1、BSI Standards PublicationBS ISO 16303:2013Water quality Determination of toxicity of fresh watersediments using Hyalella aztecaBS ISO 16303:2013 BRITISH STANDARDNational forewordThis British Standard is the UK implementation of ISO 16303:2013. The UK participation in its preparation was entrusted to
2、 TechnicalCommittee EH/3/5, Biological Methods.A list 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 Sta
3、ndards Institution 2013.Published by BSI Standards Limited 2013ISBN 978 0 580 72182 3ICS 13.060.70Compliance 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 31 December 201
4、3.Amendments/corrigenda issued since publicationDate T e x t a f f e c t e dBS ISO 16303:2013 ISO 2013Water quality Determination of toxicity of fresh water sediments using Hyalella aztecaQualit de leau Dtermination de la toxicit des sdiments deau douce vis-vis de Hyalella aztecaINTERNATIONAL STANDA
5、RDISO16303First edition2013-12-01Reference numberISO 16303:2013(E)BS ISO 16303:2013ISO 16303:2013(E)ii ISO 2013 All rights reservedCOPYRIGHT PROTECTED DOCUMENT ISO 2013All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized otherwise in any form or
6、by any means, electronic or mechanical, including photocopying, or posting on the internet or an intranet, without prior written permission. 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-121
7、1 Geneva 20Tel. + 41 22 749 01 11Fax + 41 22 749 09 47E-mail copyrightiso.orgWeb www.iso.orgPublished in SwitzerlandBS ISO 16303:2013ISO 16303:2013(E) ISO 2013 All rights reserved iiiContents PageForeword ivIntroduction v1 Scope . 12 Normative references 13 Terms and definitions . 14 Principle 25 Te
8、st environment . 35.1 Facilities 35.2 Lighting 36 Reagents, test organisms, and materials . 36.1 Test organism . 36.2 Control sediment . 36.3 Overlying water 46.4 Food 56.5 Reference substance . 67 Apparatus . 68 Treatment and preparation of samples 78.1 General . 78.2 Test sediment . 78.3 Preparati
9、on of sediment samples . 89 Test procedure 89.1 Preparing the test containers . 89.2 Introducing the organisms . 99.3 Test conditions 99.4 Renewal of overlying water. 109.5 Test observations and measurements . 1010 Expression of results .1010.1 Survival . 1010.2 Growth . 1110.3 Validity of the test
10、1111 Analysis and interpretation of results 1111.1 Data analysis 1111.2 Non-contaminant factors 1112 Reference substance 1212.1 Water-only test 1212.2 Whole sediment test 1213 Test report 13Annex A (informative) Description of Hyalella azteca.14Annex B (informative) Culturing Hyalella azteca .18Anne
11、x C (informative) Sources of Hyalella azteca 20Annex D (normative) Procedure for preparing YCT 22Annex E (informative) 42-day sediment reproduction test .23Annex F (informative) 14-day water-only survival-and-growth test24Annex G (informative) Performance data 25Bibliography .26BS ISO 16303:2013ISO
12、16303:2013(E)ForewordISO (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 f
13、or 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
14、 all matters of electrotechnical standardization.The procedures used to develop this document and those intended 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.
