Lab 3

Introduction:
Total Plate Count (TPC) is a method used to estimate the amount of microbial life in
a sample. When applied to the food industry, this can be used for general purposes
such as determining the degree of spoilage in food, or more specific applications
such as determining the amount of cells in a serving of a probiotic beverage. The
unit of measurement for TPC is the Colony Forming Unit (CFU), usually used in
conjunction with a mass or volume. After dilutions are performed, this is calculated
to estimate how many microorganisms were present in the original sample.
Objectives:
Learning how to obtain quantitative values for microorganisms in food through
dilutions and the calculation of CFU/mL or CFU/g.
Materials:
9 mL tubes of peptone water for dilution (x5)
Petri plates containing Plate Count Agar (PCA) medium (x12)
Aerobic count petrifilm (x6)
Procedure:
Samples were first diluted by factors between 10^-1 to 10^-5. These were plated
onto the PCA medium in addition to a dilution of 10^0, or undiluted sample. These
plates were then incubated at 30C for 48 hours. Petrifilms with dilution factors
between 10^0 to 10^-5 were also prepared. Each plate had 2 replicants, while
there was only one petrifilm for each dilution factor.
Results:
Dilution Factor
10^0
10^-1
10^-2
10^-3
10^-4
10^-5

Replicant 1
TNTC
TNTC
TNTC
TNTC
277
14

Replicant 2
TNTC
TNTC
TNTC
TNTC
355
13

Petrifilm
TNTC
TNTC
TNTC
TNTC
TNTC
TNTC

PCA plate CFU/serving: 10^9.87989832

This value was obtained by averaging the values of the two replicants from the 10^4 plates, which would give the average CFU/mL. Multiplying this value based on
how many mL are in a serving gives CFU/serving. The log of this value was taken
for recording purposes and then compared to the results of the other groups.
Discussion: