1、raising standards worldwideNO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAWBSI Standards PublicationBS ISO 10710:2010Water quality Growth inhibition test with the marine and brackish water macroalga Ceramium tenuicorneBS EN ISO 10710:2013BS EN ISO 10710:2013 BRITISH STANDARDNat
2、ional forewordThis British Standard is the UK implementation of EN ISO 10710:2013. It is identical to ISO 10710:2010. It supersedes BS ISO 10710:2010, which is withdrawn.The UK participation in its preparation was entrusted to Technical Committee EH/3, Water quality.A list of organizations represent
3、ed 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 Limited 2013ISBN 978 0 580 7
4、9934 1ICS 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 July 2010Amendments/corrigenda issued since publicationDate Text affected31 May 2013 This c
5、orrigendum renumbers BS ISO 10710:2010 as BS EN ISO 10710:2013EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN ISO 10710 April 2013 ICS 13.060.70 English Version Water quality - Growth inhibition test with the marine and brackish water macroalga Ceramium tenuicorne (ISO 10710:2010) Qualit de lea
6、u - Essai dinhibition de croissance sur la macro algue deaux marine et saumtre Ceramium tenuicorne (ISO 10710:2010) Wasserbeschaffenheit - Bestimmung der Wachstumshemmung auf die marine und stuarine Makroalge Ceramium tenuicorne (ISO 10710:2010) This European Standard was approved by CEN on 14 March
7、 2013. 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 standard without any alteration. Up-to-date lists and bibliographical references concerning such national standards may be obtain
8、ed 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 in any other language made by translation under the responsibility of a CEN member into its own language and notified to the CEN-
9、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 Republic, Denmark, Estonia, Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland,
10、Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, 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 Marni
11、x 17, B-1000 Brussels 2013 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN ISO 10710:2013: EBS EN ISO 10710:2013EN ISO 10710:2013 (E)EN ISO 10710:2013 (E) 3 Foreword The text of ISO 10710:2010 has been prepared by Technical Committ
12、ee ISO/TC 147 “Water quality” of the International Organization for Standardization (ISO) and has been taken over as EN ISO 10710:2013 by Technical Committee CEN/TC 230 “Water analysis” the secretariat of which is held by DIN. This European Standard shall be given the status of a national standard,
13、either by publication of an identical text or by endorsement, at the latest by October 2013, and conflicting national standards shall be withdrawn at the latest by October 2013. Attention is drawn to the possibility that some of the elements of this document may be the subject of patent rights. CEN
14、and/or CENELEC shall not be held responsible for identifying any or all such patent rights. According to the CEN-CENELEC Internal Regulations, the national standards organizations of the following countries are bound to implement this European Standard: Austria, Belgium, Bulgaria, Croatia, Cyprus, C
15、zech Republic, Denmark, Estonia, Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United King
16、dom. Endorsement notice The text of ISO 10710:2010 has been approved by CEN as EN ISO 10710:2013 without any modification. BS ISO 10710:2010ISO 10710:2010(E) ISO 2010 All rights reserved iiiContents Page Foreword iv Introduction.v 1 Scope1 2 Terms and definitions .1 3 Principle2 4 Test organisms, nu
17、trients, media, and materials.2 5 Cultivation 6 6 Apparatus.6 7 Procedure.7 7.1 Algae material 7 7.2 Preparation of dilution water8 7.3 Choice of test concentrations8 7.4 Start of test.9 7.5 Incubation.9 7.6 Measurement .9 7.7 Test with reference substances.9 8 Validity criteria.9 9 Evaluation of re
18、sults .10 9.1 Calculations .10 9.2 Determination of ErCx.10 9.3 Expression of results10 9.4 Interpretation of results 11 10 Reproducibility 11 11 Test report11 Annex A (informative) Growth pattern of Ceramium tenuicorne in 7S and 20S13 Annex B (informative) Results of ring test with Ceramium tenuico
19、rne at 20 S.16 Bibliography17 BS ISO 10710:2010ISO 10710:2010(E) ISO 2010 All rights reserved iiiContents Page Foreword ivIntroduction.v1 Scope2 Terms and definitions .13 Principle4 Test organisms, nutrients, media, and materials.25 Cultivation 6 Apparatus 67 Procedure.7 7.1 Algae material 7 7.2 Pre
