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Standard Plate
Count.....
. This method consists of growing the bacteria in a nutrient culture petridish or
(petrifilm) and counting colonies which develop. It can be used for all types of
dairy products and is generally used in the examination of Grade A raw and
pasteurized milk. This method used to determine the general quality of the milk
supply.
.Procedure..............
• Wash the hands and disinfect the stage.
• Mixing of milk sample..
• Make a series dilutions of milk sample...
• Transfer a suitable dilution to a petridish by sterile pipette...
• 5. Add a suitable amount (12 ml ) from SPC agar on the dilution and
mixing
• gently by moving the dish on different direction..
• 6. Incubate 48 hours at 32°C .
• 7. Count colonies and report results.
Note..
Choosing the plates with ( 25 - 250 ) colonies
SPC(Cfu/ml)= Average number of colonies x Reciprocal of the dilution used
.Counting: ...
mean estimate of bacterial populations and it is not exact because:-
• The agar used in method (SPC agar) is not suitable for growing all species
of bacteria...
• Incubation Degree which used in method (32°C) is not suitable for
growing all species of bacteria.
• Sharing more than one bacteria to form one colony.....
Duplicate plate dilution
Note: SPC(Cfu/ml)= Average number of colonies x Reciprocal of the dilution used
1/100 1/1000 Only one dilution yields
175 16 plates with 25-250
colonies compute the
208 17
mean for that dilution as
175+208=383 383÷2=191.5 the basis for the SPC .
191.5x100=19150….SPC=19000
1/100 1/1000 Both dilutions
yield plates with
230 28
25 to 250
246 36 colonies: Average
the mean count
230+246=476 28+36=64
for each dilution
476÷2=238 64÷2=32 as the basis for
SPC.
238x100=23800…SPC=23800 32x1000=32000 SPC=32000
23800+32000=55800÷2=27900… SPC= 28000
unless the count 1/1000 1/100
computed for the higher
42 138
dilution is more than
twice the computed for 30 162
the lower dilution .In the
latter instance , use the 42+30=72 138+162=300
lower computed count as 72÷2=36 300÷2=150
the SPC.
36x1000=36000…SPC=36000 150x100=15000… SPC=15000
1/100 1/1000 Neither dilution yields
plates with 25-250
287 23
colonies: Use the mean
263 19 count the lower dilution
as the basis for estimating
287+263=550 the SPC.
550÷2=275
275x100=27500… ESPC=28000
1/100 1/1000 Both dilutions yield only
plates with fewer than 25
18 2
colonies: Use the mean
14 0 count of the duplicates of
18+14=32 the lowest dilution to
estimate the Spc
32÷2=16
16x100=1600…….ESPC=1600
1/100 1/1000 Both dilutions yield
plates with no colonies:
0 0
Estimate the SPC as less
0 0 than (<) 1 times the
lowest dilution .
Lower than 1x100
ESPC=lower than 100
1/100 1/1000 Only one plate of one
dilution contains 25-250
272 23 colonies: When one plate
of one dilution contains 25-
248 19 250 colonies and the
duplicate contains more
272+248=520 520÷2=260 than 250 colonies , use
both plates in computing
260x100=26000………SPC=26000
the SPC.
1/100 1/1000 One plate of each
dilution contains
275 22
25-250 colonies and
240 35 the duplicate
contains more than
275+240=515 22+35=57 250 or less than 25
515÷2=257.5 57÷2=28.5 colonies :use all
four in computing
257.5x100=25750…..SPC=26000 28.5x1000=28500…SPC=29000 the SPC.
26000+29000 = 55000 ÷2 = 27.500 …SPC=28000
1/1oo 1/1000 Both plates ;one of
them(lower dilution) TNTC
TNTC 14,20,34,12 TNTC ,the other (high dilution
)with 10 - 100 colonies per
SPC(Cfu)=Average of(4 square cm)x 65 cm²xreciprocal of cm² : use the estimated
dilution count of the ESPC.
In this case take the average
= 20 x 65 x 1000 of (4 square cm²).
1/100 1/1000 Both plates ;one of
them(lower dilution) TNTC
TNTC 3,6,7,8,2,5,3,2,7,9,6,3 ,the other (high dilution )with
less than 10 colonies per
SPC(Cfu)=Average of(4 square cm)x 65 cm² x reciprocal of cm²: use the estimated count
dilution = 5 x 65 x 1000 of the ESPC. In this case take
the average of(12 square
cm²).
