total plate count (TPC)

aerobic plate count (APC)

total mesophilic plate count
Total bacterial number is an important
indication for food quality
Basic principle for TPC
Sampling size, 25 or 50 g

Solid sample
Add 9 times volume of buffer (25 g + 225
mL wt:vol; 25 mL + 225 mL vol:vol)

Homogenize
Homogenizer or stomacher

Materials and methods

Food samples
store at room temperature

Depend on the food samples to

decide the dilution range
Dilution range is from -1 to -4

Detection limit
No dilution
1 CFU/ml; 10 CFU/g

-1 dilution
10 CFU/ml; 100 CFU/g

However, we need 25 to 250 CFU in one

plate
Thus, reliable detection limit is:
No dilution
25 CFU/ml; 250 CFU/g

-1 dilution
250 CFU/ml; 2500 CFU/g


TSA

Dilutions (every group has their own)

25 g + 225 mL phosphate buffer
Homogenize for 2 min
Decimal serial dilution (1:9)

procedures
1. Homogenize sample
For milk, no homogenization

2. Decimally serial dilution

3. Place one mL of the undiluted
homogenate into a Petri-dish (0 dilution)
1. Add agar to mix the sample: pour plate

4. Place 0.1 mL of the undiluted

homogenate onto a Petri-dish (-1 dilution)
: spread plate
5. Place 0.1 mL of -1 dilution the
homogenate onto a Petri-dish (-2 dilution)

 Incubate the plates at 37C for 24 (spread)

to 48 (pour) hours
Count the colony (25-250 colony/plate)
Calculate bacterial number in food sample
Solid food: average CFU number x dilution x 10
E.g. 56 x 103x 10 = 560000 = 5.6 x 10 5 CFU/g
= 5.75 log10 CFU/g
Milk: average CFU number x dilution
E.g. 56 x 103 = 56000 = 5.6 x 104 CFU/g = 4.75
log10 CFU/mL