MICROBIAL GROWTH

By the end of the topic you should be able to

1. Describe how bacteria reproduce

2. Describe the growth curve and its phases
3. Describe factors that affect microbial growth
pH
temperature
aeration
pCO2
osmotic pressure
Microbial growth

1. The growth curve and its phases

2. Factors that affect microbial
growth
a. Physical factors:
pH, temperature, aeration,
pCO2, osmotic pressure (water activity Aw)
b. Chemical factors
inorganic ions and trace
elements, growth factors,
carbon sources, energy sources,
nutritional classes

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growth

•Majority of bacteria reproduce by a mechanism

termed binary fission.
•The time required for the doubling of cells is
known as the generation time.
Generation Time (Doubling Time)
•time required for a cell to divide
•most about 1 Hr. To 3 Hrs.
E. coli - 20 minutes

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Binary Fission - unchecked!

 E. coli - generation time of 20 min.

 20 generations (about 7 hrs.)
• 1 million cells
 30 generations ( about 10 hrs.)
• 1 billion cells
 72 generations ( about 24 hrs.)
• 1 x 1021

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 Growth in Batch Culture

• “Growth” is generally used to refer to the

acquisition of biomass leading to cell division,
or reproduction

• A “batch culture” is a closed system in broth

medium in which no additional nutrient is added
after inoculation of the broth.
• Bacteria added to fresh media typically go
through four more or less distinct phases of
growth

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 Growth in Batch Culture
• Typically, a batch culture passes through four
distinct stages:
• Lag stage
• Logarithmic (exponential) growth
• Stationary stage
• Death (decline) stage

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Growth in Batch Culture

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1. Lag Phase

 Bacteria are first introduced into an environment or

media
 Bacteria are “checking out” their surroundings
 cells are very active metabolically
 # of cells changes very little
 1 hour to several days
 Length depends on no. of factors e.g. Previous
history of inoculum

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2. Log Phase

 Rapid cell growth (exponential growth)

 Maximum growth rate possible at given genetic
potential, nature of media and env. conditions
 population doubles every generation where
 Nf = (Ni)2n
 Where Ni = initial size of pop., n no. og generations
elapsed, Nf = final pop
 microbes are sensitive to adverse conditions
• antibiotics
• anti-microbial agents

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3. Stationary Phase

 Death rate = rate of reproduction

 Secondary metabolites produced during this phase
e.g. Pigments, exotoxins, antibiotics. Bacteria -
spores
 cells begin to encounter environmental stress
• lack of nutrients
• lack of water
• not enough space
• metabolic wastes
• oxygen
• PH

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4. Death Phase

 Death rate > rate of reproduction

 Due to limiting factors in the environment
 Autolytic enzymes activated batch culture becomes
progressively less turbid as dying cells lyse.

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Mean Generation Time
and Growth Rate
• The mean generation time (doubling time) is the
amount of time required for the concentration of
cells to double during the log stage. It is
expressed in units of minutes.

• Growth rate (min-1) =

1
mean generationtime
• Mean generation time can be determined
directly from a semilog plot of bacterial
concentration vs time after inoculation

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Mean Generation Time
and Growth Rate

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 Growth in Continuous Culture
• A “continuous culture” is an open system in
which fresh media is continuously added to the
culture at a constant rate, and old broth is
removed at the same rate.
• This method is accomplished in a device called a
chemostat.
• Typically, the concentration of cells will reach an
equilibrium level that remains constant as long as
the nutrient feed is maintained.

