1、September 2011 Translation by DIN-Sprachendienst.English price group 17No 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).
2、ICS 13.080.30!$tPq“1814578www.din.deDDIN EN ISO 15952Soil quality Effects of pollutants on juvenile land snails (Helicidae) Determination of the effects on growth by soil contamination(ISO 15952:2006)English translation of DIN EN ISO 15952:2011-09Bodenbeschaffenheit Wirkungen von Schadstoffen auf Ju
3、ngtiere von Landschnecken Bestimmung der Wirkungen auf das Wachstum durch Bodenverunreinigung(ISO 15952:2006)Englische bersetzung von DIN EN ISO 15952:2011-09Qualit du sol Effets des polluants vis-vis des escargots juvniles (Helicidae) Dtermination des effets sur la croissance par contamination du s
4、ol (ISO 15952:2006)Traduction anglaise de DIN EN ISO 15952:2011-09www.beuth.deDocument comprises pagesIn case of doubt, the German-language original shall be considered authoritative.4008.11 DIN EN ISO 15952:2011-09 2 A comma is used as the decimal marker. National foreword This standard has been pr
5、epared by Technical Committee ISO/TC 190 “Soil quality” in collaboration with Technical Committee CEN/TC 345 “Characterization of soils” (Secretariat: NEN, Netherlands). The responsible German body involved in its preparation was the Normenausschuss Wasserwesen (Water Practice Standards Committee),
6、Working Committee NA 119-01-02-04 UA Biologische Verfahren of 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 112
7、67 DIN ISO 11267 ISO 11268-1 DIN ISO 11268-1 ISO 11268-2 DIN ISO 11268-2 ISO 11269-2 DIN ISO 11269-2 ISO 11274 DIN ISO 11274 ISO 11465 DIN ISO 11465 ISO 16387 DIN ISO 16387 National Annex NA (informative) Bibliography DIN ISO 10381-6, Soil quality Sampling Part 6: Guidance on the collection, handlin
8、g and storage of soil under aerobic conditions for the assessment of microbiological processes, biomass 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
9、 ISO 11267, Soil quality Inhibition of reproduction of Collembola (Folsomia candida) by soil pollutants DIN ISO 11268-1, Part 1: Determinationof acute toxicity using artificial soil substrate DIN ISO 11268-2, Part2:Determinationof effects on reproduction DIN ISO 11269-2, Soil quality Determination o
10、f the effects of pollutants on soil flora Part 2: Effects of chemicals on the emergence and growth of higher plants DIN ISO 11274, Soil quality Determination of the water retention characteristics Laboratory methods DIN ISO 11465, Soil quality Determination of dry matter and water content on a mass
11、basis Gravimetric method DIN ISO 16387, Soil quality Effects of pollutants on Enchytraeidae (Enchytraeus sp.) Determination of effects on reproduction and survival Soil quality Effects of pollutants on earthworms (Eisenia fetida/Eisenia andrei) Soil quality Effects of pollutants on earthworms (Eisen
12、ia fetida/Eisenia andrei) EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN ISO 15952 June 2011 ICS 13.080.30 English Version Soil quality - Effects of pollutants on juvenile land snails (Helicidae) - Determination of the effects on growth by soil contamination (ISO 15952:2006) Qualit du sol - Ef
13、fets des polluants vis-vis des escargots juvniles (Helicidae) - Dtermination des effets sur la croissance par contamination du sol (ISO 15952:2006) Bodenbeschaffenheit - Wirkungen von Schadstoffen auf Jungtiere von Landschnecken - Bestimmung der Wirkungen auf das Wachstum durch Bodenverunreinigung (
14、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 which stipulate the conditions for giving this European Standard the status of a national standard without any alteration. Up-to-date lists and biblio
15、graphical 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 in any other language made by translation under the responsi
16、bility 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, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greec
17、e, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and United Kingdom. EUROPEAN COMMITTEE FOR STANDARDIZATION COMIT EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR NORMUNG Managemen
18、t 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 Members. Ref. No. EN ISO 15952:2011: EContents Page Foreword3 Introduction .4 1 Scope 5 2 Normative references 5 3 Terms and definitions .6 4 Principle7 5
19、Test environment.7 6 Reagents.7 7 Apparatus .9 8 Storage and preparation of the samples.10 8.1 Soil to be tested .10 8.2 Waste material10 9 Procedure .10 9.1 Preparation of the test.10 9.2 Distribution of the test mixture 12 9.3 Introduction of the feed.12 9.4 Introduction of the biological reagent
20、.12 9.5 Handling during the tests .12 10 Reference substance.14 11 Calculations and expression of results.14 11.1 Calculations14 11.2 Expression of results 16 12 Validity of test for Helix aspersa aspersa .17 13 Test report 17 Annex A (normative) Static method 19 Annex B (informative) Breeding techn
