Professional Documents
Culture Documents
Industrial Microbiology
Bacterial growth
Binary fission
Generation time
Phases of growth
4-2
Binary fission
Figure 4.2
Generation time
The time required to for a population to double (doubling
time) in number.
Ex. Escherichia coli (E. coli) double every 20 minutes
Ex. Mycobacterium tuberculosis double every 12 to 24
hours
4-4
Principles of Bacterial Growth
Growth can be calculated
(Nt ) number of cells in population
(N0 ) original number of cells in the population
(n) number of divisions
Example
N0 = 10 cells in original population
n = 12
4 hours assuming 20 minute generation time
Nt = 10 x 212
Nt = 10 x 4,096
Nt = 40,960
Growth in Batch Culture
1. Bacteria growing in batch culture produce a growth curve with up to four
distinct phases.
2. Batch cultures are grown in tubes or flasks and are closed systems where no
fresh nutrients are added or waste products removed.
3. Lag phase occurs when bacteria are adjusting to them medium. For example,
with a nutritionally poor medium, several anabolic pathways need to be turned
on, resulting in a lag before active growth begins.
4. In log or exponential phase, the cells are growing as fast as they can, limited
only by growth conditions and genetic potential. During this phase, almost all
cells are alive, they are most nearly identical, and they are most affected by
outside influences like disinfectants.
5. Due to nutrient depletion and/or accumulation of toxic end products,
replication stops and cells enter a stationary phase where there is no net
change in cell number.
6. Death phase occurs when cells can no longer maintain viability and numbers
decrease as a proportion.
Growth in Batch Culture
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.
1
Growth rate (min-1) =
mean generation time
pH
Water availability
4-12
Temperature
Enzymes, the machinery of the cell, are influenced
by external factors and can be shown to have a
range where they function that includes an optimal
value that produces the highest activity.
The range of enzyme activity determines the range
for growth of specific bacteria, analogously leading
to a value for optimal growth rate.
In the case of temperature, bacteria are divided
into categories based on the temperature range
where they can grow and the temperature that
provides optimal growth.
4-15
Temperature
Psychrophile
0o to 18o C
Psychrotroph
20°C to 30°C
Important in food spoilage
Mesophile
25°C to 45°C
More common
Disease causing
Thermophiles
45°C to 70°C
Common in hot springs and hot water heaters
Hyperthermophiles
70°C to 110°C
Live at very high temperatures, high enough where water threatens to become
a gas
Usually members of Archaea
Found in hydrothermal vents
Oxygen requirements
• Oxygen is a very reactive molecule and can affect cells in several ways. The effect of oxygen is often determined using
thioglycollate broth, a special medium that contains a reducing agent (thioglycollate) that removes oxygen so that a
gradient occurs within the tube.
• Obligately aerobic bacteria can obtain energy only through aerobic respiration and have to have oxygen available. Thus,
they will grow only at the surface of thioglycollate broth.
• Obligately anaerobic bacteria die in the presence of oxygen and can only grow at the bottom of thioglycollate broth.
Some anaerobes are so sensitive to oxygen that even thioglycollate broth is not anoxic enough to provide suitable
anaerobic conditions.
• Microaerophiles require oxygen for growth but the 20% in air is too toxic. As a result, they grow near the top but
beneath the surface of thioglycollate broth where the oxygen concentration is typically 4 – 10%.
• Facultative anaerobes can use oxygen for aerobic respiration but can switch to fermentative metabolism in the absence
of oxygen. As a result, they will grow throughout thioglycollate broth. (Heavier growth at top.)
• Aerotolerant anaerobes are anaerobic bacteria that can grow in the presence of 4-17
air. (Not shown in diagram.)
pH
Neutrophiles grow best around neutral pH (7)
Acidophiles grow best at pH < 7
Alkophiles grow best at pH > 7
Acidotolerant grow best at pH 7 but can also grow at
lower pH
Alkotolerant grow best at pH 7 but can also grow at
higher pH
4-18
Water Activity
Organic nitrogen
Primarily from the catabolism of amino acids
Oxidized forms of inorganic nitrogen
Nitrate (NO32-) and nitrite (NO2-)
Reduced inorganic nitrogen
Ammonium (NH4+)
Dissolved nitrogen gas (N2) (Nitrogen
fixation)
Phosphate Source
Organic phosphate
Inorganic phosphate (H2PO4- and HPO42-)
Sulfur Source
Organic sulfur
Oxidized inorganic sulfur
Sulfate (SO42-)
Reduced inorganic sulfur
Sulfide (S2- or H2S)
Elemental sulfur (So)
Growth Factors
Some bacteria cannot synthesize some cell constituents
These must be added to growth environment
Referred to as growth factors
Organisms can display wide variety of factor
requirements
Some need very few while others require many
These termed fastidious
Typical molecules
Amino acids
Nucleotide bases
Enzymatic cofactors or “vitamins”
Culture Media
Complex (contains undefined components)
Chemically defined (all concentrations are known)
Selective (favors the growth of a particular organism or
group of organisms)
Differential (has reactions that give isolates different
appearance)
Anaerobic (oxygen-free)
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4-32
Characteristics of Media