How Microbes Grow?

Chapter 3:

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Content
o Bacterial Cell Division
o Cell Growth and Binary Fission

o Population growth
o The Concept of Exponential Growth

o The Microbial Growth Cycle

o Measuring Microbial Growth

o Environmental Factors Affecting Growth

o Temperature

o Acidity and Alkalinity

o Osmotic Effects on Microbial Growth

o Oxygen and Microorganisms

o Toxic Forms of Oxygen

1. Cell Growth and Binary
Fission
Bacterial growth Definition

• Bacterial growth refers to an increase in bacterial

numbers, not an increase in the size (like plants and
animals)of the individual cells.
• Bacteria normally reproduce by binary fission,
• Few bacterial species reproduce by budding; they
form a small initial outgrowth (a bud) that enlarges
until its size approaches that of the parent cell, and
then it separates
 Budding

• Some bacteria reproduce by

budding, a process in which a
small protuberance (bud)
develops at one end of the
cell, enlarges and develops into a
new cell that later separates from
the parent cell, e.g.
Rhodopseudomonas acidophila
1.budding-lactic-bacteria

(bud) parent cell

2. Some filamentous bacteria (certain actinomycetes) reproduce by
producing chains of conidiospores (an asexual spore) carried externally
at the tips of the filaments.
 binary fission,
➢ The normal reproductive method of bacteria is binary
fission in which a single cell divides into two
identical cells after developing a cross wall (transverse)
septum.
➢ It is an asexual reproductive process.
➢ Thus, bacteria increase their numbers by geometric
progression or exponential growth, i.e. doubling of
bacterial population every generation as : 1, 2, 4, 8, etc. or
20, 21, 22, 23.........2n (where n = the number of
generations).
“Binary Fission” = divide in half

Fission = cells separate, resulting

in two daughter cells which are
IDENTICAL to each other and the
original parent cell
 For exp: Escherichia coli, every 20
minutes birth of 2 identical bacteria
2. The Concept of Exponential Growth
• one cell’s division produces two cells, two cells’ divisions produce four cells,
and so on. When the number of cells in each generation is expressed as a
power of 2, the exponent tells the number of doublings (generations) that
have occurred.
Growth of Microbial Populations
 Generation Time (G)
 Time required for a bacterial cell to grow and divide
 Dependent on chemical and physical conditions

• Varies among species

• Environmental conditions

G=t/n
G = generation time (G)
t = time interval in hours or minutes
n = number of generations

➢ Under a given set of growth conditions (medium, temperature, pH,

etc.) each bacterial species has a genetically determined generation
time.
 Calculation of Generation Time
The relationship between the number of bacteria in a population
at a given time (Nt), the original number of bacterial cells in the
population (N0), and the number of divisions those bacteria have
undergone during this time (n) can be expressed by the following
equation:
Nt = N0 x 2n
Solving for ‘n’: log Nt = log N0 + nlog2
or nlog2 = log Nt - log N0
or n = log (Nt - N0 ) / log2 (=0.301)
or n = 3.3 log Nt / N0
Since G = t/n
Therefore G = t / 3.3 log Nt / N0
Growth rate Constant (µ):

➢The reciprocal of the Generation Time (1/G) is

called the “Growth Rate Constant
It is defined as the number of divisions per unit of time
( [generations/unit time]).
:

µ = n/t = 1/G
t = time (known)

n = number of divisions
 A growth curve for an exponentially increasing population,
plotted logarithmically (dashed line) and arithmetically (solid
line).
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3. The Microbial Growth Cycle
 THE BACTERIAL GROWTH CURVE
➢ When we measures the growth of a bacterial culture in a closed
system such as a flask or a fermenter and plots the logarithm of
cell number over time then one obtains a characteristic curve,
which is called a bacterial growth curve.
➢ The growth curve consists of six distinct phases, with a transition
period in between each phase:
1. Lag phase
2. Acceleration phase
3. Log (logarithmic or exponential) phase
4. Deceleration Phase
5. Stationary phase
6. Decline (death) phase
 log N
Ln N

•
•
The Lag Phase
• Death
Deceleration Phase
Phase
• 4 Accelration
• 5
•
Phase
Exponential (Log)
Phase
Stationary
•
phase temps
•
1/ Lag phase
• Definition :
Phase characterized by:
- absence of multiplications during this phase: µ = 0 multiplication / h
- Prepare to synthesize enzymes and molecules for adaptation to a new
environment

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Meaning of each phase
1/ Lag phase
• Characteristic: Variable duration of this phase

Duration dependent:
- Of the composition of the medium
- On the age of microorganisms
- From the inoculum
- the nature of the microorganism

Declining microorganisms take

longer to restore their
damaged enzymes
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1/ Lag phase

• Noticed :
• Microorganisms taken in exponential phase from an X medium
• Microorganisms reseeded in X medium

No Lag phase (zero duration)

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2/ Acceleration Phase
Definition :
▪ Phase during which the number of adapted bacteria that begin to
multiply becomes greater and greater
▪ the rate of multiplication is increasing (µ )

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3. The Log Phase (logarithmic phase, exponential phase)
➢ Acceleration Phase is followed by log phase during which binary
fission occurs and the bacteria multiply at the fastest rate
possible under the conditions provided (population doubles every
generation ). µ is constant and maximum therefore G constant and
minimum.
➢ This phase of growth is called logarithmic or exponential because the
rate of increase in cell number is by geometric progression: 1, 2, 4, 8,
etc. or 20, 21, 22, 23.........2n (where n = the number of generations).

