Department of Pharmaceutical Sciences

Pharmaceutical Microbiology I (PHR 124)

Umarah Miazi

Lecture 8: Reproduction and Growth of Bacteria

Bacterial Growth and Reproduction
Asexual reproduction is any form of reproduction in
which a single parent organism produces offspring
without combining male and female gametes.
Bacteria have four different modes of asexual
reproduction by which it can increase in number.

Bacteria can also exchange genetic materials with

other bacteria in three different ways. These ways
can also be referred as sexual reproduction of
bacteria.
Modes of Cell Division - Asexual
Reproduction
 1. Binary Fission
The most common and important mode of cell division in bacteria is
by binary fission.
Binary fission is that a parent cell divides into two identical cells. In
binary fission, the cell divides after developing a transverse septum
(cross-wall).
Binary: compounded or consisting of two things or parts;
characterized by two
Fission: dividing
Example: Binary fission occurs in Bacillus Subtilis.
 1.Binary Fission…Process
DNA Replication:
The circular nucleoid (bacterial chromosome) replicates.
Cellular DNA is replicated repeatedly to ensure each daughter cell receives a complete
genome.

Elongation & Segregation:

The nucleoid becomes elongated, forming a dumbbell-shaped structure.
Replicated nucleoids remain attached to the plasma membrane before separating.
Nucleoids migrate toward opposite poles of the cell.

Septum Formation:
The plasma membrane invaginates (pinches inward) at the midpoint.
A new cell wall forms along the septum, dividing the parent cell.

Division: Two identical daughter cells are produced, each with a copy of the original DNA.
Binary Fission
 2. Budding
Budding is a process in which a small protuberance (bud)
develops at one end of the cell.

This enlarges and eventually develops into a new cell which

separates from the parent.

Chemoheterotrophic, photoautotrophic

Example: Rhodopseudomonas acidophila and

Hyphomicrobium vulgare. Mostly, Chemoheterotrophic,
photoautotrophic follow this process. Yeast, Hydra also undergo
this process
 2. Budding …process
Genome Replication:
• The parent cell’s DNA replicates; one copy
migrates into the bud.
• Only part of the cytoplasm is shared initially.

Bud Initiation:
• A small protuberance (bud) forms at one end of
the parent cell.

Bud Growth:
• The bud enlarges as it receives organelles (e.g.,
mitochondria, ribosomes) and cytoplasm.

Separation:
A septum (cell wall partition) forms between the
parent and bud.
2. Budding
 3. Fragmentation
• The parent organism cell splits into individual fragments.
• Each fragment develops independently into a complete individual.
This type of reproduction gives rise to genetically identical
offspring. Fragmentation is also known as the splitting method.

Example: Nocardia species reproduce by fragmentation. This type of

reproduction is found in many organisms such as cyanobacteria,
Spirogyra, Fungi
3. Fragmentation
 4. Spore Formation
Endospores:
Endospores are highly resistant, dormant spores formed
within Gram-positive bacteria. These structures can survive
extreme heat, ultraviolet and gamma radiation, chemical
disinfectants.
The durability of endospores make the bacteria a critical
concern in medicine and industry.
Several endospore-forming species are dangerous pathogens,
including Clostridium botulinum. It produces the lethal
botulinum toxin causing foodborne disease, with its heat-
resistant spores. Other pathogenic examples are Bacillus
anthracis (anthrax), Clostridium tetani (tetanus).
 4. Spore Formation
Process of spore formation (sporulation)
When a Bacteria senses starvation or environmental stress, the
bacteria starts forming an endospore.
Stage I: Axial Filament Formation
The bacterial chromosome condenses into an elongated structure
aligned along the cell’s long axis.
Stage II: Asymmetric Septum Formation
The cell starts to divide two unequal compartments: a
smaller forespore (which will become the endospore) and a
larger mother cell. One complete copy of the DNA will be allocated
to each part.
Stage III: Engulfment of the Forespore
The mother cell's membrane wraps around the forespore until it
fully surrounds it. This forms a forespore with two membranes,
floating inside the mother cell.
4. Spore Formation..cont.
Process of spore formation (sporulation)
Stage IV-VI: Cortex Formation
Around the forespore, cortex and a special type of cell wall is
synthesized. This provides chemical resistance protect DNA
from heat, radiation, and enzymes.
Stage VII: Lysis and Release
Finally, the mother cell undergoes lysis, releasing the
mature endospore into the environment. The endospore is
now dormant, resistant to extreme temperatures, chemicals,
and even vacuum. It can survive for centuries until favorable
conditions trigger germination.
Bacterial Reproduction -
Sexual Reproduction
 Bacterial Reproduction/Sexual
Reproduction
In some ways, bacterial genetic material can be exchanged between
individuals. three different mechanism were later discovered for
transferring gene or genetic material from one bacterial cell to
another.
1. Transformation

