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10
VIABLE PLATE COUNTS
RESULTS AND DISCUSSION
Table 1. Summary of Results
dilution
Trial 1
Trial 2
Trial 3
134
TNTC
TNTC
Contaminated with
fungal growth
Contaminated with
fungal growth
TFTC
TFTC
Contaminated with
fungal growth
TFTC
10-6
10-7
10-8
Bacterial numbers in a broth culture of E. coli were determined using the viable
plate method. This method is an indirect measure of cell density for live bacteria. The
bacteria was inoculated in the plates using the spread plate technique which the
bacteria are evenly spread onto the surface of the medium by means of a spreader. The
plates were incubated for 24h and the number of colonies were counted. In this
procedure, each colony counted is assumed to arise from one bacterial cell.
The results of the experiment (Table 1) did not show the correlation of the serial
dilution and the number of colonies present in the agar plates. Theoretically, as the
dilution proceeds further, the number of bacterial colonies decreases. Only dilution
10-6 Trial 1 has a countable bacterial colonies. From which, the CFU ml -1 of the
original broth culture containing E. coli was calculated to be 1.3x108. Other results are
labelled to be TFTC or Too Few To Count (#colonies < 25), Too Many To Count
(#colonies >250). Some plates were contaminated with fungal growth characterized
by the large to moderate sized filamentous growth present in the plates. Errors may be
due to unsuitable culture conditions which may include inappropriate use of medium,
inadequate environment to facilitate growth (Temperature and other conditions). Error
may also arise from procedural errors such as inaccurate transfer or dilutions and
inadequate mixing of culture before inoculation. Cell clumping may also overestimate
the number of bacteria present.
STUDY QUESTIONS:
1. Why do you think it is important to be able to quantify the number of viable
bacteria in a sample?
Quantification of the number of viable bacteria in a sample is one of the methods
of measuring microbial growth which is essential clinically, particularly in the
potency,
mechanism and diagnosis of infectious diseases. Also, the total number
of viable
counts are important in dairy microbiology, food microbiology, and
water microbiology.
2.
What is a CFU?
Colony forming unit or CFU refers to each colony that can be counted in a plate.
In addition, each viable cell can yield one colony.
3.
REFERENCES
from
APPENDIX
Calculation of CFU ml-1 of original broth culture
*Only for the dilution 10-6 Trial 1 can be calculated because the other results is TNTC,
TFTC or contaminated with fungal growth.