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In this experiment, 3 types of bacteria were grown in broth media and their Optical Density was measured from time to time, the values obtained were plotted into a semi-log graph against the time to compare the growth rate and generation time of the three different bacteria: Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus.
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Universiti Tunku Abdul Rahman (Kampar Campus)
Faculty of Science, Engineering, and Technology
Bachelor of Science (Hons) Biotechnology
Year 2 Semester 1
UESB 2142 Laboratory 2A
(III) Microbiology
Lecturer: Dr. Teh Yok Lan
Student’s Name: Cheah Hong Leong
Student’s ID: 08AIB03788
Group Member: Samuel Tan Sha Loong
Experiment No. 5
Title: Growth Curve
Date: August 17, 2009
Title: Growth Curve
Objectives:
- To obtain the graphs of the microbial growth of different bacteria through optical
density measurement of the media.
- To compare the growth curve of the bacteria.
Results:
Table 1: Optical Density of Inoculated Nutrient Broth with Time of Pseudomonas aeruginosa
Time (minutes) Optical Density, OD600 (A)To, 0 0.013T1, 15 0.015T2, 30 0.017T3, 45 0.020T4, 60 0.022T5, 75 0.026T6, 90 0.031
Table 2: Optical Density of Inoculated Nutrient Broth with Time of Escherichia coli
Time (minutes) Optical Density, OD600 (A)To, 0 0.023T1, 15 0.020T2, 30 0.074T3, 45 0.085T4, 60 0.226T5, 75 0.262T6, 90 0.223
Table 3: Optical Density of Inoculated Nutrient Broth with Time of Staphylococcus aureus
Time (minutes) Optical Density, OD600 (A)To, 0 0.022T1, 15 0.028T2, 30 0.034T3, 45 0.040T4, 60 0.062T5, 75 0.091T6, 90 0.143
Note: Data in Table 1, 2, and 3 were plotted in “Semi-log Graph: Optical Density of
Inoculated Nutrient Broth versus Time” in Attachment.
Calculation of Generation Time:
Pseudomonas aeruginosa, Generation time = 45 minutes
Escherichia coli, Generation time = T0.100 – T0.050
= (45 – 24) minutes
= 21 minutes
Staphylococcus aureus, Generation time = T0.100 – T0.050
= (87 – 48) minutes
= 39 minutes
Discussion:
Based on the semi-log graph and the calculation, the generation time of
Escherichia coli was the shortest (21 minutes), followed by Staphylococcus aureus (39
minutes) and Pseudomonas aeruginosa (45 minutes). With the shortest generation time,
Escherichia coli was expected to have the greatest growth rate under similar condition
among the three microorganisms, for every 21 minutes, the cell numbers of Escherichia
coli doubled.
Theoretically, the photospectrometry measurement assumes that the bacterial cells
were distributed in the nutrient broth evenly; the optical density obtained can be
proportional to the population of the bacteria in broth only when the assumption
occurred.
However, all the three bacteria have the ability to form biofilm on the flask surface,
which can cause errors in the photospectrometry measurement regarding the total cell
mass of the microorganisms in the nutrient broth.
Stirring of the nutrient broth during the incubation was solving the problem, but
during the incubation in the laboratory practical, non simultaneous timing of the students
from different subgroups made the incubation machine had to be turned on and off
repeatedly. That might lead to the uneven distribution of the bacteria in the broth.
Moreover, sometimes the incubation machine was also did not turned on by the students
after placing the flask into it, leaving the broth unstirred for several minutes.
This might be able to explain the unusual results shown by the Escherichia coli in which
the optical density was unexpectedly dropped at time 15 minute and 90 minute.
Conclusion:
The generation time was shortest for Escherichia coli, followed by Staphylococcus
aureus and Pseudomonas aeruginosa.
References:
Cappuccino, J. G. & Sherman, N. (2008). Microbiology. A Laboratory Manual, 8th ed.,
San Francisco, CA: Pearson Benjamin Cummings.
Escherichia coli. (2009, August 30). In Wikipedia, the free encyclopedia. Retrieved
August 31, 2009, from http://en.wikipedia.org/wiki/Escherichia_coli
Madigan, M. T., Martinko, J. M., Dunlap, P. V., & Clark, D. P. (2009). Brock Biology of
Microorganisms, 12th ed., San Francisco, CA: Pearson Benjamin Cummings.
Pseudomonas aeruginosa. (2009, August 28). In Wikipedia, the free encyclopedia.
Retrieved August 31, 2009, from http://en.wikipedia.org/wiki/Pseudomonas_aeruginosa
Staphylococcus aureus. (2009, August 26). In Wikipedia, the free encyclopedia.
Retrieved August 31, 2009, from http://en.wikipedia.org/wiki/Staphylococcus_aureus
Attachment:
Semi-log Graph: Optical Density of Inoculated Nutrient Broth versus Time