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1、STDmISO 7899-1-ENGL 1998 m it85L903 083273b 7b7 W INTERNATIONAL STANDARD IS0 7899-1 :1998 TECHNICAL CORRIGENDUM 1 Published 2000-05-01 INTERNATIONAL ORGANIZATION FOR STANDARDIZATION MWYHAPOLIW OPrAHM3ALIMfl no CTA“TM3AUMM ORGANISATION INTERNATIONALE DE NORMALISATION Water quality - Detection and enu
2、meration of intestinal enterococci in surface and waste water - Part 1: Miniaturized method (Most Probable Number) by inoculation in liquid medium TECHNICAL CORRIGENDUM 1 Qualit de leau - Recherche et dnombrement des entrocoques intestinaux dans les eaux de surface et rsiduaires - Partie 1: Mthode m
3、iniaturise (nombre le plus probable) par ensemencement en milieu liquide RECTIFICATIF TECHNIQUE 1 Technical Corrigendum 1 to International Standard IS0 7899-1 :1998 was prepared by Technical Committee ISO/TC 147, Water quality, Subcommittee SC 4, Microbiological methods. Cover, page iii and page 1:
4、In the second element of the title of this part of IS0 7899, delete the words “in surface and waste watet and change the third element to give the following new title: “ Water quality - Detection and enumeration of intestinal enterococci - Part 1: Miniaturized method (Most Probable Number) for surfa
5、ce and waste water“ ICs 07.1 00.20 Ref. No. IS0 7899-1:1998/Cor.l:2000(E) O IS0 2000 - All rights reserved Printed in Switzerland Copyright International Organization for Standardization Provided by IHS under license with ISO Licensee=NASA Technical Standards 1/9972545001 Not for Resale, 04/18/2007
6、21:19:24 MDTNo reproduction or networking permitted without license from IHS -,-,- STD.ISO 7899-1-ENGL 1996 451903 0773490 827 m INTERNATIONAL STANDARD IS0 7899-1 Second edition 1998-1 1 -1 5 Water quality - Detection and enumeration of intestinal enterococci in surface and waste water - Part 1: M i
7、n at u rized method (M Ost Probable N um ber) by inoculation in liquid medium Qualit de leau - Recherche et dnombrement des entrocoques intestinaux dans les eaux de surface et rsiduaires - Partie 1: Mthode miniaturise (nombre le plus probable) par ensemencement en milieu liquide This material is rep
8、roduced from IS0 documents under International Organization for Standardization WO) Copyright License Number IHSllCC11996. Not for resale. No part of these IS0 documents may be reproduced in any form, electronic retrieval system or otherwise, except as allowed in the copyright law of the country of
9、use, or with the prior written consent of IS0 (Case postale 56. 121 1 Geneva 20, Switzerland, Fax +41 22 734 10 79). IHS or the IS0 Licensors members. Reference number IS0 7899-1 :1998( E) Copyright International Organization for Standardization Provided by IHS under license with ISO Licensee=NASA T
10、echnical Standards 1/9972545001 Not for Resale, 04/18/2007 21:19:24 MDTNo reproduction or networking permitted without license from IHS -,-,- . STD-IS0 7899-L-ENGL 1998 D Y85L903 077L491 7b3 IS0 7899-1 :1998(E) Contents 1 Scope 1 2 Normative references 1 3 Definitions 1 4 Principle 2 5 Apparatus . 2
11、 6 Sampling . 2 7 Culture media and diluents . 3 8 Procedure . 4 9 Expression of results 6 1 O Test report 7 11 Performance data . 7 Annex A (informative) Example of software for statistical analysis of MPNs . 8 Annex B (informative) Example of software for computation of MPNs . 11 Annex C (informat
12、ive) Synthetic sea salt 13 Annex D (informative) Performance characteristics of the method . 14 Annex E (normative) Quality criteria for manufacturing of the medium in microtitre plates 15 Annex F (normative) Preparation of calibration microtitre plates 17 Annex G (informative) Bibliography 19 O IS0