20、paration of dilution water8 7.3 Choice of test concentrations8 7.4 Start of test.9 7.5 Incubation.9 7.6 Measurement .9 7.7 Test with reference substances.8 Validity criteria.99 Evaluation of results .10 9.1 Calculations .10 9.2 Determination of ErCx.10 9.3 Expression of results10 9.4 Inte pretation
21、of results 1110 Reproducibility 1111 Test report11A Growth pattern of Ceramium tenuicorne in 7S and 20S13Annex B (informative) Results of ring test with Ceramium tenuicorne at 20 S.16Bibliography17 BS EN ISO 10710:2013ISO 10710:2010(E)BS ISO 10710:2010ISO 10710:2010(E) ISO 2010 All rights reserved v
22、Introduction The red macroalga Ceramium tenuicorne belongs to Ceramiaceae, Rhodophyta. The species can be used as a model organism for the near coastal ecosystem. This species is found in temperate marine waters in both the northern and southern hemispheres and is thus relevant for large areas. As p
23、rimary producers, they are a food source for many invertebrates and serve as living habitat for bacteria, invertebrates, and juvenile fish. They also serve as substrate for many oviparous fish species. ivBS EN ISO 10710:2013ISO 10710:2010(E)BS ISO 10710:2010INTERNATIONAL STANDARD ISO 10710:2010(E) I
24、SO 2010 All rights reserved 1Water quality Growth inhibition test with the marine and brackish water macroalga Ceramium tenuicorne WARNING Persons using this International Standard should be familiar with normal laboratory practice. This International Standard does not purport to address all of the
25、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 conducted according to this International Stan
26、dard be carried out by suitably trained staff. 1 Scope This International Standard specifies a method for the determination of the inhibition of growth of the macroalga Ceramium tenuicorne by substances and mixtures contained in seawater or by waste water with salinities between 4S and 32S. This met
27、hod is applicable to substances that are easily soluble in water. NOTE With modifications as described in ISO 144424and ISO 5667-162, the inhibitory effects of poorly soluble organic and inorganic materials, volatile compounds, metals, waste water, marine water samples, and elutriates of sediments c
28、an be tested. 2 Terms and definitions For the purposes of this document, the following terms and definitions apply. 2.1 algal length length from the first division to the most distant tip of the plant NOTE The algal length is expressed in millimetres. See Figure 1. 2.2 control medium combination of
29、dilution water and/or nutrient medium used in the test ISO 20079:20055, 3.6 2.3 control batch control medium including organisms used for testing ISO 20079:20055, 3.5 2.4 effective concentration ErCxconcentration of test sample at which an effect of x % is measured, if compared to the control ISO 20
30、079:20055, 3.9 BS EN ISO 10710:2013BS ISO 10710:2010ISO 10710:2010(E) 2 ISO 2010 All rights reserved2.5 growth medium mixture of natural seawater and nutrients in which algal plants are cultivated, which is used for pre-cultures NOTE Adapted from ISO 8692:20043, 3.3. 2.6 growth rate water quality pr
31、oportional rate of increase in algal length per day NOTE See Clause 9. 2.7 salinity practical salinity S seawater ratio K15of the electrical conductivity of the seawater sample, at the temperature of 15 C and a pressure of one standard atmosphere, to that of a potassium chloride solutions in which t
32、he mass fraction of KCl is 32,435 6 103, at the same temperature and pressure NOTE Adapted from Reference 14, p. 12. 2.8 test medium mixture of seawater, nutrients and test sample NOTE Adapted from ISO 8692:20043, 3.5. 2.9 test sample water quality aqueous sample, e.g. chemical substance, mixture of
33、 chemicals or waste water, for which the inhibitory effects on the growth of algae are determined NOTE Adapted from ISO 8692:20043, 3.4. 3 Principle Algal tips from monocultures of ceramium female gametophytes are grown in defined test conditions and in a defined medium containing a range of concent
34、rations of the test sample. The test solutions are incubated for a period of 7 d after which the increase in length is measured and the growth rate is calculated. The growth inhibition is determined as a reduction in growth rate, relative to control cultures grown under identical conditions. When to
35、xicity of samples is to be compared to the toxicity of other chemicals or waste waters, tests can be performed in artificial seawater. If the purpose of the testing is to assess and to predict effects in a specific receiving water body, the tests can be conducted with algae adapted to the salinity i
