1、raising standards worldwideNO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAWBSI Standards Publication11269-2:201Soil quality Determination of the effects of pollutants on soil floraPart 2: Effects of contaminated soil on the emergence and early growth of higher plants3BS EN ISOB
2、S EN ISO 11269-2:2013National forewordThis British Standard is the UK implementation of EN ISO 11269-2:2013. It is identical to ISO 11269-2:2012. It supersedes BS ISO 11269-2:2012, which is withdrawn.The UK participation in its preparation was entrusted to Technical Committee EH/4, Soil quality.A li
3、st of organizations represented on this committee can be 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. The British Standards Institution 2013. Published by BSI Standards
4、 Limited 2013ISBN 978 0 580 79337 0 ICS 13.080.30 Compliance with a British Standard cannot confer immunity from legal obligations.This British Standard was published under the authority of the Standards Policy and Strategy Committee on 29 February 2012.Amendments/corrigenda issued since publication
5、Date Text affected30 June 2013 This corrigendum renumbers BS ISO 11269-2:2012 as BS EN ISO 11269-2:2013BRITISH STANDARDEUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN ISO 11269-2 February 2013 ICS 13.080.30 English Version Soil quality - Determination of the effects of pollutants on soil flora
6、- Part 2: Effects of contaminated soil on the emergence and early growth of higher plants (ISO 11269-2:2012) Qualit du sol - Dtermination des effets des polluants sur la flore du sol - Partie 2: Effets des sols contamins sur lmergence et la croissance des vgtaux suprieurs (ISO 11269-2:2012) Bodenbes
7、chaffenheit - Bestimmung der Wirkungen von Schadstoffen auf die Bodenflora - Teil 2: Wirkung von verunreinigten Bden auf Saatauflauf und frhes Wachstum hherer Pflanzen (ISO 11269-2:2012) This European Standard was approved by CEN on 5 February 2013. CEN members are bound to comply with the CEN/CENEL
8、EC 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 bibliographical references concerning such national standards may be obtained on application to the CEN-CENELEC Management Centre or
9、 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 responsibility of a CEN member into its own language and notified to the CEN-CENELEC Management Centre has the same status as the offi
10、cial versions. CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands,
11、 Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and United Kingdom. EUROPEAN COMMITTEE FOR STANDARDIZATION COMIT EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR NORMUNG Management Centre: Avenue Marnix 17, B-1000 Brussels 2013 CEN All rights of exploitation
12、 in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN ISO 11269-2:2013: EForeword The text of ISO 11269-2:2012 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
13、 ISO 11269-2:2013 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 August 2013, and conflictin
14、g national standards shall be withdrawn at the latest by August 2013. 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
15、 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, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hu
16、ngary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom. Endorsement notice The text of ISO 11269-2:2012 has been approved by CEN as EN ISO 11269-2:2013 without
17、 any modification. iiBS EN ISO 11269-2:2013 ISO 11269-2:2013 (E) ISO 2013 All rights reservedEN iiiContents PageIntroduction .1 Scope 12 Normative references . 13 Terms and definitions . 14 Units 35 Principle . 36 Test plants . 37 Materials . 47.1 Test vessels 47.2 Soil . 48 Equipment . 69 Reference
18、 substance 610 Procedure 610.1 Experimental design . 610.2 Preparation of the pots 710.3 Preparation of the seeds . 710.4 Growth conditions . 810.5 Start of the test . 810.6 Handling during the test 811 Validity criteria . 912 Assessment of the results 912.1 Data presentation 912.2 Expression of the
19、 results 913 Statistical analysis 913.1 General . 913.2 Range-finding test .1013.3 Final test 1014 Test report . 11Annex A (informative) Additional recommended plant species based on test results gained by applying Environment Canada Test Method: EPS 1/RM/454.13Annex B (informative) Phytotoxic value
