1、May 2014Translation by DIN-Sprachendienst.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.
2、080.30!%1*“2149407www.din.deDDIN EN ISO 16387Soil quality Effects of contaminants on Enchytraeidae (Enchytraeus sp.) Determination of effects on reproduction (ISO 16387:2014);English version EN ISO 16387:2014,English translation of DIN EN ISO 16387:2014-05Bodenbeschaffenheit Wirkungen von Verunreini
3、gungen auf Enchytraeidae (Enchytraeus sp.) Bestimmung der Wirkungen auf die Reproduktion (ISO 16387:2014);Englische Fassung EN ISO 16387:2014,Englische bersetzung von DIN EN ISO 16387:2014-05Qualit du sol Effets des contaminants sur les Enchytraeidae (Enchytraeus sp.) Dtermination des effets sur la
4、survie et la reproduction (ISO 16387:2014);Version anglaise EN ISO 16387:2014,Traduction anglaise de DIN EN ISO 16387:2014-05SupersedesDIN ISO 16387:2004-09www.beuth.deIn case of doubt, the German-language original shall be considered authoritative.Document comprises 30 pages04.14 DIN EN ISO 16387:2
5、014-05 2 A comma is used as the decimal marker. National foreword This document (EN ISO 16387:2014) has been prepared by Technical Committee ISO/TC 190 “Soil quality” in collaboration with Technical Committee CEN/TC 345 “Characterization of soils” (Secretariat: NEN, Netherlands). The preliminary wor
6、k was done by Working Group WG 2 “Effects on soil fauna” of ISO/TC 190/SC 4 (lead-management: DIN, Germany). The responsible German body involved in its preparation was the Normenausschuss Wasserwesen (Water Practice Standards Committee), Working Committee NA 119-01-02-04 UA Biologische Verfahren of
7、 NA 119-01-02- AA Abfall- und Bodenuntersuchung. 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 ISO 10694 ISO 11260 DIN EN ISO 11260 ISO 11272 DIN ISO 11272 ISO 11277 DIN IS
8、O 11277 ISO 11465 DIN ISO 11465 ISO 14240-1 DIN EN ISO 14240-1 ISO 15799 DIN ISO 15799 Amendments This standard differs from DIN ISO 16387:2004-09 as follows: a) the scope has been restricted to reproduction (survival has been deleted); b) the standard has been brought into line with the state of th
9、e art; c) Clause 3 “Terms and definitions” has been revised; d) Clause 11 “Statistical analysis” has been added; e) the former Annex C “Determination of the pH value” has been deleted, reference is now made to ISO 10390; f) Annex D “Detailed description of extraction techniques” has been revised; g)
10、 the standard has been editorially revised. Previous editions DIN ISO 16387: 2004-09 DIN EN ISO 16387:2014-05 3 National Annex NA (informative) Bibliography DIN EN ISO 11260, Soil quality Determination of effective cation exchange capacity and base saturation level using barium chloride solution DIN
11、 EN ISO 14240-1, Soil quality Determination of soil microbial biomass Part 1: Substrate-induced respiration method 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 processes, biomass
12、 and diversity in the laboratory DIN ISO 10390, Soil quality Determination of pH DIN ISO 10694, Soil quality Determination of organic and total carbon after dry combustion (elementary analysis) DIN ISO 11272, Soil quality Determination of dry bulk density DIN ISO 11277, Soil quality Determination of
13、 particle size distribution in mineral soil material Method by sieving and sedimentation DIN ISO 11465, Soil quality Determination of dry matter and water content on a mass basis Gravimetric method DIN ISO 15799, Soil quality Guidance on the ecotoxicological characterization of soils and soil materi
14、als DIN EN ISO 16387:2014-05 4 This page is intentionally blank EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM January 2014 ICS 13.080.30 English Version Soil quality - Effects of contaminants on Enchytraeidae (Enchytraeus sp.) - Determination of effects on reproduction (ISO 16387:2014) Qualit du
15、 sol - Effets des contaminants sur les Enchytraeidae (Enchytraeus sp.) - Dtermination des effets sur la survie et la reproduction (ISO 16387:2014) Bodenbeschaffenheit - Wirkungen von Verunreinigungen auf Enchytraeidae (Enchytraeus sp.) - Bestimmung der Wirkungen auf die Reproduktion (ISO 16387:2014)
16、 This European Standard was approved by CEN on 12 October 2013. 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 re
17、ferences 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
18、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 Macedo
19、nia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey andUnited Kingdom. EUROPEAN COMMITTEE FOR STANDARDIZATIONCOMIT EUROPEN DE NORMALISATION EUROPIS
20、CHES KOMITEE FR NORMUNGCEN-CENELEC Management Centre: Avenue Marnix 17, B-1000 Brussels 2014 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN ISO 16387:2014 EEN ISO 16387 Contents Page Foreword . 3 Introduction . 4 1 Scope . 5 2 Nor
21、mative references. 5 3 Terms and definitions 5 4 Principle 7 5 Reagents and material . 7 6 Apparatus 10 7 Test environment . 10 8 Procedure 11 8.1 Experimental design 11 8.2 Preparation of test mixtures . 12 8.3 Addition of the biological material . 13 8.4 Test conditions and measurements . 13 8.5 R
22、eference substance . 13 9 Calculation and expression of results . 14 9.1 Calculation 14 9.2 Expression of results . 14 10 Validity of the test 14 11 Statistical analysis . 14 11.1 General 14 11.2 Range-finding test 14 11.3 Definitive test 15 12 Test report . 15 Annex A (informative) Conditions for c
23、ulture of Enchytraeus sp . 17 Annex B (informative) Test procedure using other Enchytraeus species . 19 Annex C (informative) Determination of maximum water-holding capacity 21 Annex D (informative) Detailed description of extraction techniques 22 Annex E (informative) Overview of the statistical as
