1、BRITISH STANDARDBS EN ISO 9509:2006Water quality Toxicity test for assessing the inhibition of nitrification of activated sludge microorganisms The European Standard EN ISO 9509:2006 has the status of a British StandardICS 13.060.70g49g50g3g38g50g51g60g44g49g42g3g58g44g55g43g50g56g55g3g37g54g44g3g51
2、g40g53g48g44g54g54g44g50g49g3g40g59g38g40g51g55g3g36g54g3g51g40g53g48g44g55g55g40g39g3g37g60g3g38g50g51g60g53g44g42g43g55g3g47g36g58BS EN ISO 9509:2006This British Standard was published under the authority of the Standards Policy and Strategy Committee on 31 July 2006 BSI 2006ISBN 0 580 48741 5Nati
3、onal forewordThis British Standard is the official English language version of EN ISO 9509:2006. It is identical with ISO 9509:2006. It supersedes BS EN ISO 9509:1995 which is withdrawn.The UK participation in its preparation was entrusted by Technical Committee EH/3, Water quality, to Subcommittee
4、EH/3/5, Biological methods, which has the responsibility to: A list of organizations represented on this subcommittee can be obtained on request to its secretary.Cross-referencesThe British Standards which implement international or European publications referred to in this document may be found in
5、the BSI Catalogue under the section entitled “International Standards Correspondence Index”, or by using the “Search” facility of the BSI Electronic Catalogue or of British Standards Online.This publication does not purport to include all the necessary provisions of a contract. Users are responsible
6、 for its correct application. Compliance with a British Standard does not of itself confer immunity from legal obligations. aid enquirers to understand the text; present to the responsible international/European committee any enquiries on the interpretation, or proposals for change, and keep UK inte
7、rests informed; monitor related international and European developments and promulgate them in the UK.Summary of pagesThis document comprises a front cover, an inside front cover, the EN ISO title page, the EN ISO foreword page, the ISO title page, pages ii to v, a blank page, pages 1 to 12, an insi
8、de back cover and a back cover.The BSI copyright notice displayed in this document indicates when the document was last issued.Amendments issued since publicationAmd. No. Date CommentsEUROPEAN STANDARDNORME EUROPENNEEUROPISCHE NORMEN ISO 9509July 2006ICS 13.060.70 Supersedes EN ISO 9509:1995 English
9、 VersionWater quality - Toxicity test for assessing the inhibition ofnitrification of activated sludge microorganisms (ISO 9509:2006)Qualit de leau - Essai de toxicit pour lvaluation delinhibition de la nitrification des micro-organismes desboues actives (ISO 9509:2006)Wasserbeschaffenheit - Toxizit
10、tstest zur Bestimmung derNitrifikationshemmung in Belebtschlamm (ISO 9509:2006)This European Standard was approved by CEN on 30 June 2006.CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this EuropeanStandard the status of a national
11、 standard without any alteration. Up-to-date lists and bibliographical references concerning such nationalstandards may be obtained on application to the Central Secretariat or to any CEN member.This European Standard exists in three official versions (English, French, German). A version in any othe
12、r language made by translationunder the responsibility of a CEN member into its own language and notified to the Central Secretariat has the same status as the officialversions.CEN members are the national standards bodies of Austria, Belgium, Cyprus, Czech Republic, Denmark, Estonia, Finland, Franc
13、e,Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania,Slovakia, Slovenia, Spain, Sweden, Switzerland and United Kingdom.EUROPEAN COMMITTEE FOR STANDARDIZATIONCOMIT EUROPEN DE NORMALISATIONEUROPISCHES KOMITEE FR NORMU
14、NGManagement Centre: rue de Stassart, 36 B-1050 Brussels 2006 CEN All rights of exploitation in any form and by any means reservedworldwide for CEN national Members.Ref. No. EN ISO 9509:2006: EForeword This document (EN ISO 9509:2006) has been prepared by Technical Committee ISO/TC 147 “Water qualit
15、y“ in collaboration with 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, either by publication of an identical text or by endorsement, at the latest by January 2007, and conflicting nat
16、ional standards shall be withdrawn at the latest by January 2007. This document supersedes EN ISO 9509:1995. According to the CEN/CENELEC Internal Regulations, the national standards organizations of the following countries are bound to implement this European Standard: Austria, Belgium, Cyprus, Cze
17、ch Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and United Kingdom. Endorsement notice The text of ISO 9509:2006 has
