av-mb-sop-010 (bis) sop for isolation and identification of coliforms by pour plate method

7/27/2019 Av-mb-sop-010 (Bis) Sop for Isolation and Identification of Coliforms by Pour Plate Method

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Department: Quality Assurance DOC. No. AV/MB/SOP/010

Page No. : Page 1 of 4

Effective Date: 01.06.06 Revision No. : 00

Subect SOP for isolation ! i"entification of Coli forms

RE#$S$ONS

Revision % Revise" Pages Description Date

00 Original issue &'.&(.&(

Prepare" )y Approve" )y Aut*ori+e" by

Microbiologist Management Representative actor! Manager

"#$ASS%%&'( %nternal )se Onl! "


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'.& Purpose

+o ,efine proce,-re for #oli forms b! po-r plate meto,.

,.& Scope

+is proce,-re is applicable to ,etermine #oli forms in a given sample.

-.& Responsibilities

Microbiologist

AM-alit!

.& Definition :

SOP( Stan,ar, operating proce,-re.

/.& Proce"ure

/.' +est portion initial s-spension an, ,il-tions(2%SO 6335

/., $noculation an" incubation:0

/.,.' +ae t7o sterile Petri ,ises. )sing a sterile pipettetransfer to eac ,is 1 ml of test sample if te pro,-ct is

li8-i, or 1 ml of te initial s-spension in case of oter

pro,-cts.

/.,., +ae t7o oter sterile Petri ,ises. )sing a fres sterile

pipette transfer to eac ,is 1 ml of te first ,ecimal

,il-tion 21015 of te test sample if te pro,-ct is li8-i,

or 1 ml of te first ,ecimal ,il-tion 210 *5 of te initial

s-spension in case of oter pro,-cts.

/.,.- Repeat te proce,-re 7it te f-rter ,il-tions.

/.,. Po-r abo-t 19 ml of te VRB$ me,i-m at 49:# ; 0.9:#

into eac Petri ,is./.,./ +e time elapsing bet7een te en, of te preparation of

te initial s-spension an, te moment 7en te me,i-m

is po-re, into te ,ises sall not e


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/.,.3 %nvert te prepare, ,ises an, inc-bate tem in inc-bator

set at >0:#>9:# or >:# for *4 o-rs ; * o-rs.

/.- Counting of t*e colonies

After te specifie, perio, of inc-bation co-nt -sing colon!

co-nting e8-ipment te colonies in eac ,is containing not

more tan 190 colonies.

E4pression of results:0

/. 5et*o" of calculation

/..' ?eneral case @ 'ises containing bet7een 19190

colonies

/..'.' Retain ,ises containing not more tan 190

colonies at t7o consec-tive ,il-tions. %t is

necessar! tat one of tese ,ises contains at least

19 caracteristic colonies.

/..'., #alc-late te n-mber N of #oli forms per

milliliter or per gram of pro,-ct ,epen,ing onte case -sing te follo7ing e8-ation(

6 C N 7 00000000000000000000 8n'9 &.' n, d

ere

6 C is te s-m of colonies co-nte, on all te ,ises retaine,

n' is te n-mber of ,ises retaine, in te first ,il-tions n, is te n-mber of ,ises retaine, in te secon, ,il-tions

d is te ,il-tion factor correspon,ing to te first ,il-tion.

Ro-n, te res-lt calc-late, to t7o significant fig-res.

/..'.- +ae as res-lt te n-mber of microorganisms per

milliliter or per gram of pro,-ct e


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/..,.' %f te t7o ,ises correspon,ing to te test sample2li8-i, pro,-cts5 or te initial s-spension 2oter

pro,-cts5 contain less tan 19 colonies calc-late

te aritmetic mean mof te colonies co-nte, on

bot ,ises.

/..,., Report te res-lts as follo7s(

estimate, n-mber of #oli forms per milliliter(

NE 7 m8li;ui" pro"ucts5

estimate, n-mber of #oli forms per gram(

NE 7 m4 d0'2oter pro,-cts5 7ere , is te

,il-tion factor of te initial

s-spension./..- No c*aracteristic colonies

/..-.' %f te t7o ,ises correspon,ing to te test sample

2li8-i, pro,-cts5 or te initial s-spension2oter

pro,-cts5 contain no colonies report te res-lts as

follo7s(

less tan 1 #oliform per milliliter2li8-i,

pro,-cts5

less tan 1 < ,1 #oliform per gram2oter

pro,-cts5 7ere , is te ,il-tion factor of te

initial s-spension.

(.& Documentation :

Microbiological +est Report

2.& References:%S 9401 2PAR+ 15(*00*

Prepare" )y Approve" )y Aut*ori+e" by

Microbiologist Management Representative actor! Manager

"#$ASS%%&'( %nternal )se Onl! "

av-mb-sop-010 (bis) sop for isolation and identification of coliforms by pour plate method

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