1、raising standards worldwideNO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAWBSI Standards PublicationBS ISO 14380:2011W 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 acute toxicity to Thamnocephalus platyurus(Crustacea, Anostraca)BS ISO 14380:2011 BRITISH STANDARDNation
2、al forewordThis British Standard is the UK implementation of ISO 14380:2011. The UK participation in its preparation was entrusted to T e c h n i c a l Committee EH/3/5, Biological Methods.A list of organizations represented on this committee can be obtained on request to its secretary.This publicat
3、ion does not purport to include all the necessary provisions of a contract. Users are responsible for its correct application. BSI 2011 ISBN 978 0 580 69873 6 ICS 13.060.70 Compliance with a British Standard cannot confer immunity from legal obligations.This British Standard was published under the
4、authority of the Standards Policy and Strategy Committee on 31 December 2011.Amendments issued since publicationDate T e x t a f f e c t e dBS ISO 14380:2011Water quality Determination of the acute toxicity to Thamnocephalus platyurus (Crustacea, Anostraca)Qualit de leau Dtermination de la toxicit a
5、igu envers Thamnocephalus platyurus (Crustacea, Anostraca) ISO 2011Reference numberISO 14380:2011(E)First edition2011-11-15ISO14380INTERNATIONAL STANDARDBS ISO 14380:2011ISO 14380:2011(E)COPYRIGHT PROTECTED DOCUMENT ISO 2011All rights reserved. Unless otherwise specified, no part of this publication
6、 may be reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying and microfilm, without permission in writing 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.
7、 + 41 22 749 01 11Fax + 41 22 749 09 47E-mail copyrightiso.orgWeb www.iso.orgPublished in Switzerlandii ISO 2011 All rights reservedBS ISO 14380:2011ISO 14380:2011(E) ISO 2011 All rights reserved iiiContents PageForeword ivIntroduction v1 Scope 12 Normative references .13 Terms and definitions .14 P
8、rinciple .25 Test environment .26 Reagents, test organisms and media 27 Apparatus and materials .38 Treatment and preparation of samples .48.1 Special precautions 48.2 Preparation of the stock solutions of substances to be tested 49 Procedure 49.1 Hatching of the cysts .49.2 Selection of test concen
9、trations .59.3 Preparation of the test and control solutions 59.4 Introduction of the organisms .69.5 Incubation of the test system .69.6 Measurements 710 Estimation of the LC50711 Reference test 812 Validity criteria .813 Test report .8Annex A (informative) Rapid test for determination of sublethal
10、 effects on Thamnocephalus platyurus (1 h exposure) .10Annex B (informative) Culturing of Thamnocephalus platyurus for cyst production 16Annex C (informative) Precision data19Bibliography .20BS ISO 14380:2011ForewordISO (the International Organization for Standardization) is a worldwide federation o
11、f 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 technical committee has been established has the right to be represented on that committee. In
12、ternational 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.International Standards are drafted in accordance with t
13、he 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 the member bodies for voting. Publication as an International Standard requires approval b
14、y 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 identifying any or all such patent rights.ISO 14380 was prepared by Technical Committee ISO/
15、TC 147, Water quality, Subcommittee SC 5, Biological methods.ISO 14380:2011(E)iv ISO 2011 All rights reservedBS ISO 14380:2011IntroductionThe evaluation of harmful effects on water quality has for several years involved the performance of biological tests. Crustaceans are of interest from the ecotox
16、icological point of view because they are primary consumers and a major component of the zooplankton in aquatic ecosystems.The test specified in this International Standard involves determination of the lethal effects on the fresh water fairy shrimp Thamnocephalus platyurus after 24 h exposure to th
17、e toxicant. A rapid test can also be carried out to determine sublethal effects after a very short exposure time (1 h).The beavertail fairy shrimp T. platyurus is to date already used extensively in toxicity testing for several reasons:a) this anostracan crustacean has a sensitivity to chemicals whi
18、ch is quite similar to that of the cladoceran crustacean Daphnia magna (see References 4567);b) the assays are performed with neonates hatched from dormant eggs (cysts), which bypasses the need for culturing or maintaining live stock cultures of test organisms;c) T. platyurus neonates are substantia