18、 been approved by CEN as EN ISO 9509:2006 without any modifications. EN ISO 9509:2006Reference numberISO 9509:2006(E)INTERNATIONAL STANDARD ISO9509Second edition2006-07-01Water quality Toxicity test for assessing the inhibition of nitrification of activated sludge microorganisms Qualit de leau Essai
19、 de toxicit pour lvaluation de linhibition de la nitrification des micro-organismes des boues actives EN ISO 9509:2006ii iiiContents Page Foreword iv Introduction v 1 Scope . 1 2 Normative references . 1 3 Terms and definitions. 2 4 Principle. 2 5 Reagents and materials . 3 6 Apparatus 4 7 Procedure
20、 4 8 Calculation and expression of results 5 9 Validity of results 6 10 Precision 7 11 Test report . 7 Annex A (normative) Determination of the nitrifying activity of an activated sludge. 8 Annex B (normative) Example for preparation of the test . 9 Annex C (informative) Apparatus for culturing nitr
21、ifying activated sludge . 10 Bibliography . 12 EN ISO 9509:2006iv Foreword ISO (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
22、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. International organizations, governmental and non-governmental, in liaison with ISO, also take part in the work. ISO collaborates closely with
23、the International Electrotechnical Commission (IEC) on all matters of electrotechnical standardization. International Standards are drafted in accordance with the rules given in the ISO/IEC Directives, Part 2. The main task of technical committees is to prepare International Standards. Draft Interna
24、tional Standards adopted by the technical committees are circulated to the member bodies for voting. Publication as an International Standard requires approval by 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 b
25、e the subject of patent rights. ISO shall not be held responsible for identifying any or all such patent rights. ISO 9509 was prepared by Technical Committee ISO/TC 147, Water quality, Subcommittee SC 5, Biological methods. This second edition cancels and replaces the first edition (ISO 9509:1989),
26、which has been technically revised. EN ISO 9509:2006vIntroduction Nitrification is an important process in the treatment of waste waters, since it is necessary to reduce the polluting effects of ammonium in treated discharges to receiving waters. It is further necessary to convert ammonium to nitrat
27、e in order to allow the subsequent process of denitrification (producing nitrogen gas) in the anoxic stage of the modified activated sludge process, thus considerably reducing the potential for eutrophication in the receiving waters. The nitrification process is generally performed by two separate g
28、roups of autotrophic bacterial species This International Standard describes a method for assessing the inhibition of the production of oxidized nitrogen (nitrite plus nitrate), or of the removal of ammonium, by nitrifying activated sludge. EN ISO 9509:2006blank1Water quality Toxicity test for asses
29、sing the inhibition of nitrification of activated sludge microorganisms WARNING Sewage and activated sludge contain potentially pathogenic organisms. Appropriate precautions are necessary when handling them. Toxic test substances and those with unknown properties are to be handled with care. Persons
30、 using this International Standard should be familiar with normal laboratory practice. This 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 compli
31、ance with any national regulatory conditions. IMPORTANT It is absolutely essential that tests conducted according to this standard be carried out by suitably trained staff. 1 Scope This International Standard specifies a method for assessing the short-term inhibitory effect of waters, waste waters o
32、r test substances on nitrifying bacteria in activated sludge. The inhibitory effect is estimated over an exposure period of usually 3 h or up to 24 h with weakly nitrifying sludge. The method is applicable to nitrifying activated sludge derived from domestic and synthetic sewage and also to sludges
33、from industrial and mixed domestic and industrial waste waters. The nitrifying activity of the sludge is verified by testing in the presence and absence of a specific inhibitor (e.g. N-allylthiourea; see Annex A). If the nitrification rate is within a suitable range for the test, i.e. 2 mg of nitrog
34、en per gram of suspended solid and hour to 6,5 mg of nitrogen per gram of suspended solids and hour, the sludge may be used directly. If not, adjustments are necessary (see Clause 9). The method is applicable to water-soluble, non-volatile chemicals, and to waste waters Sludges from different source