36、n the receiving water body. In this case, natural seawater from an uncontaminated site of the same properties is used. 4 Test organisms, nutrients, media, and materials Unless otherwise stated, use only reagents of recognized analytical grade and distilled or demineralized water or water of equivale
37、nt purity. BS EN ISO 10710:2013ISO 10710:2010(E)BS ISO 10710:2010ISO 10710:2010(E) ISO 2010 All rights reserved 34.1 Test organisms Use either of the following marine algal clones: a) Ceramium tenuicorne Ktzing Waern (7S clone originating from the Baltic Sea); b) C. tenuicorne Ktzing Waern (20S and
38、30S clone originating from the Oslo fjord). This alga is a widely distributed macroalgae species (phylum Rhodophyta) in estuarine and coastal areas. The strains recommended are available in unialgal, non-axenic cultures1). NOTE 1 This growth inhibition test is based on two clones, which were formerl
39、y regarded as two different species. These species were the marine Ceramium strictum Harvey sensu Kylin and the brackish water species C. tenuicorne Ktzing Waern. Complete interfertility (References 1112) and DNA data (Reference 10) have shown that the two entities belong to the same species, with C
40、. tenuicorne as the valid name. The marine clone (former C. strictum) used in this test was isolated in 1973 and originates from the Oslo fjord (20S to 25S). It has been maintained as a laboratory culture for over 30 years. The brackish water clone was isolated in 1995 and originates from the Baltic
41、 Sea, 20 km south of the Ask laboratory in northern Baltic proper (6S to 7S). Cultures can be maintained in the medium specified in Clause 5. Regular subculturing is necessary. NOTE 2 Among the red algae, changes occur between haploid and diploid generations. In the growth inhibition test, the femal
42、e gametophytic generation is used since it has an even dichotomous growth pattern and the fastest growth rate. In nature it is difficult to distinguish between male and female plants. This can be done in the laboratory where spermatangia are found on the branches of the male plants and trichogynes c
43、an be seen on the tips of the claws on the female plants. NOTE 3 The Baltic Sea clone can be adapted and used in tests in salinities between 4S and 12S. The marine clone can be used as test organism in salinities between 12S and 32S. 4.2 Natural and artificial seawater 4.2.1 General Natural seawater
44、 is used for the cultivation of the algae and either natural or artificial seawater should be used for testing. The type of seawater to be used depends on the objective of the test. When natural seawater is used, care shall be taken to ensure that it is not polluted. Take special care to avoid conta
45、mination of the water by inorganic or organic substances during preparation and storage. Equipment made of copper shall not be used. 4.2.2 Artificial seawater Prepare the stock solutions for artificial seawater according to Table 1. Start with about one third of the desired volume of water, add the
46、weighed quantities of chemicals in accordance with Table 1 and make up to volume with water. This stock solution with a salinity of 100S (equivalent to 10 % mass per volume) has a durability of at least six months, if stored in darkness at room temperature. Before use, the stock solution should be d
47、iluted with water to the desired salinity. Adjust the pH to 8,0 0,2 with 1 mol/l HCl or 1 mol/l NaOH. The artificial seawater shall be sterilized by autoclaving or sterile filtration (pore size, 0,2 m) before use. Re-check the pH after sterilization, and adjust if necessary to 8,0 0,2 with 1 mol/l H
48、Cl or 1 mol/l NaOH, before use. 1) Suitable suppliers are: a) ITM, Department of Applied Environmental Research of Science, Stockholm University, S-106 91 Stockholm, Sweden; b) University of Oslo, Department of Biology, P.O. Box 1066 Blindern, N-0316 Oslo, Norway. This information is given for the c
49、onvenience of users of this document and does not constitute an endorsement by ISO of these suppliers. BS EN ISO 10710:2013ISO 10710:2010(E)BS ISO 10710:2010ISO 10710:2010(E) 4 ISO 2010 All rights reservedTable 1 Artificial seawater with a salinity of 10 % mass per volume or 100S (adapted from Reference 13) Quantity per 1 l medium Quantity per 5 l medium Quantity per 10 l medium Substance g/l g/5 l g/10 l NaCl 70,1 351 702 Na2SO411,7 58,7 117 KCl 2,03 10,2 20,3 KBr 0,293 1,47 2,93 Na2B4O
