1、raising standards worldwideNO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAWBSI Standards PublicationBS EN ISO 15952:2011Soil quality Effects ofpollutants on juvenileland snails (Helicidae) Determination of the effects ongrowth by soil contamination(ISO 15952:2006)BS EN ISO 1595
2、2:2011 BRITISH STANDARDNational forewordThis British Standard is the UK implementation of EN ISO 15952:2011. It is identical to ISO 15952:2006.The UK participation in its preparation was entrusted to Technical Committee EH/4, Soil quality.A list of organizations represented on this committee can be
3、obtained on request to its secretary.This publication does not purport to include all the necessary provisions of a contract. Users are responsible for its correct application. BSI 2011 ISBN 978 0 580 73152 5 ICS 13.080.30 Compliance with a British Standard cannot confer immunity from legal obligati
4、ons.This British Standard was published under the authority of the Standards Policy and Strategy Committee on 31 July 2011.Amendments issued since publicationDate Text affectedEUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN ISO 15952 June 2011 ICS 13.080.30 English Version Soil quality - Effect
5、s of pollutants on juvenile land snails (Helicidae) - Determination of the effects on growth by soil contamination (ISO 15952:2006) Qualit du sol - Effets des polluants vis-vis des escargots juvniles (Helicidae) - Dtermination des effets sur la croissance par contamination du sol (ISO 15952:2006) Bo
6、denbeschaffenheit - Wirkungen von Schadstoffen auf Jungtiere von Landschnecken - Bestimmung der Wirkungen auf das Wachstum durch Bodenverunreinigung (ISO 15952:2006) This European Standard was approved by CEN on 10 June 2011. CEN members are bound to comply with the CEN/CENELEC Internal Regulations
7、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. This
8、 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 membe
9、rs are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain,
10、Sweden, Switzerland and United Kingdom. EUROPEAN COMMITTEE FOR STANDARDIZATION COMIT EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR NORMUNG Management Centre: Avenue Marnix 17, B-1000 Brussels 2011 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Membe
11、rs. Ref. No. EN ISO 15952:2011: EBS EN ISO 15952:2011EN ISO 15952:2011 (E) 3 Foreword The text of ISO 15952:2006 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 15952:2011 by Technical Co
12、mmittee 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 December 2011, and conflicting national standards shall be w
13、ithdrawn at the latest by December 2011. 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 Regulat
14、ions, 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, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta
15、, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and the United Kingdom. Endorsement notice The text of ISO 15952:2006 has been approved by CEN as a EN ISO 15952:2011 without any modification. BS EN ISO 15952:2011ISO 15952:2006(E) ISO 2006 All rights r
16、eserved iiiContents Page Foreword iv Introduction v 1 Scope . 1 2 Normative references . 1 3 Terms and definitions. 2 4 Principle. 3 5 Test environment 3 6 Reagents 3 7 Apparatus 5 8 Storage and preparation of the samples 6 8.1 Soil to be tested 6 8.2 Waste material. 6 9 Procedure 6 9.1 Preparation
17、of the test 6 9.2 Distribution of the test mixture . 8 9.3 Introduction of the feed 8 9.4 Introduction of the biological reagent 8 9.5 Handling during the tests 8 10 Reference substance 10 11 Calculations and expression of results 10 11.1 Calculations. 10 11.2 Expression of results . 12 12 Validity
18、of test for Helix aspersa aspersa 13 13 Test report . 13 Annex A (normative) Static method . 15 Annex B (informative) Breeding technique for snails 16 Annex C (informative) Example of composition of snail feed . 21 Annex D (informative) Example of table of data 22 Annex E (informative) Example of re
19、sults with Helix aspersa aspersa 23 Annex F (informative) Determination of the effects on growth by food contamination . 26 Annex G (informative) Test performance with other snail species. 30 Bibliography . 31 BS EN ISO 15952:2011ISO 15952:2006(E) iv ISO 2006 All rights reservedForeword ISO (the Int
20、ernational 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 member body interested in a subject for which a technical committee ha
21、s 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 Electrotechnical Commission (IEC) on all matters of electrotechnical
22、standardization. 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 bodie
23、s for voting. Publication 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 o
24、r all such patent rights. ISO 15952 was prepared by Technical Committee ISO/TC 190, Soil quality, Subcommittee SC 4, Biological methods. BS EN ISO 15952:2011ISO 15952:2006(E) ISO 2006 All rights reserved vIntroduction Because of the limited amount of data available concerning toxicity of contaminant
25、s on soil organisms, the problems of assessing the ecotoxicity of soils and waste are cause for serious concern at both national and international levels. Currently available tests use soil-fauna organisms restricted to annelid (earthworms and Enchytraeidae) and arthropod phyla (insects: Collembola
26、and Coleoptera). Among the latter, two standards assess acute toxicity earthworms (ISO 11268-1) and coleoptera larvae 5 and three other standards assess sublethal effects of soil contaminants on reproduction (earthworms 2, Collembola 1, Enchytraeidae 3). In the biological cycles of organisms, it app
27、ears that growth is, like reproduction, a fundamental ecophysiological parameter to be taken into consideration for the sustainability of species and ecosystems33. Snails are pertinent ecological indicators for assessing the quality of soils 15, as they are characteristic of the soil surface layer (
28、saprophagous and phytophagous) of which a large part of the biological cycle takes place in the soil (egg-laying, hatching, initial stages of development, hibernation, etc.) 6, 17, 26. During the other phases of their cycle, they eat soil and are in contact with the soil via their moist pedal sole (