➢ During log phase, the bacterial division is at a constant rate

(generation time), but varies with species, temperature and media.
➢ Microbes are most sensitive to adverse conditions and antibiotics .
4/ Deceleration Phase

• Definition: Phase during which µ gradually decreases until

it becomes zero (µ )

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 5.The Stationary Phase

➢ Stationary phase is a steady-state equilibrium where the rate of cell

growth (division) is exactly balanced by the rate of cell death, i.e.
death rate = rate of reproduction.
➢ Cell death occurs because of one of three factors: (a) exhaustion of
available nutrients and water; (b) accumulation of catabolic end
products (c) exhaustion of space – “biological space”, and (d) change
in oxygen concentration and pH.
➢ The stationary phase is not necessarily a period of quiescence.
Bacteria produce secondary metabolites, such as antibiotics and the
sporulation process starts.
 6.The Decline Phase
(death phase)
➢ Stationary phase is followed by a die-off of cells during which the
number of deaths exceeds the number of new cells formed, i.e.
Death rate > rate of reproduction.
➢ Cell death in bacteria cultures basically means that the cells are
unable to resume division following their transfer to new
environments.
➢ During the death phase, the number of viable cells decreases
geometrically (exponentially), essentially the reverse of growth
during the log phase.
➢ Deaths are due to the factors in stationary phase in addition to
lytic enzymes that are released when bacteria lyse.
Environmental Factors Affecting Growth

❖Temperature
❖Acidity and Alkalinity
❖Osmotic Effects on Microbial Growth
❖Oxygen and Microorganisms
❖Toxic Forms of Oxygen
 Effect of Temperature on Growth

• Temperature is a major environmental

factor controlling microbial growth
• Cardinal temperatures: the minimum,
optimum, and maximum temperatures
at which an organism grows.
Microorganisms can be classified into
groups by their growth temperature
optima.
❖Psychrophile: low temperature
❖Mesophile: midrange temperature
❖Thermophile: high temperature
❖Hyperthermophile: very high
temperature

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 Acidity and Alkalinity

• The pH of an environment greatly affects microbial growth .

• Some organisms have evolved to grow best at low or high pH, but most
organisms grow best between pH 6 and 8 ( neutrophiles)
• Acidophiles: organisms that grow best at low pH (<5,5)
• Some are obligate acidophiles; membranes destroyed at neutral
pH
• Stability of cytoplasmic membrane critical
• Alkaliphiles: organisms that grow best at high pH (>8,5)
• Some have sodium motive force rather than proton motive force

➢ The internal pH of a cell must stay relatively close to neutral even

though the external pH is highly acidic or basic
➢ Microbial culture media typically contain buffers to maintain constant
pH
 Osmotic Effects on Microbial Growth

✓ Typically, the cytoplasm has a higher solute concentration than the

surrounding environment, thus the tendency is for water to move into the
cell (positive water balance)
✓When a cell is in an environment with a higher external solute
concentration, water will flow out unless the cell has a mechanism to
prevent this.
❖Halophiles: organisms that grow best at reduced water potential; have a
specific requirement for NaCl.

❖Extreme halophiles: organisms that require high levels (15–30%) of NaCl

for growth

❖Halotolerant: organisms that can tolerate some reduction in water

activity of environment but generally grow best in the absence of the added
solute

❖Nonhalophile
Figure 5.25

Halotolerant Halophile Extreme

Example: Example: halophile
Staphylococcus Aliivibrio fischeri Example:
aureus Halobacterium
salinarum
Growth rate

Nonhalophile
Example:
Escherichia coli
0 5 10 15 20
NaCl (%)
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Oxygen and Microorganisms

• Aerobes: require oxygen to live

• Anaerobes: do not require oxygen and may even be killed
by exposure
• Facultative organisms: can live with or without oxygen
• Aerotolerant anaerobes: can tolerate oxygen and grow in
its presence even though they cannot use it
• Microaerophiles: can use oxygen only when it is present at
levels reduced from that in air
 Oxygen and Microorganisms

• Thioglycolate broth : Complex

medium that separates
microbes based on oxygen
requirements.
oReacts with oxygen so oxygen
can only penetrate the top of
the tube.

Growth versus oxygen concentration.

(a) aerobic, (b)anaerobic, ( c) facultative,
(d)microaerophilic, (e) aerotolerant anaerobe