9-70 MICHOBIOLOGCAL

Trr 92?1:II. MPN hinex .xo 95% Coxiloexce Lnsrs r*n Ar- T,lele 9221:III. MPN Iosx .qo 957c Cosriurrr i;sffi
Cownerroxs o Posnvr ero Necrr' Rsrtrs wHEN FM 20-ML Colqgn,toNs cr PosIw o Nc.q.rtv Rrsulrs 'lgt
Pontos An UsrP PoRrroxs ,{xs Useir

9-5r,r.Curlitincc
lo. oi
ftrbes Giving No. of Tr-rbes Giving
L:mirs (Eraer)
Positi.;e Reaciol Out l{PN Inde.r/ Positive Reactirn Out of ]\.IPN indexl
ol 5 (ll) ml- Each 10{) mL Lo*'cr Upper 10 (10 u Eacir.) 100 mL

0 <1.i 3.5 0 <1.i -:..

I Li o.osr 5.4 1 1.1 .#&
2 1.6 0.40 8.4 2 2.2 *-1:.',
3 4.6 I.\' 13 3 3.6 *.'*..
,+ 8.{J /-- I 23 4 .5.1 ;.b, :

5 >.8.0 3.4 5. 6.9 1,.5 ,,

6 g.2 i.,+',
1 12 ,r.S.,
8 16 f."&'.
2. Table ileading and Hecording of Most Probable Number I 23 &-{..',
(MPi) 10 >23 13 '::.,,':

I{ccorrl coiitbrm eixlcenirition as MPN/i00 mL. X'IPN values

lbl a larietS' oi positive rnd tregatirre tLrhe combinations are
gilen in Tabies 9221:II. III, arid IV. The sampie volunes incii- s'ithin trvo dilutions of 0.001 mL^ rvhich h. I.
clteil in Tabics 9221:11 nci III ai'e chosen especially fbl'exau.r- In Example D, the hi-chest tiili,tion u'ith i'.il . '
ining drilking waters. Table 9221:IV illustrates iiPN values tbr rnl-. *.hich is lvithir tu,o deciinai diiutions t: r,

conbinatiorts of po.itive aud ne-uative results u'hen five 1O-mL.
trre 1.{)-url,, anci five 0.1-mL s;iuri:rie portiorl volumes of non-
poiable wlrter irre tcsled. if the -sampie portion volu[res used are
those t'ound in thc tables. report ttrle v|g qrlresponding io ihe
nurnbe r ol positir.e anci negative restrlts in rhe series as XiPNl l llt)
ilil-. When thc eries o1'clecimal dilutions is clift'erent from ihat
in Tahle 92li:lY. -relec the t'{PN value fi'ori '1'able 9221:IV lbr
the conbinalioil ol'positivc rcsults *nd calcuiate according to the
iolLorvrng lbirui::
l,Ii'N/ l{ltl ilL : ( f ai:le NIPN/ 100 mL )
a crnbinaiion ol -1-.s- 1.
Il'. aiier removal ol tlie lorvest diiution rlii:: ...-
no dilLrtion lvitl.r al1 positive reactions relr.r. ::
Iorvest tno dihitions and asslgn fhe suur oi .:".
tiorls to the thid dilution. Irt Example E. :he ;.
a1i positir,e trlbes conlains 10 mL; thi-s dihi. -
the second step. Four dilutions, none of u'i',:..:
tnbes. renrai*. l,nder ihese circttmst&oces. ::- :'
renlaining dilutions cortesponding to 1 axil
tl-re third rlilrrtion, add the ilumber of posilii;
x I ()/}j dilutions (0.01 anti 0.001 ntl- salltple). to 1". : ;
rion oi' -i-J- I .

n'hcle: If no dilution has all positive tubes (Ex*ry@

1/ :: vohirr'le oi srirtipie Fcfiiun at the loq'esl sclecterl ciilution. lou.'est tr,,,o diiutions, corresportding to 1C :: :
the thild dilution. add the number {-}l r . - :
\Yhen role tiran three cliiutions ate usei,l in a clccilrlrl seriesi lemaining dilution,s (0.1. 0.0l , anrl 0.0fii r.,,
oi dl jutions. lse the lollou in,c guideline ,s to select tire three most final coiibination ol 4-3-2. 1l ihe thiti 11iir,: '
'Iable 9221:lV. Several ilius-
applr.lpriie clilutic,rrs incl i*t'er to tllan five positive tubes. then lhe selected
laiive eliirrplcs (A through G) o'combinaiions of posilives ale in Tabie 9221:IV.
siior'.'n in T'ah1e 922 1:V. [rir"st. renlove the highest clilution if the thee dilutions sclecl"ed ire oot lttr.:. ,:

