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1、 Reference number ISO 17512-1:2008(E) ISO 2008 INTERNATIONAL STANDARD ISO 17512-1 First edition 2008-12-15 Soil quality Avoidance test for determining the quality of soils and effects of chemicals on behaviour Part 1: Test with earthworms (Eisenia fetida and Eisenia andrei) Qualit du sol Essai dvite
2、ment pour contrler la qualit des sols et les effets des produits chimiques sur le comportement Partie 1: Essai avec des vers de terre (Eisenia fetida et Eisenia andrei) Copyright International Organization for Standardization Provided by IHS under license with ISO Licensee=Boeing Co/5910770001 Not f
3、or Resale, 01/31/2009 02:08:42 MSTNo reproduction or networking permitted without license from IHS -,-,- ISO 17512-1:2008(E) PDF disclaimer This PDF file may contain embedded typefaces. In accordance with Adobes licensing policy, this file may be printed or viewed but shall not be edited unless the
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8、 Web www.iso.org Published in Switzerland ii ISO 2008 All rights reserved Copyright International Organization for Standardization Provided by IHS under license with ISO Licensee=Boeing Co/5910770001 Not for Resale, 01/31/2009 02:08:42 MSTNo reproduction or networking permitted without license from
9、IHS -,-,- ISO 17512-1:2008(E) ISO 2008 All rights reserved iii Contents Page Foreword iv Introduction.v 1 Scope1 2 Normative references1 3 Terms and definitions.2 4 Principle .2 5 Reagents and materials3 6 Apparatus.4 7 Procedure.5 7.1 Appropriate concentration range 5 7.2 Testing of soils5 7.3 Test
10、ing of chemical.6 7.4 Reference substance 6 7.5 Validity criteria.6 8 Calculation and expression of results 6 9 Test report7 Annex A (informative) Test chambers 8 Annex B (informative) Example of a breeding technique for Eisenia fetida and Eisenia andrei11 Annex C (informative) Further test organism
11、s.12 Annex D (informative) Contaminants that earthworms can detect and avoid in the avoidance test.13 Annex E (normative) Testing of chemicals in the avoidance test .14 Annex F (normative) Determination of water-holding capacity.16 Annex G (informative) Comparison of the results obtained in the two
12、section chamber and six section chamber system 17 Annex H (informative) Influence of soil properties on avoidance behaviour Basis for the threshold value of 20 % 20 Annex I (informative) Data gained in “dual” tests with the same (untreated) control soil on both sides of the test vessels.22 Bibliogra
13、phy24 Copyright International Organization for Standardization Provided by IHS under license with ISO Licensee=Boeing Co/5910770001 Not for Resale, 01/31/2009 02:08:42 MSTNo reproduction or networking permitted without license from IHS -,-,- ISO 17512-1:2008(E) iv ISO 2008 All rights reserved Forewo
14、rd 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 technic
15、al 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 el
16、ectrotechnical 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 t
17、he 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 id
18、entifying any or all such patent rights. ISO 17512-1 was prepared by Technical Committee ISO/TC 190, Soil quality, Subcommittee SC 4, Biological methods. ISO 17512 consists of the following parts, under the general title Soil quality Avoidance test for determining the quality of soils and effects of
19、 chemicals on behaviour: Part 1: Test with earthworms (Eisenia fetida and Eisenia andrei) The following part is under preparation: Part 2: Test with collembolans (Folsomia candida) Copyright International Organization for Standardization Provided by IHS under license with ISO Licensee=Boeing Co/5910
20、770001 Not for Resale, 01/31/2009 02:08:42 MSTNo reproduction or networking permitted without license from IHS -,-,- ISO 17512-1:2008(E) ISO 2008 All rights reserved v Introduction Ecotoxicological test systems are applied to obtain information about the effects of contaminants in soil and are propo
21、sed to complement conventional chemical analysis (see ISO 15799). ISO 15799 includes a list and short characterisation of recommended and standardised test systems. Aquatic test systems with soil eluate are applied to obtain information about the fraction of contaminants potentially reaching the gro
22、undwater by the water path (retention function of soils), whereas terrestrial test systems are used to assess the habitat function of soils. As standardised test systems, a mortality test (ISO 11268-1) and a reproduction test (ISO 11268-2) exist to investigate the habitat function of a soil with res
23、pect to earthworms as representatives of the soil biocenosis. The reproduction test with earthworms (ISO 11268-2) is applied to detect effects resulting from sublethal concentrations. Such endpoints are preferably applied to obtain information on environmental effects. However, the reproduction test
24、 is very labour-intensive and time-consuming, needing long incubation periods with results obtained only after 56 days. As the test period and the work expense dictate the costs of a given test, it is preferable to obtain the results within a short test period and at a high level of sensitivity. Tha
25、t is especially the case for the assessment of remediated soils. This feature is offered by the avoidance test with Eisenia fetida and Eisenia andrei. Experiences gained in a laboratory comparison test with eight contaminated soils in three laboratories point out that the avoidance test is as sensit
26、ive as the reproduction test (Reference 5). However, it is not intended to use this test to replace the earthworm reproduction test. NOTE The results were compared with those of the earthworm acute and reproduction tests carried out with the same soils. The results showed that with a criterion of 80
27、 % avoidance response, a 72 % agreement of the results was achieved. Copyright International Organization for Standardization Provided by IHS under license with ISO Licensee=Boeing Co/5910770001 Not for Resale, 01/31/2009 02:08:42 MSTNo reproduction or networking permitted without license from IHS -
