CSU-008 – Microbiology

Lecture 11 – Microbial Growth

 Topics to be covered

1. Introduction to growth
2. Basic concepts of microbial growth
3. Reproduction in bacteria
4. Stages of bacteria growth
5. Factors affecting the microbial growth
 Introduction
• Living organisms grow and reproduce.
• When microbes are provided with nutrients & required environmental factors, they
become metabolically active and grow.
• Growth common refers to increase in a microbial size, population number or
both.
• Growth also results when cells simply become longer and larger
• Involves an increase in the number of cells rather than in the size of individual cells.
The change in population in bacteria involves Binary fission.
• Binary fission is a type of asexual reproduction where parents cells divides resulting in two
identical cells (duplicate its genetic material and divides into two parts).

• So Growth can be defined as an orderly increase of all chemical compounds in a

cell/population which results in increase in mass of the cell/population
 Levels of Growth

1. A cell synthesizes new components, increase its size

2. Increase number of cells in the population

Increase in cellular constituents that may result in:

➢ Increase in cell number; e.g., when microorganisms reproduce by budding or
binary fission

➢ Increase in cell size; e.g., some microorganisms have nuclear divisions that are
not accompanied by cell divisions

❖ Microbiologists usually study population growth rather than growth of individual

cells.
 Important concepts in microbial growth

Bacterial Stages of Environmental

Bacterial growth Measurement of
Division/reproduc Microbial growth factors effect
tion curve microbial growth
 1. Bacterial Division or reproduction

❖ Bacteria normally reproduce by a

method called binary fission.
❖ Binary Fission is the process of
bacteria reproduction where one cell
become two
 Generation time (Doubling time)

• The time required for a cell to divide (and

its population to double) is called the

generation time

• Specific time interval between two

subsequent binary fission is known as

generation time

• As you seen in the picture, cell’s division

produces two cells, two cells’ divisions

produce four cells, and so on.

Bacteria undergoes exponential Growth
Bacterial generation time
 Mathematical expression of growth
To calculate the generation time of individual microorganism the following
experimental data are required:
1. The number of organisms present at beginning
2. The number of organisms present at the end of a given timeinterval
3. The time interval

1. So starting with a single cell, the total population B at the end of a given time period
would be expresses as:

B=1*2n
Where 2n is the bacterial population after n generations.
 Mathematical expression of growth
2. But under practical conditions several thousands of bacteria are introduced into the
medium at 0 time and not one, so the formula nowbecomes:

Bn=B0*2n

Where Bn = no. of organisms after n generations

B0= no. of organisms at 0time
t= number of generations

Solving the equation for n, we have

log Bn = log B0 + n log 2
n= log Bn - log B0 / log 2
Thus we can calculate the number of generations if we know the initial population (B0)
and population (Bn) after time t. So generation time (G) is equal to t divided by the
no. of generations. G=t/n
Example of exponential growth
 2. Stages in Bacterial Growth

• When a few bacteria are inoculated into a liquid growth medium, it is possible to plot a
bacterial growth curve that shows the growth of cells over time.

• There are four basic phases of growth:

The Lag Log

(Exponential)

Stationary Death
 Bacterial growth curve (Monoauxic growth)
1. Batch culture (closed system)
2. Bacteria grown in a liquid medium
3. Single batch of medium
4. Nutrient get exhausted and there is decline in growth
5. When a graph is plotted as the logarithm of the no. of viable cells Vs the incubation
time
 A. Lag Phase
• Introduction of microorganism into fresh medium
• Bacteria are trying to adapt to nutrients
• No immediate increase in cell number, no increase in net mass
• Lag phase-preparing to grow in size and synthesize enzymes etc.
• Cell is synthesizing their own components
• Can be very short or even absent

Importance of lag phase-

1. Cells may be old and depleted of ATP, essential cofactors, ribosomes and these must be
synthesized before growth begins.
2. The medium may be the different than the previous one so enzymes are required
 B. Log Phase
• Also called Exponential phase
• Growth and division at maximal rate
• Also called balanced growth since all cell constituents are synthesized at constant rate.
• Population number of cells undergoing binary fission doubles at a constant interval called
generation time
• Continue as long as cells have adequate nutrients & good environment
• All studies related to their various properties are done in this phase.
 Balanced and Unbalanced growth