19、lly smaller than neonates of Daphnia magna, hence the assays require much smaller test containers, and much less bench space and incubation space;d) T. platyurus is very sensitive to cyanotoxins produced by algal blooms in eutrophicated waters (see References 89).ISO 14380:2011(E) ISO 2011 All right
20、s reserved vBS ISO 14380:2011BS ISO 14380:2011INTERNATIONAL STANDARD ISO 14380:2011(E)Water quality Determination of the acute toxicity to Thamnocephalus platyurus (Crustacea, Anostraca)WARNING Persons using this International Standard should be familiar with normal laboratory practice. This standar
21、d 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 essential that tests conduc
22、ted in accordance with this International Standard be carried out by suitably qualified staff.1 ScopeThis International Standard specifies a method for the determination of the lethal effects of toxicants to Thamnocephalus platyurus test organisms after 24 h exposure. A second method (rapid test) is
23、 described in Annex A for the determination of sublethal effects after a very short exposure time (1 h).The methods are applicable to:a) chemical substances which are soluble or which can be maintained as stable suspensions or dispersions under the conditions of the test;b) industrial or sewage effl
24、uents, treated or untreated, if appropriate after decantation, filtration or centrifugation;c) fresh waters;d) aqueous extracts;e) toxins of blue-green algae.This International Standard is not applicable to the testing of unstable chemicals (hydrolysing, absorbing, etc.) in water unless exposure con
25、centration is measured, nor to the testing of aquatic samples from the estuarine or marine environment.2 Normative referencesThe following referenced documents are indispensable for the application of this document. For dated references, only the edition cited applies. For undated references, the la
26、test edition of the referenced document (including any amendments) applies.ISO 5667-16, Water quality Sampling Part 16: Guidance on biotesting of samplesISO 5814, Water quality Determination of dissolved oxygen Electrochemical probe methodISO 10523, Water quality Determination of pH3 Terms and defin
27、itionsFor the purposes of this document, the following terms and definitions apply. ISO 2011 All rights reserved 1BS ISO 14380:20113.1control batchseries of replicates containing control solutionNOTE The role of a control batch in an experimental procedure is to demonstrate the response to the detec
28、tion system imposed collectively by compounds of the matrix used in the determination, in the absence of the subject of interest.3.2LC50concentration or dilution of the test sample which gives rise to 50 % mortality of the test organisms3.3EC50concentration or dilution of the sample which gives rise
29、 to 50 % effect on the test organisms3.4neonatenewly hatched individual3.5test batchseries of replicates filled with the same test solution4 PrincipleFreshly hatched T. platyurus larvae are exposed to a range of concentrations of the sample under analysis and the percentage mortality of the test org
30、anisms is determined after 24 h exposure, with subsequent calculation of the 24 h LC50.The test is carried out in one or two stages: a “range-finding test” to determine the range of concentrations or dilutions needed for calculation of the 24 h LC50; a “definitive test” conducted when the data of th
31、e range-finding test are not sufficient or adequate for calculation of the 24 h LC50.5 Test environmentThe test shall be carried out in the dark, in a temperature-controlled room or incubator at (25 1) C in the test containers.Maintain the atmosphere free from toxic dusts or vapours. The use of cont
32、rol solutions is a double check that the test is performed in an atmosphere free from toxic dusts and vapours.6 Reagents, test organisms and mediaUse only reagents of recognized analytical grade, unless otherwise specified.6.1 Test organisms. The test organisms are neonates of the beavertail fairy s
33、hrimp T. platyurus, which are hatched from dormant eggs (cysts) of this crustacean.Cysts of T. platyurus are obtained from laboratory cultures of the crustacean as described in Annex B or can be purchased from a specialized company1).1) MicroBioTests Inc., Mariakerke, Belgium, is an example of a sup
34、plier able to provide suitable Thamnocephalus platyurus cysts commercially. This information is given for the convenience of the users of this document and does not constitute an endorsement by ISO of this supplier.ISO 14380:2011(E)2 ISO 2011 All rights reservedBS ISO 14380:20116.2 Pure water, condu