13、 1998 or utilized in any form or by any means. electronic or mechanical. including photocopying and microfilm. without permission in writing from the publisher . I All rights reserved . Unless otherwise specified. no patt of this publication may be reproduced International Organization for Standardi
14、zation Case postale 56 CH-121 1 Genve 20 Switzerland Intemet isoiso.ch Printed in Switzerland II Copyright International Organization for Standardization Provided by IHS under license with ISO Licensee=NASA Technical Standards 1/9972545001 Not for Resale, 04/18/2007 21:19:24 MDTNo reproduction or ne
15、tworking permitted without license from IHS -,-,- IS0 STDOIS0 7899-3-ENGL 3998 m 4853903 0771492 6TT IS0 7899-1 : 1 998( E) Foreword IS0 (the International Organization for Standardization) is a worldwide federation of national standards bodies (IS0 member bodies). The work of preparing Internationa
16、l Standards is normally carried out through IS0 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. International organizations, governmental and non-governmental, in liaison with ISO,
17、also take part in the work. IS0 collaborates closely with the International Electrotechnical Commission (IEC) on all matters of electrotechnical standardization. Draft International Standards adopted by the technical committees are circulated to the member bodies for voting. Publication as an Intern
18、ational Standard requires approval by at least 75 o / o of the member bodies casting a vote. International Standard IS0 7899-1 was prepared by Technical Committee ISO/TC 147, Water quality, Subcommittee SC 4, Biological methods. This second edition cancels and replaces the first edition (IS0 7899-1
19、:1984), which has been technically revised. IS0 7899 consists of the following parts, under the general title Water quality- Detection and enumeration of intestinal enterococci in surface and waste water - Part 1: Miniaturized method (Most Probable Number) by inoculation in liquid medium - Part 2: M
20、ethod by membrane filtration Annexes E and F form an integral part of this part of IS0 7899. Annexes A, B, C, D and G are for information only. .O THEN 3360 3340 X4=X 3350 GOTO 3220 3360 X3=X 3370 GOTO 3220 3380 X5=X(2“81) 3390 PRINT“ “ IF S3-S1 =O THEN 31 30 3310 NEXT I 9 Copyright International Or
21、ganization for Standardization Provided by IHS under license with ISO Licensee=NASA Technical Standards 1/9972545001 Not for Resale, 04/18/2007 21:19:24 MDTNo reproduction or networking permitted without license from IHS -,-,- STD-IS0 7899-3-ENGL 3998 4853903 0773503 205 IS0 7899-1 : 1 998( E) 3400
22、341 O 3420 3430 4000 4010 4020 4030 4040 4050 4060 4070 4080 4090 41 O0 41 10 41 20 41 30 41 40 41 50 41 60 41 70 5000 501 O 5020 5030 5040 5050 5060 5070 6000 601 O 6020 6030 6040 6050 6060 6070 6080 6090 61 O0 61 10 61 20 61 30 61 40 61 50 61 60 61 70 61 80 61 90 6200 6210 6220 6230 6240 6250 6260
23、 6270 6280 7000 PRINT “M PN=“;X5 PRINT “FOR A SAMPLE WITH DILUTION FACTOR 1“ PRINT “ AND VOLUME 1“ RETURN REM CALCS AND PRINTS S.E. OF LOG10 (MPN) s2=0 FOR I=1 TO N X3=A(1,5) E2=X3*X IF E2=A(1,3) GOTO 6160 S4=A(I ,3)-A( I ,4) S4=S4*( LOG( S4/A( I ,3)+A( l,5)*X) s3=s3+s4 NEXT I D=2*S3 REM CHI-SQUARED
24、 TEST OF DEVIANCE V=N-1 PRINT “ “ PRINT “DEFIANCE =“;D;“ON“;V;“ D.F.“ PRINT “ “ PRINT “CHI-SQUARED SIGNIFICANCE LEVELS F0R“;V;“ D.F.“ PRINT I 5 PERCENT “;X2( 1 ,V) PRINT I 1 PERCENT “;X2(2,V) PRINT “ .1 PERCENT “ ;X2(3,V) RETURN END 10 Copyright International Organization for Standardization Provide
