1、raising standards worldwideNO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAWBSI Standards PublicationBS ISO 16191:2013W a t e r q u a l i t y D e t e r m i n a t i o n of the toxic effect of sediment on the growth behaviour of Myriophyllum aquaticumBS ISO 16191:2013 BRITISH STAN
2、DARDNational forewordThis British Standard is the UK implementation of ISO 16191:2013. The UK participation in its preparation was entrusted to TechnicalCommittee EH/3/5, Biological Methods.A list of organizations represented on this committee can be obtained on request to its secretary.This publica
3、tion 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 Limited 2013ISBN 978 0 580 71142 8 ICS 13.060.70 Compliance with a British Standard cannot confer immunit
4、y from legal obligations.This British Standard was published under the authority of the Standards Policy and Strategy Committee on 30 June 2013.Amendments issued since publicationDate T e x t a f f e c t e dBS ISO 16191:2013 ISO 2013Water quality Determination of the toxic effect of sediment on the
5、growth behaviour of Myriophyllum aquaticumQualit de leau Dtermination de leffet toxique des sdiments sur la croissance de Myriophyllum aquaticumINTERNATIONAL STANDARDISO16191First edition2013-05-01Reference numberISO 16191:2013(E)BS ISO 16191:2013ISO 16191:2013(E)ii ISO 2013 All rights reservedCOPYR
6、IGHT PROTECTED DOCUMENT ISO 2013All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized otherwise in any form or by any means, electronic or mechanical, including photocopying, or posting on the internet or an intranet, without prior written permiss
7、ion. Permission can be requested from either ISO at the address below or ISOs member body in the country of the requester.ISO copyright officeCase postale 56 CH-1211 Geneva 20Tel. + 41 22 749 01 11Fax + 41 22 749 09 47E-mail copyrightiso.orgWeb www.iso.orgPublished in SwitzerlandBS ISO 16191:2013ISO
8、 16191:2013(E) ISO 2013 All rights reserved iiiContents PageForeword ivIntroduction v1 Scope . 12 Normative references 13 Terms and definitions . 14 Principle 35 Interferences 36 Reagents 37 Apparatus . 58 Test with reference substance 69 Test organism . 610 Procedure. 610.1 Pre-culturing of Myrioph
9、yllum aquaticum for the contact test. 610.2 Preparation of the control sample . 610.3 Sampling, storage, and preparation of the test samples . 710.4 Test procedure . 710.5 Exposure conditions . 710.6 Measurements . 811 Evaluation 911.1 Growth rate r 911.2 Means per test and control vessel (rV,T, rV,
10、C) . 911.3 Means per test and control sample (rS,T, rS,C) 911.4 Inhibition I 911.5 Estimation of ErCxvalues . 911.6 Expression of results . 1012 Validity criteria .1113 Test report 11Annex A (informative) Figures .12Annex B (normative) Preparation of nutrient solution (Steinberg medium, following IS
11、O 20079)14Annex C (informative) Suppliers 16Annex D (normative) Preparation of spiked artificial sediments .18Annex E (informative) Performance data .20Bibliography .21BS ISO 16191:2013ISO 16191:2013(E)ForewordISO (the International Organization for Standardization) is a worldwide federation of nati
12、onal 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. Internat
13、ional 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.The procedures used to develop this document and those intende
14、d for its further maintenance are described in the ISO/IEC Directives, Part 1. In particular the different approval criteria needed for the different types of ISO documents should be noted. This document was drafted in accordance with the editorial rules of the ISO/IEC Directives, Part 2, www.iso.or
15、g/directives.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. Details of any patent rights identified during the development of the document will be i
16、n the Introduction and/or on the ISO list of patent declarations received, www.iso.org/patents.Any trade name used in this document is information given for the convenience of users and does not constitute an endorsement.The committee responsible for this document is ISO/TC 147, Water quality, Subco
17、mmittee SC 5, Biological methods.iv ISO 2013 All rights reservedBS ISO 16191:2013ISO 16191:2013(E)IntroductionThe contact test with Myriophyllum aquaticum described in this International Standard allows the measurement of responses of the plant to dissolved and particle-bound substances present in s
18、ediment samples within 10 d (References 345678).The test plant, Myriophyllum aquaticum (Vellosco) Verdcourt (parrot feather), is a dicotyledonous macrophyte. It is a native of the Amazon River in South America, but it has naturalized worldwide, especially in warmer climates. It has been introduced w