15、 This document was drafted in accordance with the editorial rules of the ISO/IEC Directives, Part 2 (see www.iso.org/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
16、 or all such patent rights. Details of any patent rights identified during the development of the document will be in the Introduction and/or on the ISO list of patent declarations received (see www.iso.org/patents).Any trade name used in this document is information given for the convenience of use
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18、ry informationThe committee responsible for this document is ISO/TC 147, Water quality, Subcommittee SC 5, Biological methods.iv ISO 2013 All rights reservedBS ISO 16303:2013ISO 16303:2013(E)IntroductionSediment in the aquatic environment serves as a reservoir for agricultural, industrial, and munic
19、ipal contaminants. Contaminated sediment adversely affects the benthic community directly and acts as a source of contamination for the overlying water, often negatively impacting pelagic communities as well. Sediment toxicity tests are used globally to determine and monitor the toxic effects of dis
20、crete substances or complex mixtures that might be harmful to indigenous life in the aquatic and benthic environments. This International Standard outlines procedures for conducting 14 d and/or 28 d tests for sediment toxicity, using the fresh water amphipod Hyalella azteca. The biological end point
21、s for the tests include mortality and growth. ISO 2013 All rights reserved vBS ISO 16303:2013BS ISO 16303:2013Water quality Determination of toxicity of fresh water sediments using Hyalella aztecaWARNING Persons using this International Standard should be familiar with normal laboratory practice. Th
22、is 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 practices and to ensure compliance with any national regulatory conditions.IMPORTANT It is absolutely ess
23、ential that tests conducted in accordance with this International Standard be carried out by suitably trained staff.1 ScopeThis International Standard specifies a method for the determination of toxicity to young Hyalella azteca in whole sediment based on survival and growth inhibition after 14 d an
24、d/or 28 d.The method is applicable toa) samples of contaminated whole fresh water sediment,b) chemical, industrial, or municipal sludge, or other solid wastes that may combine with fresh water sediments, andc) chemicals or preparations spiked into clean sediment.This International Standard is applic
25、able to the testing of sediment samples from the fresh water environment. Hyalella azteca can be used in the testing of brackish waters up to a maximum of 15 , with careful acclimation. This International Standard is not applicable to the testing of sediment samples from the marine and estuarine env
26、ironment with a salinity of 15 .This method is a 14 d and/or 28 d survival-and-growth test applicable to the sediment sample types described above.2 Normative referencesThe following documents, in whole or in part, are normatively referenced in this document and are indispensable for its application
27、. For dated references, only the edition cited applies. For undated references, the latest edition of the referenced document (including any amendments) applies.ISO 5814, Water quality Determination of dissolved oxygen Electrochemical probe methodISO 6059, Water quality Determination of the sum of c
28、alcium and magnesium EDTA titrimetric methodISO 10523, Water quality Determination of pH3 Terms and definitionsFor the purposes of this document, the following terms and definitions apply.3.1artificial sedimentmixture of materials that mimic the physical components of natural sedimentNote 1 to entry
29、: See 6.2.1.INTERNATIONAL STANDARD ISO 16303:2013(E) ISO 2013 All rights reserved 1BS ISO 16303:2013ISO 16303:2013(E)3.2control sedimentsediment (natural or artificial) that is used to assess the performance of the test organisms and test acceptability (i.e. clean)Note 1 to entry: The results of con
30、trol sediment testing are used for comparison to response of organisms in the contaminated test sediment(s) and for evaluating test validity.Note 2 to entry: Used routinely to assess the acceptability of a test (6.2).3.3test sedimentdiscrete portion of sediment (field collected or spiked) to be test
31、ed for possible effects on amphipods due to contamination3.4spiked sedimentsediment to which a material has been added for testing3.5reference sedimentfield collected near an area of concern with properties representing sediment conditions that closely match those of the sample(s) of the test sedime
32、nt except for the degree of contaminationNote 1 to entry: It is often selected from a site uninfluenced by the source(s) of contamination but within the general vicinity of the sites where samples of test sediment are collected. Test results provide a site-specific basis for evaluating toxicity.3.6i
33、ntermittent renewaltests in which test solutions or overlying water are renewed during the test, based on deterioration of water quality3.7overlying waterwater placed over the layer of sediment in the test containerNote 1 to entry: Also used to manipulate the sediment, if necessary (e.g. for prepari
