Discussion

Experiment 1

Experiment 1 illustrated the importance of sterilization in laboratory environments.

Sterilization is defined as a process of complete elimination or destruction of all forms of
microbial life (i.e. both vegetative and spore forms) which is carried out by various physical
and chemical methods (Mohapatra, 2017).

In this experiment, two out of three plates had nutrient agar that was sterilized using two
different methods, namely autoclaving and boiling. The third plate had nutrient agar that was
not sterilized. As expected, the result was that the autoclaved and boiled plates did not have
any bacterial growth and the nutrient agar solidified, while bacterial growth was observed in
the plate that was not sterilized. The nutrient agar that was not sterilized did not solidify.

The sterilization by autoclaving and boiling killed all microorganisms in the nutrient agar,
resulting in no bacterial growth when it was incubated, the plate that was not sterilized
contained bacteria, which then grew while being incubated.

Experiment 2 – Part 1

Experiment 2, part 1 explored the effects of boiling temperature on the growth of E.coli and
B.subtilis. The result of the experiment was that both the test tubes containing nutrient broth
that were inoculated with E.coli exhibited bacterial growth, as did the 2 test tubes containing
nutrient broth that were inoculated with B.subtilis. The control, which was the 2 test tube
containing nutrient broth only, did not exhibit any bacterial growth. An increase in
temperature will increase enzyme activity, which leads to growth in the bacteria culture.

Experiment 2 – Part 2

Experiment 2, part 2 explored the effects of different temperatures on the growth of E.coli
and B.subtilis. Single streaks of E.coli and B.subtilis were inoculated on plates with nutrient
agar and incubated overnight at different temperatures (4℃ , 27℃ , 30℃ , 37℃ )

At 4℃ , neither E.coli nor B.subtilis plates had any bacterial growth. This was to be expected
because 4℃ is an extremely low temperature and is outside of the optimal growth
temperature range of both bacteria. B.subtilis has an optimal growth temperature from 25 -
35℃ (Tanner, 2014).The central normal range of E.coli growth temperature is from 20 - 37
℃ (Farewell & Neidhardt, 1998).

At 27℃ , both E.coli and B.subtilis plates had bacterial growth. This was the expected result
because the temperature is within the optimal growth temperature range of both the bacteria.

At 30℃ , both E.coli and B.subtilis plates had bacterial growth. This was the expected result
because the temperature is within the optimal growth temperature range of both the bacteria.

At 37℃ , both E.coli and B.subtilis plates had bacterial growth. This was the expected result
because the temperature is within the optimal growth temperature range of E.coli and close
to the range of B.subtilis.
Experiment 3

Experiment 3 explored the effect of pH on the growth of E.coli and B.subtilis. Triple streaks
of E.coli and B.subtilis were inoculated on plates with nutrient agar of different pH levels and
incubated overnight.

All the plates containing acidic nutrient agar that were inoculated with E.coli and B.subtilis
had bacterial growth. This was not the expected result for E.coli as it does not grow in acidic
conditions. E.coli grows best within a pH range of 6.5-7.5 (Philip, Kern, & Goldmanns, 2018).
B.subtilis is able to grow between pH levels of 4.8-9.2 (Gauvry, Couvert, & Coroller, 2021).

All the plates containing basic or alkaline nutrient agar that were inoculated with E.coli and
B.subtilis had bacterial growth. This was the expected result because both E.coli and
B.subtilis have optimal growth pH ranges that range from neutral to basic.

All the plates containing neutral nutrient agar that were inoculated with E.coli and B.subtilis
had bacterial growth. This was the expected result because both E.coli and B.subtilis have
optimal growth pH ranges that range from neutral to basic.

Bibliography

Bibliography
Farewell, A., & Neidhardt, F. (1998). Effect of Temperature on In Vivo Protein Synthesis Capacity in
Escherichia coli. Journal of Bacteriology , 4704-4710.

Gauvry, E., Couvert, O., & Coroller, L. (2021). Effects of temperature, pH and water activity on the
growth and the sporulation abilities of Bacillus subtilis. International Journal of Food
Microbiology, 108915.

Mohapatra, S. (2017). Sterilization and Disinfection. Essentials of Neuroanesthesia, 929-944.

Philip, P., Kern, D., & Goldmanns, J. (2018). Parallel substrate supply and pH stabilization for optimal
screening of E.coli with the membrane-based fed-batch shake flask. Microbial Cell Factories,
50-62.

Tanner, B. (2014, September 1). Microorganisms available for testing - Bacillus subtilis. Retrieved
April 23, 2022, from Microchem Laboratory:
https://microchemlab.com/microorganisms/bacillus-subtilis/