24、sessment of data (NOEC determination) . 23 Bibliography 24 2DIN EN ISO 16387:2014-05 EN ISO 16387:2014 (E) ForewordThis document (EN ISO 16387:2014) has been prepared by Technical Committee ISO/TC 190 “Soil quality” in collaboration with Technical Committee CEN/TC 345 “Characterization of soils” the
25、 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 July 2014, and conflicting national standards shall be withdrawn at the latest by July 2014. Attention is draw
26、n 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 followin
27、g 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, Norw
28、ay, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom. Endorsement notice The text of ISO 16387:2014 has been approved by CEN as EN ISO 16387:2014 without any modification. 3DIN EN ISO 16387:2014-05EN ISO 16387:2014 (E)IntroductionEcotoxicologic
29、al test systems are applied to obtain information about the effects of contaminants in soil and are proposed to complement conventional chemical analysis. ISO 15799 includes a list and short characterization of recommended and standardized test systems. Aquatic test systems with soil eluate are appl
30、ied 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. For the latter, a standardized test system using Enchytraeidae (a chro
31、nic test with end-point reproduction) is proposed.This International Standard describes a method that is based on the determination of acute and sublethal effects of contaminated soils to adult Enchytraeidae of the genus Enchytraeus. Optionally, the method can be used for testing substances added to
32、 standard soils (e.g. artificial soil) for their sublethal hazard potential to Enchytraeidae.Soil-dwelling annelids of the genus Enchytraeus are ecologically relevant, i.e. they are abundant in many soils where earthworms are scarce, but can also reach high population densities in soils well inhabit
33、ed by earthworms. Enchytraeidae can be used in laboratory tests as well as in semi-field and field studies. From a practical point of view, many Enchytraeus species are easy to handle and breed, and their generation time is significantly shorter than that of earthworms the test duration for a reprod
34、uction test with Enchytraeidae is four weeks to six weeks, compared to eight weeks (12 weeks including synchronization) with earthworms. In addition, a much smaller volume of soil is needed in the enchytraeid test compared to the amount needed in earthworm tests.This International Standard has been
35、drawn up taking into consideration test procedures recommended by the Organization for Economic Cooperation and Development (see22,24).4DIN EN ISO 16387:2014-05 EN ISO 16387:2014 (E) 1 ScopeThis International Standard specifies one of the methods for evaluating the habitat function of soils and dete
36、rmining effects of soil contaminants and substances on the reproduction of Enchytraeus sp. by dermal and alimentary uptake in a chronic test. It is applicable to soils and soil materials of unknown quality, e.g. from contaminated sites, amended soils, soils after remediation, agricultural or other s
37、ites 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 soil to be tested and in a control soil. According to the objective of the study, the control and
38、 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 temperat
39、e 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 for which the vapour pressure exceeds 0,013 3 Pa at 25 C.NOTE No provision is made in the test method for monitoring the persis
40、tence of the substance under test.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 edition of the referen
41、ced 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 laboratoryISO 10390, Soil quality Determination o
42、f 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 chloride solutionISO 11277, Soil quality Determination of particle size d
43、istribution in mineral soil material Method by sieving and sedimentationISO 11465, Soil quality Determination of dry matter and water content on a mass basis Gravimetric method3 Terms and definitionsFor the purposes of this document, the following terms and definitions apply.5DIN EN ISO 16387:2014-0
44、5EN ISO 16387:2014 (E)3.1reproductionmean number of offspring per test vessel after incubation under the specified test conditionsNote 1 to entry: The test period for the determination of the reproduction (definitive test) is six weeks.Note 2 to entry: offspring = hatched juvenile enchytraeids3.2rep
45、roduction ratemean number of offspring produced per a number of adults over the test periodNote 1 to entry: The test period for the determination of the reproduction (definitive test) is six weeks.Note 2 to entry: offspring = hatched juvenile enchytraeids3.3ECxeffect concentration for x % effectconc
46、entration (mass fraction) of a test substance that causes x % of an effect on a given endpoint within a given exposure period when compared with a controlEXAMPLE 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 perio
47、d.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.3.4LOEClowest observed effect concentrationlowest test
48、substance concentration that has a statistically significant effect (probability p 2) or ionizing substances into account, 5 % of peat have proven to be sufficient for maintaining the desired structure of the artificial soil.22Prepare the artificial soil at least three days prior to start the test,
49、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. 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 da
