1、September 2011 Translation by DIN-Sprachendienst.English price group 11No 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!$tUZ“1815055www.din.deDDIN EN ISO 20963Soil quality Effects of pollutants on insect larvae Determination of acute toxicity (ISO 20963:2005)English translation of DIN EN ISO 20963:2011-09Bodenbeschaffenheit Auswirkungen von Schadstoffen auf Insektenlarven ( ) Bestimmung der akuten Toxiz
3、itt (ISO 20963:2005)Englische bersetzung von DIN EN ISO 20963:2011-09Qualit du sol Effets des polluants vis-vis des larves dinsectes ( ) Dtermination de la toxicit aigu (ISO 20963:2005)Traduction anglaise de DIN EN ISO 20963:2011-09www.beuth.deDocument comprises 18 pagesIn case of doubt, the German-
4、language original shall be considered authoritative.(Oxythyrea funesta) Oxythyrea funestaOxythyrea funesta08.11 DIN EN ISO 20963:2011-09 2 A comma is used as the decimal marker. National foreword This standard has been prepared by Technical Committee ISO/TC 190 “Soil quality” in collaboration with T
5、echnical 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), Working Committee NA 119-01-02-04 UA Biologische Verfahren. The DIN Standards c
6、orresponding 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 11260 DIN EN ISO 11260 ISO 11268-1 E DIN ISO 11268-1:2010-07 ISO 11269-2 E DIN ISO 11269-2:2011-06 ISO 11465 DIN ISO 11465 ISO/TS 2
7、0281 referred to in Subclause 9.1.1 was published in 2006-04. National Annex NA (informative) Bibliography DIN ISO 10381-6, Soil quality Sampling Part 6: Guidance on the collection, handling and storage of soil under aerobic conditions for the assessment of microbiological processes, biomass and div
8、ersity 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 EN ISO 11260, Soil quality Determination of effective cation exchange capacity and base saturation level using
9、 barium chloride solution E DIN ISO 11268-1:2010-07, Soil quality Effects of pollutants on earthworms (Eisenia fetida/Eisenia andrei) Part 1: Determination of acute toxicity E DIN ISO 11269-2:2011-06, Soil quality Determination of the effects of pollutants on soil flora Part 2: Effects of contaminat
10、ed soils on the emergence and early growth of higher plants DIN ISO 11465, Soil quality Determination of dry matter and water content on a mass basis Gravimetric method EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN ISO 20963 June 2011 ICS 13.080.30 English Version Soil quality - Effects of po
11、llutants on insect larvae (Oxythyrea funesta) - Determination of acute toxicity (ISO 20963:2005) Qualit du sol - Effets des polluants vis-vis des larves dinsectes (Oxythyrea funesta) - Dtermination de la toxicit aigu (ISO 20963:2005) Bodenbeschaffenheit - Auswirkungen von Schadstoffen auf Insektenla
12、rven (Oxythyrea funesta) - Bestimmung der akuten Toxizitt (ISO 20963:2005) 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 s
13、tandard 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 i
14、n 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, Czech
15、Republic, Denmark, Estonia, Finland, France, Germany, Greece, 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 EUROPE
16、N 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 Members. Ref. No. EN ISO 20963:2011: EContents DIN EN ISO 20963:2011-09 EN ISO 20963:2011 (E) 2
17、Page Foreword3 Introduction .4 1 Scope 5 2 Normative references 5 3 Terms and definitions .5 4 Principle6 5 Test environment.7 6 Reagents.7 7 Apparatus .8 8 Procedure .8 8.1 Test design.8 8.2 Preparation of test mixture .9 8.3 Preparation of control container10 8.4 Food addition .10 8.5 Introduction
18、 of the biological material 10 8.6 Test conditions and measurements 10 8.7 Reference substance.10 9 Expression of results 11 9.1 Calculation11 9.2 Expression of results 12 10 Validity of the test12 11 Test report 12 Annex A (informative) Example of breeding technique for Oxythyrea funesta 13 Annex B
19、 (informative) Results of the collaborative trial carried out in France.15 Bibliography 16 Foreword The text of ISO 20963:2005 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 20963:2011 b
20、y 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, and conflicting national standa
21、rds 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. According to the CEN/CENELEC In
22、ternal 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, Italy, Latvia, Lithuania, Lux
23、embourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and the United Kingdom. Endorsement notice The text of ISO 20963:2005 has been approved by CEN as a EN ISO 20963:2011 without any modification. DIN EN ISO 20963:2011-09 EN ISO 20963:2011 (E
24、) 3 Introduction This International Standard describes a method for the determination of the acute toxicity of contaminated soils and chemicals to the larvae of Oxythyrea funesta, a phytophagous coleopteran (Scarabaeidae, Cetoniinae) with wide geographic distribution (Europe, North Africa and the Mi
25、ddle East). Oxythyrea funesta has many characteristics which make it suitable for soil quality monitoring or testing effects of chemicals: ecological relevance: this type of organism contributes in many ways to soil structure by stimulating soil aeration and drainage; the first stages of development
26、, i.e. incubation of eggs, larval cycle and pupation, are underground; the larvae of Oxythyrea funesta are tolerant to modifications of the test substrate granulometry; this species can be bred under controlled conditions. DIN EN ISO 20963:2011-09 EN ISO 20963:2011 (E) 4 1 Scope This International S
27、tandard describes a method for the determination of the effects of contaminated soils and substances on the survival of the larvae of Oxythyrea funesta. The larvae are exposed to the pollutants by cuticular and alimentary uptake. For contaminated soils, the effects on the survival are determined in
28、the test soil and in a control soil. Depending on the objectives of the study, the control and dilution substrates (dilution series of contaminated soil) are either uncontaminated soil comparable to the soil sample to be tested or artificial soil substrate. Effects of substances are assessed using a