29、foot) covered with mucus and participate in the permanent exchanges with the soil (water, mineral salts, excrement and finally shell and organic matter when they die) 6, 17, 28. In addition, they constitute an important link between plants, fauna and soil microorganisms. They correspond fully to the
30、 criteria for a good biological indicator: easy to sample and identify, they are widely distributed; they accumulate contaminants 8, 10 to 14, 16, 17, 19, 21, 26, 27, 35 to 43; their ecological and physiological characteristics are well-known 6, 9, 29; and they are now easy to breed under controlled
31、 conditions 19, 23, 29. Their susceptibility to common contaminants of their environment has been demonstrated 10 to 15, 18 to 27, 32, 33, 36 to 42. This International Standard describes a method for determining the effects on survival and growth of young snails of substances, preparations, soils or
32、 waste materials added to an artificial or a natural soil. The described method is thus applicable to test contaminated soils or to compare different uncontaminated soils. The recommended species is Helix aspersa aspersa Mller (also commonly called: common garden snail, brown garden snail, garden sn
33、ail, land snail, “Petit-Gris”). Among land snails (stylommatophoran pulmonate gastropod molluscs of the Helicidae family), Helix aspersa aspersa Mller is the most ubiquitous. This palearctic species can be acclimated to regions with different types of climate: Mediterranean, oceanic temperate, midco
34、ntinental temperate and even tropical. Helix aspersa aspersa Mller is of European origin and has been introduced into all parts of the world. They are now on all continents except Antarctica 9. Indeed, in their natural environment, snails integrate the contaminants by contact (with various substrate
35、s such as soil, soil leachates, plant litter), by ingestion (of plants and soil), as well as through the respiratory tract 6, 26. So, for specific testing purposes (evaluation of the toxicity of a pesticide, for example), another test design, which is focussed on exposure via food uptake, is optiona
36、lly available (Annex F and Reference 4). BS EN ISO 15952:2011BS EN ISO 15952:2011INTERNATIONAL STANDARD ISO 15952:2006(E) ISO 2006 All rights reserved 1Soil quality Effects of pollutants on juvenile land snails (Helicidae) Determination of the effects on growth by soil contamination 1 Scope This Int
37、ernational Standard specifies a semi-static method for the determination of the effects of contaminants on growth and survival of young snails, usually Helix aspersa aspersa Mller. The animals are exposed via the cutaneous and digestive route using a test substrate (artificial or natural soil accord
38、ing to the objective of the study) to which defined amounts of the following are added: substances or preparations; soils (contaminated or of unknown quality) or waste materials. A static method may be implemented in addition to the semi-static method (optional). This method is described in Annex A.
39、 This method does not apply to volatile substances, i.e. substances for which the Henry constant, H, or the air/water partition coefficient is over 1, or for which the vapour pressure is over 0,013 3 Pa at 25 C. This test takes into account the possible change in the test substance, preparation, soi
40、l or waste material because the test mixture is prepared and renewed every 7 days during the 28-day test period. 2 Normative references The following referenced documents are indispensable for the application of this document. For dated references, only the edition cited applies. For undated referen
41、ces, 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 for the assessment of aerobic microbial processes in the laboratory ISO 10390, Soil quality Determination of pH I
42、SO 10694, Soil quality Determination of organic and total carbon after dry combustion (elementary analysis) ISO 11268-1, Soil quality Effects of pollutants on earthworms (Eisenia fetida) Part 1: Determination of acute toxicity using artificial soil substrate ISO 11269-2, Soil quality Determination o
43、f the effects of pollutants on soil flora Part 2: Effects of chemicals on the emergence and growth of higher plants ISO 11274, Soil quality Determination of the water-retention characteristic Laboratory methods ISO 11465, Soil quality Determination of dry matter and water content on a mass basis Gra
44、vimetric method EN 14735, Characterization of waste Preparation of waste samples for ecotoxicity tests BS EN ISO 15952:2011ISO 15952:2006(E) 2 ISO 2006 All rights reserved3 Terms and definitions For the purposes of this document, the following terms and definitions apply. 3.1 test substrate artifici
45、al soil or natural soil used as control and dilution substrate 3.2 matrix soil or waste material under test 3.3 test mixture mixture of the test substance, preparation or matrix with the test substrate 3.4 growth increase in the biomass, i.e. in the total fresh mass (body and shell) of the organisms
46、 and increase in the maximum shell diameter, between the start and completion of the test NOTE It is expressed in the form of a growth coefficient. 3.5 effect concentration ECxconcentration at which a specific effect is detected; x is the percentage (10, 25, 50) of this effect, e.g. growth inhibitio
47、n EXAMPLE EC50means the concentration estimated to reduce growth at the end of the test to 50 % compared to the control. 3.6 median lethal concentration LC50concentration of the substance, of the test preparation initially present, or the concentration of the matrix causing the death of 50 % of the
48、snails submitted to testing 3.7 lowest observed effect concentration LOEC lowest tested concentration at which the test substance is observed to have a statistically significant effect (p 0,05) within a given exposure time NOTE 1 The NOEC is the concentration just below the LOEC. NOTE 2 For 3.5, 3.6
49、, 3.7 and 3.8, results are given: in dry mass of test substance or preparation per dry mass of the test substrate; in mass percentage of the tested matrix in the test mixture (expressed in dry mass). BS EN ISO 15952:2011ISO 15952:2006(E) ISO 2006 All rights reserved 34 Principle Juvenile land snails (usually Helix aspersa aspersa Mller) are exposed during a period of 28 days to a test mixture containing the test substance, preparation or matrix at different co