25、d by IHS under license with ISO Licensee=NASA Technical Standards 1/9972545001 Not for Resale, 04/18/2007 21:19:24 MDTNo reproduction or networking permitted without license from IHS -,-,- 0 IS0 Annex B (i n f o r m at ive) Example of software for computation of MPNs 10 20 30 40 45 50 60 70 80 90 11
26、0 120 130 140 150 155 160 170 180 190 200 280 290 300 31 O 320 330 340 350 360 370 380 390 400 41 O 702 71 O 720 730 740 745 750 760 770 780 790 800 81 O 81 7 DIM T (20) DIM M (20) DIM P (20) L = o CLS : L = L + 1 PRINT “CALCULATION OF MPN No “;L PRINT “ . “ A = 1 PRINT INPUT “NB OF DILUTI0NS“;DI :
27、PRINT : PRINT PRINT P = o : s = : u = o FOR I = 1 TO DI PRINT “DILUTION“ ; I INPUT “NB OF POSITIVE WELLS “ ; P(i) INPUT “NB OF WELLS “ ;T(I) INPUT “WATER VOLUMEWELL (ML) .“ ;M(I) : PRINT P = P(1) + P S U NEXT I K = I NP = (P/U) 2 PM = O FOR I = 1 TO DI PM = PM + (P(1) * M (I) * EXP ( - M(I) * NP) /
28、(1 - EXP (- M(I) * NP ) NEXT I IF PM 88 THEN E(I) = 1.65E38:GOTO 760 E(I) = (EXP (M(i) * NP) - 1) NEXT I LL = o FOR I = 1 TO DI LL NEXT I = (T(I) - P(I)*M(I) + S = (M(1) TI) + U (K + K / 2 + K / 4 + K / 8 + K / 16 + K / 32 + K / 64 + K / 128 + K / 256 + K / 51 2) K (1) = (T(I) (M(1)“2) = LL + (K (I)
29、 / E (I) = (LOG (NP) - (1.96 * (1 / (NP * SQR (LL) = (LOG (NP) + (1.96 * (1 / (NP * SQR (LL) CL CU 11 Copyright International Organization for Standardization Provided by IHS under license with ISO Licensee=NASA Technical Standards 1/9972545001 Not for Resale, 04/18/2007 21:19:24 MDTNo reproduction
30、or networking permitted without license from IHS -,-,- - STD-IS0 7899-L-ENGL 3998 4853903 0773505 088 IS0 7899-1 : 1998( E) IS0 840 850 860 PRINT : PRINT 870 880 890 900 GOTO45 NP = INT (NP 100 + .5) / 100 PRINT : PRINT “ NPP = “;NP;“ / ML“ PRINT “LIMITS INF=“;INT ( EXP (CL)* 100 + .5) / 100;“ / ML
31、SUP=“ ;INT (EXP (CU) * 100 + .5) / 100;“ / ML“ PRINT : PRINT: INPUT “DO YOUWANT ANOTHER MPN (Y/N)?“;RE$ IF RE$ = “N“ THEN END Comment Ater display of the run number (L, line 50), Input the number of dilutions (line 90). For each dilution: - display the dilution rank (line 140), - input the number of
32、 positive wells or tubes (line 150), - input the number of wells inoculated with this dilution (line 155), - input the volume of water inoculated per well, in millilitres (line 160). Calculation of the MPN according to De Man 2 (lines 280-410). Calculation of the lower and upper limits of confidence
33、 interval according to De Man 2 (lines 770-817). Display - of MPN (per mi) (line 850), - the lower limit (per mi) (line 870), - the upper limit (per mi) (line 870), Question: “Another run ?“ If no: END. 12 Copyright International Organization for Standardization Provided by IHS under license with IS
34、O Licensee=NASA Technical Standards 1/9972545001 Not for Resale, 04/18/2007 21:19:24 MDTNo reproduction or networking permitted without license from IHS -,-,- Annex C (informative) Synthetic sea salt C.l Major ion composition of a convenient ocean synthetic sea salt Chloride Sodium Sulfate Magnesium
35、 Calcium Potassium Bicarbonate Borate Strontium Major ion YO Total weight 47,470 26,280 6,602 3,230 1,013 1 ,O1 5 0,491 0,015 0,001 IS0 7899-1 :1998(E) Ionic concentration at 34 g/kg salinity (mdl) 18 740 10 454 2 631 1 256 400 40 1 194 6.0 7,5 Water SOLIDS TOTAL (bo) 86,ll 13,88 34 089,50 TOTAL 99,
36、99 C.2 Example for preparation from defined substances Three basic solutions are to be made as follows: Solution A CaCIp2H20 83,6 g KC I 4335 9 SrC12.6H20 0 7 9 Distilled water to 1 O00 ml Solution B NaHC03 15,15 g Na2B407 3,O g Distilled water to 1 O00 ml Solution C MgS04.7H20 190,o g MgC12.6H20 14