2. Transduction

3. Conjugation
 1. Transformation
Some bacterial cells release DNA when they die and
break apart. This DNA can be taken up by another
bacterial cell. This is called Transformation.
 1. Transformation

• DNA is released into the environment when a bacterial cell dies.

• A living bacterial cell nearby can take in this free DNA — but only if
it's in a special condition called competence.
• The strand of DNA that enters the cell searches for a similar
sequence in the cell’s own DNA. If a match is found, it replaces that
section of the original DNA — similar to editing or updating the
genetic information.

• In nature, this process helps some bacteria change or adapt

(though it’s not very common). In labs, scientists use
transformation to insert useful genes into bacteria — for example,
adding the human insulin gene so that bacteria can produce insulin
as medicine
 2. Transduction
In this type genetic material exchange, DNA is
transferred from bacteria to another by means of
viruses (example: bacteriophage).
 3. Conjugation
It is the transferring of genetic material from one bacterium to
another by direct cell to cell contact.

A tube called the connecting bridge or pilus forms between the

donor and the recipient.

One of the two DNA in the donor bacterium is transferred into the
recipient bacterium through the tube.
3. Conjugation
Bacterial Growth
 Growth
Bacterial growth refers to an increase in cell size and cell number
(colonies).
The most common method of bacterial reproduction is binary
fission, where one cell divides into two.
This leads to exponential population growth. Thus, the increase in
population goes by geometric progression.
1 21 22 23 24 . . . . . . . 2n
Therefore, n represents the number of generations
• Each generation doubles the number of cells—one becomes two,
two become four, and so on.
• The generation time is the period required for a single cell to divide
and the population to double. It varies by species and
environmental factors like temperature.
• E. coli has a generation time of about 20 minutes.
• M. tuberculosis takes around 18 hours.
Generation Time
 Growth Curve of Bacteria
Suppose a single bacterium is inoculated in a flask of liquid culture
medium and is incubated.

It will undergo binary fission and a period of rapid growth will
follow in which the cells multiply at an exponential rate.

The growth of bacteria can be represented by curves in two ways:

Number of bacteria versus time
Logarithmic number of bacteria versus time
Growth Curve
 Phases of Bacterial Growth
There are four different phases of bacterial growth

1. The lag phase: preparing for division

2. The log phase: reproduction (relationship to disease)

3. The stationary phase: equalization

4. The decline/death phase: cell death

 Phases of Bacterial Growth
1) Lag Phase
• This is the initial stage where cells are added to fresh medium. In
this phase, cells do not divide or increase in number immediately.
• In this phase, bacteria adapt to new conditions by increasing in size
and becoming metabolically active.
• Bacteria synthesize RNA, enzymes, and other essential molecules
needed for cell division.
• At the end of the lag phase, all cells have adjusted and are ready to
divide regularly.

The length of the lag phase can vary depending on factors such as
the bacterial species, the composition of the culture medium, and
environmental conditions like temperature or pH.
 Phases of Bacterial Growth
2) log phase (logarithmic phase)

• The log phase is marked by rapid, constant cell division and

exponential population growth.
• In this phase ,bacteria exhibits uniform chemical composition,
metabolic activity, and physiological characteristics, making this
stage ideal for studying microbial metabolism.
• As nutrients are used up and waste products accumulate, growth
slows and the culture eventually enters the next phase.
• In research and industry, steady-state growth is maintained by
adding fresh medium and removing used medium to keep
bacteria in the log phase.

• During exponential growth, plotting log cell number against time

gives a straight line, showing the specific growth rate—e.g.,
cyanobacteria can double four times daily under ideal conditions.
 Stationary Phase
In this phase , the number of growing and dying cells becomes
equal. and As a result, the population size remains constant. It
happens because of the exhaustion of certain nutrients and the
accumulation of toxic byproducts produced during growth.
Additionally, mutations may occur during the stationary phase.

This state is characterized by a horizontal, linear segment in the

growth curve.
 Decline/Death Phase
In this phase, the bacteria may die faster than new cells are
produced. the number of viable cells declines exponentially.
No further cell division occurs during this phase. The death
rate follows similar kinetics to the exponential (log) growth.
This decline is primarily due to the depletion of nutrients and
the accumulation of toxic or inhibitory byproducts, such as
acids.
Some bacterial populations may die rapidly, leaving few
viable cells within 72 hours, while others can persist for
extended periods, sometimes for months or years.
Phases of Bacterial Growth
 Importance of Growth Curve: Continuos
Culture
Lag Phase
• Maintenance: Providing optimal temperature, pH, oxygen, and
nutrients prepare bacteria for division.
• Pharmaceutical Importance: Critical phase for vaccine production.
Log (Exponential) Phase
• Maintenance: Requires optimal temperature, pH, oxygen.
• Pharmaceutical Importance: Ideal for antibiotic testing, as cells are
most metabolically active and susceptible to antimicrobial agents.
Stationary Phase
• Maintenance: May involve removal of waste or replenishment of
nutrients to sustain the culture.
• Industrial Use: Important the production of antibiotics, enzymes.
Death (Decline) Phase
• Crucial for sterilization —determining how long it takes to
eliminate bacterial populations.
• Helps evaluate antimicrobial resistance.
Quantitative Measurements of Bacterial
Growth
 Bacterial Growth
Growth can be measured by the following techniques:

Direct Microscopic Count

The Plate Count Method

Membrane Filter Method

Turbidimetric Method
 Direct Microscopic Count
Bacteria can be counted easily accurately by Pertroff-Hausser
counting chamber.

This is a special slide accurately ruled into squares that are 1/400
mm2 in area.

Cover slip rests above slide.

Each square holds a volume of 1/20,000 mm³

Unstained bacterial cultures can be counted using a phase-contrast

microscope

Used to count bacteria in vaccines.

 https://www.youtube.com/watch?v=pP0xERLUhyc
Direct Microscopic Count
 The Plate Count Method
The bacterial suspension is diluted so that the number
of colonies developing on the plate will fall in the range
of 30 to 300.

After dilution, a measured amount of bacteria is

introduced in a petri dish.

Colonies are counted by illuminating them from below.

Counts are often reported as colony-forming units per

milliliter (CFU/ml)

Used to count bacteria in milk, water, etc.

The Plate Count Method

https://www.youtube.com/
watch?app=desktop&v=IlTn
LEgivts
 Membrane Filter Method
 This technique uses molecular filters.

 The filters have known uniform, porosity of predetermined size

sufficiently small to trap microorganisms.

 Useful in determining the number of bacteria in a very dilute

medium (air or water).

 Counts colonies per ml.

 The membrane with the trapped bacteria is then placed in an

appropriate medium to support growth.

 These are then counted with special media or dyes.

https://www.youtube.com/watch?v=B1AVxzccS5Q

https://www.youtube.com/shorts/l4YBIY64v7Q
Membrane Filter Method
 Turbidimetric Method
Used to estimate cell numbers in heavy suspensions.

Requires standard curve for determination.

A culture of more than 107 to 108 cells per milliliter appears turbid
to the naked eye.

A spectrophotometer can be used for turbidimetric measurements

of cell mass. It measures optical density. Simple mathematical
formulae help convert the detected turbidity to cell concentration.

Turbidimetry is a simple and rapid method for following growth.

The dead as well as the living cells contribute to turbidity.

https://www.youtube.com/watch?v=UlPsEQ0Qdqk
Turbidimetric Method