1、BSI Standards PublicationBS EN ISO 16387:2014Soil quality Effects ofcontaminants on Enchytraeidae(Enchytraeus sp.) Determination of effects onreproductionBS EN ISO 16387:2014 BRITISH STANDARDNational forewordThis British Standard is the UK implementation of EN ISO16387:2014. It supersedes BS ISO 163
2、87:2004 which is withdrawn.The UK participation in its preparation was entrusted to TechnicalCommittee EH/4, Soil quality.A list of organizations represented on this committee can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 publication does not purport to include all the necessaryprovisions of a con
3、tract. Users are responsible for its correctapplication. The British Standards Institution 2014. Published by BSI StandardsLimited 2014ISBN 978 0 580 75364 0ICS 13.080.30Compliance with a British Standard cannot confer immunity fromlegal obligations.This British Standard was published under the auth
4、ority of theStandards Policy and Strategy Committee on 31 January 2014.Amendments issued since publicationDate Text affectedEUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN ISO 16387 January 2014 ICS 13.080.30 English Version Soil quality - Effects of contaminants on Enchytraeidae (Enchytraeus s
5、p.) - Determination of effects on reproduction (ISO 16387:2014) Qualit du 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 (Enchytra
6、eus sp.) - Bestimmung der Wirkungen auf die Reproduktion (ISO 16387:2014) 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
7、 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. This European Standard exists in three official versions (English, French, German). A version
8、 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 members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czec
9、h 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 and United Kingdom. EU
10、ROPEAN COMMITTEE FOR STANDARDIZATION COMIT EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR NORMUNG CEN-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
11、 EBS EN ISO 16387:2014EN ISO 16387:2014 (E) 3 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” the secretariat of which is held by NEN. This European Standard
12、 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 drawn to the possibility that some of the elements of this docum
13、ent 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 following countries are bound to implement this European Standard: A
14、ustria, 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, Sw
15、eden, 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. BS EN ISO 16387:2014ISO 16387:2014(E) ISO 2014 All rights reserved iiiContents PageForeword ivIntroduction v1 Scope . 12 Normative r
16、eferences 13 Terms and definitions . 14 Principle 35 Reagents and material 36 Apparatus . 67 Test environment . 68 Procedure. 78.1 Experimental design 78.2 Preparation of test mixtures . 88.3 Addition of the biological material 98.4 Test conditions and measurements 98.5 Reference substance . 99 Calc
17、ulation and expression of results 109.1 Calculation . 109.2 Expression of results . 1010 Validity of the test .1011 Statistical analysis 1011.1 General 1011.2 Range-finding test . 1011.3 Definitive test 1112 Test report 11Annex A (informative) Conditions for culture of Enchytraeus sp .13Annex B (inf
18、ormative) Test procedure using other Enchytraeus species 15Annex C (informative) Determination of maximum water-holding capacity 17Annex D (informative) Detailed description of extraction techniques 18Annex E (informative) Overview of the statistical assessment of data (NOEC determination) 19Bibliog
19、raphy .20BS EN ISO 16387:2014ISO 16387:2014(E)ForewordISO (the International Organization 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 memb
20、er body interested in a subject for which a technical committee has been established 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 Ele
21、ctrotechnical Commission (IEC) on all matters of electrotechnical standardization.The procedures used to develop this document and those intended for its further maintenance are described in the ISO/IEC Directives, Part 1. In particular the different approval criteria needed for the different types
22、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).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 hel
23、d 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 and/or on the ISO list of patent declarations received (see www.iso.org/patents).Any trade name used in this document is information
24、 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 assessment, as well as information about ISOs adherence to the WTO principles in the Technical Barriers to Trade (TBT) see the foll
25、owing URL: Foreword - Supplementary information.The committee responsible for this document is ISO/TC 190, Soil quality, Subcommittee SC 4, Biological methods.This second edition cancels and replaces the first edition (ISO 16387:2004), which has been technically revised.iv ISO 2014 All rights reserv
26、edBS EN ISO 16387:2014ISO 16387:2014(E)IntroductionEcotoxicological 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 standard
27、ized test systems. Aquatic test systems with soil eluate are applied 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 t
28、he latter, a standardized test system using Enchytraeidae (a chronic 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. O
29、ptionally, the method can be used for testing substances added to 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, b
30、ut can also reach high population densities in soils well inhabited 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 significant
31、ly shorter than that of earthworms the test duration for a reproduction 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 amoun
32、t needed in earthworm tests.This International Standard has been drawn up taking into consideration test procedures recommended by the Organization for Economic Cooperation and Development (see22,24). ISO 2014 All rights reserved vBS EN ISO 16387:2014BS EN ISO 16387:2014Soil quality Effects of conta
33、minants on Enchytraeidae (Enchytraeus sp.) Determination of effects on reproduction1 ScopeThis International Standard specifies one of the methods for evaluating the habitat function of soils and determining effects of soil contaminants and substances on the reproduction of Enchytraeus sp. by dermal
34、 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 sites under concern and waste materials.Effects of substances are assessed using a standard soil, pre
35、ferably 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 dilution substrate (dilution series of contaminated soil) are either an uncontaminated soil compara
36、ble 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 temperate conditions.The method is not applicable to volatile substances, i.e. substances for which H (Henry
37、s 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 persistence of the substance under test.2 Normative referencesThe following documents, in whole or in part
38、, 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 10381-6, Soil quality Sampling Part 6: Guidance
39、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 after dry combustion (elementa
40、ry 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 sedimentationISO 11465, Soil quality Dete
41、rmination 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.INTERNATIONAL STANDARD ISO 16387:2014(E) ISO 2014 All rights reserved 1BS EN ISO 16387:2014ISO 16387:2014(E)3.1reproductionm
42、ean 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 enchytraeids3.2reproduction ratemean number of offspring
43、 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 % effectconcentration (mass fraction) of a test su
44、bstance 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 period.Note 1 to entry: The ECx is expresse
45、d 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 substance concentration that has a sta
46、tistically 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, by mixing the dry constituents listed
47、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 imme
48、diately before the test. Store the mixed artificial soil at room temperature for at least two days to equilibrate acidity. To determine pH and the maximum water holding capacity, the dry artificial soil is pre-moistened one or two days before starting the test by adding deionised water to obtain hal
49、f of the required final water content of 40 % to 60 % of the maximum water holding capacity.The total water holding capacity is determined according to Annex C, the pH is determined according to ISO 10390.NOTE 2 Allowance should be made for any water that is to be used for introducing the test substance into the soil.5.3 FoodRolled oats, preferably autoclaved (heating is also possible) before use to avoid infection with other organisms, were found to be suitable31. The first feeding is made by mixing 50 mg of ground rolled oats pe