Shift-up experiment Shift-down experiment

bacteria when transferred from bacteria when transferred from
nutritionally poor medium to richer nutritionally rich medium to poorer
medium the lag phase is longer medium the lag phase is shorter
 C. Stationary Phase
• Reproduction rate is balanced by the death rate and population remains constant
• There is exhaustion of some nutrients and accumulation of some toxic materials
• In this stage microbial death is equal to microbial growth e-g Death=growth.
• Period of equilibrium
• Metabolic activity of surviving cells slows down which stabilizes the population
• Cause of discontinuity of exponential growth is not always clear
• May play a role: exhaustion of nutrients, accumulation of waste products and harmful
changes in pH
• Chemostat – continuous culture used in industrial fermentation
 Reasons for stationary Phase

Critical
Nutrient Limited oxygen Toxic waste population
limitation availability accumulation density reached
 D. Death Phase
• Decline in the number of viable cells
• Cells dying, usually at exponential rate
• Death; loss of ability to reproduce
• In some cases, death rate slows due to accumulation of resistant cells
• Slower than log phase
• Also known as Logarithmic Decline Phase
• Continues until a small fraction of the population is diminished
• Some population dies out completely
• Others retain surviving cells indefinitely while others only retain for a few days
 3. Factors affecting the microbial growth

other
environmental
pH Temperature Gas Pressure factors
requirement
e-g radiation,
water activities
etc.
 1. pH

• Negative logarithm of the

hydrogen ion concentration

• Affect the activity & integrity of

enzymes and structural components
of a cell
 1. pH

 Optimum pH for most microbes ranges approximately from 6 to 8

 Acidophiles

 growth optimum between pH 0 and pH 5.5

 Neutrophiles

 growth optimum between pH 5.5 and pH 7

 Alkalophiles

 growth optimum between pH8.5 and pH 11.5

2. Temperature

 organisms exhibit distinct

cardinal growth temperatures

 minimum

 maximum

 optimum
 Temperature based microbes
❖ Pychrophiles
✓Grow well at 0ºC and growth at 15 ✓It is moderate temperature loving microbes
ºC or lower Examples: Staphylococcus aureus,
✓This are cold loving microbes Escherichia coli
✓It cannot grow above 20º C
Examples: Arthrobacter sp., ❖Thermophiles
Psychrobacter sp., Pseudomonas. ✓The growth range is 45 ºC and 80 ºC
✓Optimum temperature between 55-60 ºC
❖ Psychrotroph ✓It is heat loving microbes
✓ It can grow at 0-7ºC Examples: Thermoplasma acidophilum,
✓The optimum temperature between Thermus aquaticus.
20-30 ºC
✓Maximum temperature around at ❖ Hyperthermophiles
35 ºC ✓Optimum growth is observed between 80 ºC
Examples: Bacillus, Clostridium, and 100 ºC
Microbacterium ✓It may grow at 120 ºC temperature
Examples:
❖Mesophiles Pyrolobus fumarii (113 ºC) and Pyrococcus
✓It grow optimum temperature furiosus (100°C), first discovered in Italy near
around at 20-40 ºC a volcanic vent.
 3. Gas Requirement

Depends the kinds of microorganism, some microorganism need oxygen

while others not need oxygen to grow
 4. Pressure

• Microbes obtain almost all their nutrients in solution from

surrounding water barophile organisms
• Adapted to life under high pressure (found in ocean floors and deep
sea environment)
• Require or grow more rapidly in the presence of increased pressure
• They are a type of extremophile (living in very harsh environments)
• Those that can live at high pressure and in less extreme
environment are referred to as barotolerant
• Example: bottom dwellers in the ocean (Halomonas salaria,
Shewanella violacea, Moritella japonica, etc.)
 4. Measurement of Microbial Growth

Can measure changes in number of cells in a Microbial growth to determine growth

population; Can measure changes in mass of rates and generation times can be
population measured by different methods.
 Methods used to count the number of cells
There are two types:-
1 Total count/Direct cell counts:-

a) Counting chambers method/ Haemocytometer method

b) Breed method/direct microscopic method
2 Viable cell counts/Indirect method
a) Plating methods (spread, pour plate)
b) Membrane filtration methods
Dr. Dimple Tanwar
School of Bioengineering and Food Technology
Shoolini University Village Bhajol, Solan (H.P)
7833078548 (Mob No.)
dimpletanwar1@[Link]