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Bacteria
Bacteria divide using the process of binary fission during which one cell will divide into two
identical cells
The process is as follows:
• The single, circular DNA molecule undergoes DNA replication

• Any plasmids present undergo DNA replication
• The parent cell divides into two cells, with the cytoplasm roughly halved between the two
daughter cells
• The two daughter cells each contain a single copy of the circular DNA molecule and a
variable number of plasmids
Bacterial growth curve

The growth of a bacterial population follows a specific pattern over time; this is known as a growth
curve
There are 4 phases in the population growth curve of a microorganism population
Lag phase
The population size increases slowly as the
microorganism population adapts to its new
environment and gradually starts to
reproduce
Exponential phase (Log Phase).

With high availability of nutrients and plenty
of space, the population moves into
exponential growth; this means that the
population doubles with each division
This phase is also known as the log phase

Stationary phase
The population reaches its maximum as it is limited by its environment, e.g. a lack of resources
and toxic waste products.
During this phase the number of microorganisms dying equals the number being produced by
binary fission and the growth curve levels off. Rate of reproduction = Rate of death.
Death phase
Due to lack of nutrients and a build up of toxic waste build up, death rate exceeds rate of
reproduction and the population starts to decline
This phase is also known as the decline phase
N.B: Generation time is the time span between two successive bacterial cell divisions.
Exponential growth rate constants (k)

The following formula can be used to calculate the exponential growth rate constant:
N0 = The number of organisms at the start.
Nt = The number of organisms at end of the time interval (t).
T = the time for which the colony has been growing
• To use this equation the exponential growth rate constant k must be calculated. This refers
to the number of times the population doubles in a given time period.
Worked example
A bacterial colony started with 2 individuals and after 3 hours of growing there were 926
bacteria in the colony.
1. Calculate the exponential growth rate constant of this colony..(k)
Scan and watch till 2:45..or all

if you want side information!
Core practical 13: Investigate the rate of growth of microorganisms in a liquid
culture, taking into account the safe and ethical use of organisms.
Growing organisms such as bacteria in a liquid culture involves several steps and requires
aseptic techniques to prevent contamination.
1. Selecting a suitable growth medium:
- A growth medium is a nutrient-rich solution that provides essential nutrients for the organisms
to grow. (Such as..Glucose)
2. Inoculating the culture:
- A small amount of the desired organism (Bacteria, yeast..etc) is transferred into the glucose
solution in the sterile flask.
-Place the flask on a magnetic stirrer, stopper the flask with sterile cotton wool. The magnetic
stirrer allows the density of microorganisms to remain consistent throughout the flask.
- Aseptic techniques involve working in a clean environment and using sterile tools, such as pipettes
and flasks, to prevent contamination.
- Sterilization can be achieved by autoclaving, which uses high-pressure steam to kill any existing
microorganisms.
3. Incubation:
- The inoculated culture is placed in an incubator set at the optimal temperature (20°C) and
conditions for the organism's growth for suitable time (10 hours).
- Incubation allows the organisms to multiply and form visible colonies or turbidity in the liquid
culture.
4. Monitoring and maintenance:
- The liquid culture should be periodically monitored for growth, which can be visually assessed
by turbidity or by measuring optical density several methods can be employed.
Using a Colorimeter:
1. Take samples at regular intervals during the growth process.
2. Transfer the samples into cuvettes.
3. Insert the cuvette into the colorimeter.

4. The colorimeter emits light through the sample and measures the intensity of
transmitted or scattered light.
5. The data logger records the readings obtained from the colorimeter.
6. The recorded data can be used to determine the turbidity of the sample, which
correlates with the rate of growth of microorganisms. As number of bacteria cells
increases, turbidity increases, the less light is transmitted.
Using a Light Source and Light Sensor Connected to Data Logger:
1. Take samples at regular intervals during growth.
2. Place the sample in a container suitable for measurement (e.g., test tube).
3. Position the light source on one side of the container and the light sensor on the opposite
side.
4. The light emitted by the source passes through the sample and reaches the sensor.
5. The light sensor detects changes in light intensity caused by scattering or absorption
by microorganisms.
6. Connect the light sensor to a data logger to record the measurements over time.
7. Analyze the recorded data to determine turbidity and relate it to microbial growth rate.
The more cells present, the higher the turbidity, the less light is transmitted.
Using a Hemocytometer and Light Microscope:
1. Dilute an aliquot of the culture to achieve an appropriate cell concentration for counting.
2. Place a small volume (usually 10 μL) of diluted culture into the hemocytometer.
3. Position the hemocytometer on the stage of a light microscope.
4. Focus the microscope to visualize the grid lines on the hemocytometer.
5. Count the number of cells within specified grid areas under a suitable magnification.
6. Repeat the counting process for multiple grid areas and calculate the average cell count.
7. The cell count can be used to estimate turbidity and determine the rate of microbial
growth.

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Bacteria

• The single, circular DNA molecule undergoes DNA replication

Bacterial growth curve

Exponential phase (Log Phase).

This phase is also known as the log phase

Exponential growth rate constants (k)

N0 = The number of organisms at the start.

Nt = The number of organisms at end of the time interval (t).

T = the time for which the colony has been growing

1. Calculate the exponential growth rate constant of this colony..(k)

Scan and watch till 2:45..or all

1. Selecting a suitable growth medium:

2. Inoculating the culture:

4. Monitoring and maintenance:

1. Take samples at regular intervals during the growth process.

2. Transfer the samples into cuvettes.

3. Insert the cuvette into the colorimeter.

1. Take samples at regular intervals during growth.

3. Position the hemocytometer on the stage of a light microscope.

4. Focus the microscope to visualize the grid lines on the hemocytometer.

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