28、,-,- Copyright International Organization for Standardization Provided by IHS under license with ISO Licensee=Boeing Co/5910770001 Not for Resale, 01/31/2009 02:08:42 MSTNo reproduction or networking permitted without license from IHS -,-,- INTERNATIONAL STANDARD ISO 17512-1:2008(E) ISO 2008 All rig
29、hts reserved 1 Soil quality Avoidance test for determining the quality of soils and effects of chemicals on behaviour Part 1: Test with earthworms (Eisenia fetida and Eisenia andrei) 1 Scope This part of ISO 17512 specifies a rapid screening method for evaluating the habitat function of soils and th
30、e influence of contaminants and chemicals on earthworm behaviour. The sublethal test is a rapid method that reflects the bioavailability of contaminant mixtures in natural soils and substances spiked into soils to Eisenia fetida and Eisenia andrei. The avoidance behaviour of the worms is the measure
31、ment endpoint of the test. This test is not intended to replace the earthworm reproduction test. Two different designs (a two section unit and a six section unit) have been developed and successfully applied. Both designs are applicable to either single-concentration (e.g. for assessing the quality
32、of a field soil) or multi-concentration (e.g. for assessing the toxicity of a spiked chemical) tests. In both cases, the earthworms are allowed to make the initial choice on which compartment, control and a treatment in the two section test vessel between right and left side; in the six section test
33、 vessel between the (3 + 3) alternating compartments, to enter. 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 documen
34、t (including any amendments) applies. ISO 10390, Soil quality Determination of pH ISO 11268-2:1998, Soil quality Effects of pollutants on earthworms (Eisenia fetida) Part 2: Determination of effects on reproduction ISO 11269-2, Soil quality Determination of the effects of pollutants on soil flora Pa
35、rt 2: Effects of chemicals on the emergence and growth of higher plants ISO 11465, Soil quality Determination of dry matter and water content on a mass basis Gravimetric method ISO 15799, Soil quality Guidance on the ecotoxicological characterization of soil and soil materials Copyright Internationa
36、l Organization for Standardization Provided by IHS under license with ISO Licensee=Boeing Co/5910770001 Not for Resale, 01/31/2009 02:08:42 MSTNo reproduction or networking permitted without license from IHS -,-,- ISO 17512-1:2008(E) 2 ISO 2008 All rights reserved 3 Terms and definitions For the pur
37、poses of this document, the following terms and definitions apply. 3.1 avoidance behaviour tendency (of an organism) to avoid the test soil while preferring the control soil 3.2 habitat function ability of soils/soil materials to serve as habitat for micro-organisms, plants and soil-living animals a
38、nd their interactions ISO 15799:2003 NOTE Ecotoxicological tests as indicators for the habitat function provide information concerning the respective test parameter, e.g. acute test for survival, or chronic tests for reproduction. 3.3 limited habitat function habitat function (3.2) is limited if on
39、average 80 % of worms are found in the control soil (indication as an impact on behaviour) 3.4 effective concentration ECx concentration at which a specific effect is detected where x is a percentage (10, 25, 50) of this effect; e.g. avoidance EXAMPLE In this part of ISO 17512, an EC50 means the con
40、centration of a substance or mixture of substances in soil that is estimated to cause a behavioural response in 50 % of the test earthworms. 4 Principle Ten adult earthworms (species Eisenia fetida or Eisenia andrei) are exposed at the same time to a control soil and a contaminated soil or a soil co
41、ntaining test substances. Test soil and control soil are placed into each test vessel and the earthworms are thus presented with a choice between the test soil and the control soil. Two test-vessel designs are available: a) a two section test vessel; and b) a six section test vessel. After an incuba
42、tion period of two days, the number of worms is determined in all sections of the vessels. Individual studies (e.g. testing boric acid in one of the two designs in different laboratories) or comparative investigations (testing the same chemical or soil in the same laboratory, e.g. Reference 8) have
43、in some cases shown different results. Recently, both designs were validated in interlaboratory tests in Canada (Reference 2) and France; however, no international ring test using both designs in parallel has been performed so far. Therefore, for the time being, the choice of the design is up to the
44、 experimenter. When doing so, practical considerations like costs of the units as well as the amount of waste produced should also be taken into consideration. Copyright International Organization for Standardization Provided by IHS under license with ISO Licensee=Boeing Co/5910770001 Not for Resale
45、, 01/31/2009 02:08:42 MSTNo reproduction or networking permitted without license from IHS -,-,- ISO 17512-1:2008(E) ISO 2008 All rights reserved 3 5 Reagents and materials 5.1 Boric acid reference toxicant, recommended. H3BO3 has been used historically as a soil chemosterilant and is an effective no
46、n-selective biocide (relative molecular mass: 61,81). Earthworms can detect and avoid sublethal concentrations that adversely affect reproduction. Boric acid satisfies the following criteria that attest to its suitability as a reference toxicant: a) it is effective at relatively low concentrations t
47、hat are not strongly influenced by the nature of the substrate; b) it is relatively stable and persistent so that concentrations do not change rapidly over the duration of the test; c) it is reasonably water soluble or miscible in water, does not volatilise readily, and can be readily mixed with soi
48、ls; d) there is a standard method for measuring boric acid concentrations in soil; e) it represents a minimal hazard to technicians and it is free of disposal problems. 5.2 Biological material, consisting of adult earthworms of the species Eisenia fetida or Eisenia andrei (individual mass: between 300 mg and 600 mg). Synchronisation of breeding of the organisms for this test is not necessary. An example of how to breed compost worms is given in Annex