sruai1esl salupie t'clume) ii it has all negative tnbes aucl at least then solnething il the serial dilution rr'a:
L)t1c reir"ilining cliiution has a negaiil'e lLrbe. Next. rell1ove the t1.re usual methods fbr caicularing the NIP\
1or,, e st clihrtiolr {ii}fgest san.rpie r,ohime) if it has all positir e tubes not rpp1!'. If a nerv sample cannot be
int1 li least one lenraiuiitg rlilution has a positive tube. Accord- va1e is still desired, ilse the higirest dil',:,.
1nq io these g*irJeiines, ihe three cliluticns ill Example A are positive tubc ancl the ilvo dilutions il:t;:-:.'-:
-eiccierl b't remr:r'al of the highest (0.001-r:tl-) ar.rcl the lou'est tllrcc \clcclc(l tlilutiorr". In Ertttllrlc C. :' -
(i0-ruLl diiulionr. (the outcome froni the highest fhree tlil.:.' .:'
lf ihe lorvest riiiution cloes lot have ill positive tubcs. and 9221:fY because 6 is greater than 5. :.'.
seleral o the higl-rest dilutitns have 11 negatlvc tubes. then according to the above -tuidelines. u'cu1 :: --
ieniove the- irighest egative dihiitts {Eran-rple B). set of selected dilutions is no in Tatie -l-
l'lore thrn lhree dililtions ntay remain after renlovai ol the iiillou.ing formula io caicuiate the IlP\
lur,',est dilution rvith a11 positive tul-.es i:nd high diliitions u'ith all
negaiive tiles. In this casc. ii th* irighest dihitior.r r','ith all
positive ubes is q,itirin i*'tr dilutions of the highest clilution r",.ith 230.1 I
log,,,i
-!,.. ;
r-r lllrsitive iubes. ihea rsL'the highest ciilillion lvith r/t.] positive
1
-.!
A-l

ir.ri:ci iinrl tle ivr'.r irr- rrierliatell' lorver ciilutions. in Erample C, .-'" -
\
thc iiigticsl ililutit'in lviih ali positivc tubes i-s 0'1 mL' rvl.rich is
3t8t2017 Laboratory Methods > BAM Appendix 2: Mosi Probable Number from Serial Dllutions

Table 1Yxtue t
S*r 3 tc"{h*s #**h s? *.{, S"*t. iss}* S"**t # ir.x*flLxe, tfu* MPI** $}*r #rsr* mc?{ *$ *r**xr*
e.*n 4*,"1 r.* ir: (e 's* s
fl

Pos. Tubes Conf. lim. Pos. tubes Conf. Iim.

I

q.10 0.01 I 0.001

*Li*3 MPN/g Low I High 0.10 ; 0.0r 0.001 MPNIg High

U <3.0 9.5 n 21 4.5

0 3.0 0.15 9.6 1 28 8.7 94

0 I 3.0 0.15 11 2 35 8.7 g4

0 1 6.1 1"2 1B 0 29 8.7 g4

2 6.2 1.2 18 1 Jb 8.7 94

ole 9.4 J.b 38 0 23 4.6 94

1i 0 3.6 0.17 1B 1 3B 8.7 110

1i0 7.2 1.3 18 2 64 17 180

110 11 3.6 38 0 43 I 180

1 1 7.4 1.3 2A 1 1 75 17 200

1 1 11 3.6 3B 1 2 120 37 420

1 2 11 3.6 42 1 ? 160 40 420

1 2 15 4_5 42 0 o? 18 , 420

1 J 16 4.5 42 2 1 150 37 r 42A

210 9.2 1.4 38 2 2 210 4A $0

210 14 3.6 42 ? 290 90 : 1,000
2 2 2A 4.5 42 J 0 240 42 I 1,000
2 1 0 15 3.7 42 1 460 90 . 2,000
AN
2 1 1 20 4.5 +L J 2 1100 180 i 4,100

2 1 2 27 8.7 943 3 J >1 100 420 ,

https:/lwwwfda. gov/food/foodscienceresearch/la boratorymethod slucm 1 09656. htm 1t1

,:

.:

Tabla de valores de NMP para combinaciones de

ar
kEFI; tubos positivos y negativos usando 5 porciones
de 10 mL, I mLy 0,1 mL en 100 mL de muestra ffi

Positivos -

0-24 3,7 0,70 10

a-2-1 5,5 1,8 15

1-0-2 6,0 , 1,8 ' -iE ' " ''

1-1-2 8,1 3,4 22

l-3{ 9,3 3,4 22

?.-l .j- 6,S,, , 1,8 , t5

z'ta 1 - ., -,-",9,1---.-- ---, -- -* 1,9 17

2-3{' 1? 4,1 26

3,1{ 11 S,S i.6

3-2-1 17 o,e 40

TITT'ID
ffi -rt*
!
 '. I

;---"j
1

i
_ ..1

,i
..t

4-3'3S.t4f00
4.3{
'**-l 27 9,9 70
, 33 . 10 70
4-3-2 39 14 100
100

120
100
4-5.1 48 15 120
54{ 23 6,8 7A
5.0-1 31 10 7A

, 4-3 8 22 15O

6.2$ 49 . 15
5-2-1 7A 22 17A
5.2.2 94 34 230 :i
.:.,.....-,-]

5-2-g pa 36 250
5-24 150 6J

5.3{} 79 o,
s3-1 110 34 250
5t-? 140 52 400, :

5-3-3 fiA 7A 400

, 513-4 | 21 : 70 400 :

wz 220 7A 44A
d-? , 280 : 100 710 l

ID *i&J;-
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