ISO-15952-2006.pdf

Reference number ISO 15952:2006(E) © ISO 2006 INTERNATIONAL STANDARD ISO 15952 First edition 2006-02-15 Soil quality Effects of pollutants on juvenile land snails (Helicidae) Determination of the effects on growth by soil contamination Qualité du sol Effets des polluants vis-à-vis des escargots juvéniles (Helicidae) Détermination des effets sur la croissance par contamination du sol Copyright International Organization for Standardization Provided by IHS under license with ISO Licensee=Aramco HQ/9980755100 Not for Resale, 04/11/2007 06:17:15 MDTNo reproduction or networking permitted without license from IHS -,-,- ISO 15952:2006(E) PDF disclaimer This PDF file may contain embedded typefaces. In accordance with Adobe's licensing policy, this file may be printed or viewed but shall not be edited unless the typefaces which are embedded are licensed to and installed on the computer performing the editing. In downloading this file, parties accept therein the responsibility of not infringing Adobe's licensing policy. The ISO Central Secretariat accepts no liability in this area. Adobe is a trademark of Adobe Systems Incorporated. Details of the software products used to create this PDF file can be found in the General Info relative to the file; the PDF-creation parameters were optimized for printing. Every care has been taken to ensure that the file is suitable for use by ISO member bodies. In the unlikely event that a problem relating to it is found, please inform the Central Secretariat at the address given below. © ISO 2006 All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying and microfilm, without permission in writing from either ISO at the address below or ISO's member body in the country of the requester. ISO copyright office Case postale 56 CH-1211 Geneva 20 Tel. + 41 22 749 01 11 Fax + 41 22 749 09 47 E-mail copyrightiso.org Web www.iso.org Published in Switzerland ii © ISO 2006 All rights reserved Copyright International Organization for Standardization Provided by IHS under license with ISO Licensee=Aramco HQ/9980755100 Not for Resale, 04/11/2007 06:17:15 MDTNo reproduction or networking permitted without license from IHS -,-,- ISO 15952:2006(E) © ISO 2006 All rights reserved iii Contents 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 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 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 results 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 Copyright International Organization for Standardization Provided by IHS under license with ISO Licensee=Aramco HQ/9980755100 Not for Resale, 04/11/2007 06:17:15 MDTNo reproduction or networking permitted without license from IHS -,-,- ISO 15952:2006(E) iv © ISO 2006 All rights reserved Foreword ISO (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 member 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 Electrotechnical Commission (IEC) on all matters of electrotechnical 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 bodies 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 or all such patent rights. ISO 15952 was prepared by Technical Committee ISO/TC 190, Soil quality, Subcommittee SC 4, Biological methods. Copyright International Organization for Standardization Provided by IHS under license with ISO Licensee=Aramco HQ/9980755100 Not for Resale, 04/11/2007 06:17:15 MDTNo reproduction or networking permitted without license from IHS -,-,- ISO 15952:2006(E) © ISO 2006 All rights reserved v 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 available 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 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 ecosystems 33. Snails 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.) 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, 17, 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 to 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 Standard 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. The recommended species is Helix aspersa aspersa Müller (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 Müller is the most ubiquitous. This palearctic species can be acclimated to regions with different types of climate: Mediterranean, oceanic temperate, midcontinental temperate and even tropical. Helix aspersa aspersa Müller 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 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 (evaluation 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). Copyright International Organization for Standardization Provided by IHS under license with ISO Licensee=Aramco HQ/9980755100 Not for Resale, 04/11/2007 06:17:15 MDTNo reproduction or networking permitted without license from IHS -,-,- Copyright International Organization for Standardization Provided by IHS under license with ISO Licensee=Aramco HQ/9980755100 Not for Resale, 04/11/2007 06:17:15 MDTNo reproduction or networking permitted without license from IHS -,-,- INTERNATIONAL STANDARD ISO 15952:2006(E) © ISO 2006 All rights reserved 1 Soil quality Effects of pollutants on juvenile land snails (Helicidae) Determination of the effects on growth by soil contamination 1 Scope This International 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 Müller. 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 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. 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, soil 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 references, 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 ISO 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 of 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 Gravimetric method EN 14735, Characterization of waste Preparation of waste samples for ecotoxicity tests Copyright International Organization for Standardization Provided by IHS under license with ISO Licensee=Aramco HQ/9980755100 Not for Resale, 04/11/2007 06:17:15 MDTNo reproduction or networking permitted without license from IHS -,-,- ISO 15952:2006(E) 2 © ISO 2006 All rights reserved 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 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 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 ECx concentration at which a specific effect is detected; x is the percentage (10, 25, 50) of this effect, e.g. growth inhibition EXAMPLE EC50 means the concentration estimated to reduce growth at the end of the test to 50 % compared to the control. 3.6 median lethal concentration LC50 concentration 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 NOTE 1 The NOEC is the concentration just below the LOEC. NOTE 2 For 3.5, 3.6, 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). Copyright International Organization for Standardization Provided by IHS under license with ISO Licensee=Aramco HQ/9980755100 Not for Resale, 04/11/2007 06:17:15 MDTNo reproduction or networking permitted without license from IHS -,-,- ISO 15952:2006(E) © ISO 2006 All rights reserved 3 4 Principle Juvenile land snails (usually Helix aspersa aspersa Müller) are exposed during a period of 28 days to a test mixture containing the test substance, preparation or matrix at different concentrations. The test mixture is freshly prepared and renewed every 7 days. According to the objectives, the test mixture may be prepared with artificial soil (6.3.2) or with a suitable natural soil (6.3.3). The snails are fed during the test with uncontaminated food. The effects on growth (fresh mass and shell diameter) and on survival are measured after 28 days of exposure (optionally, effects could be measured every 7 days during 28 days). The results obtained during testing are compared with thos