Exercise 4 – Bacterial Growth

1. Which stage of growth would we expect to see the highest concentration of bacterial cells in broth?

a. lag
b. log/exponential
c. stationary
d. death

2. Which stage of growth is most likely seen in a colony?

a. log
b. stationary
c. death
d. all of these

3. Which of the following act as a growth factor for bacteria in broth? (Hint: think about growth factors for humans.)

a. correct pH
b. oxygen availability
c. presence of necessary vitamins
d. all of these

4. One ml of a broth culture is mixed with 9 ml of sterile water. Once mixed, 1 ml of that diluted mixture is placed in another test tube
with 9 ml of sterile water. The original broth culture is now diluted times.

a. 10
b. 100
c. 9
d. 90

5. One factor that will slow down rapid growth of bacteria in broth is:

a. increase in waste products

b. decrease in nutrients
c. decrease in oxygen availability
d. all of these

6. Addition of salt or sugar to foods prevents spoilage due to microbial growth because it changes the:

a. oxygen concentration
b. osmotic pressure
c. concentration of trace elements
d. pH

A. CRITICAL THINKING

1. What factors may be the cause for an extended lag phase in a bacteria growth curve?

Temperature, initial cellular mean mass, maintenance energy, effects of inoculum

2. If you altered the conditions under which bacterial growth normally occurs i.e., increase the temperature of incubating a
culture from 35 to 40°C, what effect would this have on the bacterial growth curve of the organism
under study?

3. How might the bacterial growth curve change if a facultative anaerobe was first monitored for growth when grown
in the presence of an O2 environment and, during its log (exponential) growth phase, the organism was suddenly
placed in an anaerobic environment?

A facultative anaerobe is actually aerobe & prefers to grow in O2 environment but is able to carry on metabolism even when O2 is
absent. (Aerobe is of two types Obligate aerobes & facultative anaerobes, whereas anaerobe is of two types obligate anaerobe &
facultative aerobe.)
So when they are suddenly transferred into an anaerobic environment they'll need some time to adapt in that new environment. As lag
phase indicates the adaptation phase, growth curve will show another lag phase & then it'll be followed by the log phase as usual, but
it'll be less steeper as not every bacteria could adapt to metabolize in an O2 free environment. It is due delayed enzyme synthesis in
new bacterial cells & also death of some bacteria.

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 4. A bacterial growth curve was determined in this exercise for E. coli by measuring the absorbance of the culture
at 20-minute intervals. When the curve showed plateaus (lag and stationary phases) was the absorbance obtained
proportional to the number of bacteria present in the cuvette tube?

B. FILL-IN-THE-BLANK

1. The phase of the bacterial growth curve where cells are dying at rapid rates is called the death phase.

2. Bacterial cells are most likely to be affected by antibiotics when they are in this phase of the bacterial growth curve: growth

3. If 1 ml of a diluted culture (1:1000) was added to 9 ml of water, the dilution made is: 1:10,000

4. An organism that strictly requires an atmosphere of CO2 for growth is called a .

5. List five physical factors required by bacteria for growth: pH, temperature, moisture, oxygen concentration, osmotic pressure

6. Identify three nutritional factors required by most bacteria for growth: carbon, nitrogen, sulfur, phosphorus, trace elements

7. A microbe that requires serum components to be added to the growth medium is called fastidious

8. What is a countable number of colonies on a bacterial plate? Must average 30-300 colonies/plates

9. A dilution made whereby the liquid transferred from tube to tube is uniform and the total volume in each tube remains uniform is
called serial dilution.

10. Measurement of a bacterial growth curve is referred to as a semiquantitative (quantitative or semiquantitative) procedure.

C. MATCHING

You may use each growth phase more than once to match with the characteristics of growth identified.

Growth Phase
a. Lag
b. Log/Exponential
c. Stationary
d. Death

Characteristic of Growth
C – 1. No net increase in cells has occurred.
D – 2. Cells experience autolysis.
A – 3. Cells acclimate to the environment.
B – 4. Cellular generations have occurred in culture.
C – 5. Binary fission is maximal.
C – 6. Cells are producing toxic by-products of metabolism.
D – 7. Cells that are sporeformers will generate spores at this phase.
A – 8. Cells are gathering nutrients and synthesizing macromolecules for cell division.
C & D – 9. No increase in absorbance occurs at these growth phases.
D – 10. Antibiotics probably have their greatest killing effectiveness at this phase of growth.

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