37、7,O g Distilled water to 1 O00 ml The diluent is made by adding to 960 ml distilled water 10 ml of solution A, 10 ml of solution B, 20 ml of solution C, and then 14,9 g sodium chloride, mixing until completely dissolved and setting the pH to 7,5 It 0,2. The diluent is distributed to containers of de
38、sired volumes, and sterilized by autoclaving at (121 It 3) “C for 15 min. 13 Copyright International Organization for Standardization Provided by IHS under license with ISO Licensee=NASA Technical Standards 1/9972545001 Not for Resale, 04/18/2007 21:19:24 MDTNo reproduction or networking permitted w
39、ithout license from IHS -,-,- STD=ISO 7899-3-ENGL 3998 H 4853903 0773507 950 IS0 7899-1 1998( E) IS0 Annex D (i n f o r m at i ve) Performance characteristics of the method The performances of repeatability (r) and reproducibility (R) calculated according to IS0 5725-2 as part of interlaboratory tes
40、ts have shown: On bathing waters (spiked samples, with 100 French laboratories, on three occasions in 1995 and 1996) (one sea water and two fresh waters, without significant difference): i) At the level 1 O0 intestinal enterococci / 100 ml i) At the level 400 intestinal enterococci / 1 O0 ml r 2- 3,
41、6 R -J 5,2 r = 2,l R 2- 3,7 and, for information only, during tests (in 1993 and 1994) limited to nine laboratories and four samples (naturally contaminated): b) On river waters containing between 2,2 x 1OJand 1,5 x 104 intestinal enterococci / 100 ml: r = 1,5 R = 2,7 c) On sewage waters containing
42、between 1,9 x 104 and 5,l x 106 intestinal enterococci / 100 ml: f = 2,6 R 2- 3,9 14 Copyright International Organization for Standardization Provided by IHS under license with ISO Licensee=NASA Technical Standards 1/9972545001 Not for Resale, 04/18/2007 21:19:24 MDTNo reproduction or networking per
43、mitted without license from IHS -,-,- 0 IS0 STDmISO 7899-3-ENGL 3998 4853903 0773508 897 IS0 7899-1 :1998(E) Annex E (normative) Quality criteria for manufacturing of the medium in microtitre plates E.l General For each of the criteria which follow, a quality control shall be made on each batch of m
44、anufactured microtitre plates. The microtitre plates to be tested are taken at random or in a systematic way to constitute a sample in accordance with IS0 3951, respecting the general control level No. II of the normal control. The threshold of positivity of a microtitre plate is defined as being th
45、e fluorescence level leading to a positive reading without ambiguity, to the eye, under a Wood light (366 nm). It shall be measured in application of the protocol of annex F. The quality criteria to be respected are given in E.2 to ES. The batch is rejected if any of the criteria is not respected. E
46、.2 Background noise Absence of positive well in each microtitre plate of the sample, after inoculation with sterile special diluent and incubation of 48 h at (44 I 0,5) O C . The medium background noise of the sample shall be inferior to 25 YO of the positivity threshold defined above and the variat
47、ion coefficient shall be inferior to 10 %. E . 3 Average level of fluorescence This is the geometric mean of the fluorescence signal obtained from the 96 wells of a microtitre plate inoculated uniformly with 200 pl per well of a suspension of E. faecalis CCM 2541 containing 500 microorganisms per ml
48、 of Special Diluent (7.2.1), and incubated for 48 h at (44 I 0,5) “C. The average signal so obtained shall be at least twice the threshold of positivity and the variation coefficient shall be inferior to 10 %. E.4 Fertility Fertility is calculated as the ratio of the number of germs observed with the batch of microtitre plates under test to the number of microorganisms expected with a stable reference material (target value). The