BS-6068-PART-4-SECTION-4.3-1989.pdf
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1、BRITISH STANDARD BS 6068-4.3: 1989 Water quality Part 4: Microbiological methods Section 4.3 Detection and enumeration (most probable number) of faecal streptococci by the multiple tube technique UDC 556.11:628.1.03:628.191:614.777:579.68.083.1:579.862.1.087.23:543.39.062 Licensed Copy: sheffieldun
2、sheffieldun, na, Tue Dec 05 01:26:50 GMT+00:00 2006, Uncontrolled Copy, (c) BSI BS 6068-4.3:1989 This British Standard, having been prepared under the direction of the Environment and Pollution Standards Policy Committee, was published under the authority of the Board of BSI and comes into effect on
3、 29 September 1989 BSI 07-1999 The following BSI references relate to the work on this standard: Committee reference EPC/44 Draft for comment 83/54980 DC ISBN 0 580 17295 3 Foreword This Section of BS 6068, which has been prepared under the direction of the Environment and Pollution Standards Policy
4、 Committee, is related to ISO 7899-1:1984 “Water quality Detection and enumeration of faecal streptococci Part 1: Method by enrichment in a liquid medium”. ISO 7899-1 was prepared by subcommittee 4, Microbiological methods, of Technical Committee 147, Water quality, of the International Organization
5、 for Standardization (ISO) as a result of discussion in which the UK participated. This Section of BS 6068 differs from ISO 7899-1:1984 in that the catalase test has been removed, the incubation conditions in the isolation stage have been changed, and the scope has been expanded. This British Standa
6、rd is being published in a series of Parts subdivided into Sections that will generally correspond to particular international standards. Sections are being, or will be, published in Parts 1 to 7 which, together with Part 0, are listed below. Part 0: Introduction; Part 1: Glossary; Part 2: Physical,
7、 chemical and biochemical methods; Part 3: Radiological methods; Part 4: Microbiological methods; Part 5: Biological methods; Part 6: Sampling; Part 7: Precision and accuracy. NOTEThe tests described in this Section of BS 6068 should only be carried out in laboratories with suitable facilities and b
8、y suitably qualified persons with an appropriate level of microbiological expertise. Standard microbiological procedures should be followed throughout. A British Standard does not purport to include all the necessary provisions of a contract. Users of British Standards are responsible for their corr
9、ect application. Compliance with a British Standard does not of itself confer immunity from legal obligations. Summary of pages This document comprises a front cover, an inside front cover, pages i and ii, pages 1 to 4, an inside back cover and a back cover. This standard has been updated (see copyr
10、ight date) and may have had amendments incorporated. This will be indicated in the amendment table on the inside front cover. Amendments issued since publication Amd. No.Date of issueComments Licensed Copy: sheffieldun sheffieldun, na, Tue Dec 05 01:26:50 GMT+00:00 2006, Uncontrolled Copy, (c) BSI B
11、S 6068-4.3:1989 BSI 07-1999i Contents Page ForewordInside front cover 0Introduction1 1Scope1 2Definitions1 3Principle1 4Diluents, culture media and reagents1 5Apparatus2 6Sampling2 7Procedure2 8Expression of results3 9Test report3 Appendix A Further microbiological information relevant to water exam
12、ination for faecal streptococci4 Publications referred toInside back cover Licensed Copy: sheffieldun sheffieldun, na, Tue Dec 05 01:26:50 GMT+00:00 2006, Uncontrolled Copy, (c) BSI ii blank Licensed Copy: sheffieldun sheffieldun, na, Tue Dec 05 01:26:50 GMT+00:00 2006, Uncontrolled Copy, (c) BSI BS
13、 6068-4.3:1989 BSI 07-19991 0 Introduction The presence and extent of faecal pollution is an important factor in assessing the quality of a body of water. Examination of samples for the presence of faecal streptococci1), which normally inhabit the intestines of man and other warm-blooded animals, ca
14、n provide an indication of such pollution. Opinions vary in different countries as to which streptococcal species should be included in the term “faecal streptococci”, however in this Section of BS 6068 they are restricted to those possessing Lancefields Group D antigen. Although serological confirm
15、ation is not included in the method, the selectivity of the media used is such as to give a reasonable estimate of the numbers of Group D streptococci present. 1 Scope This Section of BS 6068 describes a method for the detection and enumeration of faecal streptococci in water samples by culture in a
16、 liquid medium in multiple tubes and the calculation of their most probable numbers in the sample. The method can be applied to all types of water, including turbid water. It is not suitable for the analysis of large volumes of water containing few faecal streptococci (see BS 6068-4.4). NOTEThe titl
17、es of the publications referred to in this standard are listed on the inside back cover. 2 Definitions For the purposes of this Section of BS 6068, the following definitions apply. 2.1 presumptive faecal streptococci organisms capable of aerobic growth at 37 0.5 C in a liquid culture medium containi
18、ng glucose and sodium azide with the production of acid within 48 h 2.2 faecal streptococci presumptive faecal streptococci as defined in 2.1 which can also grow at 44 0.25 C in the presence of bile salts and sodium azide, and hydrolyse aesculin 3 Principle Test portions of the sample, and/or diluti
19、ons of it, are inoculated into a series of containers of a selective liquid culture medium containing glucose and sodium azide. The containers are incubated for 24 h and 48 h at 37 C and examined. Confirmatory tests are then carried out on each culture showing acid production. The cultures are subcu
20、ltured to a confirmation medium and incubated at 44 C for up to 6 h to demonstrate growth in the presence of bile salts and sodium azide, and the hydrolysis of aesculin. Statistical tables are used to calculate the most probable number of presumptive faecal streptococci and confirmed faecal streptoc
21、occi likely to be present in 100 mL of the sample from the numbers of containers giving positive confirmatory results. 4 Diluents, culture media and reagents WARNING. All selective media described in this Section of BS 6068 contain sodium azide. As this substance is highly toxic and mutagenic, preca
22、utions should be taken to avoid contact with it, especially by the inhalation of fine dust during the preparation of commercially available dehydrated complete media. Azide-containing media should not be mixed with strong inorganic acids, as toxic hydrogen azide (HN3) may be produced. Solutions cont
23、aining azide can also form explosive compounds when in contact with metal pipework and sinks. Azides can be decomposed safely by an excess of a nitrite solution. 4.1 Basic material For the preparation of culture media and diluents use chemicals of a recognized analytical grade and ingredients of uni
24、form quality. For the preparation of culture media use glass distilled or deionized water, free from substances which may inhibit bacterial growth, or water of equivalent purity that complies with grade 3 of BS 3978. Alternatively use commercially available dehydrated culture media. Prepare such med
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