the LOEC and NOEC (3.7) have been selected.3.7NOECno observed effect concentrationtest soil percentage immediately below the LOEC or, highest tested concentration of a test substance which, when compared to the control, has no
48、statistically significant lethal or other effect such as mass alteration (error probability: p 2, where Kow is the partitioning coefficient (octanol/water) or ionizing substances into account, 5 % of peat have proven to be sufficient for maintaining the desired structure of the artificial soil.4 ISO
49、 2012 All rights reservedBS EN ISO 11268-1:2015ISO 11268-1:2012(E)NOTE 2 It has been demonstrated that Eisenia fetida can comply with the validity criteria, even as regards reproduction, when tested in field soils with lower organic carbon content (e.g. 2,7 %) 17, and experience shows that this can be achieved in artificial soil with 5 % peat. It is therefore not necessary, before using such a soil in a definitive test, to demonstrate the suitability of the artificial soil in complying with the validity