19、orldwide for use in indoor and outdoor aquaria. For its use as test organism, its capability for emersed growth (no additional liquid as supernatant is needed), its strong regeneration potential, and its vegetative growth are harnessed in the contact test. Furthermore, Myriophyllum aquaticum grows w
20、ithout generating side shoots during the test period, which facilitates handling in the laboratory. However, it should be ensured that no live plant material is lost from the laboratory.Myriophyllum aquaticum can be affected by phytotoxic substances present in sediments (e.g. dredged material). The
21、subsequent inhibition of growth is calculated from the parameter (fresh mass) by a number of defined calculation methods. ISO 2013 All rights reserved vBS ISO 16191:2013BS ISO 16191:2013Water quality Determination of the toxic effect of sediment on the growth behaviour of Myriophyllum aquaticumWARNI
22、NG Persons using this International Standard should be familiar with normal laboratory practice. This International Standard does not purport to address all of the safety problems, if any, associated with its use. It is the responsibility of the user to establish appropriate safety and health practi
23、ces and to ensure compliance with any national regulatory conditions. It shall be ensured that no plant material can elude the laboratory.IMPORTANT It is absolutely essential that tests conducted according to this International Standard be carried out by suitably trained staff.1 ScopeThis Internatio
24、nal Standard specifies a method for determining the toxicity of environmental samples on the growth of Myriophyllum aquaticum. The method described is applicable to natural fresh water sediment and artificial sediment.2 Normative referencesThe following documents, in whole or in part, are normativel
25、y referenced in this document and are indispensable for its application. For dated references, only the edition cited applies. For undated references, the latest edition of the referenced document (including any amendments) applies.ISO 5667-15, Water quality Sampling Part 15: Guidance on the preserv
26、ation and handling of sludge and sediment samplesISO 5667-16, Water quality Sampling Part 16: Guidance on biotesting of samplesISO 10523, Water quality Determination of pHISO 11465, Soil quality Determination of dry matter and water content on a mass basis Gravimetric methodISO 20079, Water quality
27、Determination of the toxic effect of water constituents and waste water on duckweed (Lemna minor) Duckweed growth inhibition testOECD 218, OECD Guidelines for the testing of chemicals Sediment-water Chironomid toxicity test using spiked sediment3 Terms and definitionsFor the purposes of this documen
28、t, the following terms and definitions apply.3.1artificial sedimentdefined artificial sedimentSOURCE: ISO 10872:2010,2definition 3.3, modifiedNote 1 to entry: See 6.9.INTERNATIONAL STANDARD ISO 16191:2013(E) ISO 2013 All rights reserved 1BS ISO 16191:2013ISO 16191:2013(E)3.2chlorosisloss of pigments
29、 (yellowing of plant tissue)SOURCE: ISO 20079:2005, definition 3.3, modified3.3control sampleartificial sediment pre-treated according to the need of this test that serves as negative control to which the effect in the respective test material is comparedSOURCE: ISO 10872:2010,2definition 3.6, modif
30、ied3.4effective concentrationErCxconcentration of a substance in a test sample (ECx) at which an effect of x % is measured, if compared to the controlSOURCE: ISO 20079:2005, definition 3.9, modifiedNote 1 to entry: To unambiguously denote an EC value deriving from growth rate, it is proposed to use
31、the symbol “ErCx”.3.5emersed growthmorphological habitus of aquatic macrophytes, growing above the water surface3.6head-whorlapical part of a Myriophyllum plantNote 1 to entry: See Figure A.1.3.7necrosislocalized dead plant tissue (i.e. brown or white)SOURCE: ISO 20079:2005, definition 3.16, modifie
32、d3.8nutrient solutionsolution of nutrients and micronutrients in water which are essential for the growth of Myriophyllum SOURCE: ISO 20079:2005, definition 3.17, modified3.9pre-cultureculture of Myriophyllum aquaticum used for acclimatization of test plants to the test conditions and for the growin
33、g of the plants to be used as whorls at test startSOURCE: ISO 20079:2005, definition 3.19, modifiedNote 1 to entry: See Figure A.2.3.10replicateone of a selected number of test vessels (containing sample material from one sample and test organisms)Note 1 to entry: Each vessel is tested.Note 2 to ent
34、ry: The replicates mentioned in this International Standard contain sample material (e.g. natural sediment) and three whorls of Myriophyllum aquaticum.2 ISO 2013 All rights reservedBS ISO 16191:2013ISO 16191:2013(E)3.11test samplediscrete portion of a sample (e.g. sediment or artificial sediment)SOU
