ISO/TS 19036:2006/Amd.1:2009(E) | i Microbiology of food and animal feeding stuffs — Guidelines for the estimation of measurement uncertainty for quantitative determinations \ AMENDMENT 1: Measurement uncertainty for low counts Pago 1, Clause 1, paragraphs 3 and 4 Delete paragraphs 3 and 4 and insert, “This Technical Speciation is not applicable to enumeration using a most probable number technique. \. I. this Technical Specfeation, MU is estimated using a simplifed approach taking inte accourt the Poisson istabution and is then applicable to any resui, including low" counts andior "low" numbers of organisms.” Pee 4, 4.1, paragraph 2 Bc ote "(4.2)." and insert"(4.2), combined with a component due to Poisson distibution.* P00 4,43 Dolote “= 2r4" from the equation Pago § 52.1, paragraph 4, sonience 1 Delete “given tat ow tras are no considered nee" and insert provided tat resus based en lw counts te nol vein the ealculins (eos 3). Page 7, 53 ‘Addiat the beginning of the subclause: ‘Experiments should be performed to ensuro that cuffcionty large numbers of ecuniod colonies can be used for the calculations. Enumeration results based on less than 10 counted colonies should be excluded, Enumerations resuils wih 10 to 30 counted colries may be inchided only i the standard deviation of eproduciilty. sg, hats osing estimated is expected to be higher than 02 logy (cfu) oF 0,2 logy (um), oxperinntal protec! for be inrlaberalryslerdand assess he uncertainty) and atte use os tarda (ee Clas 8)ISO/TS 19036:2006/Amd.1:2009(E) ‘ Page 10 After Clause 7, Insert a new Clause 8, “8 Calculation of expanded uncertainty 8.1 Introduction It is assumed that the number of colony-forming unis in Petri dishes follows a Poisson distibution. This random errr is taken into account in the estimation ofthe expanded uncertainty described in 8.2. NOTE The calaulatons described fo the asimaion of the intalaberatory standard deviation of reproductity (see 53) neglect the random ertor due to Poisson distibutin, which means that they should exclude enumeration results Dated on low numbers o courted colores. 22 Calculation 82.1 General case Denoting the test result y=logygr, then the expanded uncertainly, U, with a coverage factor ef 2 (corresponding approximately to a confidence level of 95 %) can be calculated using Equation (1): wisest U=2 Ha a | C where Me Se Is the standard deviation of reproducibility - 0,189 61/5C is the variance component due to the Poisson distribution. in which ZC is the sum of the total numbers of colonies counted on all plates. = NOTE —_Temumeratris dered by using» teeacal nopary ofthe Paso datbsen (eaualy othe expeciton ‘end the variance, which immediately leads to'an estimated Poisson component coefident of variaion, CV = 1NZC), and ‘the epprosimation thatthe Poisson variance comporent on a logarthric scale is approximately equal to the coefcient of Spear pee (017 wt snarl ont: sal Bd, er revo fo Noga)? = 0.188 61 x (CV? when a decimat ‘sealois used. ‘Measurement uncertainty sccording to Equation (1) depends both on the reproducibility stancard deviation estimated from an experiment with high counts, sq and on the total plate count for the semple under investigation, ZC. itis recommended, forthe sake of simplicity, to use Equation (1) wherever possible, 8.22. Différentiation between low and high counts (optional) For high counts, the second term under the square rec, the Poisson term depending on ZC, can be ignored o (1) simplifies to: 3 ep te eB FO) @ISO/TS 19036:2006/Amd.1:2009(E) Once shi tir 'nhas been estimated, Cyn, can be either calculated from Equation (2) or taken from Table 8.1, ‘TWO cases can be differentiated: # EC>Ciq use Equation (2) to derive U: 4 EC < Cim use Equation (1) to derive U. NOTE Calculation of Cig is not necessary when Equation (1) is used in all cases.” Page 10 Delete Clause 8, and insert, “a Expression of measurement uncertainty in the test reports Once the measurement uncertainty has been derived as explained in Clause 8, it may be expressed in the report, together with the test resuit, as an interval on the decimal logarithmic scale (see Note to 5.3) or as natural values (cfu per gram or cfu per milli), of as a percentage, as illustrated by the following possibilities. The test result can be reported according to one of the following possibilities: 2) interval for log result: y£U [l0G4q (cfulg)] or YEU [10949 (Cfulm))); 8 9) decimal logarithmic result estimate with limits: yllogyg (cfulg)) * y- U. y+ Ujor yHlogya (otuimn] Uy + Ut 4 result estimate with absolute limits: xcfulg (107-4, 107+Y4 or * full foy-¥, 107+4; ) Tesult estimate with relative limits: xcfulg [-{1—10-Y) x 100%, +(-4 + 