21、ique for snails .20 Annex C (informative) Example of composition of snail feed 26 Annex D (informative) Example of table of data.27 Annex E (informative) Example of results with Helix aspersa aspersa .28 Annex F (informative) Determination of the effects on growth by food contamination 31 Annex G (i
22、nformative) Test performance with other snail species35 Bibliography 36 DIN EN ISO 15952:2011-09 EN ISO 15952:2011 (E) 2 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 tak
23、en over as EN ISO 15952:2011 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 endorsement, at the latest by December 2011, a
24、nd conflicting national standards shall be withdrawn 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. A
25、ccording 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, France, Germany, Greece, Hungary, Iceland, Ireland,
26、 Italy, Latvia, Lithuania, Luxembourg, Malta, 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. DIN EN ISO 1595
27、2:2011-09 EN ISO 15952:2011 (E) 3 Introduction Because of the limited amount of data available concerning toxicity of contaminants 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 availa
28、ble tests use soil-fauna organisms restricted to annelid (earthworms and Enchytraeidae) and arthropod phyla (insects: Collembola 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
29、of soil contaminants on reproduction (earthworms 2, Collembola 1, Enchytraeidae 3). In the biological cycles of organisms, it appears that growth is, like reproduction, a fundamental ecophysiological parameter to be taken into consideration for the sustainability of species and ecosystems33. Snails
30、are pertinent ecological indicators for assessing the quality of soils 15, as they are characteristic of the soil surface layer (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
31、.) 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 (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, 1
32、7, 28. In addition, they constitute an important link between plants, fauna and soil microorganisms. They correspond fully to the 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 t
33、o 43; their ecological and physiological characteristics are well-known 6, 9, 29; and they are now easy to breed under controlled 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 Sta
34、ndard describes a method for determining the effects on survival and growth of young snails of substances, preparations, soils or 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.
35、 The recommended species is Helix aspersa aspersa Mller (also commonly called: common garden snail, brown garden snail, garden snail, land snail, “Petit-Gris”). Among land snails (stylommatophoran pulmonate gastropod molluscs of the Helicidae family), Helix aspersa aspersa Mller is the most ubiquito
36、us. This palearctic species can be acclimated to regions with different types of climate: Mediterranean, oceanic temperate, midcontinental 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 continent
37、s except Antarctica 9. Indeed, in their natural environment, snails integrate the contaminants by contact (with various substrates 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 (evalua
38、tion of the toxicity of a pesticide, for example), another test design, which is focussed on exposure via food uptake, is optionally available (Annex F and Reference 4). DIN EN ISO 15952:2011-09 EN ISO 15952:2011 (E) 4 1 Scope This International Standard specifies a semi-static method for the determ
39、ination 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 according to the objective of the study) to which defined amounts of th
40、e 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. This method does not apply to volatile substances, i.e. substanc
41、es 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, soil or waste material because the test mixture is prepared and rene
42、wed 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 references, the latest edition of the referenced document (including any
43、 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 ISO 10694, Soil quality Determination of organic and total carbon
44、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 of the effects of pollutants on soil flora Part 2: Effects of chem
45、icals 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 Gravimetric method EN 14735, Characterization of waste Preparation o
46、f waste samples for ecotoxicity tests DIN EN ISO 15952:2011-09 EN ISO 15952:2011 (E) 5 3 Terms and definitions For the purposes of this document, the following terms and definitions apply. 3.1 test substrate artificial soil or natural soil used as control and dilution substrate 3.2 matrix soil or wa
47、ste 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 and increase in the maximum shell diameter, between the start and completion of the
48、 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 inhibition EXAMPLE EC50means the concentration estimated to reduce growth at the end of the t
49、est 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 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