35、RCE: ISO 10872:2010,2definition 3.14, modified3.12whorlarrangement of leaves that radiate from a single point and surround the stemNote 1 to entry: See Figure A.1.4 PrincipleMyriophyllum aquaticum whorls are exposed to test samples over a period of 10 d. The growth of Myriophyllum aquaticum in a tes
36、t sample is compared with its growth in the control sample. Phytotoxic effects are quantified as growth inhibition (%) relative to the control growth.5 InterferencesIn case of problems with Myriophyllum control growth using artificial sediment, the respective components should be checked, first to e
37、xclude contamination with, for example, heavy metals (kaolin) or suitability of peat (if the recommended peat is not used).6 ReagentsUse, as far as possible, reagents of recognized analytical grade.6.1 Water, distilled or deionized water or water of equivalent purity, conductivity 0,710 0,710 to 0,5
38、00 0,500 to 0,355 0,355 to 0,250 40,250 to 0,180 340,180 to 0,125 500,125 to 0,090 11 0,090 11) This information is given for the convenience of users of this document and does not constitute an endorsement by ISO of this supplier.16 ISO 2013 All rights reservedBS ISO 16191:2013ISO 16191:2013(E)For
39、peat: Lithuanian peat moss2)Klasmann-Deilmann GmbH Georg-Klasmann-Str. 2-10 49744 Geeste Germany Tel: +49 (0)59 37-31-0 Fax: +49 (0)59 37-31-279 www.klasmann-Lithuanian peat moss (data sheet 12/2007)Structure extra fine (0 mm to 5 mm)pH value (H2O, volume fraction 1:2,5) 3,0 to 3,5Salt level (volume
40、 fraction 1:3,6) 150 mg/lNitrogen 50 mg/lPhosphate 30 mg/lPotassium 30 mg/lMagnesium 80 mg/lOrganic substances in dry mass (mass fraction in %) 94 to 99Ash in dry mass (mass fraction in %) 1 to 6Water capacity (volume fraction in %) 80 to 852) This information is given for the convenience of users o
41、f this document and does not constitute an endorsement by ISO of this supplier. ISO 2013 All rights reserved 17BS ISO 16191:2013ISO 16191:2013(E)Annex D (normative) Preparation of spiked artificial sedimentsD.1 GeneralThe spiking procedure described here follows the OECD Guideline 218. All test conc
42、entrations are calculated as concentration of the test substance in the sediment (mg/kg sediment dry mass).If dilution series will be performed, the different concentration(s) of the test substance will be prepared by spiking a defined solution of the test substance into the sediment, not via diluti
43、on of the sediment. Only the stock solution of the test substance will be diluted in the appropriate concentration steps.If scarcely soluble substances which are dissolved in an organic solvent are to be investigated, a solvent control is needed.Split up the original sediment sample into the require
44、d number of portions by taking into account the number of concentration levels and solvent controls needed for the test.A control sample (see 3.3 and 6.9.1) is needed for all test series and is performed according to 6.9.1.D.2 Spiking procedureUse freshly prepared (6.9.1) artificial sediment.Determi
45、ne the dry mass of the sediment (ISO 11465).Calculate the required amount of spiking substance needed for the test concentration.D.2.1 Preparation of the sediment aliquot for spikingWeigh out 10 % (wet mass) of the required amount of sediment and dry it according to ISO 11465. If necessary, pulveriz
46、e the dried material, so that it consists of possibly small particles. Keep in mind to compensate the lost water content later (see below).NOTE Experience has shown that drying the sediment aliquot 24 h at 105 C is sufficient.D.2.2 Preparation of the test substance solution for spikingDissolve the c
47、hosen test substance in a suitable solvent e.g. water (6.1), nutrient solution (6.6), or organic solvent (e.g. 2-propanone), depending on the solubility of the test substance in as small a volume as possible maximum 4 % (volume/wet mass) of the required amount of wet sediment.D.2.3 Spiking of the pr
48、epared sediment aliquotAdd the test substance solution to the dried sediment aliquot (D.2.1). Let the solvent evaporate under a fume hood until it has been totally removed from the sediment aliquot up to 3 d in case of inorganic solvents e.g. water (6.1). If lumps or crusts form during drying, crush
49、 them with a stainless-steel spatula or, if they are persistent, in a mortar.D.2.4 Spiking of the total sedimentWeigh out 90 % (wet mass) of the required amount of sediment.18 ISO 2013 All rights reservedBS ISO 16191:2013ISO 16191:2013(E)To compensate the lost water content of the 10 % sediment aliquot (see D.2.1), add the respective aliquot of water (6.1) needed for compensation, and thoroughly mix the dried, spiked sediment aliquot (D.2.3), the aliquot of compensat