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 Factors that Influence Growth
• Growth vs. Tolerance
• “Growth” is generally used to refer to the acquisition of
biomass leading to cell division, or reproduction
• Many microbes can survive under conditions in which
they cannot grow
• The suffix “-phile” is often used to describe conditions
permitting growth, whereas the term “tolerant”
describes conditions in which the organisms survive,
but don’t necessarily grow
• For example, a “thermophilic bacterium” grows under
conditions of elevated temperature, while a
“thermotolerant bacterium” survives elevated
temperature, but grows at a lower temperature
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• Obligate (strict) vs. facultative
– “Obligate” (or “strict”) means that a given
condition is required for growth
– “Facultative” means that the organism can
grow under the condition, but doesn’t require it
– The term “facultative” is often applied to sub-
optimal conditions
– For example, an obligate thermophile requires
elevated temperatures for growth, while a
facultative thermophile may grow in either
elevated temperatures or lower temperatures

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 Requirements for Growth
• Physical
• Chemical

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Factors that Influence Growth
OXYGEN CONCENTRATION
Bacteria can be classified based on their oxygen requirements

• Strict aerobes: Require oxygen for growth (~20%)

• Strict anaerobes: Grow in the absence of oxygen; cannot
grow in the presence of oxygen. Lack both superoxide
dismutase and catalase
• Facultative anaerobes: Grow best in the presence of oxygen,
but are able to grow (at reduced rates) in the absence of
oxygen
• Aerotolerant anaerobes: Can grow equally well in the
presence or absence of oxygen, produce superoxide
dismutase, not catalase
• Microaerophiles: Require reduced concentrations of oxygen
(~2 – 10%) for growth
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Growth of different motile bacteria in stab
cultures in sloppy agar

Obligate aerobes

• B. subtilis

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Obligate anaerobes
• Lack both superoxide
dismutase and
catalase
• e.g. Clostridium
rubrum

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Facultative anaerobes
• Grow both in O2 and
in CO2 but do better
with oxygen
• e.g. E. coli

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Microaerophile
• Grows in slmall
amounts (2 10%) of
oxygen
• e.g. Mycobacterium
tuberculosis

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Oxygen is lethal to some organisms

 All organisms produce superoxide ( O2-)

 Superoxide is toxic to cells (steals electrons)

 Superoxide must be neutralized

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Superoxide dismutase

 O2- + O2- + 2 H+ -------> H2O2 + O2

 Hydrogen peroxide is also toxic to cells and it must

be neutralized

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Catalase

 2 H2O2 --------> 2 H2O + O2

 Obligate Anaerobes lack:

• Superoxide dismutase ( SOD )
• Catalase

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Factors that Influence Growth

TEMPERATURE
- primary effect – enzyme activity
organisms exhibit 3 cardinal temperatures (minimal, maximal, optimal growth
temps.

• Most bacteria grow throughout a range of approximately

20 Celsius degrees, with the maximum growth rate at a
certain “optimum temperature”
• Psychrophiles: Grows well at 0ºC; optimally between
0ºC – 15ºC
• Psychrotrophs: Can grow at 0 – 10ºC; optimum
between 20 – 30ºC and maximum around 35ºC
• Mesophiles: Optimum around 20 – 45ºC
• Moderate thermophiles: Optimum around 55 – 65 ºC
• Extreme thermophiles (Hyperthermophiles):
Optimum around 80 – 113 ºC
• Stenothermal – narrow range of cardinal growth temps
• Eurythermal – wide range of cardinal growth temps

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 Factors that Influence Growth
• pH
• Acidophiles:
• Grow optimally between ~pH 0 and 5.5
• Neutrophiles
• Grow optimally between pH 5.5 and 8
• Alkalophiles
• Grow optimally between pH 8 – 11.
• Despite wide variations in habitat pH, the internal pH of most
microorganisms is maintained near neutrality either by
proton/ion exchange or by internal buffering.

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 Factors that Influence Growth
• Salt concentration
• Halophiles require elevated salt concentrations to grow;
often require 0.2 M ionic strength or greater and may
some may grow at 1 M or greater; example,
Halobacterium
• Osmotolerant (halotolerant) organisms grow over a
wide range of salt concentrations or ionic strengths; for
example, Staphylococcus aureus

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Classification of microorganisms based on the
carbon and energy sources
Chemical factors affecting growth

inorganic ions and trace

elements, growth factors,
carbon sources, energy sources,
nutritional classes

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