20、s for reference compounds: sodium trichloro-acetate and boric acid .15Annex C (informative) Recommended method for the measuring of the water-holding capacity of the soil .16Annex D (informative) Recommendations for nutrient supply of soils 17Bibliography .18ivBS EN ISO 11269-2:2013 ISO 11269-2:2012
21、 (E)iv IntroductionThis part of ISO 11269 describes a procedure for evaluating the quality of soils of different origin carrying unknown contaminations. The evaluation of the effects on plant growth is based on emergence and inhibitory effects on early growth of at least two species of higher plants
22、. Guidance for assessing potential effects of substances on seedling emergence and growth is given in OECD Guideline 20814.This part of ISO 11269 refers closely to ISO 22030 and is based on:a) results from the German research project “Entwicklung eines innovativen und technischen Instrumentariums zu
23、r Optimierung der kotoxikologischen Bewertung von Bden im Hinblick auf Sanierungsziele und Schutzerfordernisse”;b) discussions within the joint project “Ecotoxicological Test Batteries” forming part of the BMBF Joint Research Group “Processes for the Bioremediation of Soil”23;c) results from the BMB
24、F Joint Research Group ERNTE “Erprobung und Vorbereitung einer praktischen Nutzung kotoxikologischer Testsysteme”17;d) ring-test results of “Ecotoxicological Characterisation of Waste Results and Experiences from an International Ring Test”8.Plant growth can be influenced strongly by soil properties
25、 such as texture, pH or levels of nutrients. When testing natural soils either reference soils (uncontaminated soils with the same properties as the test soil) or standard soils are used as mixing and control substrate. In the latter case, variations in plant growth can result from either soil conta
26、minants or differences in soil properties like nutrients and texture. Therefore, results from soil testing can less easily be interpreted than results from testing of chemicals .BS EN ISO 11269-2:2013 ISO 11269-2:2012 (E)Soil quality Determination of the effects of pollutants on soil flora Part 2: E
27、ffects of contaminated soil on the emergence and early growth of higher plantsWARNING Contaminated soils may contain unknown mixtures of toxic, mutagenic, or otherwise harmful chemicals or infectious micro-organisms. Occupational health risks may arise from dust or evaporated chemicals during handli
28、ng and incubation. Furthermore, test plants might take up chemicals from the soil and safety measures should also be considered when handling the test plants.1 ScopeThis part of ISO 11269 describes a method to assess the quality of an unknown soil and the soil habitat function by determining the eme
29、rgence and early growth response of at least two terrestrial plant species compared to reference or standard control soils. It is applicable to soils of unknown quality, e.g. from contaminated sites, amended soils or soils after remediation.2 Normative referencesThe following referenced documents ar
30、e 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 sto
31、rage of soil under aerobic conditions for the assessment of microbiological processes, biomass and diversity in the laboratoryISO 10390, Soil quality Determination of pHISO 10694, Soil quality Determination of organic and total carbon after dry combustion (elementary analysis)ISO 11260, Soil quality
32、 Determination of effective cation exchange capacity and base saturation level using barium chloride solutionISO 11268-1, Soil quality Effects of pollutants on earthworms Part 1: Determination of acute toxicity to Eisenia fetida/Eisenia andreiISO 11268-2, Soil quality Effects of pollutants on earthw
33、orms Part 2: Determination of effects on reproduction to Eisenia fetida/Eisenia andreiISO 11277, Soil quality Determination of particle size distribution in mineral soil material Method by sieving and sedimentationISO 11465, Soil quality Determination of dry matter and water content on a mass basis
34、Gravimetric methodISO 22030, Soil quality Biological methods Chronic toxicity in higher plants3 Terms and definitionsFor the purposes of this document, the following terms and definitions apply.INTERNATIONAL STANDARD 1BS EN ISO 11269-2:2013 ISO 11269-2:201 (E)23.1emergenceappearance of the coleoptil
35、e or cotyledon above the soil3.2contaminantsubstance or agent present in the soil as a result of human activityISO 15176:2002283.3hormesisimprovement of seedling emergence, growth or survival (or other response of the test plants) at low concentrations of chemicals or mixtures of soil that are toxic