34、ng formulated sediment or mixtures of spiked sediment).3.8growthincrease in dry weight of test organisms during the experiment and expressed as mean dry weight per surviving amphipod4 PrincipleYoung fresh water amphipods, Hyalella azteca, aged 2 d to 9 d and ranging in age by 1 d to 2 d, are exposed
35、 in groups of 10 organisms to a contaminated sediment or a test-chemical spiked sediment for 14 d and/or 28 d.1 2 3The end points for the test are percent mortality and growth inhibition assessed relative to organisms exposed concurrently to control sediment. The test is performed in glass container
36、s with the ratio of sediment to water (volume:volume) being either 1:1,75 or 1:4 (e.g. 100 ml of sediment with 175 ml of overlying water or 100 ml of sediment with 400 ml of overlying water). Comparative testing of the two recommended sediment-to-water ratios showed no significant difference in test
37、 results using Hyalella azteca.4One advantage of the 1:4 sediment-to-water ratio is greater overlying water volume for chemical analysis. The exposure is primarily static unless renewal is triggered by deterioration of water in the control treatment (e.g. shifting pH affecting the form of background
38、 ammonia).2 ISO 2013 All rights reservedBS ISO 16303:2013ISO 16303:2013(E)A long-term test option (i.e. 42 d) for whole sediment toxicity testing using Hyalella azteca is described in Annex E. End points of this long-term test include survival (Days 28, 35, and 42), growth (Days 28 and 42), and repr
39、oduction (number of young per female produced from Days 28 to 42).A water-only method using Hyalella azteca is also described in Annex F. This method is a 14-day test of survival and growth using young amphipods exposed to samples of industrial or sewage effluents, fresh waters (e.g. receiving water
40、), aqueous extracts, or chemical substances which are soluble or which can be maintained as stable suspensions or dispersions under the conditions of the test.5 Test environment5.1 FacilitiesThe test facility shall be well ventilated, isolated from physical disturbances, and free from dust and fumes
41、. Tests shall be carried out in a temperature-controlled room or chamber that maintains a temperature of (23 2) C in the test containers.5.2 LightingAll test containers shall receive direct, overhead illumination that provides normal laboratory lighting (i.e. 100 lx to 1 000 lx) at the air/water int
42、erface. Illumination should be uniform and shall have a day/night cycle (photoperiod) of 16 h of daylight and 8 h of darkness.6 Reagents, test organisms, and materialsUse only reagents of recognized analytical grade, unless otherwise specified.6.1 Test organismHyalella azteca is an epibenthic sedime
43、nt-burrowing detritivore that lives in close contact with the surficial 1 cm or 2 cm of fresh water sediments. They reside in temperate lakes, ponds, and slow-flowing streams and are widely distributed on the North and South American continents.1 2 3Young Hyalella azteca are obtained from laboratory
44、 cultures maintained under the conditions of temperature, photoperiod, and food identical to those in the test. The species identification should be confirmed by qualified personnel experienced in identifying fresh water amphipods using the distinguishing taxonomic features described in Annex A and
45、in previous publications.2 56.1.1 Life stage and sizeAmphipods used for the test shall be between the ages of 2 d and 9 d and shall not vary in age by more than 2 d. A method for culturing Hyalella azteca and for obtaining known-age test organisms is provided in Annex B. If growth is expressed as me
46、an size at the end of the test, a mean length of organisms should be determined at test initiation.6.1.2 SourceAll amphipods used in a test shall be derived from the same population and source. Sources of animals to be used to establish cultures include government or private laboratories which are c
47、ulturing Hyalella azteca for sediment toxicity tests or a reputable biological supply company.1 2 3A list of possible sources of Hyalella azteca is provided in Annex C.6.2 Control sedimentEach sediment toxicity test shall include a control with a minimum of five replicate test containers containing
48、control sediment. Responses of organisms exposed to control sediment during a test provide measurements for determining test validity (see 10.3), evidence of the health and normal behaviour of ISO 2013 All rights reserved 3BS ISO 16303:2013ISO 16303:2013(E)the test organisms, and a basis for interpr
49、eting data derived from the test sediments. Control sediment is either natural sediment or artificial (i.e. formulated) sediment.6.2.1 Natural sedimentNatural sediment taken from a fresh water or slightly brackish ( 1 cm) and indigenous organisms by hand sorting using tweezers or similar instruments. Wet sieving the sediment is not recommended because water-soluble contaminants and fine non-settling clay particles could be lost. Each sediment sample is homogenized in the laboratory before use,1 2 1
