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
• 培養基
– 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: 混稀 pour plate
4. Place 0.1 mL of the undiluted homogenate onto
a Petri-dish (-1 dilution) 塗抹: 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 105 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