36、 when applied at higher levels in comparison to the control123.4lowest observed effect rate or effect concentrationLOEClowest tested percentage of a test soil in a reference or a standard control soil or concentration of a substance at which a statistically significant effect is observedNOTE The LOE
37、C is expressed as a percentage of the test-soil dry mass per soil-mixture dry mass. All test mixtures above the LOEC have a harmful effect equal to or greater than that observed at the LOEC. If this condition cannot be satisfied, an explanation should be given for how the LOEC and NOEC (3.5) have be
38、en selected.3.5no observed effect concentrationNOECtest-soil percentage immediately below the LOEC, which when compared to the control has no statistically significant effect (r 2) or ionizing substances into account, 5 % of peat have proven to be sufficient for maintaining the desired structure of
39、the artificial soil. In this case, the respective percentages of the constituents are modified as follows: peat, 5 %; clay, 20 %; sand 75 %).NOTE 2 pH (KCl) is measured in a mixed sample in a 1 M solution of potassium chloride (KCl) or a 0,01 M solution of calcium chloride (CaCl2).The artificial soi
40、l is prepared, at least three days prior to starting the test, by mixing the dry constituents listed above thoroughly in a large-scale laboratory mixer. A portion of the deionized water required to obtain half of the final water content of 40 % to 60 % of the maximum water-holding capacity is added
41、while mixing is continued. The mixed artificial soil shall be stored at room temperature for at least two days to equilibrate acidity. The amount of calcium carbonate required might vary, depending on properties of the individual batch of sphagnum peat and should be determined by measuring the pH of
42、 subsamples immediately before the test. The total water-holding capacity is determined according to Annex C, the pH is determined according to ISO 10390.Allowance should be made for any water that is to be used for introducing the test substance into the soil.To obtain a dilution series, the test s
43、oil is mixed with the control soil thoroughly (either manually or by using a hand mixer). The homogeneity of the mixture is checked visually.5BS EN ISO 11269-2:2013 ISO 11269-2:2012 (E)8 EquipmentStandard laboratory equipment including the following materials is required.8.1 Controlled environmental
44、 chamber, phytotron, plant growth room or greenhouse suitable to maintain the specified conditions.8.2 Balance (0,1 mg).8.3 Balance for heavier loads (e.g. 10 kg) for preparation of soil mixtures.8.4 Sieve, stainless steel, with mesh size 4 mm.8.5 Glass-fibre wicks ( 1 mm).9 Reference substanceIt is
45、 recommended that a reference substance be tested to demonstrate the uniformity of the laboratory test conditions and the response of the particular batch of seeds. Sodium trichloroacetate or boric acid is suggested as the reference substance. A reference test should be carried out regularly and if
46、any major changes in operating procedures are introduced, for example, change of phytotron/growth room/greenhouse, change of soil or change of watering regime, etc. Examples of phytotoxic values for two reference compounds are given in Annex B.10 Procedure10.1 Experimental designA sample of field-co
47、llected test soil may be tested at a single concentration (typically 100 %) or evaluated for toxicity in a multi-concentration test whereby a series of concentrations (dilutions) are prepared by mixing defined quantities with a control soil.Depending on the knowledge of relevant response levels, a p
48、reliminary range-finding test may precede the final test. Each final test consists of a series of soil mixtures (treatments). Each treatment is replicated at least four times, i.e. four test pots containing a number of test plants are used.To avoid any masking of the phytotoxic effect expression by
49、nutrient deficiency, all treatment groups including control soils are amended by fertilizers (10.6.3) after seedlings have emerged. It has been shown that effects of pollutants on plant growth are pronounced in soils with optimum nutrient supply67172225.To assure functioning of the watering system (see 10.6.2), it should be checked whether the test soil sucks water via wicks sufficiently. Water repellence or poor water transport can occur with very sandy soils, soils highly contaminated with hydro