35、s respond differently to a given concentration of an inhibitor mainly due to reaction between the inhibitor and components of the sludge. This results in a partial neutralisation of the toxic effect. Also, since the test lasts only hours, any inhibitory effects may diminish or increase over a longer
36、 period, e.g. in the continuous activated sludge system (see ISO 5667-16). 2 Normative references The following referenced documents are indispensable for the application of this document. For dated references, only the edition cited applies. For undated references, the latest edition of the referen
37、ced document (including any amendments) applies. ISO 5667-16, Water quality Guidance on biotesting of samples ISO 6777, Water quality Determination of nitrite Molecular absorption spectrometric method ISO 7150-1, Water quality Determination of ammonium Part 1: Manual spectrometric method EN ISO 9509
38、:20062 ISO 11733, Water quality Determination of the elimination and biodegradability of organic compounds in an aqueous medium Activated sludge simulation test 3 Terms and definitions For the purposes of this document, the following terms and definitions apply. 3.1 nitrification oxidation of ammoni
39、um compounds by bacteria NOTE Usually the intermediate product is nitrite and the end product nitrate ISO 6107-1:2004, 49 3.2 test material pure chemicals, clearly defined mixtures of chemicals, chemical products, waste waters and treated waste waters 3.3 activated sludge accumulated biological mass
40、 (floc) produced in the treatment of waste water by the growth of bacteria and other microorganisms in the presence of dissolved oxygen ISO 6107-1:2004, 2 3.4 concentration of suspended solids of an activated sludge amount of solids obtained by filtration or centrifugation of a known volume of activ
41、ated sludge and drying at about 105 C to constant mass ISO 9888:1999, 3.4 3.5 toxic range range of concentration of a test material over which 0 % to 100 % inhibition occurs ISO 8192:1) 3.6 EC50effective concentration of the test material giving a calculated or interpolated inhibition of nitrificati
42、on of 50 %, compared with a blank control 4 Principle The percentage inhibition of nitrification by various concentrations of the test material is calculated by assessing the difference in concentration of oxidized nitrogen (nitrite plus nitrate) produced, or of ammonium utilized, under standard con
43、ditions by the oxidation of ammonium salts after the parallel aeration of a nitrifying sludge in the presence and absence of test material. 1) To be published. (Revision of ISO 8192:1986) EN ISO 9509:200635 Reagents and materials 5.1 Deionized water, for the preparation of defined stock solutions. F
44、or washing procedures, tap water is suitable as well. Make sure that the water is free from chemicals which may inhibit nitrification processes (e.g. Cu2+ions). 5.2 Nitrifying activated sludge. Collect a sufficient volume of a nitrifying activated sludge from a local waste water treatment plant, or
45、from a laboratory-grown sludge (see Annex C), in which nitrification is known to be occurring. According to the purpose of the test, sludge may be collected from plants treating predominantly domestic sewage, mixed domestic industrial waste water or solely industrial waste water; the source of the s
46、ludge and the treated waste water should be reported since the results of the test often depend of the origin of the sludge used (see Reference 6). Maintain the sludge in an aerobic condition. Since the toxicity to nitrification may change with time of storage (see Reference 1), assessments should b
47、e made as soon as possible after collection and preferably within 24 h (see ISO 5667-16). Instead of using activated sludge from a waste water plant, nitrifying sludge can be grown in the laboratory (see Annex C). Although the sludge may be used as collected, it is preferable to wash the sludge to r
48、emove any inhibitors and nitrate present, before re-suspending in chlorine-free, nitrate-free tap water. This washing procedure may be carried out by centrifuging or settling and is optional. Centrifuge (e.g. 10 000 min1for 5 min) or settle the sludge and discard the supernatant liquid. Wash the res
49、idue with a volume of tap water equal to the original volume, re-centrifuge or settle and again discard the supernatant liquid. Finally, re-suspend the centrifuged or settled sludge in an appropriate volume of tap water to give the required concentration of mixed liquor suspended solids (e.g. 3 g/l) and aerate until use. 5.3 Full medium. Dissolve 5,04 g of sodium hydrogen carbonate, NaHCO3, and 2,65 g of ammonium sulfate, (NH4)2SO4, in 1 l of water (5.1). NOTE This medium, when diluted 1:10 (1 + 9) with water (5.1), contain
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