29、 defined artificial soil substrate. This International Standard is not applicable to volatile substances, i.e. substances for which Henrys constant or the air/water partition coefficient is greater than 1, or for which the vapour pressure exceeds 0,001 33 Pa at 25 C. NOTE This method does not take i
30、nto account the possible degradation of the substances or pollutants during the test. 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
31、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 11268-1, Soil quality Ef
32、fects of pollutants on earthworms (Eisenia fetida) Part 1: Determination of acute toxicity using artificial soil substrate ISO 11269-2:1), Soil quality Determination of the effects of pollutants on soil flora Part 2: Effects of chemicals on the emergence and growth of higher plants 3 Terms and defin
33、itions For the purposes of this document, the following terms and definitions apply. 1) To be published. (Revision of ISO 11269-2:1995) DIN EN ISO 20963:2011-09 EN ISO 20963:2011 (E) 5 3.1 lethal concentration 50 LC 50 median lethal concentration of test substance or percent dilution of contaminated
34、 soil, which kills 50 % of the test organism within the test period 3.2 lethal concentration x LCx concentration of the test substance or percent dilution of contaminated soil which kills x % of the test organism within the test period NOTE x is the percentage (10, 20, 25) of this effect. 3.3 Lowest
35、 Observed Effect Concentration LOEC lowest tested concentration of test substance or contaminated soil at which a statistically significant effect is observed compared with the control NOTE All test concentrations above the LOEC have a harmful effect equal or greater than those observed at the LOEC.
36、 3.4 No Observed Effect Concentration NOEC highest tested concentration of test substance or contaminated soil at which no statistically significant effect is observed compared with the control NOTE The NOEC is the test concentration immediately below the LOEC. 3.5 test mixture mixture of the test s
37、ubstance with the test substrate, mixture of contaminated soil with the test substrate or mixture of contaminated soil with an uncontaminated soil comparable to the soil sample to be tested 3.6 diapause interruption of metabolism during egg, larva, pupa or imago development 4 Principle Larvae of Cet
38、oniinae (species Oxythyrea funesta) are exposed to a dilution range of contaminated soil or to a range of concentrations of test substance. The mortality of larvae is determined after 10 days. Test mixtures are prepared at the start of the test and are not renewed within the test period. The results
39、 obtained from the test are compared with a control and are used to determine the concentration which causes mortality of 50 % of the larvae (LC 5010days). The test is conducted in two steps: a preliminary test to determine appropriate dilution/concentration range in the final test; the definitive t
40、est to determine the dilutions/concentrations causing between 10 % and 90 % mortality, which yields the test result. DIN EN ISO 20963:2011-09 EN ISO 20963:2011 (E) 6 It may also be possible to determine the effects of contaminated soils or substances on the growth of larvae (optional). The increase
41、in mass within the test period allows this criterion to be considered as complementary to mortality, in order to assess the effects of contaminated soils or substances. 5 Test environment Tests shall be performed at a temperature of (26 1) C in complete darkness. 6 Reagents 6.1 Biological material T
42、he species used in the test is Oxythyrea funesta (Scarabaeidae, Cetoniinae). Third-instar larvae with a fresh mass within the range 100 mg to 200 mg are required to perform the test. The larvae shall be healthy, without any bites or other visible injuries. NOTE Depending on the breeding conditions,
43、described in Annex A, larvae approximately two weeks old are suitable for the test. Larvae of similar size shall be selected. The difference in mass between the smallest and the largest larva within a single test container shall not exceed 50 mg. Eliminate the particles of breeding substrate stuck t
44、o the integument using, for example, a soft brush before weighing the larvae. It is also possible to leave the animals to move along on slightly moist paper in order to eliminate the breeding substrate stuck to the integument. Synchronisation of breeding is necessary. An example of breeding techniqu
45、e for Oxythyrea funesta is given in Annex A. 6.2 Test substrate The mass of substrate used per glass container (7.1) shall be equivalent to 300 g (dry mass). The substrate, called artificial soil, shall have the following composition (in accordance with ISO 11268-1): sphagnum peat, air-dried, finely
46、 ground and with no visible plant remains: 10 % (expressed on a dry mass basis); kaolinite clay containing not less than 30 % kaolinite: 20 % (expressed on a dry mass basis); industrial quartz sand (dominant fine sand with more than 50 % particle size between 0,05 mm and 0,20 mm): 70 % (expressed on
47、 a dry mass basis). Add pulverised calcium carbonate (CaCO3), of recognised analytical grade, as necessary to bring the pH of the wetted substrate to 6,0 0,5 (commonly between 0,5 % and 1 % of the mass of the dry ingredients). Prepare the artificial soil by mixing the dry constituents listed above t
48、horoughly in a large-scale laboratory mixer. The amount of calcium carbonate required can vary, depending on properties of the individual batch of sphagnum peat, and should be determined by weighing subsamples immediately before the test. Store the mixed artificial soil at room temperature. To deter
49、mine pH and the maximum water-holding capacity, pre-moisten the dry artificial soil at least two days before starting the test by adding deionized water to obtain half of the required final water content of 50 % of the maximum water-holding capacity. DIN EN ISO 20963:2011-09 EN ISO 20963:2011 (E) 7 Determine the water-holding capacity in accordance with ISO 11269-2:, Annex A, and pH in accordance with ISO 10390. If the measured pH is not within