35、ctivity below 10 S/cm.6.3 Test medium, prepared by dissolving the following mineral substances in 1 l of pure water (6.2):NaHCO396 mgCaSO42H2O 60 mgMgSO460 mgKCl 4 mgThis test medium corresponds to a synthetic water of moderate hardness, i.e. containing CaCO3at concentrations of 80 mg/l to 100 mg/l
36、(see Reference 13). Thus prepared, the medium has a pH of 7,6 0,3.When stored in a refrigerator at (4 2) C in the dark, the solution can be used for several months.Aerate the test medium until the dissolved oxygen concentration has reached the air saturation value and until the pH has stabilized. If
37、 necessary, adjust the pH to 7,6 0,3 using sodium hydroxide or hydrochloric acid solutions. The concentration of the acid or base required shall be selected so that the volume to be admixed is as small as possible. Bring the temperature of the test medium up to (25 1) C prior to use.6.4 Hatching med
38、ium. An eightfold dilution of the test medium (6.3) with pure water (6.2).6.5 Reference substance. Potassium dichromate (K2Cr2O7) is the recommended reference chemical.7 Apparatus and materialsUsual laboratory apparatus and glassware and in particular the following.7.1 Temperature-controlled room or
39、 chamber.7.2 Hatching Petri dishes, small Petri dishes, diameter 5 cm, in glass or in inert plastic material.7.3 Test containers. Disposable microplates made from chemically inert material, comprising wells with a capacity 1 ml. For example, 24 (4 6) well microplates with a well diameter of approxim
40、ately 16 mm are suitable.7.4 Pipette for sampling the test organisms, with a sufficient diameter for capturing the animals while allowing sampling of only a small volume of medium.Micropipettes of inert plastic material with a bulb at the end are very suitable for the operations.7.5 Stereomicroscope
41、 with incident (bottom) illumination, with a magnification of at least eight times and, if possible, a continuous magnification.7.6 Light source, providing a range of light intensity in the hatching Petri dish of 3 000 lx to 4 000 lx.7.7 Sample collecting bottles, as specified in ISO 5667-16.ISO 143
42、80:2011(E) ISO 2011 All rights reserved 3BS ISO 14380:20118 Treatment and preparation of samples8.1 Special precautionsSpecial precautions are required for sampling, transportation, storage and treatment of water, effluent, or aqueous extract samples to be tested.Sampling, transportation and storage
43、 of the samples should be performed as specified in ISO 5667-16.Carry out the toxicity test as soon as possible, ideally within 12 h of collection. If this time interval cannot be met, cool the sample to 0 C to 5 C and test the sample within 24 h. If it is not possible to perform the test within 72
44、h, the sample may be frozen and maintained deep-frozen (below 18 C) for testing within 2 months of collection, provided that characteristics are known to be unaffected by freezing. At the time of testing, homogenize the sample to be analysed by shaking manually, and, if necessary, allow to settle fo
45、r 2 h in a container, and sample by drawing off (using a pipette) the required quantity of supernatant, maintaining the end of the pipette in the centre of the section of the test tube and halfway between the surface of the deposited substances and the surface of the liquid.If the raw sample of the
46、decanted supernatant is likely to interfere with the test (due to the presence of residual suspended matter, protozoa, microorganisms, etc.), filter or centrifuge the raw or decanted sample.The sample obtained by either of these methods is the sample submitted to testing.Measure the pH (as specified
47、 in ISO 10523) and the dissolved oxygen concentration (as specified in ISO 5814) and record these values in the test report.If the aim of the test is to assess the acute toxicity without considering the pH effects, the test may also be carried out after adjustment of the pH value to 7,6 0,3 with hyd
48、rochloric acid or sodium hydroxide solutions. Proceed, if appropriate, as indicated above, for the separation of the suspended matter formed following the adjustment of the pH. Mention any pH adjustment in the test report.8.2 Preparation of the stock solutions of substances to be testedPrepare the s
49、tock solution of the substance to be tested by dissolving a known quantity of substance in a specified volume of test medium (6.3) at the time of use. However, if the stock solution of the substance is stable under certain conditions, it may be prepared in advance and stored under these conditions.For substances sparingly soluble in the test medium, refer to the specifications given in ISO 5667-16.9 Procedure9.1 Hatching of the cysts9.1.1 GeneralT. platyurus cysts shall be hatched under the condit