10%) x 100 %] or ae FH1 - 107%) x 100 %, +(-1 ee 4100 %). arithm, y Is, like pH, dimensionless, and has none. To used, 10g,9 (clulg) or 1099 (cfulml) can be added in\ ‘The test result is 100 000 cfulg, Le. y= 5,00 [logyq (cfu/g], with ZC = 110 (dilution ~9: 102 colonies; dilution ~4: 8 colonies), ISO/TS 19036:2006/Amd.1:2009(E) Thus the expanded uncertainty, U, with a coverage factor of 2 (95 % confidence level) is, using Equation (1): + U=2)0,15? est The test result may be reported according to one of the following possibilities: 0.31 5.0 £03 [ony (tua): 5,0 flogyg (chug) [4,7; 5,3}; 110-108 eg (4,9 « 10%; 2,0 x 1055 1,0 « 108 cfulg | 51 %; +100 %}, NOTE Applicable ony iftwo formulae are used (see 8.2.2): Cyq= 78. Then, a8 C= 110> Cyq= 78, simplified Equation (2) for high counts U = 254 = 0,30 could have been used. EXAMPLE 2 ‘The standard deviation of reproducibility, sis 0,25 (logy (cfu/g)) The test result is 260 clulg, i.e. y= 2,45 flog49 (cfulg)), with EC = 31 (dilution ~1, 1 mi on three plates: 9 + 9 +9 colonies, dilution -2: 4 colonies). ‘Thus the expanded uncertainty, U, with a coverage factor of 2 (95 % confidence level) is, using Equation (1): fos? aise a5 ‘The test result may be reported according to one of the following possibilities: 24 408: NOD» (cfu/g)I; 24 ody (ura 1.8 3.0% 280 cfulg (85; 930}, 280 ctulg [-70 %; +230 %). NOTE Applicable only two formulae ae used (See 82.2) Cy 28. Then, a8 2C= 31> Cq228,simpiied Equain (2 or igh counts U2 44 = 0,50 could have been used.ISO/TS 19036:2006/Amd.1:2008(E) ‘The test resuit may be reported according to one of the following possibilies: 2,0.£.0,3 (10949 (cfulg)|: . 2.0 [logy (chug) (1,7; 2.31; 100 cfulg (46; 220}; i 100 efulg [-54 %: +120 %). ‘ NOTE Applicable only if two formulse are used (see 8.2.2): C,, = 144, Then, a8 EC= 11 Cyq = 38, simplified Equation (2) for high counts, U= 2 : 0.22 =0,44 apples. ‘Ageneral rule, applicable only for results with £C'> 36, may'be staled according to one of the follawing possibilities: fag result + 0,44 [logyg (cfu/g)I; log result (log, (cfu/g) {log result -0,44; log result + 0,44); result chulg [10a endl O44; 4qlonreust+ 0-44), result cfulg [-64 %; +175 Sel. 2£C < 36, or if single universally applicable equation is preferred, ue Equation (1) “Page 16 ‘add Annex B (overleaf) before the bibliography.Annex B (informative) Values of Cj, and relative limits Once sq is known, the value of Cy is obtained using Equation (3), Cyq values are also given in Table B.1 for Be TASS Of te. Moreover, once U is known (using either Equatit® (1) oF Equation (2)), the relative limits FO TAT AUE 100%. 1(-1 4 10% x 100%] may help the exprestion er eecre (See Clause 9). These relative limits are also given in Table 8.1 for some values of U Table B.1 — Values of Cim and relative limits Standard deviation * Standard deviation of reproducibility, Uncertainty | Relative limits? | of reproduc ibility Uncertainty | Relative limits % % fe Gm | | tower | upper 26 Gt] YY tower | upp O08 n7aso] aoe | -s | a5 051 7] 102 | -90 | ser 0,02 4365| 004 | -9 | v0 052 6 | 104 [| -st | 4006 003 [1940] 06 =13 | +15 053 6 1,06 -91_ | +1048 2.04 fost] 008 | —17 | 20 ost [6 |. 108 | -02 [rite [a0 soe | oi | 21 | 26 055 6] ito | 22 | o150 0,06 405 | oiz | -24 | 32 oss [el] 12 2 | +1218 0.07 356 | 014 | 28 | 338 057 s| saa] -03 | +1280 _ 008 a3] ose | 31 | a5 ose s| ste | 03 [ms 0,09 216 | - ote | 34 | 451 059 5 1se | -03 [ata 0.10 178 | 0,20 | -s7 | +50 060 s {120 | -o4 [+1405 | ~40 | +66 0.61 6 1,22 -94 | +1560 ~42 | 474 2 5 -45_ | +02 063 4 45 | ast 064 4 -s0 | +100 | oes 4 52 | +100 068 4 -s4 | sit0 087 4 85 -s6 | +120 4] 130~ | -96 [s2i97 | 58 | +140 4] 130 | -96 | 2205 so | +151 5 4} 1407 | -96 | zara 3] 142” | -6 | 2530 3] aa | -06 | 2654. ; 3{ sao | -e7 | i270 | 3 | 148 | -97 | 2920 a) 3, 7 3ISOITS 19036:2008/Amd.1:2009(E) Table 8.1 (continued) Standard deviation of reproducibility Uncertainty | Relative limits? ‘oreproduetbityy Uncertainty | Relative limits? % % [ew] vf] oe fat] | er oe 0.27 24 [05a | 71 | v2ar oy? 3 9.28 a | ose | -72 | 263 078 3 0,20 21 | 058 | —74 | +200 0,79 3 ae. +296 0,80 3a 031 +317 0.81 3. 032 1337 0.82 3 0,33 oss | -7a | ~357 0.83) 3|_ [ ose | “088 79 | +379 oa [2 038 14 [070 | -20 | +401 |. os 2 0.36 13 [072 | si | +025 086 2 037 13 | ova | 22 [ +450 087 2 038 wz] o7e | -0 | +475 088 2 039 1 [o7e | -a3 | +503 089 2 2 2 [192 | -90 | sos07 2| 194 |-s86| eat0 2 | 196 | -s86| +7 148 2 | 188 | -967 | +7466 2 [190 | -967| +7643 2| 192 | -98e| se2t8 2 | 194 [-98.9| +8610 2 | 196 | -98| 49020 2 |_.19¢ |-99.0| 9450 2 | 200 [-990| +9900 Gini the nit for ZC above which the simplifcation U2, % possibie, : > acne ona of ern chu cl ran nc of